The effects of photobiomodulation therapy on Staphylococcus aureus infected surgical wounds in diabetic rats. A microbiological, histopathological, and biomechanical study.
- 1Full professor, Molecular Biology Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran. Design and supervised all phases of the study.
- 2PhD, Young Researchers and Elites Club, Science and Research Branch, Islamic Azad University, Tehran, Iran. Conception and design of the study; acquisition, analysis and interpretation of data; statistical analysis; manuscript writing.
To evaluate the effects of photobiomodulation therapy (PBMT) at 685 nm on diabetic wound healing in rats suffering from bacterial infection induced by Staphylococcus aureus (S. aureus).
Thirty streptozotocin-induced diabetic rats were allocated into two groups: control and PBMT. A 4-cm full-thickness linear-incision was made on the dorsal midline and was contaminated with S. aureus. The wounds in the PBMT group were irradiated daily for 5 consecutive days, starting 3 days after the induction and always in the mornings.
The result revealed that PBMT resulted in a significant decrease in S. aureus CFU in the PBMT group in comparison to the control group (P<0.05). The length of wounds, in the 2nd and 3rd weeks, in the PBMT group were significantly shorter compared to the control group (P<0.05). PBMT caused a significant increase in the histological parameters in comparison to the control group (P<0.05). Moreover, PBMT significantly increased the breaking strength of the surgical scars produced in the skin of the PBMT group when compared to the control group (P<0.05).
Photobiomodulation therapy may be useful in the management of wound infection through a significant bacterial growth inhibition and an acceleration of wound healing process.
Ultra-low power laser stimulation impairs the adhesion of Staphylococcus aureus to primary human cells, and interferes with the expression of staphylococcal pathogenic factors.
Lasers are commonly used in several fields of medicine as a complementary therapy for internal medicine, surgery and also diagnostics. The efficacy of ultra-low level laser therapy (ULLLT) at power levels around 0.15 mW/cm2 has been demonstrated both in in vitro experiments and in the clinical environment. This work used an ULLLT laser source to analyze its efficacy on Staphylococcus aureus adhesion to cells and on its ability to produce pathogenic factors. Laser stimulation succeeded in impairing the binding of S. aureus to primary human cells in culture and in inhibiting the expression of coagulase, one of the main staphylococcal pathogenic factors. The importance of the extracellular matrix (ECM) and the modification of the ECM redox potential in these activities were also evidenced.
UVC light prophylaxis for cutaneous wound infections in mice.
Wellman Center for Photomedicine, Massachusetts General Hospital, Boston, Massachusetts, USA.
UVC light has long been known to be highly germicidal but has not been much developed as a therapy for infections. This study investigated the potential of UVC light for the prophylaxis of infections developing in highly contaminated superficial cutaneous wounds. In vitro studies demonstrated that the pathogenic bacteria Pseudomonas aeruginosa and Staphylococcus aureus were inactivated at UVC light exposures much lower than those needed for a similar effect on mammalian keratinocytes. Mouse models of partial-thickness skin abrasions infected with bioluminescent P. aeruginosa and S. aureus were developed. Approximately 10(7) bacterial cells were inoculated onto wounds measuring 1.2 by 1.2 cm on the dorsal surfaces of mice. UVC light was delivered at 30 min after bacterial inoculation. It was found that for both bacterial infections, UVC light at a single radiant exposure of 2.59 J/cm(2) reduced the bacterial burden in the infected mouse wounds by approximately 10-fold in comparison to those in untreated mouse wounds (P < 0.00001). Furthermore, UVC light increased the survival rate of mice infected with P. aeruginosa by 58.3% (P = 0.0023) and increased the wound healing rate in mice infected with S. aureus by 31.2% (P < 0.00001). DNA lesions were observed in the UVC light-treated mouse wounds; however, the lesions were extensively repaired by 48 h after UVC light exposure. These results suggested that UVC light may be used for the prophylaxis of cutaneous wound infections.
Photomed Laser Surg. 2012 May;30(5):281-5. Epub 2012 Apr 17.
In vitro analysis of bacterial morphology by atomic force microscopy of low level laser therapy 660, 830 and 904 nm.
Department of Physical Therapy, FAMINAS-School of Minas, Minas Gerais, Brazil.
The objective of this study was to analyze the bacterial morphology by atomic force microscopy (AFM) after the application of low-level laser therapy (LLLT) in in vitro culture of Staphylococcus aureus ATCC 29213.
Infections caused by S. aureus are among the highest occurring in hospitals and can often colonize pressure ulcers. LLLT is among the methods used to accelerate the healing of ulcers. However, there is no consensus on its effect on bacteria.
MATERIALS AND METHODS:
After being cultivated and seeded, the cultures were irradiated using wavelengths of 660, 830, and 904 nm at fluences of 0, 1, 2, 3, 4, 5, and 16 J/cm(2). Viable cells of S. aureus strain were counted after 24 h incubation. To analyze the occurrence of morphological changes, the topographical measurement of bacterial cells was analyzed using the AFM.
The overall assessment revealed that the laser irradiation reduced the S. aureus growth using 830 and 904 nm wavelengths; the latter with the greatest inhibition of the colony-forming units (CFU/mL) (331.1±38.19 and 137.38±21.72). Specifically with 660 nm, the statistical difference occurred only at a fluence of 3 J/cm(2). Topographical analysis showed small changes in morphological conformity of the samples tested.
LLLT reduced the growth of S. aureus with 830 and 904 nm wavelengths, particularly with 904 nm at a fluence of 3 J/cm(2), where the greatest topographical changes of the cell structure occurred.
Photomed Laser Surg. 2009 Apr;27(2):221-6.
Blue 470-nm light kills methicillin-resistant Staphylococcus aureus (MRSA) in vitro.
Enwemeka CS, Williams D, Enwemeka SK, Hollosi S, Yens D.
School of Health Professions, Behavioral, and Life Sciences, New York Institute of Technology, Old Westbury, New York 11568-8000, USA. Enwemeka@nyit.edu
BACKGROUND DATA: In a previous study, we showed that 405-nm light photo-destroys methicillin-resistant Staphylococcus aureus (MRSA). The 390-420 nm spectral width of the 405-nm superluminous diode (SLD) source may raise safety concerns in clinical practice, because of the trace of ultraviolet (UV) light within the spectrum. OBJECTIVE: Here we report the effect of a different wavelength of blue light, one that has no trace of UV, on two strains of MRSA–the US-300 strain of CA-MRSA and the IS-853 strain of HA-MRSA–in vitro. MATERIALS AND METHODS: We cultured and plated each strain, and then irradiated each plate with 0, 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 25, 30, 35, 40, 45, 50, 55, or 60 J/cm2 of energy a single time, using a 470-nm SLD phototherapy device. The irradiated specimens were then incubated at 35 degrees C for 24 h. Subsequently, digital images were made and quantified to obtain colony counts and the aggregate area occupied by bacteria. RESULTS: Photo-irradiation produced a statistically significant dose-dependent reduction in both the number and the aggregate area of colonies formed by each strain (p < 0.001). The higher the dose the more bacteria were killed, but the effect was not linear, and was more impressive at lower doses than at higher doses. Nearly 30% of both strains was killed with as little as 3 J/cm2 of energy. As much as 90.4% of the US-300 and the IS-853 colonies, respectively, were killed with an energy density of 55 J/cm2. This same dose eradicated 91.7% and 94.8% of the aggregate area of the US-300 and the IS-853 strains, respectively. CONCLUSION: At practical dose ranges, 470-nm blue light kills HA-MRSA and CA-MRSA in vitro, suggesting that a similar bactericidal effect may be attained in human cases of cutaneous and subcutaneous MRSA infections.
Appl Environ Microbiol. 2009 Apr;75(7):1932-7. Epub 2009 Feb 6.
Inactivation of bacterial pathogens following exposure to light from a 405-nanometer light-emitting diode array.
Maclean M, MacGregor SJ, Anderson JG, Woolsey G.
The Robertson Trust Laboratory for Electronic Sterilisation Technologies, University of Strathclyde, 204 George Street, Glasgow, Scotland. email@example.com
This study demonstrates the susceptibility of a variety of medically important bacteria to inactivation by 405-nm light from an array of light-emitting diodes (LEDs), without the application of exogenous photosensitizer molecules. Selected bacterial pathogens, all commonly associated with hospital-acquired infections, were exposed to the 405-nm LED array, and the results show that both gram-positive and gram-negative species were successfully inactivated, with the general trend showing gram-positive species to be more susceptible than gram-negative bacteria. Detailed investigation of the bactericidal effect of the blue-light treatment on Staphylococcus aureus suspensions, for a range of different population densities, demonstrated that 405-nm LED array illumination can cause complete inactivation at high population densities: inactivation levels corresponding to a 9-log(10) reduction were achieved. The results, which show the inactivation of a wide range of medically important bacteria including methicillin-resistant Staphylococcus aureus, demonstrate that, with further development, narrow-spectrum 405-nm visible-light illumination from an LED source has the potential to provide a novel decontamination method with a wide range of potential applications.
Photomed Laser Surg. 2006 Dec;24(6):684-8
In vitro bactericidal effects of 405-nm and 470-nm blue light.
Guffey JS, Wilborn J.
Physical Therapy Plus, Bauxite, Arkansas 72011, USA. firstname.lastname@example.org
OBJECTIVE: The aim of this study was to determine the bactericidal effect of 405- and 470-nm light on two bacteria, Staphylococcus aureus and Pseudomonas aeruginosa, in vitro. BACKGROUND DATA: It is well-known that UV light kills bacteria, but the bactericidal effects of UV may not be unique since recent studies indicate that blue light produces a somewhat similar effect. The effects of blue light seem varied depending on wavelength, dose and the nature of the bacteria, hence this study. METHODS: Two common aerobes, Staphylococcus aureus and Pseudomonas aeruginosa, and anaerobic Propionibacterium acnes were tested. Each organism was treated with Super Luminous Diode probes with peak emission at 405 and 470 nm. Treatment was timed to yield 1, 3, 5, 10, and 15 Jcm2 doses. Colony counts were performed and compared to untreated controls. RESULTS: The 405-nm light produced a dose dependent bactericidal effect on Pseudomonas aeruginosa and Staphylococcus aureus (p < .05), achieving as much as 95.1% and nearly 90% kill rate for each, respectively. The 470-nm light effectively killed Pseudomonas aeruginosa at all dose levels, but only killed Staphylococcus aureus at 10 and 15 J cm2. With this wavelength, as much as 96.5% and 62% reduction of Pseudomonas aeruginosa and Staphylococcus aureus was achieved, respectively. Neither of the two wavelengths proved bactericidal with anaerobic Propionibacterium acnes. CONCLUSION: The results indicate that, in vitro, 405- and 470-nm blue light produce dose dependent bactericidal effects on Pseudomonas aeruginosa and Staphylococcus aureus but not Propionibacterium acnes.
Photomed Laser Surg. 2006 Dec;24(6):680-3.
Effects of combined 405-nm and 880-nm light on Staphylococcus aureus and Pseudomonas aeruginosa in vitro.
Guffey JS, Wilborn J.
Physical Therapy Plus, Bauxite, Arkansas 72011, USA. email@example.com
OBJECTIVE: The aim of this study was to determine the effect of a combination of 405-nm blue light and 880-nm infrared light on Staphylococcus aureus and Pseudomonas aeruginosa in vitro. BACKGROUND DATA: Reports indicate that certain wavelengths and treatment parameters of light promote the growth of bacteria, but our earlier study indicates that light at specific wavelengths and intensities are bactericidal for specific organisms (1). METHODS: Two common aerobes, Staphylococcus aureus and Pseudomonas aeruginosa were tested because of their frequent isolation from skin infections and wounds. Each organism was treated simultaneously with a combination of 405-nm and 880-nm light emitted by a cluster of Super Luminous Diodes (SLDs). Doses of 1, 3, 5, 10, and 20 Jcm2 were used. Colony counts were performed and compared to untreated controls using Student t tests and one-way ANOVA with Tukey and Scheffe post hoc analyses. RESULTS: The results revealed significant dose-dependent bactericidal effects of the combined blue and infrared light on Staphylococcus aureus (F 4,94 = 5.38, p = 0.001) and Pseudomonas aeruginosa (F 4,95 = 21.35, p < 0.001). With P. aeruginosa, the treatment reduced the number of bacteria colonies at all doses, achieving statistical significance at 1, 3, and 20 J cm2 doses and reducing bacterial colony by as much as 93.8%; the most effective dose being 20 J cm2. Irradiation of S. aureus resulted in statistically significant decreases in bacterial colonies at all dose levels; the most decrease, 72%, was also achieved with 20 Jcm2. CONCLUSION: Appropriate doses of combined 405-nm and 880-nm phototherapy can kill Staphylococcus aureus and Pseudomonas aeruginosa in vitro, suggesting that a similar effect may be produced in clinical cases of bacterial infection.
J Antimicrob Chemother. 1997 Dec;40(6):873-6.
Killing of methicillin-resistant Staphyococcus aureus in vitro using aluminium disulphonated phtalocyanine, a light-activated antimicrobial agent.
Griffiths MA, Wren BW, Wilson M.
Department of Microbiology, Eastman Dental Institute for Oral Health Care Sciences, University of London, UK.
The aim of this study was to determine whether 16 epidemic methicillin-resistant strains of Staphylococcus aureus (EMRSA) could be killed by the light-activated antimicrobial agent aluminium disulphonated phthalocyanine (AIPcS2). EMRSA suspensions were irradiated with light from a laser diode in the presence of AIPcS2 and survivors enumerated. All strains were susceptible to killing, the bactericidal effect being dependent on the AIPcS2 concentration and the light dose. AIPcS2 rendered the bacteria light-sensitive almost immediately and killing was unaffected by the growth phase of the organism. Scavengers of singlet oxygen and free radicals protected the bacteria from killing. These results imply that light-activated antimicrobial agents may be useful in eliminating EMRSA from wounds or carriage sites.
Med Microbiol. 1995 Jan;42(1):62-6
Killing of methicillin-resistant Staphylococcus aureus by low-power laser light.
Wilson M, Yianni C.
Department of Microbiology, Eastman Dental Institute for Oral Health Sciences, University of London.
The purpose of this study was to determine whether a methicillin-resistant strain of Staphylococcus aureus (MRSA) could be sensitised by toluidine blue O (TBO) to killing by light from a low-power helium/neon (HeNe) laser. Suspensions containing c. 10(10) cfu of MRSA were irradiated with light from a 35 mW HeNe laser (energy dose: 0.5-2.1 J) in the presence of TBO (1.6-12.5 micrograms/ml) and the survivors were enumerated. The kills attained depended on both the light energy dose and concentration of TBO employed. A 4.47 log10 reduction in the viable count was achieved with a TBO concentration of 12.5 micrograms/ml and a light dose of 2.1 J (energy density 43 J/cm2). MRSA were susceptible to killing by the laser light within 30 s of exposure to the TBO. The results of this study have demonstrated that MRSA can be rapidly sensitised by TBO to killing by HeNe laser light and that killing depends on the light energy dose and sensitiser concentration.
Lethal photosensitisation of Staphylococcus aureus.
Wilson M, Pratten J.
Department of Microbiology, Institute of Dental Surgery, University of London, Great Britain.
The purpose of this study was to determine whether toluidine blue O (TBO) could sensitise Staphylococcus aureus to killing by light from a low-power helium/neon (HeNe) laser. Suspensions of the organism were irradiated with light from a HeNe laser in the presence and absence of TBO and the survivors enumerated. A 95% reduction (9 x 10(7) cfu) in the viable count was achieved following irradiation with 0.88 J of HeNe laser light in the presence of 12.5 micrograms/ml TBO whereas no significant reductions in viability were found when suspensions were exposed to this dose of laser light in the absence of TBO. With higher doses (3.5 J) of laser light statistically significant kills (3 x 10(7) cfu) were obtained in the absence of TBO implying the presence of an endogenous photosensitiser in the organism.