Radioimmunotherapy of methicillin-resistant Staphylococcus aureus in planktonic state and biofilms.

BACKGROUND: Implant associated infections such as periprosthetic joint infections are difficult to treat as the bacteria form a biofilm on the prosthetic material. This biofilm complicates surgical and antibiotic treatment. With rising antibiotic resistance, alternative treatment options are needed to treat these infections in the future. The aim of this article is to provide proof-of-principle data required for further development of radioimmunotherapy for non-invasive treatment of implant associated infections. METHODS: Planktonic cells and biofilms of Methicillin-resistant staphylococcus aureus are grown and treated with radioimmunotherapy. The monoclonal antibodies used, target wall teichoic acids that are cell and biofilm specific. Three different radionuclides in different doses were used. Viability and metabolic activity of the bacterial cells and biofilms were measured by CFU dilution and XTT reduction. RESULTS: Alpha-RIT with Bismuth-213 showed significant and dose dependent killing in both planktonic MRSA and biofilm. When planktonic bacteria were treated with 370 kBq of 213Bi-RIT 99% of the bacteria were killed. Complete killing of the bacteria in the biofilm was seen at 185 kBq. Beta-RIT with Lutetium-177 and Actinium-225 showed little to no significant killing. CONCLUSION: Our results demonstrate the ability of specific antibodies loaded with an alpha-emitter Bismuth-213 to selectively kill staphylococcus aureus cells in vitro in both planktonic and biofilm state. RIT could therefore be a potentially alternative treatment modality against planktonic and biofilm-related microbial infections.

Photobiomodulation plus Adipose-derived Stem Cells Improve Healing of Ischemic Infected Wounds in Type 2 Diabetic Rats.

In this study, we sought to investigate the impact of photobiomodulation and adipose-derived stem cells (ADS), alone and in combination, on the maturation step of wound healing in an ischemic infected delayed healing wound model in rats with type 2 diabetes mellitus (DM2). We randomly divided 24 adult male rats into 4 groups (n = 6 per group). DM2 plus an ischemic delayed healing wound were induced in all rats. The wounds were infected with methicillin-resistant Staphylococcus aureus. Group 1 was the control (placebo) group. Group 2 received only photobiomodulation (890 nm, 80 Hz, 0.324 J/cm2, and 0.001 W/cm2). Group 3 received only the allograft ADS. Group 4 received allograft ADS followed by photobiomodulation. On days 0, 4, 8, 12, and 16, we performed microbiological examination (colony forming units, [CFU]), wound area measurement, wound closure rate, wound strength, and histological and stereological examinations. The results indicated that at day 16, there was significantly decreased CFU (Analysis of variance, p = 0.001) in the photobiomodulation + ADS (0.0 ± 0.0), ADS (1350 ± 212), and photobiomodulation (0.0 ± 0.0) groups compared with the control group (27250 ± 1284). There was significantly decreased wound area (Analysis of variance, p = 0.000) in the photobiomodulation + ADS (7.4 ± 1.4 mm2), ADS (11 ± 2.2 mm2), and photobiomodulation (11.4 ± 1.4 mm2) groups compared with the control group (25.2 ± 1.7). There was a significantly increased tensiometeric property (stress maximal load, Analysis of variance, p = 0.000) in the photobiomodulation + ADS (0.99 ± 0.06 N/cm2), ADS (0.51 ± 0.12 N/cm2), and photobiomodulation (0.35 ± 0.15 N/cm2) groups compared with the control group (0.18 ± 0.04). There was a significantly modulated inflammatory response in (Analysis of variance, p = 0.049) in the photobiomodulation + ADS (337 ± 96), ADS (1175 ± 640), and photobiomodulation (69 ± 54) treatments compared to control group (7321 ± 4099). Photobiomodulation + ADS gave significantly better improvements in CFU, wound area, and wound strength compared to photobiomodulation or ADS alone. Photobiomodulation, ADS, and their combination significantly hastened healing in ischemic methicillin-resistant Staphylococcus aureus infected delayed healing wounds in rats with DM2. Combined application of photobiomodulation plus ADS demonstrated an additive effect.

Impact of Photobiomodulation and Condition Medium on Mast Cell Counts, Degranulation, and Wound Strength in Infected Skin Wound Healing of Diabetic Rats.

Background: Numerous people suffer from diabetes mellitus (DM) and resultant diabetic foot ulcers (DFU), which lack effective treatment. Photobiomodulation (PBM) has accelerated wound healing in diabetic animals and patients in some studies. However, there is scant information on the number and activation state of skin mast cells (MCs) in PBM-treated diabetic wounds. Objective: We intend to assess the influence of the number of MCs and degranulation in the remodeling step of an infected wound model on wound strength and its microbial flora in a type 1 DM (T1DM) rat model by administration of PBM, condition medium (CM) derived from human bone marrow mesenchymal stem cells (hBMMSCs), and the combination of PBM+CM. Methods: We prepared CM by culturing hBMMSCs. T1DM was induced in 72 rats and, after 1 month, we created one excisional wound in each rat. All wounds were infected with methicillin-resistant Staphylococcus aureus (MRSA). We divided the rats into four groups: (n = 18): (i) control; (ii) PBM; (iii) CM, and (iv) PBM+CM. On days 4, 7, and 15, we conducted microbiological, tensiometrical, and stereological analyses. The type of MCs (T1MCs, T2MCs, or T3MCs) and total number of MCs (TOMCs) were counted by light microscopy. Results: On day 15, the PBM+CM, PBM, and CM groups had significantly increased wound strength compared with the control group. There was a significant decrease in colony-forming units (CFU) at all time points in the PBM+CM and PBM groups. The PBM+CM and PBM groups had more stable MCs (T1MCs), less significant degranulated MCs (T2MCs), less significant disintegrated MCs (T3MCs), and less significant TOMCs compared with the control group at all time points. Conclusions: PBM+CM and PBM treatments significantly increased the healing process in an ischemic and MRSA-infected wound model of T1DM rats. PBM+CM and PBM significantly decreased both TOMCs and their degranulation, and significantly decreased CFU.

Improvement in infected wound healing in type 1 diabetic rat by the synergistic effect of photobiomodulation therapy and conditioned medium.

We investigated the effects of photobiomodulation therapy (PBMT) and conditioned medium (CM) of human bone marrow mesenchymal stem cells (hBM-MSC) individually and/or in combination on the stereological parameters and the expression of basic fibroblast growth factor (bFGF), hypoxia-inducible factor (HIF-1α), and stromal cell-derived factor-1α (SDF-1α) in a wound model infected with methicillin-resistant Staphylococcus aureus (MRSA) in diabetic rats. CM was provided by culturing hBM-MSCs. Type 1 diabetes mellitus (T1DM) was induced in 72 rats, divided into four groups, harboring 18 rats each: group 1 served as a control group, group 2 received PBMT, group 3 received CM, and group 4 received CM + PBMT. On days 4, 7, and 15, six animals from each group were euthanized and the skin samples were separated for stereology examination and gene expression analysis by real-time polymerase chain reaction. In the CM + PBMT, CM, and PBMT groups, significant decreases were induced in the number of neutrophils (1460 ± 93, 1854 ± 138, 1719 ± 248) and macrophages (539 ± 69, 804 ± 63, 912 ± 41), and significant increases in the number of fibroblasts (1073 ± 116, 836 ± 75, 912 ± 41) and angiogenesis (15 230 ± 516, 13 318 ± 1116, 14 041 ± 867), compared with those of the control group (2690 ± 371, 1139 ± 145, 566 ± 90, 12 585 ± 1219). Interestingly, the findings of the stereological examination in the CM + PBMT group were statistically more significant than those in the other groups. In the PBMT group, in most cases, the expression of bFGF, HIF-1α, and SDF-1α, on day 4 (27.7 ± 0.14, 28.8 ± 0.52, 27.5 ± 0.54) and day 7 (26.8 ± 1.4, 29.6 ± 1.4, 28.3 ± 1.2) were more significant than those in the control (day 4, 19.3 ± 0.42, 25.5 ± 0.08, 22.6 ± 0.04; day 7, 22.3 ± 0.22, 28.3 ± 0.59, 24.3 ± 0.19) and other treatment groups. The application of PBMT + CM induced anti-inflammatory and angiogenic activities, and hastened wound healing process in a T1 DM model of MRSA infected wound.

Demonstration of the bactericidal effects of the boron neutron capture reaction.

This pilot study represents a paradigm shift, using BNCT for the treatment of bacterial overgrowth on surgically implanted medical devices. In this study, titanium diboride disks were inoculated with S. aureus and irradiated in a thermal neutron beam. After a delivery of 2.6 × 1012 n/cm2 the surviving fraction of S. aureus on an irradiated disk was 3.1 × 10-5 when compared with non-irradiated controls. This pilot study demonstrates proof of principle of boron neutron capture therapy for infection control (BNCIC).

The effects of photobiomodulation therapy on Staphylococcus aureus infected surgical wounds in diabetic rats. A microbiological, histopathological, and biomechanical study.

PURPOSE: 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). METHODS: 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. RESULTS: 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). CONCLUSION: Photobiomodulation therapy may be useful in the management of wound infection through a significant bacterial growth inhibition and an acceleration of wound healing process.

The effects of photobiomodulation therapy on Staphylococcus aureus infected surgical wounds in diabetic rats. A microbiological, histopathological, and biomechanical study

ABSTRACT PURPOSE: 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). METHODS: 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. RESULTS: 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). CONCLUSION: Photobiomodulation therapy may be useful in the management of wound infection through a significant bacterial growth inhibition and an acceleration of wound healing process.

The effects of low level laser therapy on Staphylococcus aureus infected third-degree burns in diabetic rats.

PURPOSE: To characterize the effects of low-level laser (LLL) on third-degree burn wounds which were infected with Staphylococcus aureus (S. aureus) in diabetic rats. METHODS: Thirty streptozotocin-induced diabetic rats were divided into two groups: the control and the LLL groups. Third-degree burns were induced using a heated metal rod, and then, were contaminated with S. aureus. The wounds in the LLL group were irradiated with a LLL (685nm) daily for five consecutive days, starting three days after the induction. The wound area was measured at 3, 5, 8, 14 and 21 days after burning. At the end of trial, the skin samples were harvested. RESULTS: Reduction in wound areas in the LLL and control groups were significantly different only on the 21st day (p<0.05). The mean bacterial numbers in the LLL group were significantly lower (p<0.05) than those in the control group. The number of macrophages, new blood vessels, fibroblast, and elevated collagen deposition in the LLL group significantly increased compared to the control group (p<0.05). The mean breaking strength of scars in the control group was significantly lower (p<0.05) than that of the LLL group. CONCLUSION: The low-level laser improved the healing of S. aureus third-degree burn infections in diabetic rats.

The effects of low level laser therapy on Staphylococcus aureus infected third-degree burns in diabetic rats

PURPOSE: To characterize the effects of low-level laser (LLL) on third-degree burn wounds which were infected with Staphylococcus aureus (S. aureus) in diabetic rats. METHODS: Thirty streptozotocin-induced diabetic rats were divided into two groups: the control and the LLL groups. Third-degree burns were induced using a heated metal rod, and then, were contaminated with S. aureus. The wounds in the LLL group were irradiated with a LLL (685nm) daily for five consecutive days, starting three days after the induction. The wound area was measured at 3, 5, 8, 14 and 21 days after burning. At the end of trial, the skin samples were harvested. RESULTS: Reduction in wound areas in the LLL and control groups were significantly different only on the 21st day (p<0.05). The mean bacterial numbers in the LLL group were significantly lower (p<0.05) than those in the control group. The number of macrophages, new blood vessels, fibroblast, and elevated collagen deposition in the LLL group significantly increased compared to the control group (p<0.05). The mean breaking strength of scars in the control group was significantly lower (p<0.05) than that of the LLL group. CONCLUSION: The low-level laser improved the healing of S. aureus third-degree burn infections in diabetic rats.

The efficiency of cross-linking methods in eradication of bacteria is influenced by the riboflavin concentration and the irradiation time of ultraviolet light.

PURPOSE: To determine bacterial eradication using numerous riboflavin concentrations and different ultraviolet light A (UVA) radiant and exposure time in an experimental model. METHODS: Dilutions of Staphylococcus epidermidis were mixed with riboflavin at varying concentrations (0.007­0.09%). Effects on bacterial growth were evaluated after 0, 3, 6, 30 and 60 min of UVA exposure (irradiance 30 and 3 mW/cm2). Standard settings of UVA were compared with high-power UVA approach. Different fluid thicknesses of the exposed dilutions were also examined to improve the model. RESULTS: Bacterial eradication (%) was increased after 60 compared with 30 min of UVA exposure for concentrations of 0.03­0.07% but not for 0.09% riboflavin.There was a significant difference between the efficacy between 0.03 and 0.09% and eradication dropped from 80%to 50%(p = 0.01).A correlation could be calculated for the amount of riboflavin at 60 min of UVA and the ability to kill bacteria(p = 0.01). The antibacterial effect was more pronounced when the tested bacterial suspension thickness was reduced. High-power UVA method was less potent in microbial elimination, eradicating only 60%of bacteria after 6 min versus 97­99%after 60 min in the low-power setting, compared with respective controls (p = 0.02). CONCLUSIONS: In these in vitro experiments, a longer UVA exposure time in combination with lower riboflavin levels were found to be favourable in killing bacteria as compared to the standard cross-linking settings. Further studies are needed to evaluate the clinical relevance of these findings.

X-Ray treatment of carbuncles and furuncles (boils): a historical assessment.

The goal of this article was to assess the historical role of radiotherapy in the treatment of selected inflammatory diseases. The specific research involved a literature-based assessment of the use of x-rays during the first half of the 20th century for the treatment of furuncles and carbuncles, the potentially serious staphylococcus infections. X-Rays were reported to be effective as a treatment at relatively low dose, about 10-20% of the skin erythema dose, which often quickly and profoundly reduce pain and accelerate the resolution/healing of the furuncles and carbuncles. These findings were based on considerable clinical experience that was generally reported in the form of case studies. The mechanism of x-ray-induced reduction of inflammation and acceleration of healing was suggested to result from a combination of immune alterations that enhanced phagocytosis as well as via an anti-localization effect on the pathogenic organism that facilitates their destruction.

Low level laser therapy (AlGaInP) applied at 5J/cm2 reduces the proliferation of Staphylococcus aureus MRSA in infected wounds and intact skin of rats.

BACKGROUND: Laser therapy is a low cost, non-invasive procedure with good healing results. Doubts exist as to whether laser therapy action on microorganisms can justify research aimed at investigating its possible effects on bacteria-infected wounds. OBJECTIVE: To assess the effect of low intensity laser on the rate of bacterial contamination in infected wounds in the skin of rats. METHODS: An experimental study using 56 male Wistar rats. The animals were randomly divided into eight groups of seven each. Those in the "infected" groups were infected by Staphylococcus aureus MRSA in the dorsal region. Red laser diode (AlGaInP) 658nm, 5J/cm2 was used to treat the animals in the "treated" groups in scan for 3 consecutive days. Samples were drawn before inoculating bacteria and following laser treatment. For statistical analysis we used the nonparametric Wilcoxon (paired data) method with a significance level of p <0.05. RESULTS: The statistical analysis of median values showed that the groups submitted to laser treatment had low bacterial proliferation. CONCLUSION: The laser (AlGaInP), with a dose of 5J/cm2 in both intact skin and in wounds of rats infected with Staphylococcus aureus MRSA, is shown to reduce bacterial proliferation.

Low level laser therapy (AlGaInP) applied at 5J/cm2 reduces the proliferation of Staphylococcus aureus MRSA in infected wounds and intact skin of rats* / Laser de baixa intensidade (AlGaInP) aplicado com 5J/cm2 reduz a proliferação de Staphilococcus aureus MRSA em feridas infectadas e em pele íntegra de ratos

Laser therapy is a low cost, non-invasive procedure with good healing results. Doubts exist as to whether laser therapy action on microorganisms can justify research aimed at investigating its possible effects on bacteria-infected wounds. To assess the effect of low intensity laser on the rate of bacterial contamination in infected wounds in the skin of rats. An experimental study using 56 male Wistar rats. The animals were randomly divided into eight groups of seven each. Those in the "infected" groups were infected by Staphylococcus aureus MRSA in the dorsal region. Red laser diode (AlGaInP) 658nm, 5J/cm2 was used to treat the animals in the "treated" groups in scan for 3 consecutive days. Samples were drawn before inoculating bacteria and following laser treatment. For statistical analysis we used the nonparametric Wilcoxon (paired data) method with a significance level of p <0.05. The statistical analysis of median values showed that the groups submitted to laser treatment had low bacterial proliferation. The laser (AlGaInP), with a dose of 5J/cm2 in both intact skin and in wounds of rats infected with Staphylococcus aureus MRSA, is shown to reduce bacterial proliferation. .

Fundamentos: A terapia a laser é um procedimento de baixo custo, não invasiva e com bom desempenho na cicatrização. As dúvidas existentes quanto a sua ação sobre microrganismos justifica a realização de pesquisas visando investigar os possíveis efeitos em feridas infectadas por bactérias. Avaliar o efeito do laser de baixa intensidade sobre a taxa de contaminação bacteriana em feridas infectadas na pele de ratos. Estudo experimental, utilizando 56 ratos machos Wistar. Os animais foram distribuídos aleatoriamente em oito grupos de sete animais. Nos animais dos grupos lesionados foi realizada uma incisão na região dorsal.Os animais dos grupos infectados foram infectados por Staphylococcus aureus MRSA. os animais dos grupos tratados foram tratados com laser de Diodo vermelho (AlGaInP) 658nm, 5J/cm2 em varredura, durante 3 dias consecutivos. Foi colhida uma amostra antes de inocular as bactérias e outra após o tratamento com laser. Para a análise estatística foram utilizados os testes não paramétricos de Wilcoxon (dados pareados). Considerando como significante p<0,05. Através da análise estatística das medianas, observou-se que os grupos submetidos ao laser apresentavam uma proliferação bacteriana menor. O laser (AlGaInP), com uma dose de 5J/cm2, tanto em feridas quanto em pele íntegra de ratos infectados por Staphilococcus aureus MRSA, se mostrou capaz de reduzir a proliferação bacteriana. .

Laser-assisted nasal decolonization of Staphylococcus aureus, including methicillin-resistant Staphylococcus aureus.

OBJECTIVES: Methicillin-sensitive Staphylococcus aureus (MSSA) and methicillin-resistant S aureus (MRSA) contribute to 25% of nosocomial infections, increasing complications, health care cost, and growing antibiotic resistance. Nasal decolonization (ND) can reduce the staphylococcal infection rate. A new method of laser therapy (LT) MRSA ND was tested. STUDY DESIGN: This is a prospective, nonrandomized study. METHODS: Following institutional review board approvals, 25 patients colonized with MSSA/MRSA were allocated to 4 treatment arms; low-power, dual-wavelength 870-/930-nm laser alone (GR1); low-power, dual-wavelength laser followed by erythromycin (E-mycin) cream (GR2); low-power, dual-wavelength laser followed by peroxide irrigation (GR3); and high-power 940-nm laser alone (GR4). Quantitative cultures were obtained before and after in all arms. Laser therapy was performed via a laser fiber diffuser, delivering 200 to 600 J/cm² to each naris circumferentially. Patient's distribution was 3 in GR1, 14 in GR2, 4 in GR3, and 4 in GR4 (last 10 recruited to GR4). RESULTS: Nasal decolonization for GR1, GR2, GR3, and GR4 was 1 of 3, 13 of 14, 2 of 4, and 4 of 4, respectively. Because LT + E-mycin cleared all first 3 patients of MRSA and MSSA, all remaining patients were treated with LT + Er with over 90% of patients clearing. No adverse events or discomfort were reported. CONCLUSIONS: First human study using LT and topical E-mycin in ND is presented. Laser therapy can eradicate MRSA and potentially resensitization of bacteria to the antimicrobial effect of erythromycin. Although decolonization was maintained at 4 weeks posttreatment, further studies can determine the LT long-term effect.

Antiinflammatory effect of low-level laser therapy on Staphylococcus epidermidis endophthalmitis in rabbits.

A rabbit model of endophthalmitis was established to evaluate the antiinflammatory effect of low-level laser therapy (LLLT) as an adjunct to treatment for Staphylococcus epidermidis endophthalmitis. Rabbits were randomly divided into three groups to receive intravitreal injections into their left eye: group A received 0.5 mg vancomycin (100 µl), group B received 0.5 mg vancomycin + 0.2 mg dexamethasone (100 µl), and group C received 0.5 mg vancomycin (100 µl) and continuous wave semiconductor laser irradiation (10 mW, λ = 632 nm) focused on the pupil. Slit lamp examination and B-mode ultrasonography were conducted to evaluate the symptoms of endophthalmitis. Polymorphonuclear cells and tumour necrosis factor alpha (TNF-α) in aqueous fluid were measured at 0 h, and 1, 2, 3, 7 and 15 days. A histology test was conducted at 15 days. B-mode ultrasonography and histology revealed that groups B and C had less inflammation than group A at 15 days. Groups B and C had fewer polymorphonuclear cells and lower levels of TNF-α in aqueous fluid than group A at 2, 3 and 7 days (P < 0.05). There was no significant difference between groups B and C (P > 0.05). There was no significant difference between groups A, B and C at 15 days (P > 0.05). As an adjunct to vancomycin therapy to treat S. epidermidis endophthalmitis, LLLT has an antiinflammatory effect similar to that of dexamethasone.

Laser irradiation effect on Staphylococcus aureus and Pseudomonas aeruginosa biofilms isolated from venous leg ulcer.

Chronic wounds, including diabetic foot ulcers, pressure ulcers and venous leg ulcers, represent a significant cause of morbidity in developed countries, predominantly in older patients. The aetiology of these wounds is probably multifactorial, but the role of bacteria in their pathogenesis is still unclear. Moreover, the presence of bacterial biofilms has been considered an important factor responsible for wounds chronicity. We aimed to investigate the laser action as a possible biofilm eradicating strategy, in order to attempt an additional treatment to antibiotic therapy to improve wound healing. In this work, the effect of near-infrared (NIR) laser was evaluated on mono and polymicrobial biofilms produced by two pathogenic bacterial strains, Staphylococcus aureus PECHA10 and Pseudomonas aeruginosa PECHA9, both isolated from a chronic venous leg ulcer. Laser effect was assessed by biomass measurement, colony forming unit count and cell viability assay. It was shown that the laser treatment has not affected the biofilms biomass neither the cell viability, although a small disruptive action was observed in the structure of all biofilms tested. A reduction on cell growth was observed in S. aureus and in polymicrobial biofilms. This work represents an initial in vitro approach to study the influence of NIR laser treatment on bacterial biofilms in order to explain its potentially advantageous effects in the healing process of chronic infected wounds.

Enhancement of cutaneous immune response to bacterial infection after low-level light therapy with 1072 nm infrared light: a preliminary study.

We investigated the photobiomodulation effects of 1072 nm infrared light on the natural immune response involved in anti-bacterial and wound healing processes. Thirty mice infected with MRSA on the skin were divided into two groups. The experimental group was treated with 1072 nm infrared light (irradiance: 20 mW/cm(2), fluence: 12 J/cm(2) for 10 min) at 2, 4, 8, 12, 24 h, 3 and 5 days after inoculation and the control group with sham light. Serial changes of the mRNA levels of TLR2, IL-1ß, TNF-α, IL-6, iNOS, MCP-1, TGF-ß, bFGF and VEGF were studied by real time RT-PCR and those of the expression level of VEGF, bFGF, TGF-ß and NF-κB by immunohistochemistry. The mRNA levels of the cytokines involved in the early phase of anti-bacterial immune response (IL-1ß, TNF-α, IL-6, MCP-1) increased significantly in the 1072 nm group, peaking between 12 and 24 h post-inoculation. These levels normalized after 3-5 days. Immunohistochemistry revealed a notably stronger expression of VEGF in the 1072 nm group from 8-h post-inoculation to 5-day post-inoculation. We concluded that 1072 nm infrared light had a photobiomodulation effect which resulted in an enhanced biological immune response to the bacterial infection by MRSA and also increased the expression of VEGF to a significant level.

Photodynamic therapy using intra-articular Photofrin for murine MRSA arthritis: biphasic light dose response for neutrophil-mediated antibacterial effect.

BACKGROUND AND OBJECTIVE: Bacterial arthritis does not respond well to antibiotics and moreover multidrug resistance is spreading. We previously tested photodynamic therapy (PDT) mediated by systemic Photofrin® in a mouse model of methicillin-resistant Staphylococcus aureus (MRSA) arthritis, but found that neutrophils were killed by PDT and therefore the infection was potentiated. STUDY DESIGN/MATERIALS AND METHODS: The present study used an intra-articular injection of Photofrin® and optimized the light dosimetry in order to maximize bacterial killing and minimize killing of host neutrophils. MRSA (5 × 10(7) CFU) was injected into the mouse knee followed 3 days later by 1 µg of Photofrin® and 635-nm diode laser illumination with a range of fluences within 5 minutes. Synovial fluid was sampled 6 hours or 1-3, 5, and 7 days after PDT to determine MRSA colony-forming units (CFU), neutrophil numbers, and levels of cytokines. RESULTS: A biphasic light dose response was observed with the greatest reduction of MRSA CFU seen with a fluence of 20 J cm(-2), whereas lower antibacterial efficacy was observed with fluences that were either lower or higher. Consistent with these results, a significantly higher concentration of macrophage inflammatory protein-2, a CXC chemokine, and greater accumulation of neutrophils were seen in the infected knee joint after PDT with a fluence of 20 J cm(-2) compared to fluences of 5 or 70 J cm(-2). CONCLUSION: PDT for murine MRSA arthritis requires appropriate light dosimetry to simultaneously maximize bacterial killing and neutrophil accumulation into the infected site, while too little light does not kill sufficient bacteria and too much light kills neutrophils and damages host tissue as well as bacteria and allows bacteria to grow unimpeded by host defense.

The use of 808-nm light therapy to treat experimental chronic osteomyelitis induced in rats by methicillin-resistant Staphylococcus aureus.

BACKGROUND DATA: In vivo and in vitro studies have reported that laser energy in differing wavelengths and irradiation regimes has a potential bactericidal effect on Staphylococcus aureus. OBJECTIVE: The purpose of this study was to investigate whether a light wavelength of 808 nm in varying doses has an effect on chronic osteomyelitis induced experimentally in the rat tibia. METHODS: Intramedullary cavities were surgically created in the left tibias of 39 adult Wistar albino rats. Five randomly selected subjects were injected with a sterile saline solution, and methicillin-resistant S. aureus (MRSA) was used to induce osteomyelitis in the remaining rats. After 3 weeks, rats with evidence of osteomyelitis were treated with debridement alone (n = 7), with debridement plus laser irradiation to induce photoeradication (n = 21), or were not treated at all [negative control, (n = 6)]. Active irradiation was performed using an 808 nm, 100 mW continuous-wave diode laser with a beam spot size of 0.7854 cm(2) (irradiance = 127.3 mW/cm(2)). Laser treatment commenced immediately after debridement surgery and was applied daily for 5 consecutive days. Irradiation lasted 60 secs (6 J at 7.64 J/cm(2): n = 7), 120 secs (12 J at 15.29 J/cm(2): n = 7), or 180 secs (18 J at 22.93 J/cm(2): n = 7). Rats in the sham and negative control groups were killed 21 days post-induction surgery, and those in the treatment groups were killed after 42 days. Following killing, tibias were removed and analyzed histopathologically, radiographically, and microbiologically. RESULTS: Histopathological analysis showed that infection levels had decreased by 37%, 67%, 81%, and 93% in the groups treated by debridement or by debridement plus 7.64, 15.29, and 22.93 J/cm(2) light therapy, respectively, compared to the negative control group. Osteomyelitis-induced rats had the highest bacteria count (5 × 10(5)). Bacterial counts fell to 1.6 × 10(4), 4.3 × 10(2), 5.5 × 10(1), and 3.3 × 10(0) in groups treated by debridement or by debridement plus 7.64, 15.29, and 22.93 J/cm(2) light therapy, respectively, compared to the negative control group. CONCLUSIONS: Within the limitations of this study, laser phototherapy with the appropriate irradiation parameters appears to be a promising adjunct and/or alternative technique to pharmacological agents in the treatment of osteomyelitis. The 808 nm 100 mW (127.3 mW/cm(2)) laser device used in this study achieved a maximum effect with an irradiation time of 180 secs, delivering 18 J at an energy density of 22.93 J/cm(2).