Respiratory Disease

Vopr Kurortol Fizioter Lech Fiz Kult. 2016 Mar-Apr;93(2):9-12.

The influence of low-intensity laser radiation on the functional activity of neutrophils in the patients presenting with community-acquired pneumonia.

[Article in Russian]
Burduli NM1, Gabueva AA1.

Author information

  • 1State budgetary educational institution of higher professional education ‘North-Ossetian State Medical Academy’, Russian Federation Ministry of Health, Republic of North Ossetia – Alania, Vladikavkaz, Russia.

Abstract

in English, Russian

AIM:

The objective of the present study was to elucidate the influence of low-intensity laser radiation on the results of the nitroblue tetrazolium (NBT) test in the patients presenting with community-acquired pneumonia.

PATIENTS AND METHODS:

A total of 100 patients with community-acquired pneumonia were available for the examination of whom 70 were treated with intravenous low-intensity laser irradiation of blood (ILIB) by means of the ILIB-405 technique during 7 days. The functional activity of neutrophils was estimated from their ability to reduce nitroblue tetrazolium in both spontaneous and stimulated NBT-tests.

RESULTS:

The analysis of the data obtained in the study has demonstrated the significant improvement of the results of the NBT tests in the group of patients receiving the ILIB treatments regardless of whether its content was originally elevated or reduced.

CONCLUSION:

The inclusion of intravenous low-intensity laser irradiation of blood in the combined treatment of the patients with community-acquired pneumonia appreciably promotes normalization of the bactericidal activity of neutrophils.

Ter Arkh. 2016;88(3):32-5.

Pathogenetic effects of low-intensity laser therapy for chronic obstructive pulmonary disease.

[Article in Russian]
Aksenova IZ1, Burduli NM1.

Author information

  • 1North Ossetian State Medical Academy, Ministry of Health of Russia, Vladikavkaz, Republic of North Ossetia-Alania, Russia.

Abstract

AIM:

To study the impact of low-intensity laser therapy (LILT) on typical pathological processes of impairments in platelet aggregation and microcirculation (MC) in patients with chronic obstructive pulmonary disease (COPD).

MATERIALS AND METHODS:

A photo optical aggregatometric method was used to investigate platelet aggregation function in patients with COPD and MC was estimated by laser Doppler flowmetry over time.

RESULTS:

There were muitidirectional changes in platelet aggregation function with a predominance of hyperaggregation, as well as a significant improvement in aggregation indicators during LILT; the latter was shown to have a correcting impact on MC disorders in patients with COPD in the presence of the spastic and stagnant-stasic types of MC, which were prevalent in the majority of patients.

CONCLUSION:

Laser therapy used in the combination treatment of patients with COPD promotes correction of the typical pathological processes.

 
PLoS One. 2015; 10(8): e0136942.
Published online 2015 Aug 31. doi: 10.1371/journal.pone.0136942
Human Tubal-Derived Mesenchymal Stromal Cells Associated with Low Level Laser Therapy Significantly Reduces Cigarette Smoke-Induced COPD

Jean Pierre Schatzmann Peron,1 Auriléia Aparecida de Brito,2 Mayra Pelatti,3 Wesley Nogueira Brandão,1 Luana Beatriz Vitoretti,2 Flávia Regina Greiffo,2 Elaine Cristina da Silveira,2 Manuel Carneiro Oliveira-Junior,3 Mariangela Maluf,4 Lucila Evangelista,5 Silvio Halpern,5 Marcelo Gil Nisenbaum,5 Paulo Perin,4 Carlos Eduardo Czeresnia,5 Niels Olsen Saraiva Câmara,6 Flávio Aimbire,7 Rodolfo de Paula Vieira,3 Mayana Zatz,2 and Ana Paula Ligeiro de Oliveira3,*
Yuanpu Peter Di, Editor
1Neuroimmune Interactions Laboratory, Department of Immunology, Institute of Biomedical Sciences, University of Sao Paulo, São Paulo, SP, Brazil
2Laboratory of Pulmonary and Exercise Immunology–LABPEI, Nove de Julho University (UNINOVE), São Paulo, SP, Brazil
3Division of Human Genome Research Center, Biosciences Institute, University of São Paulo, São Paulo, SP, Brazil
4CEERH—Specialized Center for Human Reproduction, Division of Reproductive Medicine, São Paulo, SP, Brazil
5Division of Reproductive Medicine—Célula Mater, São Paulo, SP, Brazil
6Laboratory of Transplantation Immunobiology, Department of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, SP, Brazil
7Department of Science and Technology, Federal University of São Paulo, São José dos Campos, SP, Brazil
University of Pittsburgh, UNITED STATES
Competing Interests: The authors have declared that no competing interests exist.
Conceived and designed the experiments: JPSP PP CEC APLdO. Performed the experiments: MM LE SH MGN PP CEC JPSP AAdB MP ECdS WNB LBV FRG MCO-J. Analyzed the data: JPSP NOSC FA RdPV MZ APLdO. Contributed reagents/materials/analysis tools: JPSP NOSC FA RdPV MZ APLdO. Wrote the paper: JPSP MZ APLdO.* E-mail: moc.liamg@oriegilpa
Author information ? Article notes ? Copyright and License information ?
Received 2015 Feb 2; Accepted 2015 Aug 11.
 
Copyright notice
This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited
This article has been corrected. See PLoS One. 2015 September 25; 10(9): e0139294.Abstract
Cigarette smoke-induced chronic obstructive pulmonary disease is a very debilitating disease, with a very high prevalence worldwide, which results in a expressive economic and social burden. Therefore, new therapeutic approaches to treat these patients are of unquestionable relevance. The use of mesenchymal stromal cells (MSCs) is an innovative and yet accessible approach for pulmonary acute and chronic diseases, mainly due to its important immunoregulatory, anti-fibrogenic, anti-apoptotic and pro-angiogenic. Besides, the use of adjuvant therapies, whose aim is to boost or synergize with their function should be tested. Low level laser (LLL) therapy is a relatively new and promising approach, with very low cost, no invasiveness and no side effects. Here, we aimed to study the effectiveness of human tube derived MSCs (htMSCs) cell therapy associated with a 30mW/3J—660 nm LLL irradiation in experimental cigarette smoke-induced chronic obstructive pulmonary disease. Thus, C57BL/6 mice were exposed to cigarette smoke for 75 days (twice a day) and all experiments were performed on day 76. Experimental groups receive htMSCS either intraperitoneally or intranasally and/or LLL irradiation either alone or in association. We show that co-therapy greatly reduces lung inflammation, lowering the cellular infiltrate and pro-inflammatory cytokine secretion (IL-1?, IL-6, TNF-? and KC), which were followed by decreased mucus production, collagen accumulation and tissue damage. These findings seemed to be secondary to the reduction of both NF-?B and NF-AT activation in lung tissues with a concomitant increase in IL-10. In summary, our data suggests that the concomitant use of MSCs + LLLT may be a promising therapeutic approach for lung inflammatory diseases as COPD.Introduction
According to the World Health Organization (WHO), more than five million deaths per year are the result of direct use of tobacco, whereas more than 600 000 second hand smokers also perish from cigarette exposure (World Health Organization, 2014). According to the US Centers for Disease Control and Prevention, cigarette smoking is the dominant risk factor for the development of chronic obstructive pulmonary disease (COPD) and emphysema [1,2]. A wide range of dangerous agents are found in cigarette smoke and, aside from solid particles, it includes more than 4000 chemicals, of which at least 250 are known to be highly harmful and more than 50 are tumorigenic [2]. Altogether, these factors may induce airway inflammation, cellular recruitment, lung fibrosis, mucus hypersecretion and also cancer [2,3]. Among the features of cigarette smoke-induced pathology, parenchymal fibrosis and emphysema may be considered the most characteristic and also the most deleterious, resulting in a significant organic impairment and restricted life quality of the patients [3,4].Lung inflammation induced by cigarette smoke is characterized by an initial phase of inflammatory cells recruitment to the parenchymal space, matrix metalloproteinase (MMPs) activation, extracellular matrix degradation and tissue damage [3,4]. This is followed by the intense release of pro-inflammatory cytokines, such as IL-1 [5] and IL-6 [3], chemokines as CCL2 and CXCL1 and also lipid mediators, as PGI and LTB4, as reviewed [2]. These molecules are able to recruit more inflammatory cells, as neutrophils, macrophages and also CD4 and CD8 T lymphocytes of the Th1 and Tc1 IFN-?-secreting subtypes, respectively, perpetuating lung tissue damage [6]. Chronic exposure to cigarette noxious agents leads to the perpetuation of the inflammatory and fibrotic processes, with more and more infiltrating cells recruited, progressive elastin and collagen degradation, followed by parenchymal destruction and finally the establishment of lung emphysema [3,7]. Thus, campaigns to prevent cigarette smoking and to alert the population about its morbidity are extremely important. However the need for effective and also less expensive therapeutic approaches for the treatment of cigarette smoke-induced emphysematous patients is unquestionable. In this context, mesenchymal stromal cells (MSCs) therapy seem very promising, not only due to its regenerative capacity and immunomodulatory features, but also due to its low cost and relatively easy manipulation, as reviewed [4].

Adult MSCs are typically defined as undifferentiated multipotent cells endowed with the capacity for self-renewal and the potential to differentiate into several distinct cell lineages, as recently reviewed [8]. These cells may be obtained from different organs and tissues, as bone marrow [9,10], skeletal muscle [11], adipose tissue [12,13], dental pulp [14], umbilical cord [15,16], fallopian tubes [17] and other tissues. It is noteworthy that MSCs are well tolerated in vivo, either under syngeneic [18–19] or xenogenic conditions [20, 21, 22]. Concerning its immunomodulatory function, MSCs are known to: i) secrete anti-inflammatory cytokines; ii) express reduced levels of MHC and costimulatory molecules; iii) express the tryptophan-depleting enzyme indoleamine-2,3-dioxigenase [23] iv) induce T regulatory cells [18] and many others. Altogether, these mechanisms may be useful tools for the treatment of several chronic inflammatory diseases such as multiple sclerosis, arthritis, diabetes, lupus, and also COPD and emphysema [24–26]. Moreover, the possibility to use associated approaches able to boost the overall response must be investigated.

In this sense, Low Level Laser Therapy (LLLT) seem to be an interesting approach, specially due to its non-invasiveness, lack of secondary effects, low cost [27] and ability to promote stem-cell proliferation in vitro [28]. LLLT has already shown interesting results for different human diseases, as oral mucositis [29], coronary disease [30] and also in experimental models, as for muscle dystrophy [31], asthma [32] and articular inflammation in mice [33]. Actually, our group has recently demonstrated that LLLT is able to reduce lung inflammation both during the asthma model [32] as well as secondary to gut ischemia Among the possible mechanisms proposed, we have recently demonstrated that alveolar macrophages irradiated with LLL had augmented AMPc synthesis, and this is able to reduce NF-?B activation and thus IL-6 and IL-1 secretion [21]. In this context, we decided to evaluate the effectiveness of associating human tubal mesenchymal stromal cells (htMSCs) and LLLT using the murine model of cigarette smoke-induced COPD. It is worthy to mention that we have previously reported that htMSCs display stemness properties and also miogenic and adipogenic capacity to differentiate [17].

Here, our results demonstrate that the co-therapy with htMSCs and LLLT are very effective in lowering important pulmonary inflammatory parameters, as cytokine secretion, cellular infiltrate of leukocytes, mucus secretion, collagen deposition and the activation of important transcription factors, as NF-?B and NF-AT. Altogether, we show that the association of stem cells therapy with LLL irradiation are clinically beneficial, and thus, may be considered as further interesting therapeutic approach.

Materials and Methods
Cigarette Smoke–Induced Emphysema and Treatment

Female C57BL/6 mice (6–8 weeks) were exposed to cigarette smoke during 75 days by whole body exposure using an adapted protocol from Biselli et all [34]. All experiments were performed on day 76. Briefly, animals were exposed to 14 commercially available cigarettes (containing Alcatrão 13mg, nicotine 1,10 mg and carbon monoxide 10 mg each) divided on 7 cigarettes in the morning and 7 cigarettes in the afternoon and each session lasted for 30 minutes. LLL irradiation with diode laser (30mW/3J at 660 nm) was performed twice a day from day 60 to day 75 (one hour after each cigarette exposure) during 180 seconds on the skin over the right upper bronchus with a spot size of 0.785 cm2. htMSCs (1×106 cells) were infused on days 60 and 67. Experimental groups were divided in 7 groups, as follow: 1) Basal 2) COPD 3) COPD + LLL 4) COPD + MSCs (i.p) 5) COPD + MSCs (i.n) 6) COPD + MSCs (i.p.) + LLL 7) COPD + MSCs (i.n.) + LLL. We declare that all experiments were approved by the UNINOVE animal research committee (Comitê de Ètica no uso de Animais—CEUA—#AN005/2013) and human research committee (Comitê de Ética em Pesquisa com Seres Humanos (CEPSH) of University of Sao Paulo (# CEP-IBUSP-106/2010).

htMSCs Isolation and Culture

Human tubal tissue was obtained from women (age 35–53 years) submitted to hysterectomy or tubal ligation/resection surgery. Samples were collected during the proliferative phase from fertile women who had not undergone exogenous hormonal treatment within the last three months. htMSCs experiments were approved by both the ethics committee of the Biosciences Institute of the University of São Paulo and the UNINOVE–Universidade Nove de Julho. Each sample was collected in HEPES-buffered Dulbecco Modified Eagle Medium / Hams F-12 (DMEM/F-12; Invitrogen, Carlsbad, CA) with 100 IU/mL penicillin (Invitrogen) and 100 IU/mL streptomycin (Invitrogen, Carlsbad, CA), maintained at 4°C and processed within 24 hours period. All samples were washed twice in phosphate saline buffer (PBS, Gibco, Invitrogen, Carlsbad, CA), finely minced with a scalpel and transferred to a 50 mL tube, and dissolved with collagenase IV (Sigma-Aldrich) at 0,1% diluted in PBS (Invitrogen) for 15 minutes, at 37°C, in a water bath. Further, samples were washed with 10 mL of DMEM/F-12 and added with 15 ml of pure TripLE Express, (Invitrogen, Carlsbad, CA) for 15 minutes at 37°C in a water bath. Subsequently, TripLe was inactivated with DMEM/F-12 supplemented with 20% FBS, 100 IU/mL penicillin and 100 IU/mL streptomycin, and pelleted by centrifugation at 400g for five minutes at room temperature. Supernatant was removed with a sterile Pasteur pipette. Cells were then plated in DMEM/F-12 (5mL) supplemented with 20% FBS, 100 IU/mL penicillin and 100 IU/mL streptomycin in plastic flasks (25cm2), and maintained in incubator with controlled humidified atmosphere of 5% CO2 at 37°C. The medium used for expansion was initially changed every 72 hours and routinely replaced twice a week thereafter. Data concerning osteogenic, chondrogenic and adipogenic differentiation, as well as cellular phenotype were previously published by our group [17] demonstrating the stem cell potential of this population. Human tubal tissue was obtained after written and signed informed consent approved by Comitê de Ética em Pesquisa com Seres Humanos (CEPSH) of University of Sao Paulo (# CEP-IBUSP-106/2010) and maintained at the Division of Human Genome Research Center, Biosciences Institute, University of São Paulo, São Paulo–SP—Brazil.

Bronchoalveolar Lavage Fluid Cells and Cytospin

After euthanasia, bronchoalveolar lavage fluid was obtained as previously described [35]. Briefly, 1.5 mLs of PBS as intra-tracheally injected in the lungs and 1 mL was re-collected and centrifuged at 450 g 4°C during 5 minutes. Supernatants were discarged and the pellet suspended in the desired amount of PBS 2% FBS. Total cell count was performed in Newbauer chambers and differential cell count after cytospin protocol. For cytospin, aliquots of 100 ?L were centrifuged at 300 g for 5 minutes. Samples were stained by May-Grunwald-Giemsa method and 300 cells per sample were counted on a blind fashion.

Lung Mononuclear Cells and Flow Cytometry

Lungs were obtained after right heart perfusion of 10 mLs of cold PBS. The tissue was minced with scissors and incubated for 45 minutes at 37°C with 2,5 mg/mL of Colagenase D (Roche) in HBSS. To stop collagenase, cells were suspended in Ca2+/Mg2+ free HBSS and centrifuged at 450 g and 4°C for 5 minutes. Cellular pellet was suspended in 5–6 mLs of 37% Percoll (GE) in HBSS and gently laid over 5–6mLs of 70% Percoll in HBSS using 15 mLs conical falcon tubes. Samples were centrifuged at 950 g and 4°C for 30 minutes without breaks. Cell-containing ring was collected from the percoll gradient interface and suspended in PBS 2% FBS and centrifuged again at 450 g and 4°C for 5 minutes. Further, cells were suspended in PBS 2% FBS, counted and used as desired. For flow cytometry analysis, 5×105 cells were first incubated with 0,25 ?g of anti-CD16/32 at 4°C for 20 minutes to avoid inespecific binding. Cells were then stained with 0,25 ?g of anti-CD4 APC and 0,25 ?g of anti-CD8 PE in 25 ?L of PBS 2% FBS for 20 minutes at 4°C. In the next step, cells were washed twice with 200 ?L of PBS 2% and finally suspended in paraformaldehyde 1%. Cells were acquired in the flow cytometer Accuri C6 (BD Biosciences). 5×103 T CD4+ events were collected after elimination of cell doublets by FSH-height x FSC-area plot analysis.

Cytokine Secretion

BAL lavage fluid was centrifuged at 450 g and 4°C during 5 minutes. Supernatants were used to evaluate the presence of IL-1?, IL-6, IL-10, TNF-?, IFN-? and KC by the ELISA method according to manufacturer instruction (R&D System).

Histomorphometric Analysis of Inflammation, Collagen and Mucus

After perfusion, lung samples were maintained in formaldehyde 4% for inclusion in paraffin. Slices were stained with Siriud Red for collagen detection or Periodic Acid Schiff (PAS) for the detection of mucus. Were analyzed 15 airways per each animal under a 400X magnification. H&E staining was used for the analysis of peribronchial infiltrate and mean linear intercept measurement [36]. Morphological features were analyzed using the Image Pro Plus 4.5 (Media Cybernetics, Rockville, MD, USA).

Peribronchial Inflammation

The area between the airway basal membrane and the airway adventitia was quantified using the software Image Pro-Plus and the number of mononuclear and polymorphonuclear cells was quantified in this area according to the morphological criteria. The results were expressed as the number of mononuclear and polymorphonuclear cells per square millimeter.

Mucus Production and Collagen Fibers Deposition

For the analysis of mucus production, the epithelium area of the whole airway, 15 airways per mouse were quantified. The positive stained area in the epithelium area was quantified and the results were expressed as the percentage of positive epithelial area. For the analysis of collagen fibers deposition in the airways wall, the area between the outlayer of epithelium until airway adventitia were measured and the amount of positive stained area was calculated. The results were expressed as percentage aof collagen fibers in the airway wall [37–39].

Immunohistochemistry

For immunohistochemistry analysis, all samples were previously submitted to deparaffinization. Endogenous peroxidase activity was blocked with H2O2 3% three times during 10 minutes. Samples were then washed and blocked with bovine serum albumin 10% during 1 hour and then incubated with primary rabbit anti-mouse IgG antibodies: i) anti-NF?B (Santa Cruz, CA) at 1:500; ii) anti-NF-AT 1:500 (Santa Cruz, CA) and iii) anti-IL-10 (Santa Cruz, CA) at 1:500 during 2 hours at room temperature. Samples were washed twice with TBS- BSA 10% and then incubated with secondary antibody goat anti-Rabbit IgG at 1:1000 for 1 hour. After washing samples twice with BSA 2% diaminobenzidine (DAB) was added. Finally samples were washed again and counter stained with H&E. Slides were analyzed under light microscope on a blind fashion.

Statistical Analysis

All analysis were performed using the GraphPad Prism software (GraphPad Software, Inc). For parametric analysis we used one-way ANOVA followed by Tukey post-test whereas for non-parametric analysis, we used Kruskal-Wallis followed by Dunn´s post-test. All groups were compared to COPD control group. Differences were considered significant when p<0,05.

Results
htMSCs and LLLT Reduce Bronchoalveolar Lavage Fluid Cellularity in the Lungs of COPD Animals

To evaluate whether htMSCs and/or LLLT were effective in reducing lung inflammation after cigarette smoke-induced COPD, we first analyzed BAL fluid cellularity from all experimental groups for total and differential cell counts. Interestingly, a significant reduction in total cell counts, macrophages and neutrophils was observed only when htMSCs were associated with LLL therapy, although a trend is observed in all other groups, even for LLL alone (Fig 1A, 1B and 1C). However, we had a discrepant result for lymphocyte counts, as we observed that only htMSCs by i.p. delivery was able to reduce its amount. Association with LLLT did not changed the result. It is worthy to mention that the lowest amount of total cells (Fig 1A), macrophages (Fig 1B) and neutrophils (Fig 1C) was observed in the group of htMSCs associated with LLLT, either i.p or i.n, demonstrating that association with LLLT is more important for these features than the route of htMSCs delivery. It is also noteworthy that such reduction in BAL cellularity reached levels no different from naïve controls. Moreover, intraperitoneal delivery of htMSCs seemed more effective than intranasal delivery in reducing neutrophil and lymphocyte infiltration.

Fig 1
Fig 1
Reduced cellular infiltration in the lungs of htMSC and LLLT treated COPD animals.
Next, we performed flow cytometry analysis in the BAL fluid from all experimental groups to evaluate changes in the frequency of CD4+ and CD8+ T cells in the lungs. This could be indicative, either of a reduced T cell activation in the lymph nodes, or an impairment in cell migration to the target organ. In fact, corroborating findings from Fig 1 for polymorphonuclear and total cells, the frequency of both T CD4+ (Fig 2A and 2B) and T CD8+ (Fig 2C and 2D) cells were significantly reduced irrespective of the therapeutic approach used, i.e, htMSCs alone or in association with LLLT. However, no differences were observed, when lung tissue was analyzed for perivascular infiltrate by immunohistochemistry (S1 Fig).

Fig 2
Fig 2
Reduction of T CD4+ and T CD8+ T cells infiltration in the lungs of htMSC and/or LLLT treated animals.
htMSCs and/or LLLT Reduce Inflammatory Cytokines in the BAL Fluid of COPD Animals

We next analyzed the amount of cytokines as IL-1?, IL-6, TNF-?, IFN-? and the chemockine KC, to evaluate whether reduced cellularity in the lungs of our experimental culminated also in reduction of inflammatory mediators. Our results show that IL-1? was reduced when htMSCs either ip. or i.n. were associated with LLLT, although htMSCs i.n alone also reached significance (Fig 3C). The same pattern was observed for TNF-? (Fig 3A). KC, an important chemokine for the recruitment of neurotrophils, was reduced in all groups irrespective of the treatment (Fig 3F). Interestingly, IL-6 was reduced only when htMSCs were associated with LLL (Fig 3B) whereas no difference was observed for IFN-? (Fig 3D) Very interestingly, the anti-inflammatory cytokine IL-10 was up-regulated only with LLL irradiation alone (Fig 3E).

Fig 3
Fig 3
Reduced cytokine secretion in the lungs of htMSC and/or LLLT treated animals.
htMSCs and/or LLLT Reduced Mucus Secretion and Alveolar Enlargement in the Lungs of COPD Animals

Another important feature in the pathophysiology of COPD is the intense secretion of mucus, resulting in reduced airway lumen, airflow and breathlessness. Thus, we next evaluated whether either htMSCs or LLL therapies, associated or not, were able to reduce the amount of mucus observed in the airways. In fact, our data demonstrated significantly decreased amount of mucus in the bronchi of all experimental animals under therapeutic protocols, with the exception of the LLLT alone. Significant differences reached from 10 to 15-fold reduction when experimental groups were compared to COPD control. On the other hand, when associated with htMSCs, either nasally or intraperitoneally, LLL therapy restored mucus secretion to basal levels, with no difference from naïve animals (Fig 4A and 4B). We next decided to verify some possible effects of htMSCs and LLLT in the enlargement of lung parenchyma, which reflects the destruction of the alveolar septa. As indicated by S2 Fig, mean linear intercept measures were significantly reduced in the COPD + MSC(ip)+LLLT group when compared to control animals, indicating reduced lung damage.

Fig 4
Fig 4
Reduced mucus secretion in the lungs of htMSC and/or LLLT treated animals.
Collagen Deposition Was Reduced Only When htMSCs Were Associated with LLL

Aside from mucus secretion, collagen deposition is also considered an important marker for COPD. Therefore, we also evaluated the amount of peribronchial collagen deposition through Sirius Red methodology (Fig 5A and 5B). Very surprisingly, a significant collagen decrease of around 4-fold was observed only when LLLT was associated with htMSCs intranasally. Although there was a trend for the htMSCs (i.p) + LLLT, none of the other groups reached statistical significance (Fig 5B).

Fig 5
Fig 5
Reduced collagen deposition only in the lungs of htMSC i.n and LLL treated animals.
htMSCs and/or LLLT Reduces NF-?B and NF-AT Transcription Factors Expression in the Lungs of COPD Animals

Most pro-inflammatory cytokines evaluated, such as IL-1, IL-6 and TNF-?, have their transcription under the control of the transcription factor NF-?B. Moreover, these cytokines may also signal through NF-?B after engagement with its receptors cognate recptors. NF-?B is found in a great variety of cell types, as macrophages and neutrophils, but also T lymphocytes. Thus, with the aim to correlate the overall reduction in BAL cellularity and pro-inflammatory cytokine secretion with a reduction in NF-?B expression, we sought to evaluate its expression by immunohistochemistry of the perialveolar space of COPD animals (Fig 6). Corroborating the reduction in pro-inflammatory cytokine secretion (Fig 3C), all groups displayed significantly reduced NF-?B staining in the target tissue when compared to the COPD control group (Fig 6B). Differences vary from 2 to 3-fold depending on the treatment.

Fig 6
Fig 6
Reduced total NF-B staining in the lungs of htMSC i.n and/or LLLT treated animals.
Another transcription factor also important for the activation of the immune cells is the NF-AT, which is mostly expressed on T cells, both CD4+, CD8+ and NK cells, and also in the lung tissue, as shown by the literature [40]. NF-AT is the main transcription factor involved in the synthesis of IL-2 and its active form is the non-phosphorilated state, as reviewed [41]. It was very surprising to notice that LLLT by itself was already able to significantly reduce NF-AT activation (Fig 7A). This reduction was no different from that observed when LLL was associated with htMSCs i.n. (Fig 7A).

Fig 7
Fig 7
Reduced total NF-AT staining in the lungs of htMSC i.n and/or LLLT treated animals.
LLL Therapy Boosts IL-10 Secretion in Lung Tissue

In order to confirm the data obtained by the ELISA method, and concerning its importance as an anti-inflammatory factor, we also evaluated by immunohistochemistry the amount of IL-10 in lung tissue of the experimental groups. Interestingly, LLL irradiation per se was able to increase the amount of epithelial IL-10 (S3 Fig) whereas htMSCs i.p. was not able to so. Consistently, this was also observed only when htMSCs were associated with LLL therapy, although htMSCs i.n. alone could also up-regulate this cytokine (S3 Fig).

Discussion
In the present research we show that the treatment with htMSCs associated with LLL irradiation significantly reduced lung inflammation in mice with cigarette smoke-induced COPD. Many of the evaluated parameters, such as BAL cellularity, pro-inflammatory cytokine secretion, perivascular infiltrate and the presence of inflammatory transcription factors, as NF-?B and NF-AT were significantly reduced. This was associated with a better maintenance of tissue integrity when compared to COPD untreated controls, evidenced by reduced mucus secretion, collagen deposition and tissue damage. It is worthy to mention that these features are greatly responsible for tecidual destruction and further decline in patient´s life quality, as it greatly reduces airflow, lung complacence and lowering pO2 [2,42]. Moreover, our findings also demonstrate that the route of administration of htMSCs, i.e. intraperitoneal vs. intranasal, were both effective in suppressing the disease, although with some peculiarities. For instance, intranasal delivery was more effective in reducing the presence of NF-AT as well as collagen deposition when associated with LLL therapy, which was not observed after intra-peritoneal injection. This might be explained by a local regenerative/suppressive mechanism, as the cells were directly delivered to the lungs. For intra-peritoneal route, however, immune modulation on lymphoid organs, specially mediastinal lymph nodes is more likely.

Cigarette smoking-associated diseases, which may result in severe decrease of life quality represent an important social and economical burden, as billions of dollars are spent each year in the treatment of emphysematous patients worldwide. Despite the constant campaigns against tobacco, it is still the fourth leading cause of death in the United States [43]. Therefore, the need for a more efficient and yet cheaper therapeutic approach in the management of COPD patients is unquestionable. Cell therapy with MSCs has shown clinical benefits by us and other groups in conditions such as experimental autoimmune encephalomyelitis (EAE)[22], multiple sclerosis [44], chronic renal failure [13], ischemic cerebral stroke [9], myocardial stress [45] and even for COPD. In fact, several of our findings were corroborated by previous research, in which lung inflammation, weight loss and lung integrity were ameliorated after adipose-tissue MSCs treatment [26]. On the other hand, a recent clinical trial on MSCs have not shown promising results, as COPD patients treated with MSCs (Prochymal) had not shown improvement of lung function, as shown for forced expiratory volume (FEV1) and forced volume capacity (FVC). Interestingly however, it seemed that, corroborating our findings, lung inflammation was decreased, inferred by the lower level of C-reactive protein [46]. Our report however, although using an experimental model, is the first to associate htMSCs with LLL irradiation, considered a promising approach [27].

COPD is a chronic and obstructive disease of the lungs, result of the chronic exposure of the airways to the noxious agents found in cigarette smoke and droplets. It has been established that lung destruction directly correlates with the amount of total particulate matter found in each cigar [7]. Initial exposure leads to cellular infiltrate of neutrophils and blood-derived monocytes secreting pro-inflammatory and pro-fibrotic cytokines, as IL-1, IL-6, IL-12, TNF-? and TGF-?, along with chemokines, lipid mediators and several other molecules [2,3]. This is greatly accompanied by extracellular matrix degradation by MMP-1 (matrixmetalloproteinase-1) secreted by alveolar macrophages causing tissue destruction and emphysema [3]. Chronic exposure leads to intense T CD4 and T CD8 lymphocyte infiltration, and IFN-? is probably the most important T cell-derived cytokine. It is relevant that IFN-?-secreting T CD8 cells are also very important in the late phase of the disease [47]. Due to its pro-inflammatory function, and in association with other cytokines, as TNF-?, TGF-?, IFN-? induces important activation of both immune and parenchymal cells of the lung and finally leading to fibrosis and parenchymal destruction [2,3]. In this context, therapeutic approaches with the capacity to dampen such activation could greatly contribute to the maintenance of lung integrity and both MSCs and LLL therapy independently had already proven it [10,32]. However, we did not observe differences for IFN-?.

A growing body of evidence have shown several different immunosuppressive mechanisms of MSCS, such as: lowering the expression of MHC and costimulatory molecules; up-regulation of indoleamine-2,3-dioxigenase [19,48] and FAS-L expression [49]; expansion of Tregs [48]; block of IFN-? and IL-17 secretion [22], reduces tecidual caspase-3 activation [10] and many others, as reviewed [4]. Altogether, these mechanisms greatly impair immune cells activation, further avoiding or reducing tissue inflammation and destruction. However, although we may only speculate the mechanisms used by the htMSCs + LLL irradiation to suppress COPD, it is suitable to think that many of the aforementioned mechanisms may be taking place in our system.

Corroborating our findings, several other groups have already addressed the capacity of MSCs in modulating chronic diseases, including COPD. Bone marrow-derived MSCs were shown to significantly reduce tissue destruction in COPD mice by a mechanisms dependent on VEGF. Consistently, the group observed among many other cytokines, reduction in IL-1? and IL-6, in accordance with our data. More relevant was the fact that lung function, including inspiratory capacity and vital capacity were significantly higher when compared to control animals, which is again discrepant from data obtained in humans [10].

Adipose tissue-derived MSCs (ASCs) transplantation was also shown to be effective in improving COPD in mice reducing lung infiltration of inflammatory cells, as neutrophils and macrophages reduced associated with parenchymal destruction, as evaluated by active caspase-3 [12]. In line with our findings, the group also demonstrated reduced lung infiltration of inflammatory cells, as total cells, neutrophils and macrophages.

Associated with its immunosuppressive activity, it is also possible that MSCs exert some regenerative/reparative function in situ. For instance, intra-tracheal delivery of bone marrow-derived stem cells after hyperventilation-induced injury in rats greatly improved lung recovery, reducing cellular infiltrate of neutrophils and lymphocytes, pro-inflammatory cytokine secretion, associated with normal alveolar space [50]. In accordance, we have observed that intra-nasally treated animals have also displayed a significant reduction in several inflammatory markers, as cellular infiltrate and cytokine secretion. Moreover, we also observed amelioration in alveoli enlargement in animals treated with htMSCs(ip) + LLL, evidenced by mean linear intercept, but unfortunately lung function was not assessed by us. However, it is noteworthy that collagen deposition, a very significant factor for tissue fibrosis and lung emphysema, was significantly reduced when htMSCs were associated with LLLT. This may indicate that LLLT boosts htMSCs regerative capacity, however we may not describe the mechanisms so far.

LLLT started to be studied in the late 60`s, when reports had shown its use in improving hair growing in rats. Several other reports soon followed using different models, as wound healing and muscle regeneration. On the other hand, the effect of laser therapy is poorly understood. We have previously observed that the in vitro proliferative capacity of dental pulp derived mesenchymal stem-cells increased with low intensity laser application [28]. Interestingly, recent reports have shown that LLL therapy exert its function by increasing intracellular AMPc and thus suppressing important inflammatory transcription factors as NF-?B [51], which is consistent with our findings. In fact, the intriguing capacity of LLL therapy in suppressing the immune response has been previously shown by us and others, as mentioned. LLLT irradiation had the capacity to reduce joint inflammation in rats treated locally with papain. Consistently, irradiated groups demonstrated reduced cellular infiltration and also reduction in IL-1? and IL-6, corroborating our findings [33]. In the context of lung disease, we have previously published that LLLT reduces lung inflammation using OVA-induced lung inflammatory disease [32]. In this case, several parameters, as cellular infiltrate of the lungs, cytokine secretion, mucus secretion and collagen deposition were significantly reduced after irradiation and antigen challenge. As expected, many of these results were reproduced in the present research, and also it indicated that htMSCs and LLLT may act by an additive manner.

Aside from overall reduction in lung inflammation, another interesting observation was the reduction of the transcription factors NF-?B and NF-AT which could be secondary to the reduced infiltration of macrophages / neutrophils and lymphocytes in lung tissue, respectively. Moreover, we have also detected lower levels of IL-1? in BAL fluid, which is known to signal through IL-1r and thus induce NF-?B activation. However, we may not exclude the possibility that LLLT could also directly abrogate NF-AT activation, as observed when it was used alone. It is plausible to think that LLLT somehow changes intracellular Ca2+ levels, consequently modulating NF-AT activation by calcineurin / calmodulin. It is relevant that NF-AT, which may also be found in lung arteries, is responsible for up-regulating the expression of smooth muscle ?-actin and myosin heavy chain after hypoxia, a very important mechanism for chronic hypoxia-induced pulmonary vascular remodeling [52].

In summary, our results clearly indicate that, aside from lack of toxicity and other complications, both htMSCs and LLLT were shown to be safe for the treatment of COPD in an experimental model. More important, however, is the fact that although htMSCs and LLLT could act independently, some pathological parameters were effectively reduced when both therapies were associated. It may indicate an important additive effect that may be responsible for the overall reduction in lung inflammation and tissue destruction in cigarette smokers. Our results of the combined use of htMSCs and LLLT were effective in reducing inflammatory immune response and further overall destruction of lung parenchyma in experimental COPD. Thus, we encourage other groups to keep focus on the potential of MSCs in COPD, and also to consider the relevance of associating it with LLLT. This is not only to reinforce our data, but mainly to refine the therapeutic scheme with the aim to reach a more translational approach.

Conclusions
Our research highlights the important suppressive capacity of htMSCs in reducing overall lung inflammation during COPD in mice. Besides, we also observed a beneficial additive effect when htMSCs and LLLT are associated. Reduced lung cellularity and cytokine secretion, mucus production, collagen deposition and transcription factors activation are among the downregulated parameters. Interestingly, our data also show that this phenomenom is very consistent, irrespective of the MSCs administration via, i.e. intraperitoneal or intranasal. In summary, our data highlights the possibility of using this approach for the treatment of chronic inflammatory lung diseases.

Supporting Information
S1 Fig

Analysis of polymorphonuclear and mononuclear perivascular cells in the lungs of htMSC and/or LLLT treated animals.
COPD animals were submitted to therapeutic protocols as described in materials and methods. Further, all animals were euthanized and lungs were obtained for histomorphometric analysis. In (A) mononuclear and (B) polymorphonuclear cells per mm2 of tissue. Data representative from two experiments. n = 5–8 animals per group. One-way ANOVA.

(TIF)

Click here for additional data file.(125K, tif)
S2 Fig

Reduction in alveolar destruction in the lungs of htMSC and/or LLLT treated animals.
COPD animals were submitted to therapeutic protocols as described in materials and methods. Further, all animals were euthanized and lungs were obtained for mean linear intercept in central and peripheral measures of both lungs. In A) representative graphs and B) photomicrographs of haematoxylin- and eosin-stained pulmonary parenchyma. Data representative of two experiments. n = 5–8 animals per group. One-way ANOVA.

(TIF)

Click here for additional data file.(348K, tif)
S3 Fig

Increased IL-10 staining in the lungs of htMSC i.n and/or LLLT treated animals.
COPD animals were submitted to therapeutic protocols as described in materials and methods. Further, all animals were euthanized, lungs obtained and sections were stained with anti-IL-10. In A) representative graphs and B) photomicrographs of immunohistochemistry stained sections. Data representative of two experiments. n = 5–8 animals per group. One-way ANOVA.

(TIF)

Click here for additional data file.(366K, tif)
Go to:
Acknowledgments
We would like to thank Vanessa Roza da Silva for her great support with maintaining the experimental animals and Prof. Regiane Albertini for kindly sharing the LLL irradiator. We would like to thank also Eliane Gomes for technical support.
:
Funding Statement
Financial support from FAPESP (Fundação de Apoio à Pesquisa do Estado de São Paulo) – grants # 2011/18703-2 for Peron JPS and 2012/16498-5 for Ligeiro de Oliveira AP.

References
1. World Health Organiation. 2014. Available: http://www.who.int/mediacentre/factsheets/fs339/en/.
2. Morse D, Rosas IO. Tobacco smoke-induced lung fibrosis and emphysema. Annu Rev Physiol. 2014;76:493–513. doi: 10.1146/annurev-physiol-021113-170411 [PubMed]
3. Postma DS, Reddel HK, Ten Hacken NHT, van den Berge M. Asthma and Chronic Obstructive Pulmonary Disease: Similarities and Differences. Clin Chest Med. 2014;35(1):143–56. doi: 10.1016/j.ccm.2013.09.010 [PubMed]
4. Inamdar AC, Inamdar A a. Mesenchymal stem cell therapy in lung disorders: pathogenesis of lung diseases and mechanism of action of mesenchymal stem cell. Exp Lung Res. 2013; 39(8):315–27. doi: 10.3109/01902148.2013.816803 [PubMed]
5. Di Stefano A, Caramori G, Barczyk A, Vicari C, Brun P, Zanini A, et al. Innate immunity but not NLRP3 inflammasome activation correlates with severity of stable COPD. Thorax. 2014;1 doi: 10.1136/thoraxjnl-2012-203062 [PMC free article] [PubMed]
6. Longhini-Dos-Santos N, Barbosa-de-Oliveira VA, Kozma RH, Faria CA De, Stessuk T, Frei F, et al. Cell therapy with bone marrow mononuclear cells in elastase-induced pulmonary emphysema. Stem Cell Rev 2014;9(2):210–8. [PubMed]
7. John G, Kohse K, Orasche J, Reda A, Schnelle-Kreis J, Zimmermann R, et al. The composition of cigarette smoke determines inflammatory cell recruitment to the lung in COPD mouse models. Clin Sci. 2014;126(3):207–21. doi: 10.1042/CS20130117 [PMC free article] [PubMed]
8. Clark EA, Kalomoiris S, Nolta JA FF. Concise review: MicroRNA function in multipotent mesenchymal stromal cells. Stem Cells. 2014;32(5):1074–82. [PubMed]
9. Zhang X, Du F, Yang D, Yu C, Huang X, Liu W, et al. Transplanted bone marrow stem cells relocate to infarct penumbra and co-express endogenous proliferative and immature neuronal markers in a mouse model of ischemic cerebral stroke. BMC Neurosci 2010;11(1):138. [PMC free article] [PubMed]
10. Guan X-J, Song L, Han F-F, Cui Z-L, Chen X, Guo X-J, et al. Mesenchymal stem cells protect cigarette smoke-damaged lung and pulmonary function partly via VEGF-VEGF receptors. J Cell Biochem. 2013;114(2):323–35. doi: 10.1002/jcb.24377 [PubMed]
11. Tamaki T, Hirata M, Soeda S, Nakajima N, Saito K, Nakazato K, et al. Preferential and comprehensive reconstitution of severely damaged sciatic nerve using murine skeletal muscle-derived multipotent stem cells. PLoS One 2014;9(3):e91257 doi: 10.1371/journal.pone.0091257 [PMC free article] [PubMed]
12. Schweitzer KS, Johnstone BH, Garrison J, Rush NI, Cooper S, Traktuev DO, et al. Adipose stem cell treatment in mice attenuates lung and systemic injury induced by cigarette smoking. Am J Respir Crit Care Med 2011;183(2):215–25. doi: 10.1164/rccm.201001-0126OC [PMC free article] [PubMed]
13. Semedo P, Correa-Costa M, Antonio Cenedeze M, Maria Avancini Costa Malheiros D, Antonia dos Reis M, Shimizu MH, et al. Mesenchymal stem cells attenuate renal fibrosis through immune modulation and remodeling properties in a rat remnant kidney model. Stem Cells. 2009;27(12):3063–73. doi: 10.1002/stem.214 [PubMed]
14. Kerkis I, Ambrosio CE, Kerkis A, Martins DS, Zucconi E, Fonseca S a S, et al. Early transplantation of human immature dental pulp stem cells from baby teeth to golden retriever muscular dystrophy (GRMD) dogs: Local or systemic? J Transl Med. 2008;6:35 doi: 10.1186/1479-5876-6-35 [PMC free article] [PubMed]
15. Zucconi E, Vieira NM, Bueno CR, Secco M, Jazedje T, Costa Valadares M, et al. Preclinical studies with umbilical cord mesenchymal stromal cells in different animal models for muscular dystrophy. J Biomed Biotechnol. 2011;2011:715251 doi: 10.1155/2011/715251 [PMC free article] [PubMed]
16. Secco M, Bueno C, Vieira NM, Almeida C, Pelatti M, Zucconi E, et al. Systemic delivery of human mesenchymal stromal cells combined with IGF-1 enhances muscle functional recovery in LAMA2 dy/2j dystrophic mice. Stem Cell Rev. 2013;9(1):93–109. doi: 10.1007/s12015-012-9380-9 [PubMed]
17. Jazedje T, Perin PM, Czeresnia CE, Maluf M, Halpern S, Secco M, et al. Human fallopian tube: a new source of multipotent adult mesenchymal stem cells discarded in surgical procedures. J Transl Med. 2009;7:46 doi: 10.1186/1479-5876-7-46 [PMC free article] [PubMed]
18. Jumah MA Al, Abumaree MH. The Immunomodulatory and Neuroprotective Effects of Mesenchymal Stem Cells (MSCs) in Experimental Autoimmune Encephalomyelitis (EAE): A Model of Multiple Sclerosis (MS). 2012;9298–331. [PMC free article] [PubMed]
19. Croitoru-Lamoury J, Lamoury FMJ, Caristo M, Suzuki K, Walker D, Takikawa O, et al. Interferon-? regulates the proliferation and differentiation of mesenchymal stem cells via activation of indoleamine 2,3 dioxygenase (IDO). PLoS One. 2011;6(2):e14698 doi: 10.1371/journal.pone.0014698 [PMC free article] [PubMed]
20. Park M, Kim Y-H, Woo S-Y, Lee HJ, Yu Y, Kim HS, et al. Tonsil-derived Mesenchymal Stem Cells Ameliorate CCl4–induced Liver Fibrosis in Mice via Autophagy Activation. Sci Rep. 2015;5:8616 doi: 10.1038/srep08616 [PMC free article] [PubMed]
21. Lima FM De, Aimbire F, Miranda H, Vieira RDP. Low-Level Laser Therapy Attenuates the Myeloperoxidase Activity and Inflammatory Mediator Generation in Lung Inflammation Induced By Gut Ischemia and Reperfusion?: A Dose-Response Study. 2014;5(2):63–70. [PMC free article] [PubMed]
22. Peron JPS, Jazedje T, Brandão WN, Perin PM, Maluf M, Evangelista LP, et al. Human endometrial-derived mesenchymal stem cells suppress inflammation in the central nervous system of EAE mice. Stem Cell Rev. 2012;8(3):940–52. doi: 10.1007/s12015-011-9338-3 [PubMed]
23. Jones BJ, Brooke G, Atkinson K, McTaggart SJ. Immunosuppression by placental indoleamine 2,3-dioxygenase: a role for mesenchymal stem cells. Placenta. 2007;28(11–12):1174–81. [PubMed]
24. Stessuk T, Ruiz MA, Greco OT, Bilaqui A, Ribeiro-Paes MJDO, Ribeiro-Paes JT. Phase I clinical trial of cell therapy in patients with advanced chronic obstructive pulmonary disease: follow-up of up to 3 years. Rev Bras Hematol Hemoter. 2013;35(5):352–7. doi: 10.5581/1516-8484.20130113 [PMC free article] [PubMed]
25. Bluestone J a, Thomson AW, Shevach EM, Weiner HL. What does the future hold for cell-based tolerogenic therapy? Nat Rev Immunol. 2007. August;7(8):650–4. [PubMed]
26. Schweitzer KS, Johnstone BH, Garrison J, Rush NI, Cooper S, Traktuev DO, et al. Adipose stem cell treatment in mice attenuates lung and systemic injury induced by cigarette smoking. Am J Respir Crit Care Med. 2011;183(2):215–25. doi: 10.1164/rccm.201001-0126OC [PMC free article] [PubMed]
27. Lin F, Josephs SF, Alexandrescu DT, Ramos F, Bogin V, Gammill V, et al. Lasers, stem cells, and COPD. J Transl Med. 2010;8:16 doi: 10.1186/1479-5876-8-16 [PMC free article] [PubMed]
28. Eduardo CP, Fernanda FP, Bueno DF, Freitas PM, Marques MM, Passos-Bueno MR ZM. Stem cell proliferation under low intensity laser irradiation: a preliminary study. Lasers Surg Med. 2008;40(6). [PubMed]
29. Oton-Leite AF, Elias LSA, Morais MO, Pinezi JCD, Leles CR, Silva MAGS, et al. Effect of low level laser therapy in the reduction of oral complications in patients with cancer of the head and neck submitted to radiotherapy. Spec Care Dentist. 2012;33(6):294–300. doi: 10.1111/j.1754-4505.2012.00303.x [PubMed]
30. Plass C a, Wieselthaler GM, Podesser BK, Prusa AM. Low-level-laser irradiation induces photorelaxation in coronary arteries and overcomes vasospasm of internal thoracic arteries. Lasers Surg Med. 2012;44(9):705–11. doi: 10.1002/lsm.22075 [PubMed]
31. Leal-Junior ECP, de Almeida P, Tomazoni SS, de Carvalho PDTC, Lopes-Martins RÁB, Frigo L, et al. Superpulsed Low-Level Laser Therapy Protects Skeletal Muscle of mdx Mice against Damage, Inflammation and Morphological Changes Delaying Dystrophy Progression. PLoS One. 2014;9(3):e89453 doi: 10.1371/journal.pone.0089453 [PMC free article] [PubMed]
32. Silva VR, Marcondes P, Silva M, Villaverde AB, Castro-Faria-Neto HC, Vieira RP, et al. Low-level laser therapy inhibits bronchoconstriction, Th2 inflammation and airway remodeling in allergic asthma. Respir Physiol Neurobiol.2014;194:37–48. doi: 10.1016/j.resp.2014.01.008 [PubMed]
33. Alves AC, Vieira R, Leal-Junior E, dos Santos S, Ligeiro AP, Albertini R, et al. Effect of low-level laser therapy on the expression of inflammatory mediators and on neutrophils and macrophages in acute joint inflammation. Arthritis Res Ther. 2013;15(5):R116 [PMC free article] [PubMed]
34. Biselli PJC, Lopes FDTQS, Moriya HT, Rivero DHRF, Toledo a. C, Saldiva PHN, et al. Short-term exposure of mice to cigarette smoke and/or residual oil fly ash produces proximal airspace enlargements and airway epithelium remodeling. Brazilian J Med Biol Res. 2011;44(5):460–8. [PubMed]
35. Lino-dos-Santos-Franco A, Domingos HV, Oliveira APL De, Breithaupt-Faloppa AC, Peron JPS, Bolonheis S, et al. Differential effects of formaldehyde exposure on the cell influx and vascular permeability in a rat model of allergic lung inflammation. Toxicol Lett. 2010;197(3):211–8. [PubMed]
36. Hizume DC, Toledo AC, Moriya HT, Saraiva-Romanholo BM, Almeida FM, Arantes-Costa FM, et al. Cigarette smoke dissociates inflammation and lung remodeling in OVA-sensitized and challenged mice. Respir Physiol Neurobiol [Internet]. Elsevier B.V.; 2012;181(2):167–76. [PubMed]
37. Vieira RP, Claudino RC, Duarte ACS, Santos ÂBG, Perini A, Faria Neto HCC, et al. Aerobic exercise decreases chronic allergic lung inflammation and airway remodeling in mice. Am J Respir Crit Care Med. 2007;176(9):871–7. [PubMed]
38. Vieira RP, Müller T, Grimm M, Von Gernler V, Vetter B, Dürk T, et al. Purinergic receptor type 6 contributes to airway inflammation and remodeling in experimental allergic airway inflammation. Am J Respir Crit Care Med. 2011;184(2):215–23. doi: 10.1164/rccm.201011-1762OC [PubMed]
39. Silva RA, Vieira RP, Perini A, Mauad T, Martins MA. inflammation and remodelling in an. 2010;35(5):994–1002. [PubMed]
40. Rastrick JMD, Stevenson CS, Eltom S, Grace M, Davies M, Kilty I, et al. Cigarette smoke induced airway inflammation is independent of NF-?B signalling. PLoS One. 2013;8(1):e54128 doi: 10.1371/journal.pone.0054128 [PMC free article] [PubMed]
41. Vashishta A, Habas A, Pruunsild P, Zheng J-J, Timmusk T, Hetman M. Nuclear factor of activated T-cells isoform c4 (NFATc4/NFAT3) as a mediator of antiapoptotic transcription in NMDA receptor-stimulated cortical neurons. J Neurosci. 2009;29(48):15331–40. doi: 10.1523/JNEUROSCI.4873-09.2009 [PMC free article] [PubMed]
42. Kubo H. Concise review: clinical prospects for treating chronic obstructive pulmonary disease with regenerative approaches. Stem Cells Transl Med. 2012;1(8):627–31. doi: 10.5966/sctm.2012-0065 [PMC free article] [PubMed]
43. Rennard SI. Looking at the Patient—Approaching the Problem of COPD Improving Online Access to Medical Information for Low-Income Countries. 2004;965–6. [PubMed]
44. Einstein O, Grigoriadis N, Mizrachi-Kol R, Reinhartz E, Polyzoidou E, Lavon I, et al. Transplanted neural precursor cells reduce brain inflammation to attenuate chronic experimental autoimmune encephalomyelitis. Exp Neurol. 2006;198(2):275–84. [PubMed]
45. Figeac F, Lesault P-F, Le Coz O, Damy T, Souktani R, Trébeau C, et al. Nanotubular crosstalk with distressed cardiomyocytes stimulates the paracrine repair function of mesenchymal stem cells. Stem Cells. 2014;32(1):216–30. doi: 10.1002/stem.1560 [PubMed]
46. Weiss DJ, Casaburi R, Flannery R. A Placebo-Controlled, Randomized Trial of. 2014; [PubMed]
47. Barnes PJ. Review series The cytokine network in asthma and chronic obstructive pulmonary disease. 2008;118(11). [PMC free article] [PubMed]
48. Ge W, Jiang J, Arp J, Liu W, Garcia B, Wang H. Regulatory T-cell generation and kidney allograft tolerance induced by mesenchymal stem cells associated with indoleamine 2,3-dioxygenase expression. Transplantation. 2010. December 27;90(12):1312–20. doi: 10.1097/TP.0b013e3181fed001 [PubMed]
49. Atsuta I, Liu S, Miura Y, Akiyama K, Chen C, An Y, et al. Mesenchymal stem cells inhibit multiple myeloma cells via the Fas/Fas ligand pathway. Stem Cell Res Ther. 2013;4(5):111 [PMC free article] [PubMed]
50. Curley GF, Ansari B, Hayes M, Devaney J, Masterson C, Ryan A, et al. Effects of intratracheal mesenchymal stromal cell therapy during recovery and resolution after ventilator-induced lung injury. Anesthesiology. 2013;118(4):924–32. doi: 10.1097/ALN.0b013e318287ba08 [PubMed]
51. Wu JY, Chen CH, Wang CZ, Ho ML, Yeh ML, Wang YH. Low-Power Laser Irradiation Suppresses Inflammatory Response of Human Adipose-Derived Stem Cells by Modulating Intracellular Cyclic AMP Level and NF-KB Activity. PLoS One. 2013;8(1):1–9. [PMC free article] [PubMed]
52. De Frutos S, Spangler R, Alò D, Bosc LVG. NFATc3 mediates chronic hypoxia-induced pulmonary arterial remodeling with alpha-actin up-regulation. J Biol Chem. 2007;282(20):15081–9. [PMC free article] [PubMed]

 
Vopr Kurortol Fizioter Lech Fiz Kult. 2015 Mar-Apr;92(2):9-11.

The influence of low-intensity laser irradiation of blood on the lactoferrin level in the patients presenting with community-acquired pneumonia.

[Article in Russian]
Gabueva AA, Burduli NM.

Abstract

The objective of the present study was to estimate the influence of low-intensity laser irradiation of blood on the levels of lactoferrin in the patients presenting with community-acquired pneumonia.

MATERIALS AND METHODS:

All the patients were divided into two groups. Those comprising the control group received only medicamental therapy. The standard treatment of the patients in the second group was supplemented by intravenous laser irradiation of blood (IVLIB-405 technique). Each irradiation session performed in a continuous mode lasted 5-7 minutes, with the total treatment course consisting of 7 daily procedures. Lactoferrin was determined before and after the treatment by the enzyme immunoassay with the use of “Lactoferrin Strip” kits (“Vektor-Best- Yug” JSC, Russia).

RESULTS:

Analysis of the results of the study revealed a significant decrease in the blood lactoferrin level down to the normal range in the patients treated with the use of laser therapy.

CONCLUSION:

The application of intravenous laser irradiation of blood for the combined treatment of the patients with community-acquired pneumonia was accompanied by normalization of the blood lactoferrin level and the improvement of the clinical course of the disease.

Lasers Med Sci. 2014 Nov 21. [Epub ahead of print]

Phototherapy with combination of super-pulsed laser and light-emitting diodes is beneficial in improvement of muscular performance (strength and muscular endurance), dyspnea, and fatigue sensation in patients with chronic obstructive pulmonary disease.

Miranda EF1, de Oliveira LV, Antonialli FC, Vanin AA, de Carvalho PD, Leal-Junior EC.

Author information

  • 1Post-Graduate Program in Biophotonics Applied to Health Sciences, Nove de Julho University, Rua Vergueiro, 235, 01504-001, São Paulo, SP, Brazil.

Abstract

Phototherapy is an electrophysical intervention being considered for the retardation of peripheral muscular fatigue usually observed in chronic obstructive pulmonary disease (COPD). The objective of this study was to evaluate the acute effects of combination of super-pulsed laser and light-emitting diodes phototherapy on isokinetic performance in patients with COPD. Thirteen patients performed muscular endurance tests in an isokinetic dynamometer. The maximum voluntary isometric contraction (MVIC), peak torque (PT), and total work (TW) of the non-dominant lower limb were measured in two visits. The application of phototherapy or placebo (PL) was conducted randomly in six locations of femoral quadriceps muscle by using a cluster of 12 diodes (4 of 905 nm super-pulsed lasers, 0.3125 mW each; 4 of 875 nm LEDs, 17.5 mW each; and 4 of 640 nm LEDs, 15 mW each, manufactured by Multi Radiance Medical™). We found statistically significant increases for PT (174.7?±?35.7 N?·?m vs. 155.8?±?23.3 N?·?m, p?=?0.003) and TW after application of phototherapy when compared to placebo (778.0?±?221.1 J vs. 696.3?±?146.8 J, p?=?0.005). Significant differences were also found for MVIC (104.8?±?26.0 N?·?m vs. 87.2?±?24.0 N?·?m, p?=?0.000), sensation of dyspnea (1 [0-4] vs. 3 [0-6], p?=?0.003), and fatigue in the lower limbs (2 [0-5] vs. 5 [0.5-9], p?=?0.002) in favor of phototherapy. We conclude that the combination of super-pulsed lasers and LEDs administered to the femoral quadriceps muscle of patients with COPD increased the PT by 20.2 % and the TW by 12 %. Phototherapy with a combination of super-pulsed lasers and LEDs prior to exercise also led to decreased sensation of dyspnea and fatigue in the lower limbs in patients with COPD.

Vopr Kurortol Fizioter Lech Fiz Kult. 2014 Jul-Aug;(4):3-6.

The application of chromo- and laserotherapy for the treatment of the patients presenting with chronic obstructive pulmonary disease and concomitant arterial hypertension.

[Article in Russian]
Nikitin AV, Marks SI.

Abstract

This article gives evidence of the effectiveness of chromo- and laser therapy (using infrared and green wavelenth radiation) in combination with basal pharmacotherapy in the patients presenting with combined pathology. The analysis of the data obtained indicates that the proposed approach makes it possible to accelerate normalization of the clinical characteristics, reduce arterial pressure, improve the parameters of external respiration, and increase the duration of remission periods.

Photomed Laser Surg. 2010 Dec;28(6):763-71.

Low-Level Laser Therapy Associated to N-Acetylcysteine Lowers Macrophage Inflammatory Protein-2 (MIP-2) mRNA Expression and Generation of Intracellular Reactive Oxygen Species in Alveolar Macrophages.

de Lima FM, Villaverde AB, Albertini R, de Oliveira AP, Neto HC, Aimbire F.

1 Institute of Research and Development , IP&D, UNIVAP, São José dos Campos, São Paulo, Brazil .

Abstract

Abstract Objective: The aim of this work was to investigate the low-level laser therapy (LLLT) effect on alveolar macrophages (AM) activated by oxidative stress and lipopolysaccharide (LPS).

Background data: LLLT has been reported to actuate positively relieving the late and early symptoms of airway and lung inflammation. It is not known if the increased MIP-2?mRNA expression and intracellular reactive oxygen species (ROS) generation observed in acute lung inflammation (ALI) can be influenced by LLLT.

Materials and Methods: Rat AM cell line (AMJ2-C11) was cultured with LPS or H(2)O(2) and laser irradiated. MIP-2?mRNA and ROS production in the AM were evaluated by Real Time-PCR and the 2′,7′-dichlorofluorescin diacetate (DCFH-DA) respectively. The NF-?B protein in the AM was measured by the enzyme linked immunoassay method. To investigate the antioxidant effect of laser, the AM were prebathed with N-acetylcysteine (NAC) and then irradiated with laser. LLLT was also studied in the presence of an inhibitor of NF-?B (BMS 205820). In addition, the effect of LLLT on NF-?B protein was investigated.

Results: LLLT attenuated the MIP-2?mRNA expression and intracellular ROS generation after LPS or H(2)O(2). When the AM were pretreated with NAC, the laser effect was potentiated. BMS 205820 suppresses the effect of LLLT on MIP-2?mRNA expression and ROS generation, stimulated by LPS or H(2)O(2). On NF-?B transcription factor, both the LLLT and NAC reduced this protein in the AM exposed to LPS or H(2)O(2). The synergistic effect between LLLT and NAC on the reduction the NF-?B was also evidenced.

Conclusion: Results indicate that there is a synergistic action of LLLT with NAC on MIP-2?mRNA expression from LPS- or H(2)O(2)-stimulated AM, and that both ROS intracellular generation and NF-kB signaling seem to be involved.

J Transl Med. 2010 Feb 16;8(1):16. [Epub ahead of print]

Lasers, stem cells, and COPD.

Lin F, Josephs SF, Alexandrescu DT, Ramos F, Bogin V, Gammill V, Dasanu CA, De Necochea-Campion R, Patel AN, Carrier E, Koos DR.

ABSTRACT: The medical use of low level laser (LLL) irradiation has been occurring for decades, primarily in the area of tissue healing and inflammatory conditions. Despite little mechanistic knowledge, the concept of a non-invasive, non-thermal intervention that has the potential to modulate regenerative processes is worthy of attention when searching for novel methods of augmenting stem cell-based therapies. Here we discuss the use of LLL irradiation as a photoceutical for enhancing production of stem cell growth/chemoattractant factors, stimulation of angiogenesis, and directly augmenting proliferation of stem cells. The combination of LLL together with allogeneic and autologous stem cells, as well as post-mobilization directing of stem cells will be discussed.

Ter Arkh. 2010;82(3):36-8.

Changes in plasma hemostatic parameters under intravascular laser irradiation of blood in patients with community-acquired pneumonia.

[Article in Russian]

Burduli NM, Pilieva NG.

Abstract

AIM: To study the time course of changes in the activity of the protein C system and other hemostatic parameters under intravascular laser irradiation of blood (ILIB) in patients with community-acquired pneumonia (CAP). SUBJECTS AND METHODS: One hundred and forty patients aged 17 to 62 years (mean 39.5 +/- 8.4 years) with CAP were examined. A control group (n = 40) received conventional drug therapy; the study group (n = 100) had a course of ILIB in addition to conventional therapy. RESULTS: Before treatment, the patients with CAP were observed to have a lower protein C system activity and the signs of hypercoagulation that were eliminated by ILIB. CONCLUSION: ILIB is an effective method in correcting hemocoagulative disorders in patients with CAP.

Lasers Med Sci. 2009 Jul;24(4):567-76. Epub 2008 Nov 12.

Effect of low level laser therapy on bronchial hyper-responsiveness.

Aimbire F, de Lima FM, Costa MS, Albertini R, Correa JC, Iversen VV, Bjordal JM.

Institute of Research and Development, University of Vale do Paraíba (UNIVAP), São José dos Campos, SP, Brazil. aimbire@univap.br

The objective of this study was to investigate whether low level laser therapy (LLLT) could reduce bronchial hyper-responsiveness (BHR) induced by tumour necrosis factor-alpha (TNF-alpha) modulating the metabolism of inositol phosphate (IP) in bronchial smooth muscle cells (BSMCs). The study was on 28 Wistar rats, randomly divided into four groups. Irradiation (1.3 J/cm(2)) was administered 5 min and 4 h after bronchial smooth muscle (BSM) had been suspended in TNF-alpha baths, and the contractile response-induced calcium ion (Ca(2+)) sensitization was measured. The BSMCs were isolated, and the IP accumulation was measured before and after TNF-alpha immersion in the groups that had been irradiated or not irradiated. BSM segments significantly increased contraction 24 h after TNF-alpha immersion when exposed to carbachol (CCh) as Ca(2+), but it was significantly reduced by 64% and 30%, respectively, after laser treatment. The increase in IP accumulation induced by CCh after TNF-alpha immersion was reduced in the BSMCs by LLLT. The dose of 2.6 J/cm(2) reduced BHR and IP accumulation in the rats’ inflammatory BSMCs.

Vopr Kurortol Fizioter Lech Fiz Kult. 2009 Mar-Apr;(2):19-22.

Low-intensity laser radiation in the combined treatment of patients with obstructive bronchitis.

[Article in Russian]

Kashanskaia EP, Fedorov AA.

This study included 89 patients with chronic obstructive bronchitis (COB) that were exposed to harmful occupational factors. The control group consisted of 30 healthy subjects. Chronic obstructive bronchitis is characterized by serious disturbances in the cardiorespiratory system that manifest themselves in the form of bronchogenic pneumosclerosis with moderate lung emphysema and pulmonary hypertension associated with impaired general resistance. Simultaneously, activity of lipid peroxidation reactions decreases. Application of low-intensity laser radiation in combination with other therapeutic modalities for the treatment of patients with chronic obstructive bronchitis accelerates elimination of clinical symptoms, increases its efficiency, promotes drainage function of the bronchi, facilitates normalization of the patient’s immune status, and contributes to the optimization of lipid peroxidation processes.

Klin Lab Diagn. 2009 Mar;(3):13-4.

Morphometry and electrophoretic mobility of red blood cells from patients with asthma in the intravenous blood laser irradiation.

[Article in Russian]

Sarycheva TG, Tsybzhitova EB, Popova OV, Aleksandrov OV.

The morphometry and electrophoretic mobility of red blood cells from patients with infection-dependent asthma were comparatively studied prior to and following treatment. The patients who had underwent intravenous laser irradiation of blood (ILIB) in addition to conventional therapy had better morphofunctional parameters of red blood cells, by restoring their normal forms, decreasing their transitional ones, and increasing their electrophoretic mobility to normal values. Those who received traditional drug therapy showed no considerable morphofunctional changes of erythrocytes. Thus, in asthmatic patients, the changes in the morphology and function of red blood cells may suggest their membranous structural changes for whose correction ILIB should used.

Lasers Surg Med. 2009 Jan;41(1):68-74.

Low level laser therapy (LLLT): attenuation of cholinergic hyperreactivity, beta(2)-adrenergic hyporesponsiveness and TNF-alpha mRNA expression in rat bronchi segments in E. coli lipopolysaccharide-induced airway inflammation by a NF-kappaB dependent mechanism.

Mafra de Lima F, Costa MS, Albertini R, Silva JA Jr, Aimbire F.

Institute of Research and Development (IP&D), São Paulo, Brazil.

BACKGROUND AND OBJECTIVES: It is unknown if the decreased ability to relax airways smooth muscles in asthma and other inflammatory disorders, such as acute respiratory distress syndrome (ARDS), can be influenced by low level laser therapy (LLLT) irradiation. In this context, the present work was developed in order to investigate if LLLT could reduce dysfunction in inflamed bronchi smooth muscles (BSM) in rats. STUDY DESIGN/MATERIALS AND METHODS: A controlled ex vivo study was developed where bronchi from Wistar rat were dissected and mounted in an organ bath apparatus with or without a TNF-alpha. RESULTS: LLLT administered perpendicularly to a point in the middle of the dissected bronchi with a wavelength of 655 nm and a dose of 2.6 J/cm(2), partially decreased BSM hyperreactivity to cholinergic agonist, restored BSM relaxation to isoproterenol and reduced the TNF-alpha mRNA expression. An NF-kappaB antagonist (BMS205820) blocked the LLLT effect on dysfunction in inflamed BSM. CONCLUSION: The results obtained in this work indicate that the LLLT effect on alterations in responsiveness of airway smooth muscles observed in TNF-alpha-induced experimental acute lung inflammation seems to be dependent of NF-kappaB activation.

Inflammation. 2008 Jun;31(3):189-97. Epub 2008 Apr 18.

Low level laser therapy (LLLT) decreases pulmonary microvascular leakage, neutrophil influx and IL-1beta levels in airway and lung from rat subjected to LPS-induced inflammation.

Aimbire F, Ligeiro de Oliveira AP, Albertini R, Corrêa JC, Ladeira de Campos CB, Lyon JP, Silva JA Jr, Costa MS.

Instituto de Pesquisa & Desenvolvimento-IP&D, Universidade do Vale do Paraíba-UNIVAP, Av. Shishima Hifumi, 2911, CEP: 12244-000, São José dos Campos, São Paulo, Brazil. aimbire@univap.br

BACKGROUND AND OBJECTIVE: Low level laser therapy (LLLT) is a known anti-inflammatory therapy. Herein we studied the effect of LLLT on lung permeability and the IL-1beta level in LPS-induced pulmonary inflammation. STUDY DESIGN/METHODOLOGY: Rats were divided into 12 groups (n = 7 for each group). Lung permeability was measured by quantifying extravasated albumin concentration in lung homogenate, inflammatory cells influx was determined by myeloperoxidase activity, IL-1beta in BAL was determined by ELISA and IL-1beta mRNA expression in trachea was evaluated by RT-PCR. The rats were irradiated on the skin over the upper bronchus at the site of tracheotomy after LPS. RESULTS: LLLT attenuated lung permeability. In addition, there was reduced neutrophil influx, myeloperoxidase activity and both IL-1beta in BAL and IL-1beta mRNA expression in trachea obtained from animals subjected to LPS-induced inflammation. CONCLUSION: LLLT reduced the lung permeability by a mechanism in which the IL-1beta seems to have an important role.

Med Pregl. 2008 Mar-Apr;61(3-4):123-30.

Innovative approach to laser acupuncture therapy of acute obstruction in asthmatic children.

[Article in English, Serbian]

Nedeljkovi? M, Ljustina-Pribi? R, Savi? K.

Institut za onkologiju Vojvodine, Sremska Kamenica. nedeljkovic@neobee.net

A prospective randomized study encompassed 100 patients with asthma in the stage of acute bronchial obstruction, 7 to 17 years of age, of both sexes and with the similar rate of attack severity. The patients were spirometrically monitored (FVC, FEV1, PEF and FEF 25-75%) by a pediatrician-pulmonologist for 12 days. Fifty patients were subjected to conservative drug therapy, whereas other 50 patients additionally underwent the Su Jok therapy according to the 6-Ki principles, applying laser ray at the acupuncture points in the hand. After 12 days of treatment the group, in which both therapy methods were applied, reached the range of referent values for all investigated parameters. In the same period of time the group undergoing only conservative drug therapy retained values below the reference ones for the given age for 3 (FEV1, PEFR and FEF 25-75%) out of 5 investigated parameters. The obtained results differed from the group subjected to the combined therapy with the high rate of statistical significance. On the basis of the obtained results the efficacy of the 6-Ki technique of Su Jok therapy was confirmed as the therapeutic method supplementary to the conservative drug therapy in the treatment of acute bronchial obstruction in asthmatic children, provided that therapy is carried out in stationary conditions and under constant spirometric surveillance of the patient.

Ter Arkh. 2007;79(3):44-8.

Intravascular laser irradiation of blood in the treatment of patients with bronchial asthma.

[Article in Russian]

Farkhutdinov UR.

AIM: To investigate action of intravascular laser blood irradiation (ILBI) on production of active oxygen forms (AOF) in patients with bronchial asthma (BA). MATERIAL AND METHODS: The trial included 59 BA patients aged 20 to 60 years (mean age 40.2 +/- 3.1 years). AOF generation in whole blood was registered with luminol-dependent chemiluminescence (CL). Basic therapy was given to 42 patients. ILBI was added to basic therapy in 17 patients. RESULT:. CL of whole blood in BA patients depended on severity of inflammation. BA patients with intensive CL exposed to ILBI retained free radical oxidation defects and the disease symptoms. In low intensity of blood CL, ILBI activated A OF generation and raised treatment effectiveness. CONCLUSION: ILBI raises AOF production in the whole blood of BA patients. CL registration can be used for validation of ILBI administration in BA patients and control of effectiveness of laser therapy.

Bull Exp Biol Med. 2007 Aug;144(2):238-40.

Effect of laser radiation on production of reactive oxygen species in the blood of patients with chronic obstructive pulmonary disease.

Effect of laser radiation on production of reactive oxygen species in the blood of patients with chronic obstructive pulmonary disease.

Farkhutdinov UR, Farkhutdinov ShU.

Department of Pulmonology, Clinical Hospital No. 21, Ufa.

The effect of laser radiation on generation of reactive oxygen species in the whole blood from patients with chronic obstructive pulmonary disease was studied by in vitro recording of luminol-dependent chemiluminescence. Laser irradiation of the blood from patients with increased production of reactive oxygen species decreased the microbicidal potential of cells. In patients with low generation of reactive oxygen species and normal potential of cells, laser exposure increased production of O2 metabolites. Laser radiation had little effect on chemiluminescence of the blood in patients with low generation of reactive oxygen species and decreased functional activity of cells.

Vopr Kurortol Fizioter Lech Fiz Kult. 2007 Jul-Aug;(4):9-11.

Effect of low intensive helium-neon laser on superoxide-producing and methemoglobin-restoring activity of cytochrome b558III of red cell membranes in experiment.

[Article in Russian]

Simonian RM, Simonian GM, Simonian MA, Sekoian ES.

It is established that low-energy He-Ne laser without changes from the side of NADPH-dependent superoxide-producing activity of b558III cytochrome isolated from erythrocytic membranes (EM) promotes b558III activity decrease in EM. The level of metHb-restoring activity increases during irradiation by He-Ne laser. The form of optical spectrum of b558III cytochrome absorption in the visible spectrum area does not undergo essential changes after laser irradiation while optical spectral index (A412/A530) is reduced a little. It is supposed that in the basis of photochemical mechanisms of He-Ne laser protector action in oxidative stress lays its ability to increase EM stability due to a decrease of superoxide-producing and an increase of metHb-restoring activity of b558III cytochrome.

Klin Med (Mosk). 2007;85(7):48-50.

The dynamics of microcirculation parameters in patients with pneumonia receiving intravenous irradiation of blood as a part of complex treatment.

[Article in Russian]

Burduli NM, Pilieva NG.

The aim of the study was to evaluate the condition of the microcirculatation under the influence of low-intensity laser irradiation as a part of complex treatment of patients with pneumonia. The subjects were 62 patients with pneumonia divided into two groups. The 30 patients of the control group received only conventional medication therapy, while the 32 patients of the main group received a course of laser irradiation of blood (LIB) in addition to conventional medication. Positive dynamics in microcirculation was more prominent in the main group. In conclusion, LIB may be considered to be an effective method to correct microcirculatory disturbances in patients with pneumonia.

Lik Sprava. 2007 Apr-May;(3):42-6.

Patients with tuberculosis associated with chronic non-specific lung diseases

[Article in Ukrainian]

Nikolaieva OD.

159 patients have been observed to assess the efficiency of laseropuncture use in a complex treatment of patients with lung tuberculosis and chronic bronchitis. Disbalance in renal meridian (R), urinary bladder (V) and insufficiency of the energy in colon meridian (60.3%) were observed in patients with tuberculosis associated with chronic bronchitis. Medium deviations of electro-skin conductivity from the physiological gape in meridians of GI, IG, F, V, R in patients with tuberculosis associated with chronic bronchitis considerably differ from those data obtained from patients with only tuberculosis. Obtained data testify more severe disorders of energy balance in meridians of patients having except tuberculosis other associated diseases. Medium parameters of deviations from the physiological gape considerably decrease after the course of laseropuncture compared with those patients treated traditionally. Improve of the course of the chronic bronchitis was noted in patients who underwent laseropuncture.

EMLA Laser Health J 2007;2:46-67
European Medical Laser Association (EMLA)

Treatment of asthma and allergic rhinitis with low level laser and singulair in children

C. Ailioaie 1,3, L-M. Ailioaie 2,3, D.A. Chiran 1,3
1. University of Medicine and Pharmacy, Faculty of Medicine, Iassy, Romania
2. “Al. I. Cuza” University, Dept. of Medical Physics, Iassy, Romania
3. Laser Clinic, Iossy, Romania

112 patients aged 9-18 years, with asthma and allergic rhinitis, were divided into 3 groups. Group 1 received laser therapy, as follows: acupoints (0.3 J/point), scanning technique (2 J/cm 2 ), local regional ganglions (3 J/cm 2 ), nose and oral mucous membranes (0.4 J/cm 2 ) (670 and 830 nm, 25 mW -300 mW, CW and pulsed mode), three times/weekly, 12 days/monthly, during 3 months, together with classically drugs and 10 mg Singulair once daily. Group 2 was administrated placebo laser and Singulair, following the same protocol. Group 3 was treated only with classically drugs.

Low level laser therapy (LLLT) and Singulair have significantly reduced the symptoms, the duration and the dose of the corticosteroids, and have prolonged the disease remission better than other modality trials with side effects.

LLLT and Singulair improved the lung function and the quality of life in asthmatic patients with allergic rhinitis, too.

Probl Tuberk Bolezn Legk. 2007;(8):50-3.

Efficiency of a combination of haloaerosols and helium-neon laser in the multimodality treatment of patients with bronchial asthma.

[Article in Russian]

Faradzheva NA.

A hundred and thirty-eight patients with infection-dependent bronchial asthma, including 73 with moderate persistent asthma and 65 with severe persistent one, were examined. Four modes of a combination of traditional (drug) therapy (DT) and untraditional (halotherapy (HT) and endobronchial helium-neon laser irradiation (ELI) one were used. The efficiency of the treatment performed was evaluated, by determining the time course of clinical symptoms of the disease on the basis of scores of their magnitude and the patients’ condition. The findings indicated that in moderate persistent asthma, both HT and ELI in combination with DT exerted an equal therapeutic effect, which provided a good and excellent condition in 83.3% of cases. In severe persistent asthma, such a condition was achieved in 93.75% of cases only when multimodality treatment involving DT, HT, and ELI had been performed.

Ter Arkh. 2007;79(3):44-8.

Intravascular laser irradiation of blood in the treatment of patients with bronchial asthma.

Farkhutdinov UR.

AIM: To investigate action of intravascular laser blood irradiation (ILBI) on production of active oxygen forms (AOF) in patients with bronchial asthma (BA). MATERIAL AND METHODS: The trial included 59 BA patients aged 20 to 60 years (mean age 40.2 +/- 3.1 years). AOF generation in whole blood was registered with luminol-dependent chemiluminescence (CL). Basic therapy was given to 42 patients. ILBI was added to basic therapy in 17 patients.RESULT:. CL of whole blood in BA patients depended on severity of inflammation. BA patients with intensive CL exposed to ILBI retained free radical oxidation defects and the disease symptoms. In low intensity of blood CL, ILBI activated A OF generation and raised treatment effectiveness. CONCLUSION: ILBI raises AOF production in the whole blood of BA patients. CL registration can be used for validation of ILBI administration in BA patients and control of

Lasers Surg Med. 2006 Sep;38(8):773-8.

Low level laser therapy partially restores trachea muscle relaxation response in rats with tumor necrosis factor alpha-mediated smooth airway muscle dysfunction.

Aimbire F, Bjordal JM, Iversen VV, Albertini R, Frigo L, Pacheco MT, Castro-Faria-Neto HC, Chavantes MC, Labat RM, Lopes-Martins RA.

Institute of Research and Development-IP&D/UNIVAP Av., Shsima Hifumi 2911, 12240-000 São José dos Campos, São Paulo, Brazil.

Abstract

BACKGROUND AND OBJECTIVE: It is unknown if the decreased ability to relax airway smooth muscles in asthma and other inflammatory airways disorders can be influenced by low level laser therapy (LLLT) irradiation. To investigate if LLLT could reduce impairment in inflamed trachea smooth muscles (TSM) in rats.

STUDY DESIGN/MATERIALS AND METHODS: Controlled rat study where trachea was dissected and mounted in an organ bath apparatus with or without a TNF-alpha solution.

RESULTS: Low level laser therapy administered perpendicularly to a point in the middle of the dissected trachea with a wavelength of 655 nm and a dose of 2.6 J/cm(2), partially restored TSM relaxation response to isoproterenol. Tension reduction was 47.0 % (+/-2.85) in the laser-irradiated group compared to 22.0% (+/-2.21) in the control group (P < 0.01). Accumulation of cAMP was almost normalized after LLLT at 22.3 pmol/mg (+/-2.1) compared to 17.6 pmol/mg (+/-2.1) in the non-irradiated control group (P < 0.01).

CONCLUSION: Low level laser therapy partially restores the normal relaxation response in inflamed TSM and normalizes accumulation of cAMP in the presence of isoproterenol.

Ter Arkh. 2006;78(3):39-40.

Clinical efficacy of target low-intensity laser radiation on the adrenal projection region in patients with bronchial asthma.

[Article in Russian]

Nikitin AV, Titova LA.

AIM: To study efficacy of infrared laser radiation on the adrenal projection region in patients with bronchial asthma (BA). MATERIAL AND METHODS: The adrenal region of BA patients was exposed to infrared laser radiation. The effect was stated after blood count, examination of the urine and sputum, blood biochemical tests, blood hydrocortisone assay. RESULTS: Positive clinical response to laser therapy manifested with improvement of clinical, device and spirometric indices in BA patients. Blood levels of hydrocortisone normalized. CONCLUSION: Infrared laser radiation of adrenal projection region reduced the number of asphyxia attacks, improved bronchial drainage, shortened the disease duration. The method is well tolerated, safe, easy to make and suggested for use in combined treatment of BA.

Ter Arkh. 2006;78(3):41-4.

Outpatient use of laser therapy in bronchial asthma

[Article in Russian]

Ostronosova NS.

AIM: To analyse effects of outpatient laser therapy on the course of different forms of bronchial asthma (BA). MATERIAL AND METHODS: The examination has covered 466 patients with BA divided into three groups: group 1 patients received monotherapy with low-intensive laser radiation (LILR); group 2 – LILR plus conventional drug therapy (CDT); group 3 – CDT only. RESULTS: Group 1 and 2 patients achieved improvement after 2-3 sessions. Asphyxia attacks were less frequent and severe. These attacks disappeared to session 7-10, 10-12 in patients with exogenic, endogenic and mild, moderate mixed BA, respectively. The patients recovered sensitivity to sympathomimetics and methylxantines. Glucocorticosteroids (GCS) doses, hospital stay and temporary disability duration decreased. These two groups achieved remission within 0.5-1 year in 50% patients. Remission for 1-2 years was observed in 40% patients while in group. 3 – in 14%. CONCLUSION: The addition of laser therapy improves a BA course, results of outpatient treatment and rehabilitation, recovers bronchial sensitivity to sympathomimetics and xantine derivatives, reduces GCS dose and duration of hospital stay and disability in BA patients.

Vopr Kurortol Fizioter Lech Fiz Kult. 2006 Mar-Apr;(2):8-10.

Low-intensity laser radiation in therapy of bronchial asthma

[Article in Russian]

Ostronosova NS.

Efficacy of low-intensity laser radiation (LR) was studied by changes in a histamine level in peripheral blood and external respiration function (ERF). A total of 228 patients with exogenic bronchial asthma (EBA) were divided into two groups: 167 patients with nonhormone dependent asthma (group 1) and 61 patients with hormone-dependent bronchial asthma (group 2). Low-intensity LR lowered a histamine level and improved ERF in both group 1 and 2. A definite negative correlation was found between histamine content and ERF parameters. Laser therapy alone is effective in mild EBA. For moderate and severe EBA and hormone-dependent bronchial asthma a combination of conventional drug therapy with low-intensity LR is recommended.

Ter Arkh. 2004;76(3):20-3

Efficacy of noninvasive hemolaserotherapy in patients with bronchial asthma and concomitant rhinosinusitis.

[Article in Russian]

Nikitin AV, Treshchalina IuB.

AIM: To study efficacy of low-intensive infrared laser radiation impact on the tympanic membrane in patients with bronchial asthma (BA) and concomitant rhinosinusitis (RS). MATERIAL AND METHODS: 78 patients with moderate BA of a mixed type and concomitant chronic RS were divided into 3 groups: group 1 patients received medication plus infrared laser radiation of the tympanic membrane and paranasal sinuses; group 2 patients were exposed to supravenous laser radiation (0.63 mcm); group 3 received pharmacotherapy alone. The effect of the treatment was assessed by spirometry, peakflowmetry and paranasal sinuses findings. RESULTS: The highest response was achieved in group 1 which manifested with positive changes in clinical, device and spirometric data on BA and x-ray data on RS courses. The least effective treatment was observed in group 3. CONCLUSION: Use of infrared laser radiation of the tympanic membrane and paranasal sinuses projection in the treatment of BA patients with RS is effective, nontoxic and easy to use both in hospitals and outpatient departments.

Med Pregl. 2004 Jan-Feb;57(1-2):13-7.

The role of physical rehabilitation in the treatment of exudative pleurisy.

[Article in Serbian]

Milojevi? M, Kuruc V.

INTRODUCTION: Exudates are due to a variety of diseases, the major and most common ones being tuberculosis, nonspecific inflammation and malignancy. They are usually treated conservatively, sometimes combined with surgery and physical treatment. Physical therapy includes positional exercises, breathing exercises and biostimulation.

AIM OF THE STUDY: The study was aimed to find out the following: 1) Is lung function improved by physical therapy; 2) Can adhesions be diminished and mobility of the affected hemidiaphragm improved by physical treatment; 3) Is there a direct positive correlation between physical treatment and obtained improvement, or the same can be achieved in patients receiving medicamentous treatment only; 4) What are the effects of some factors we cannot influence (sex, age, effusion level, position of adhesions) on lung function and diaphragm mobility improvement, that is on the efficiency of physical treatment; 5) How do the factors we can influence (the time interval before initiating the treatment and its duration) affect improvement of the same lung function parameters, that is treatment efficacy?

MATERIAL AND METHODS: Physical treatment of patients with exudative pleurisy was accomplished at the Department of Rehabilitation in our Institute and it consisted of directed breathing exercises and laser biostimulation. Its effects were examined in a group of 175 patients, who received both conservative and physical treatment, and results were compared with the control group patients, treated only conservatively (with antibiotics, antituberculotics, corticosteroids). RESULTS: Comparative analysis confirmed a significant improvement of lung function parameters (VC, FEV1, PEF) as well as of hemidiaphragm mobility on the affected side of the thorax in favour of the examined group. The severity of the lung function and diaphragm mobility impairments have been found to be in correlation with the localization of adhesions, whereas the degree of improvement correlated with the time interval before the treatment initiation, as well as with its duration.

DISCUSSION AND CONCLUSION: The applied physical therapy resulted in: 1) significant improvement of all examined lung function parameters in the examined group, which was not registered in the control group; 2) significant improvement of the diaphragm mobility in general; 3) factors such as sex, age and effusion level have no effects on the physical treatment results; 4) treatment results are affected by the time interval passed before the treatment initiation and its duration, as well as the localization of adhesions; anterior adhesions affected lung function and diaphragm mobility least, posterior ones more, while the influence of lateral adhesions was most significant. It is finally concluded that physical treatment should necessarily be included in the treatment of exudative pleurisy.

Med Pregl. 2003 Nov-Dec;56(11-12):516-20.

Laser biostimulation in the treatment of pleurisy

[Article in Serbian]

Milojevi? M, Kuruc V.

Institut za plu?ne bolesti, Sremska Kamenica, Medicinski fakultet, Novi Sad. drmilojevic@yahoo.com

INTRODUCTION: Low-intensity lasers have been utilized in medicine in two ways: for local stimulation and for stimulation of acupuncture points. Literature data reveal that this method has been indiscriminately applied in psychiatry, rheumatology, gynecology, dermatology, otorhinolaryngology, in diverse acute and chronic pains, inflammations, vascular disorders, angina pectoris, bronchial asthma. Most commonly reported clinical effects are analgesia, spasmolytic and anti-inflammatory effects, as well as faster wound and bone healing. MATERIAL AND METHODS: This prospective study analyses effects of laser biostimulation on patients with pleurisy. The analysis included 25 patients treated at the Institute of Lung Diseases in Sremska Kamenica during 2000, 2001 and 2002. Apart from conservative treatment, these patients were treated with laser biostimulation of acupuncture points and local region for ten days. During treatment, changes of present clinical signs, general symptoms, radiological findings, as well as changes of some relevant biochemical parameters were recorded. RESULTS: Results were compared with the control group which included the same number of patients, who differed from the examined group only by not being exposed to laser biostimulation. The examined group of patients with pleuritis presented with quicker resorption of pleural effusion, less pleural adhesions, more significant decrease of clinical symptoms, especially pain, as well as more significant increase of cortisol and immunoglobulin A and decrease of circulating immune complexes (CIC), leukocytes and sedimentation rate than the control group. DISCUSSION: Mechanisms of laser biostimulation in treatment of pleurisy were described in detail and the obtained results were correlated to those reported by other authors. CONCLUSION: 1. Patients with pleurisy undergoing laser stimulation presented with faster resorption of effusion and remission of the subjective symptoms, as well as significant decrease of biochemical acute inflammation parameters in the peripheral blood and therefore with faster recovery. 2. In patients with pleurisy laser treatment increases regenerative mechanisms of the pleural surface, thus decreasing the quantity of formed adhesions and resulting in better mobility of the diaphragm.

Med Pregl. 2003 Sep-Oct;56(9-10):413-8.

Low power laser biostimulation in the treatment of bronchial asthma.

[Article in Serbian]

Milojevic M, Kuruc V.

Institut za plucne bolesti, Sremska Kamenica, Medicinski fakultet, Novi Sad. drmilojevic@yahoo.com

INTRODUCTION: Modern concept of acupuncture is based on the fact there are designated locations on the surface of human body, which are related to integrative systems of an organism by means of sensory nerves, correlating and synchronizing organ functioning, depending on external and internal conditions, by means of nervous and neurohumoral regulation of metabolic and regenerative processes, including also mobilisation of immunological, protective and antistress reactions. Apart from standard needle acupuncture, other methods of stimulating acupuncture points are also applied. Due to invention of low power lasers, irradiation laser acupuncture has been introduced into routine medical practice, characterised by painless and aseptic technique and outstanding clinical results.

MATERIAL AND METHODS: The investigation was aimed at defining therapeutic effects of low power laser irradiation by stimulating acupuncture points or local treatment of asthma. A prospective analysis included 50 patients treated at the Institute of Pulmonary Diseases in Sremska Kamenica during 2000, 2001 and 2002. Together with conservative treatment of present disease, these patients were treated with laser stimulation of acupuncture points in duration of ten days. During treatment changes of functional respiratory parameters were recorded. Results were compared with those in the control group. The control group consisted of the same number of patients and differed from the examination group only by not using laser stimulation.

RESULTS: Patients with bronchial asthma presented with significant improvement (p < 0000,5) of all estimated lung function parameters just 30 minutes after laser stimulation. Improvements achieved on the third and the tenth day of treatment were significantly higher (p < 0.001 to p < 0.00005) in the examination group in comparison with the control group. Further investigation confirmed that improvement of measured lung function parameters was significantly higher in younger patients, in patients whose disease lasted shorter, as well as in women. Patients with asthma, who were treated every three months for a one year period, presented with significantly lower frequency and intensity of attacks.

DISCUSSION: The mechanism of laser stimulation activity in treatment of bronchial asthma is explained in detail, correlating our results to those obtained by other authors.

CONCLUSIONS: A ten-day course of low-power laser stimulation of acupuncture points in patients with bronchial asthma improves both the lung function and gas exchange parameters. Positive effects of laser treatment in patients with bronchial asthma are achieved in a short time and they last long, for several weeks, even months. Successive laser stimulation in asthmatics prolongs periods of remission and decreases the severity of asthmatic attacks. Better positive effects of laser stimulation are achieved in younger asthmatics, in those with shorter disease history and in female patients. There is a negative correlation between effects of laser stimulation and patients’ age and disease history. However, these characteristics do not affect response rate and speed but positive laser stimulation effects are achieved in a shorter period in female asthmatics.

Ter Arkh. 2002;74(3):25-8.

Efficacy of low intensity laser irradiation and sodium nedocromil in the complex treatment of patients with bronchial asthma.

[Article in Russian]

Landyshev IuS, Avdeeva NV, Goborov ND, Krasavina NP, Tikhonova GA, Tkacheva SI.

AIM: To study efficiency of low-intensity laser radiation (LILR) and sodium nedokromil (tailed) in combined treatment of bronchial asthma (BA).

MATERIAL AND METHODS: The choice of the treatment depended on the activity of bronchial inflammation and the presence of contraindications. Laser was used on the skin in the area of the lung and great vessels projection, endobronchially. Tailed was given in inhalations and irrigations of the tracheobronchial tree during therapeutic fibrobronchoscopy. These methods were used in combined treatment of 220 BA patients.

RESULTS: Combined use of LILR and tailed proved highly effective and safe in BA. Cytological markers of cell reactions of the bronchopulmonary system on the action of LILR were revealed.

CONCLUSION: Availability, good reproducability, cost-effect efficacy and safety make LILR one of the most beneficial nonpharmacological treatments for bronchial asthma.

The treatment of bronchial asthma with LLLT in attack-free period in children.

Ailioaie C, Ailioaie L.

98 patient aged 10-18 years, diagnosed with moderate or severe asthma were divided into three groups. -Group 1: 35 patients received laser acupuncture using extrameridian acupuncture points plus scanning. Lasers used were 670 and 830 nm, 50 and 200 mW respectively, in continous mode. Treatment was given twice daily 10 days per month, 3 months in total. No other therapy was given. -Group 2: 33 patients inhaled Salmeterol xinofoat 2 x 25, twice daily for 3 months. -Group 3: 30 patients were treated with Theophylline retard 15 mg/kg every 12 hr for 3 months. Results: A noticeable improvement in the clinical, functional and immunological characteristics were observed in 83% of the patients in group 1, 70 in group 2 and 53 in group 3. There were no side effects in the laser group.

Effects of laser therapy on lipids and antioxidants in blood of patients with bronchial asthma.

[Article in Russian]

Rakitina DR, Urias’ev OM, Garmash VIa, Ivanova MV, Krasnovid NI, Lebedev AV.

Laser therapy was assessed for effects on lipoperoxides and free radical catchers in blood lipids of patients with bronchial asthma (BA). When a group of 52 BA patients was compared to healthy donors by dienic conjugates, vitamin E and overall lipid-soluble antioxidants levels in the whole blood and plasma, these appeared higher in the asthmatics. Combination of laser therapy with conventional treatment returned these parameters close to normal.

Ter Arkh. 2002;74(3):40-3.

Parameters of membrane permeability, microcirculation, external respiration, and trace element levels in the drug-laser treatment of pneumonia.

[Article in Russian]

Amirov NB.

AIM: To study effects of laser therapy in combined treatment of pneumonia on external respiration function, membrane permeability, microcirculation and serum trace elements. MATERIAL AND METHODS: 142 pneumonia patients were randomly divided into two groups: 96 patients treated with drugs and laser radiation (the study group) and 46 patients treated with drugs only (control group). RESULTS: In the study group there was more pronounced reduction in cell membrane permeability, a rise in concentrations of iron and chromium in the blood serum, improvement of microcirculation. These changes closely correlated with those in immunity, external respiration function. CONCLUSION: Laser therapy is an effective method of pneumonia treatment and can be included in relevant combined schemes.

THERAPY IN PATIENTS WITH CHRONIC OBSTRUCTIVE BRONCHITIS (COB)

Nikitin A.V., Evstratov A.Yu., Esaulenko I.E.

Medical Academy, Voronezh, Russia

A variety of mechanisms of laser irradiation in non-specific immunity stimulation have been reported lately. One of the factors contributing to the frequent exacerba-tion’s of COB is the presence of the secondary immunological insufficiency in this category of patients. The aim of the present study is to analyze the results of different schemes of the low intensity laser therapy use and its influence on cellular immunity and the clinical course of the disease in patients with COB during the period of exac-erbation. 100 patients, 78 male and 22 female, the mean age was 51, with the diagnosis of COB and respiratory insufficiency I-II, were divided into 4 equal groups at random. Patients of the first group underwent cndovascular laser irradiation of the blood by the helium-neon laser installation “ALOK-I” with the irradiation power of 2mW; the second group received the contact treatment on the projection of the main bronchi, intcrscapular area by the infra-red laser installation “Mustang-017” with the power of 8-10W. The third group received the combination of cndovascular laser irradiation with the contact one, The course of laser therapy in all group lasted for 15 days and was combined with conventional therapy. The control group of patients underwent only conventional therapy including antibactcrial drugs, vitamins, physiotherapy. The indices analysis of the immunity cellular link in the dynamics of laser therapy and conventional therapy in these groups has shown the evident positive dynamics of the initially decreased T-lymphocytcs (CD,J, T-suprcssors (CDg^), B-lympliocytcs (CD,^) (p<0,05). The best results were observed in patients who had received the combination of cndovascular laser irradiation with the contact one (p<0,05). In the control group the dynamic of the studied indices was positive, but not strongly marked (p>0,05).

Vopr Kurortol Fizioter Lech Fiz Kult. 2001 May-Jun;(3):15-8.

Laser therapy of elderly patients with pneumonia

[Article in Russian]

Lutai AV, Egorova LA, Shutemova EA.

The aim of the study was to evaluate the efficiency of laser therapy included into the treatment of pneumonia in the elderly. A follow-up included the analysis of their clinical status and external respiratory function, pulmonary blood flow, and immunological parameters in 2 matched groups of pneumonia patients aged 60 to 72 years. Low-intensity laser therapy (transcutaneous sliding contact procedure) was used as part of routine treatment in one of the groups. The findings demonstrate that non-drug treatment had an undeniably positive impact. There was an earlier regress of clinical symptoms and a sound recovery of functional parameters. In the absence of side effects of this method, these data allow infrared laser therapy to be recommended for rehabilitation of elderly patients with pneumonia.

LOW-LEVEL LASER THERAPY IN PATIENTS WITH PNEUMONIA

Kustova N.L., Yemelyanova L.A.

Urals State Medical Academy, Yekaterinburg, Russia

We compared the effectiveness of complex treatment including transcutaneous low-level laser radiation of chest by He-Ne laser /66 patients/, AsGa laser/68 patients/, magnet-laser therapy/66 patients/ and decimeter-wave therapy /62 patients/. We used individual laser radiation dosing method according to V.M. Lisienko and R.I Mintz / 1987/. All three types of laser therapy have beneficial influence on inflammation proc-ess in patients with pneumonia, having positive effects on laboratory and rentge-nological symptoms. After such treatment patients recover two times faster, especially after infrared laser radiation or its combination with magnettherapy. Structural optical serum properties evaluation with the help of polarization microscopy and refractome-try methods showed that there are many liquid crystals of different types in the begin-ning of pneumonia. There is feedback between serum refraction index and severity of pneumonia, existence of complications. After treatment serum refraction index aver-age values returned to normal in all four groups of patients. Laser therapy improves immune status of patients. Magnct-lasertherapy and infra-red laser radiation are the most effective, including cases of lingering disease. Individual laser radiation dosage stimulates phagocytosis. According to the above-mentioned complex treatment of pneumonia can include low-level laser radiation.

USE OF LOW POWER LASER LIGHT IN TREATMENT OF A CHRONIC PURULENT BRONCHITIS

G.G. Prosorova , A.F. Anoshkina, S.A. Afendulov

Physician in Chief, Medical Centre of the Novolipetsk, Lipetsk, Russia

Transplantation of autologic leukocytic suspension extracorporally cured with a he-lium-neon laser “TPLA” of l =o 632,8 nm and an exit light conductor power of 1,5 mW was made use of in treatment of 53 pts suffering with CPB. Curing was applied during a 7 minutes period. Transplantation of AL suspension was carried out during a sanitation fibroscopy (FN3). The treatment was conducted up to a normalisation in an cndoscoimages state and elimination of clinical and laboratory in-flainmatory signs. The results were compared to those of the conventionally treated 32 CPB pts (antibiotics , broncholytics, FBS with an infusion of mycosolvin and furagin). The following phe-nomena were revealed: 1) an improvement in the blood cells phagocytic activity under an application of HNL light cured autolcukosuspcnsion. An increase in Ph% from 71,8 up to 88,8 (p<0,05), in PhN from 5,3 up to 8,9 (p<0,05), in PhCV from 0,76 up to 1,01 (p<0,05) was evidenced, besides, these post treatment values in the experimental group didn’t differ from those of healthy persons (p>0,05 ); 2) a reduction in a number of FBS sessions from 5-7 conducted during the con vcntional treatment down to 3-4 ones carried out with LL- cured AL suspension. 3) a reduction in the quantity of anti-biotics administered: while carrying out transplantation of autoleukosuspension cured with HN-lascr light we could completely reject antibiotics administration and only 1 pt had to be subjected to a second course of administering antibactcria] preparations, whereas during .the conventional treatment antibiotics were administered to all the pts, and 47,1% of them had to be subjected to a double course of antibiotics therapy; 4) a reduction in an average hospital stay period from 30,2 down to 23,8 days. Thus, an in-troduction of transplantation of HNLL-curcd AL-suspcnsion into the CPB pts therapy causes an increase in the organism’s non-specific resistant-resistance, minimises a mcdicamcntous load upon a pt and cuts down a duration of a hosoital stay period.

THE EFFECTIVENESS OF A LASER BEAM THERAPY IN A COMPLEX HEALTH RESORT TREATMENT OF PATIENTS WITH BRONCHIAL ASTHMA

L.I. Pogosyan, S.A. Molchanov, T.V. Daraeva, V.A. Novozhilova, T.N. Prozorova, V.l.Dmitrienko

Kislovodsk, Russia

Rehabilitation and health improvement of patients with chronic, non-specific lung dis-eases at the health resort of Kislovodsk with the help of a wide application of the natu-ral physical factors alongside with the laser beam therapy is of great importance. The observation covered 79 patients with bronchial asthma (BA) of the age range from 32 to 56 with an atoimages and infcctious-atoimages form under a mild and moderate flow. At the initial stage the prevailing majority of the observed patients demonstrated a consider-able deviation from the norm concerning the parameters of external breathing function (EBF-83,1%), more than half of the patients-that of the level of adaptation reactions (Garkavi-Kvakina-Ukolova), as well as of the immunity status (V.G.Vogralik, M.V.Vogralik), 73,4% and 62,6% accordingly, and almost half of the patients – that of the peroxide oxidation of lipids (POL -42,8%) . A group of 54 patients (the Main group, MG) received an infra-red, continuous wave laser therapy by means of the “AZOR-2K” device, alongside with the general complex spa treatment (natural fac-tors, carbonic mineral baths, massage, electro- and curative physical therapy, etc.), which the Reference group (RG) of 25 patients received. The total aggregated doze of laser energy per session was 12 J , the course of treatment – 10-15 procedures. As a re-sult, the overwhelming majority of the patients (98,4%) enjoyed an improvement of clinical, laboratory and functional parameters , group-wise : MG – 99,1%; RG -92,3%. A differential analysis of the results obtained in the MG itself has revealed a higher, although unreliable percent of correction of the acquired immunodeficiency and of op-timisation of adaptation processes in comparison with a degree of positive dynamics concerning other parameters (EBF, POL). Taking into consideration previous results the observations demonstrated that the introduction of the laser therapy into the com-plex health resort treatment enhances for certain the effectiveness of the treatment of patients with BA. We should specifically mention the positive effect of the physical factors, in combination with the laser therapy, on the level of adaptation reactions and their immunocorrcctivc influence with patients, suffering from BA complicated by the secondary immunodeficiency.

COMPARATIVE EFFECTIVENESS DETOXIFICATION TRANSFUSION OF THERAPY (TT) AND INTRAVENOUS OF A LASER IRRADIATON BLOOD (ILIB) IN COMPLEX TREATMENT OF PATIENTS BY A PULMONARY TUBERCULOSIS

P.I.Pitcyco, E.I.Krivoshapova

Kharkov Institute of Physicians Advanced Training, Kharkov. Ukraine

Two groups of the patients infiltrotivc by a pulmonary tuberculosis in a phase of de-cay, MBT+, (discovered Koch’s bacillus), comparable on a floor, age, pronounced in-toxication and syndrome, extent of process in lungs and regime of chemotherapy. I groups 45 patients receiving TT (albumen 5%- 100 ml alternated with hacmodesum 200 ml, daily ?10), II groups – 68 patients, which was carried out ILIB(wavclcngth 0,63 urn, capacity of radiation on an output flexible monofibcr conductor of light 2 mW, duration of a procedure 15 minutes, course 10 sessions). The efficiency of ther-apy was estimated on a reduction intoxication, which degree was defined on a level and structure middle molecules (MM) ( presence of pathological fractions and dis-polypcptidcmy) in whey of blood. MM were studied prior to the beginning and after realisation of rates TT and ILIB by a method hclium-chromatography on hcle “Toyop-carl HW-40F” (Japan). A normal level and structure MM (361 ± 14 cond I. unit) was established with inspection 40 healthy donors. Is established, that at the patients from I groups after realisation of a course TT the general level MM on the average has de-creased from 858 ± 29 unit till 769 ± 31 unit (p<0,05), the frequency dispolypcptid-cmy (from 75 ± 7,3 % till 33 ± 6,7 % has decreased; p<0,001) and pathological fractions (from 60 ± 7,2 % to 40 ± 7,2 %; p>0,05). After a course ILIB investigated parameters also have decreased: a level MM from 920 ± 36 unit to 584 ± 23 unit (p<0,001), frequency dispolypeptidcmy from 78 ± .6,2% to 31 ± 6,8% (p<0,001) and pathological fractions from 67 ± 6,7% till 24 ± 6,4 % (p<0,001). From the given data it is visible, that the course ILIB in comparison with TT allows more and level of pathological fractions (24 ± 6,4 % against 40 ± 7,2 %).

SORT-SANATORIUM TREATMENT OF PATIENTS SUFFERING FROM LUNG DISEASES WITH THE HELP OF LOW-INTENSIVE LASER ERADIATION

I. Shuvalova, I. Klimcnko

Sanatorium <Pnicpcr>, Yalta, Ukraine

With the view of improvement of effectiveness rate of resort-sanatorium treatment of patients suffering from chronic non-specific lungs diseases (CNLD) we have used low-intensive laser eradiation. 160 patients suffering from CNLD have been exam-ined, including 50 patients suffering from bronchial asthma of mild and average sever-ity, 60 patients suffering from chronic obstructive bronchitis, and 50 patients suffering from chronic non-obstructive functionally unstable bronchitis. All the patients have been divided into two identical groups according to their sex, age and nature of the disease. 40 patients from the first group received traditional course of treatment con-sisting of clima-totherapy, massage, exercise therapy. Apart from the above mentioned measures, the course of treatment of the second group (120 patients) employed laser therapy. The treatment was provided by dint of scanning the thorax with Helium-Neon laser eradiation with the power density equal to 80-100 mW/cm2. Besides, acupunc-ture areas were exposed to infrared eradiation with power rate equal to 4-5 W and fre-quency rate equal to 150-80 Hz. The course of treatment consisted of 7-12 everyday treatment measures. The patients who received the laser therapy improved the function of external respiration (FER) in 93.7 % of cases. An immunomodelting effect and re-gress of phlogistic processes have also been observed. The patients from the reference group improved FER only in 37.5% of cases; there were no essential changes in the state of phligistric processes and of immunity either. The analysis of the actual results of treatment demonstrated the fact that the remedial complex making use of the laser therapy produces a visible clinical effect (considerable improvement – in 43.3% and improvement -in 56.7% of cases). 89% of the patients from the reference group expe-rienced improvement, and 11% of them were discharged without any changes. Thus, laser therapy is a strong efficiency-advancing factor of the integrated treatment of pa-tients suffering from (CNLD).

THE EFFECTIVENESS OF BIOCONTROLLED IMPULSE LASER IRRADIATION IN CHILDREN BRONCHOPULMONARY DISEASES

Chcrnyishcva L.A., Khan M.A., Scrcda E.V, Semenova N.Y. SRI of pediatrics of RAMS, Moscow

Laser therapy is being widely introduced in cases of respiratory organs diseases, as a result of medical supcrsaturation of the sick child’s organism, the growing resistance to medicaments, the development of toxic and allergic reactions. In the complex therapy of 80 sick children with chronic bronchopulmonary disease the biocontrollcd impulse laser radiation of ALT “Mustang-BlO” with auto-synchronisation of radiation parame-ters with the patient’s rhythm of pulse and respiration was used. The action was carried out through the contact with rcflcxo-scgmcntal zone (D,-D^) and the zone of the pathologic focus projection in the period of exacerbation and incomplete remission of the diseases according to the standard procedure. The effectiveness was estimated with regard to clinic, functional. X-ray-bronchial and cytochcmical research before, after a single procedure and after a course of treatment. The carried research showed individ-ual sensitivity of children to the established individual dose of the action, 18.75% of sick children after the first single procedure demonstrated a certain, compared to the control group, improvement of external respiration. During the course of treatment most patients (85%0 showed the lessening or liquidation of physical changes in lungs, the decreasing activity of the inflammatory process, the improvement of cytochcmical characteristics. Cat-amncsis study, carried out in 6 and 12 months showed the stability of the therapeutic effect. The obtained data broaden the possibilities of pathogenic therapy of chronic bronchopulmonary children diseases.

THE EFFECIENCY OF THE LASER THERAPY IN THE TREATMENT OF BRONCHIAL ASTHMA SUFFERED BY CHILDREN

I.M. STRELTSOV

Volgograd Regional Children’s Clinic , Volgograd, Russia

In the recent years there is a considerable increase of bronchial asthma diseases, the tendency of the aggravation of this disease being more and more noticeable herewith . There is also a tendency for more and more resistance to the therapy conducted . That is why the bronchial asthma treatment still remains one of the most crucial and viable issues in the contemporary medicine . According to the literature data and facts , laser therapy has been successfully applied for more than 10 years . We have applied this method to 49 children suffering from bronchial asthma. Laser therapy was effected by the infrared semiconductor laser AL-09 . All the patients were split into 3 groups . The first group comprised 19 children , who have had intradcrmal laser therapy. The 16 pa-tients of the second group have received I/V (intravenous) laser therapy . The rest 14 children of the third group have received laser therapy combined with plasmapharcsis . The sample group consisted of 17 children who have had Sodium Gromoglicatc treat-ment combined with symptomatic means . The efficiency of the treatment effected was assessed in accordance with the 3 point system, 2 and 3 points being considered satis-factory. After the treatment has been completed the satisfactory results breakdown looks as follows : the first group – 56.2% ; the second group – 75% ; the third group -85.7% of the cases . In control group the positive results arc registered only at 29.4% of the children. All the abosc said shows that laser therapy proved to be an effective method of the bronchial asthma treatment for children, I/V laser application being more effective than its intravenous application. The major therapeutic effect of the combination of laser therapy with plasmaphcrcsis is , probably , connected with the application of plasmaphcrcsis . Laser application during the bronchial asthma treat-ment for children allows one to considerably reduce the medication pressure on the human body.

Ultrastruct Pathol. 2000 May-Jun;24(3):183-9.

Ultrastucture of the blood and lymphatic capillaries of the respiratory tissue during inflammation and endobronchial laser therapy.

Polosukhin VV.

Laboratory of Ultrastructural Research, Russian Academy of Medical Sciences, Novosibirsk, Russia. vpolosuk@unmc.edu

For wide application of low-energy laser irradiation in the pulmonary clinic, study of the structural basis of the therapeutic effect is necessary. The aim of this research is to describe the structural changes of the blood and lymphatic capillaries in the respiratory tissues during inflammation and following laser biostimulation. Comparative ultrastructural study was carried out on 127 open respiratory biopsy specimens from 45 patients with infectious-destructive lung diseases. These patients were divided into two groups, depending on tactic of pre-operative therapy: patients treated by only traditional anti-inflammatory measures and patients receiving additional laser therapy. Heightened permeability of the blood capillary endotheliocytes was noted as the initial stage in the development of the inflammatory reaction. Intensification of the process of permeability is accompanied by interstitial edema, deformation of the interalveolar septa, and structural disorganization of alveolar epithelium cells. Local lesions of microcirculation result in tissue hypoxia and induce processes of fibrosis. Laser biostimulation promotes reversion of the inflammatory process and stabilizes fibroplastic processes. Basic principles of pathogenetic therapy were stated. It was shown that low-energy laser irradiation satisfies these requirements as an additional method in the therapy of destructive lung diseases.

Indian J Chest Dis Allied Sci. 2003 Jan-Mar;45(1):19-23.

Rose of gallium arsenide laser irradiation at 890 nm as an adjunctive to anti-tuberculosis drugs in the treatment of pulmonary tuberculosis.

Puri MM, Arora VK.

Department of Tuberculosis and Chest Diseases, LRS Institute of Tuberculosis and Allied Diseases, Sri Aurobindo Marg, New Delhi, India.

BACKGROUND: Tuberculosis is a global emergency with about nine million people developing disease every year. The long duration of treatment has emerged as a major obstacle in the control of tuberculosis. There is a need for development of new drugs and or shortened therapy. METHODS: The present study was carried out to explore whether any benefit could be achieved by the addition of low level energy laser therapy (LLLT) to the conventional anti-tubercular chemotherapy. One-hundred-thirty new sputum smear positive patients of pulmonary tuberculosis were enrolled to evaluate the bio-stimulatory effects of Gallium Arsenide laser irradiation at 890 nm, as an adjuvant therapy. These patients were randomly divided into two groups to receive either LLLT or sham irradiation (control) concomitantly with anti-tuberculosis chemotherpy. RESULTS: The patients treated with semiconductor laser as an adjuvant therapy along with anti-tuberculosis drugs had a faster clearance of tubercle bacilli from the sputum as compared to the control group (P value at :45 days=0.1392, 60 days=0.0117, 75 days=0.00805, 90 days=0.00739). CONCLUSIONS: These findings provide preliminary evidence that low level laser therapy with Gallium Arsenide laser may be a promising adjunctive therapy for patients with tuberculosis. Faster conversion of sputum should prevent the development of resistant mutants.

The Cochrane Database of Systemic Reviews. 2002.

Low level laser therapy for treating tuberculosis.

Vlassov V V, Pechatnikov L M, MacLehose H G.

The authors have made a thorough literature search for studies using laser therapy as an adjunct therapeutic modality in the treatment of tuberculosis. These studies come from the former Soviet states and India. The studies in Russian language have generally only been available as Medline abstract and they have been vague on the details. Now, for the first time, Russian researchers have evaluated the original texts. Laser therapy has been used in many ways. Acupuncture points, irradiation over the organ, blood irradiation, puncture into the lungs, irradiation into the trachea and into the urinary bladder. Laser types used have also differed a lot; HeNe, nitrogen, GaAs, Nd:YVO4 and at powers ranging from 2 to 200 mW. The weak spot in previous Cochrane reviews on laser therapy has been the lack of dosage analyses. No such analysis has been made in the current study, but with the different therapeutic approaches used, such an analysis is not possible in this case. The reviewers have not found any randomised or quasi-randomised studies, but an evaluation of the quality of the studies has been performed. There is a lack of relevant information on procedures in many studies and some contradictory statements. All in all, the reviewers come to the conclusion that laser therapy is currently being used to treat tuberculosis without evidence of its benefits and harms.

Probl Tuberk. 2002;(8):16-8.

Efficiency of supra-venous blood laser radiation used in the treatment of disseminated pulmonary tuberculosis in adolescents.

[Article in Russian]

Rusakova LI, Dobkin VG, Ovsiankina ES.

In 19 of 40 adolescent patients with disseminated pulmonary tuberculosis, supravenous blood laser radiation was used in the complex treatment 2-3 weeks after the initiation of chemotherapy. The use of this type of laser therapy enhanced the efficiency of the treatment, accelerated positive changes of tuberculosis by 2.5-3.5 months, as evidenced by clinical and laboratory parameters, led to a smooth course of tuberculosis to develop less pronounced residual changes in the lung.

Cochrane Database Syst Rev. 2002;(3):CD003490.

Low level laser therapy for treating tuberculosis.

Vlassov VV, Pechatnikov LM, MacLehose HG.

Russian Branch, The Cochrane Collaboration, PO Box 54, Moscow, Russia, 127238. vlassov@cochrane.ru

BACKGROUND: The main treatment for tuberculosis is antituberculous drugs. Low energy laser therapy is used as an adjunct to antituberculous drugs, predominantly in the former Soviet Union and India. OBJECTIVES: To assess the benefits and harms of low level laser therapy for treating tuberculosis in randomized and quasi-randomized controlled trials. To seek information about potential benefits or harms from observational studies. SEARCH STRATEGY: We searched the Cochrane Infectious Diseases Group specialized trials register (up to June 2001), the Cochrane Controlled Trials Register (Issue 1, 2001), MEDLINE (1966 to December 2001), EMBASE (1988 to December 2001), CINAHL (up to November 2001), PEDro (up to November 2001), the Science Citation Index (up to December 2001), National Centre for Science Information at the Indian Institute of Science (15 April 2002), electronic catalogue of the Central Medical Library (Moscow; 1988 to January 2002), the internet using ‘Google’ (21 January 2002), and reference lists of articles. We contacted relevant organizations and researchers. SELECTION CRITERIA: (1) Randomized and quasi-randomized controlled trials comparing low level laser therapy with no low level laser therapy in people with tuberculosis. We also conducted a subsidiary analysis of of the potential benefits and harms from observational studies. DATA COLLECTION AND ANALYSIS: Two reviewers independently assessed trial quality and extracted data. We contacted study authors for additional information. Adverse event information was collected from the studies. MAIN RESULTS: No randomized or quasi-randomized controlled trials met the inclusion criteria for the review. The potential benefits and harms from 29 observational studies involving over 3500 people are described. REVIEWER’S CONCLUSIONS: We have not identified any well designed trials using low level laser therapy (LLLT) to treat tuberculosis. Therefore, the use of LLLT to treat tuberculosis is not supported by reliable evidence.

Probl Tuberk. 2002;(8):16-8.

Efficiency of supra-venous blood laser radiation used in the treatment of disseminated pulmonary tuberculosis in adolescents.

[Article in Russian]

Rusakova LI, Dobkin VG, Ovsiankina ES.

In 19 of 40 adolescent patients with disseminated pulmonary tuberculosis, supravenous blood laser radiation was used in the complex treatment 2-3 weeks after the initiation of chemotherapy. The use of this type of laser therapy enhanced the efficiency of the treatment, accelerated positive changes of tuberculosis by 2.5-3.5 months, as evidenced by clinical and laboratory parameters, led to a smooth course of tuberculosis to develop less pronounced residual changes in the lung.

COMPARATIVE EFFECTIVENESS DETOXIFICATION TRASFUSION OF THERAPY (TT) AND INTRAVENOUS OF A LASER IRRADIATION BLOOD (ILIB) IN COMPLEX TREATMENT OF PATIENTS BY A PULMONARY TUBERCULOSIS

P.I.Pitcyco, E.I.Krivoshapova

Kharkov Institute of Physicians Advanced Training, Kharkov. Ukraine

Two groups of the patients infiltrotivc by a pulmonary tuberculosis in a phase of de-cay, MBT+, (discovered Koch’s bacillus), comparable on a floor, age, pronounced in-toxication and syndrome, extent of process in lungs and regime of chemotherapy. I groups 45 patients receiving TT (albumen 5%- 100 ml alternated with hacmodesum 200 ml, daily ?10), II groups – 68 patients, which was carried out ILIB(wavclcngth 0,63 urn, capacity of radiation on an output flexible monofibcr conductor of light 2 mW, duration of a procedure 15 minutes, course 10 sessions). The efficiency of ther-apy was estimated on a reduction intoxication, which degree was defined on a level and structure middle molecules (MM) ( presence of pathological fractions and dis-polypcptidcmy) in whey of blood. MM were studied prior to the beginning and after realisation of rates TT and ILIB by a method hclium-chromatography on hcle “Toyop-carl HW-40F” (Japan). A normal level and structure MM (361 ± 14 cond I. unit) was established with inspection 40 healthy donors. Is established, that at the patients from I groups after realisation of a course TT the general level MM on the average has de-creased from 858 ± 29 unit till 769 ± 31 unit (p<0,05), the frequency dispolypcptid-cmy (from 75 ± 7,3 % till 33 ± 6,7 % has decreased; p<0,001) and pathological fractions (from 60 ± 7,2 % to 40 ± 7,2 %;
p>0,05). After a course ILIB investigated parameters also have decreased: a level MM from 920 ± 36 unit to 584 ± 23 unit (p<0,001), frequency dispolypeptidcmy from 78 ± .6,2% to 31 ± 6,8% (p<0,001) and pathological fractions from 67 ± 6,7% till 24 ± 6,4 % (p<0,001). From the given data it is visible, that the course ILIB in comparison with TT allows more and level of pathological fractions (24 ± 6,4 % against 40 ± 7,2 %).

Minerva Med. 1983 Jun 30;74(27):1683-8.

Antiasthma therapy with laser radiation of the stellate ganglia

[Article in Italian]

Bence L, Dussert G.

The authors present a 42-case series of bronchial asthma treated with laser therapy. The technique consists in the use of a diode laser with transcutaneous irradiation projected anteriorly into stellar ganglion. The clinical cases are presented together with the therapeutical results, pointing out how children are more receptive to this therapy and outlining the importance of timing and hour of treatment.