This PDF file contains the front matter associated with SPIE Proceedings Volume 9695, including the Title Page, Copyright information, Table of Contents, and Conference Committee listing.

The abstract is not available

Photobiomodulation with blue light is used for several treatment paradigms such as neonatal jaundice, psoriasis and back pain. However, little is known about possible side effects concerning melanoma cells in the skin. The aim of this study was to assess the safety of blue LED irradiation with respect to proliferation of melanoma cells. For that purpose we used the human malignant melanoma cell line SK-MEL28. Cell proliferation was decreased in blue light irradiated cells where the effect size depended on light irradiation dosage. Furthermore, with a repeated irradiation of the melanoma cells on two consecutive days the effect could be intensified. Fluorescence-activated cell sorting with Annexin V and Propidium iodide labeling did not show a higher number of dead cells after blue light irradiation compared to non-irradiated cells. Gene expression analysis revealed down-regulated genes in pathways connected to anti-inflammatory response, like B cell signaling and phagosome. Most prominent pathways with up-regulation of genes were cytochrome P450, steroid hormone biosynthesis. Furthermore, even though cells showed a decrease in proliferation, genes connected to the cell cycle were up-regulated after 24h. This result is concordant with XTT test 48h after irradiation, where irradiated cells showed the same proliferation as the no light negative control. In summary, proliferation of melanoma cells can be decreased using blue light irradiation. Nevertheless, the gene expression analysis has to be further evaluated and more studies, such as in-vivo experiments, are warranted to further assess the safety of blue light treatment.

As the interest in skin was increased, number of studies on skin care also have been increased. The reduction of skin density is one of the symptoms of skin aging. It reduces elasticity of skin and becomes the reason of wrinkle formation. Low level laser therapy (LLLT) has been suggested as one of the effective therapeutic methods for skin aging as in hasten to change skin density. This study presents the effect of a minimally invasive laser needle system (MILNS) (wavelength: 660nm, power: 20mW) in skin density. Rabbits were divided into three groups. Group 1 didn't receive any laser stimulation as a control group. Group 2 and 3 as test groups were exposed to MILNS with energy of 8J and 6J on rabbits' dorsal side once a week, respectively. Skin density of rabbits was measured every 12 hours by using an ultrasound skin scanner.

This work aimed the assessment of the biochemical changes during bone mineralization induced by laser and LED irradiation in an animal model of bone repair using a spectral model based on Raman spectroscopy. Six groups were studied: Clot, Laser (λ780 nm, 70 mW), LED (λ850 nm ± 10 nm, 150 mW), Biomaterial (biphasic synthetic microgranular hydroxyapatite (HA) + β-tricalcium phosphate), Laser + Biomaterial and LED + Biomaterial. When indicated, defects were further irradiated at 48 h interval during 2 wks, 20 J/cm² per session. At 15th and 30th days, femurs were dissected and spectra of the defects were collected. Raman spectra were submitted to a model to estimate the relative amount of collagen, phosphate HA and carbonate HA, by using spectra of pure collagen, biomaterial and basal bone, respectively. At 15th days, the use of biomaterial associated to phototherapy reduced the collagen formation, whereas the amount of carbonate HA was not different in all groups. The phosphate HA was higher in the groups that received biomaterial grafts. At 30th days, it was observed an increase of collagen for the group Laser + Biomaterial, and a reduction in the carbonate HA for the LED + Biomaterial. The phosphate HA was higher for the groups LED + Biomaterial and Laser + Biomaterial, while decreased for the group Biomaterial. These results indicated that the use of Laser and LED phototherapies improved the repair of bone defects grafted with the biomaterial by increasing the collagen deposition and phosphate HA.

The abstract is not available

Distinct lineages of human dermal fibroblasts play complementary roles in skin rejuvenation and wound healing, which makes them a target of phototherapy. However, knowledge about differential responses of specific cell lineages to different light parameters and moreover the actual molecular targets remain to be unravelled. The goal of this study was to investigate the impact of a range of parameters of light on the metabolic activity, collagen production, and cell migration of distinct lineages of human dermal fibroblasts. A rational approach was used to identify parameters with high therapeutic potential. Fibroblasts exhibited both inhibitory and cytotoxic change when exposed to a high dose of blue and cyan light in tissue culture medium containing photo-reactive species, but were stimulated by high dose red and near infrared light. Cytotoxic effects were eliminated by refreshing the medium after light exposure by removing potential ROS formed by extracellular photo-reactive species. Importantly, distinct lineages of fibroblasts demonstrated opposite responses to low dose blue light treatment when refreshing the medium after exposure. Low dose blue light treatment also significantly increased collagen production by papillary fibroblasts; high dose significantly retarded closure of the scratch wound without signs of cytotoxicity, and this is likely to have involved effects on both cell migration and proliferation. We recommend careful selection of fibroblast subpopulations and their culture conditions, a systematic approach in choosing and translating treatment parameters, and pursuit of fundamental research on identification of photoreceptors and triggered molecular pathways, while seeking effective parameters to address different stages of skin rejuvenation and wound healing.

Prompted by a study of traumatic brain injury (TBI) in a model system of cultured astrocytes, we discovered that low level laser illumination (LLL) at 660nm elevates the level of intracellular Ca²⁺. The coherence of the illumination was not essential since incoherent red light also worked. For cells bathed in low Ca²⁺ saline so that influx was suppressed, the Ca²⁺ level rose with no significant latency following illumination and consistent with a slow leak of Ca²⁺ from storage such as from the endoplasmic reticulum and/or mitochondria. When the cells were bathed in normal Ca²⁺ saline, the internal Ca²⁺ rose, but with a latency of about 17 seconds from the beginning of illumination. Pharmacologic studies with ryanodine inhibited the light effect. Testing the cells with fluid shear stress as used in the TBI model showed that mechanically induced elevation of cell Ca²⁺ was unaffected by illumination.

Exposure to 2.88 J/cm² of red light induces an adaptive response against a lethal pulse of 2.0 μm laser radiation in hTERT-RPE cells in vitro, but not in a knockdown mutant for vascular endothelial growth factor c (VEGF-C). The generally accepted initiation sequence for photobiomodulation is that absorption of red light by cytochome c oxidase (CCOX) of the electron transport chain increases the binding affinity of CCOX for O₂ vs. nitric oxide (NO). This results in displacement of NO by O₂ in the active site of CCOX, thereby increasing cellular respiration and intracellular ATP. We've previously reported that red-light exposure induces a small, but consistently reproducible, increase in NO levels in these cells. But the relative importance of NO and oxidative phosphorylation is unclear because little is known about the relative contributions of NO and ATP to the response. However, if NO dissociation from CCOX actually increases oxidative phosphorylation, one should see a corresponding increase in oxygen consumption. A Seahorse Extracellular Flux Analyzer was used to measure oxygen consumption rates (OCR) in normal and mutant cells as a proxy for oxidative phosphorylation. Both basal respiration and maximum respiration rates in normal cells are significantly higher than in the mutant. The normal cells have a significant amount of “excess capacity,” whereas the VEGF-C(KD) have little or none. The OCR in exposed normal cells is lower than in unexposed cells when measured immediately after exposure. The exposures used for these experiments had no effect on the OCR in mutant cells.

Tibial defect is very common musculoskeletal disorder which makes patient painful and uncomfortable. Many studies about bone regeneration tried to figure out fast bone healing on early phase. It is already known that low level laser therapy (LLLT) is very convenient and good for beginning of bone disorder. However, light scattering and absorption obstruct musculoskeletal therapy which need optimal photon energy delivery. This study has used an interstitial laser probe (ILP) to overcome the limitations of light penetration depth and scattering. Animals (mouse, C57BL/6) were divided into three groups: laser treated test group 1 (660 nm; power 10 mW; total energy 5 J) and test group 2 (660 nm; power 20 mW; total energy 10 J); and untreated control group. All animals were taken surgical operation to make tibial defect on right crest of tibia. The test groups were treated every 48 hours with ILP. Bone volume and X-ray attenuation coefficient were measured on 0, 14th and 28th day with u-CT after treatment and were used to evaluate effect of LLLT. Results show that bone volume of test groups has been improved more than control group. X-ray attenuation coefficients of each groups have slightly different. The results suggest that LLLT combined with ILP may affect on early phase of bone regeneration and may be used in various musculoskeletal disease in deep tissue layer.

Chronic wounds represent a significant burden to patients, health care professionals, and health care systems, affecting over 40 million patients and creating costs of approximately 40 billion € annually. We will present a medical device for photo-stimulated wound care based on a wearable large area flexible and disposable light management system consisting of a waveguide with incorporated micro- and nanometer scale optical structures for efficient light in-coupling, waveguiding and homogeneous illumination of large area wounds. The working principle of this innovative device is based on the therapeutic effects of visible light to facilitate the self-healing process of chronic wounds. On the one hand, light exposure in the red (656nm) induces growth of keratinocytes and fibroblasts in deeper layers of the skin. On the other hand, blue light (453nm) is known to have antibacterial effects predominately at the surface layers of the skin. In order to be compliant with medical requirements the system will consist of two elements: a disposable wound dressing with embedded flexible optical waveguides for the light management and illumination of the wound area, and a non-disposable compact module containing the light sources, a controller, a rechargeable battery, and a data transmission unit. In particular, we will report on the developed light management system. Finally, as a proof-of-concept, a demonstrator will be presented and its performances will be reported to demonstrate the potential of this innovative device.

Cancer has become a public health problem worldwide. Radiotherapy may be a treatment to a number of types of cancer, frequently using gamma-radiation with sources such as ¹³⁷Cs and ⁶⁰Co, with varying doses, dose rates, and exposure times to obtain a better as a stimulant for cell proliferation and tissue healing process. However, its effects on cancer cells are not yet well elucidated. The purpose of this work was to evaluate the effects of the LPL on breast cancer cultures after ionizing radiation. The breast cancer-MDA-MB-231 cells were gamma irradiated by a ⁶⁰Co source, with dose of 2.5 Gy. After 24h, cells were submitted to LPL irradiation using a red laser emitting at λ= 660 nm, with output power of 40 mW and exposure time of 30 s and 60 s. The plates were uniformly irradiated, with energy of 1.2 J and 2.4 J, respectively. Cell viability was analyzed using the exclusion method with trypan blue. Our results show that breast cancer cells submitted to LPL after ionizing radiation remained 95 % viable. No statistically significant differences were observed between laser and control untreated cells, (P > 0.05). These findings suggest that LPL did not influenced cancer cells viability.

Nitromedicine is a new medical treatment paradigm, focused on increasing nitric oxide (NO) bioavailability and modulating redox-signaling pathways combined with phototherapy, electrotherapy and stem cell therapy. It has been known since the discovery of the biological role of NO in the 1980s, that supplying NO donors such can have many beneficial effects in different conditions by stimulating stem cells and modulating the immune response, but there also exists a substantial risk of side-effects with long-term use. Excess NO can inhibit mitochondrial metabolism by binding to cytochrome c oxidase (CCO) and can also produce reactive nitrogen species (Peroxynitrite) by interacting with reactive oxygen species (ROS). To avoid these potential damaging side-effects we propose to combine the use of NO donors with three additional components. Firstly we believe that addition of antioxidants such as hydrogen sulfide donors, polyphenols and vitamins can neutralize ROS and RNS. Secondly we believe that application of appropriate wavelengths and dosages of light (blue, red or near infrared depending on the exact condition being treated) will dissociate NO from CCO (and other storage sites) thus restoring mitochondrial ATP production and stimulating healing in many situations. Thirdly delivering electrons to the body might help to saturate the free radicals with electrons, eliminate underlying oxidative stress, stabilize mitochondria, prevent further formation of pathological free radicals and increase the nitric oxide bioavailability. This combination therapy may be applied to treat a large variety of oxidative stressed related diseases such as degenerative diseases, immunological diseases, chronic infectious diseases, cancers and a broad range of unmet medical needs involving chronic inflammation with an emphasis on pain management.

Studies have shown that using high fluences of Low Intensity Laser Irradiation (HF-LILI) produce apoptotic effects on normal and neoplastic cells. This study aimed to determine whether HF-LILI induce cell death in lung CSCs. Lung CSCs were isolated using the stem cell marker CD 133, characterized using flow cytometry, and applied in experiments which included treatment with LILI at wavelengths of 636, 825 and 1060 nm with fluences ranging from 5 J/cm² to 40 J/cm². Viability and proliferation studies, using Alamar blue assay and adenosine triphosphate luminescence (ATP), indicated an increase when treating lung CSCs with low fluences of 5 - 20 J/cm² and a decrease in viability and proliferation as well as an increase in apoptosis when applying a fluence of 40 J/cm² indicated by flow cytometry using Annexin V and propidium iodide (PI) dyes. Results indicate that LILI, when treating lung CSCs, can induce either a bio-stimulatory or bio-inhibitory effect depending on the wavelength and fluence used. This study indicated successful apoptotic induction of lung CSCs. Future experiments should be able to conclude the exact mechanism behind HF-LILI, which can be used in the targeted treatments of CSC elimination, implementing HF-LILI in the same manner as PDT in the absence of a photosensitizer.

Beside biomaterials, Laser phototherapy has shown positive effects as auxiliary therapy in bone repair process, especially when involving large bone losses. The aim of this histological study was to evaluate, by light microscopy, the influence of laser phototherapy on the repair of complete tibial fractures in rabbits treated or not with semi-rigid internal fixation and Mineral Trioxide Aggregate - MTA graft. Twelve Rabbits were randomly divided into four groups with three animals each. After general anesthesia, complete fractures were created in one tibia with a carborundum disk. All animals (groups I-IV) had the fracture stabilized with semi-rigid fixation (wire osteosynthesis - WO). Group I was routinely fixed with WO; groups II and IV fracture was filled by blood clot and MTA implant. In Groups III and IV fracture was filled by blood clot and further irradiated with laser (λ780 nm, 70 mW, CW, Φ = 0.04 cm², 20.4 J/cm², per session, t = 300s, 142.8 J/cm² per treatment). The phototherapy protocol was applied immediately after the surgery and repeated each 48 hours during 15 days. Animal death occurred on the 30th postoperative day. After removal of the specimens, the samples were routinely processed, stained with HE and evaluated by light microscopy. Histologically, the group treated with MTA graft and irradiated with laser showed the fracture filled by a more organized and mature trabecular bone, when compared with all other groups. From the results of the present study, it may be concluded that the association of Laser phototherapy + MTA graft in fractures treated with WO improved bone repair when compared with fractures treated only with WO.

LED photobiomodulation is known mostly for its restorative effects on skin and joints. While providing LED photobiomodulation aesthetic treatments of the face, not only the skin condition was improved, but a subjective well-being effect was observed, obvious both in photographs of the treated areas and in patient behaviour. This has been supported by studies showing the beneficial effects of transcranial lasers and LEDs on neurological and psychological conditions, providing great insight. LED therapy can now be used as a standalone procedure to regulate neuronal function. To measure such neurological outcomes in humans, we developed a visual analog scale questionnaire with the purpose of having a convenient tool for the assessment of quality of life following facial LED photobiomodulation.We also gauged patients' emotional state regarding overall aesthetic improvement.

The use of curcumin as antimicrobial agent has been suggested and this effect may be potentialized by appropriate light. This study evaluated the effect of PACT using blue LED (λ450ηm ± 5ηm, 220mW and spot of 0.785 cm²) associated to Curcumin at different concentrations (75, 37.5, 18.75, 9.37 and 4.68 μg /mL). Microorganisms from the oral mucosa and the posterior region of the tongue were collected and inoculated into test tubes containing 8mL of TSB medium. For these assays were performed 16 readings. In the assays were used culture plate of 24 wells. To each well was added 400 μL of the suspension containing the microorganisms. Suspensions without curcumin were placed in eight wells. Elsewhere, curcumin was applied varying concentrations with pre-irradiation time of 5 min. After stirring, 200 μL aliquot was taken from each well and the readings were immediately carried out by a spectrophotometer (SPECTRA MAX). Assessments of turbidity were performed following CLSI standard methods. After 1 hour of incubation in a bacteriological oven, 200 μL aliquot was removed from the remaining wells for a second reading. The results showed a decrease of total microorganisms in the most of test groups. The best result of the PACT was with 75 μg/mL, showing 81% of inhibition. It is concluded that PACT with blue LED associated to Curcumin could be a potential mechanism for controlling microorganism proliferation on the oral cavity.

Candida albicans plays an important role in triggering infections in HIV+ patients. The indiscriminate use of antifungals has led to resistance to Candida albicans, which requires new treatment alternatives for oral candidiasis. Low-level laser therapy promotes a considerable improvement in the healing of wounds and in curing illnesses caused by microorganisms. The aim of the present study was to assess the effect of laser radiation on the cell proliferation of Candida albicans in immunosuppressed patients. Six Candida albicans strains that had been isolated from immunosuppressed patients were divided into a control group and experimental groups, which received eight sessions of laser therapy (InGaAlP, λ685nm, P = 30mW, CW, Φ~6 mm and GaAlAs, λ830nm, P = 40mW, CW, Φ~6 mm) using dosimetries of 6J/cm², 8J/cm², 10J/cm² and 12J/cm² for each wavelength and power. The results were not statistically significant (Kruskal Wallis, p > 0.05), although the proliferation of Candida albicans was lower in some of the experimental groups. The dosimetry of 6J/cm² (GaAlAs, λ830nm, P = 40mW) provided lower mean scores than the other groups for the growth of Candida. Further studies are required to confirm whetehr laser therapy is a viable option in the treatment of fungal infections.

This study was to evaluate clinical and microbiological effectiveness of photodynamic therapy (PDT) in the treatment of periodontal disease in kidney-transplanted patients. Eight kidney transplanted patients treated at Paulista University were arranged in two groups: SRP performed scaling and root planning by ultrasound; SRP+PDT- in the same patient, which was held to PDT in the opposite quadrant, with 0.01% methylene blue and red laser gallium aluminum arsenide, wavelength 660 nm, power 100 mW. There was reduction in probing pocket depth after 45 days and 3 months regardless the group examined; plaque and bleeding index showed improvement over time, regardless the technique used, and bleeding index in the SRP+PDT group was lower when compared with the baseline the other times. There was no difference in the frequency of pathogens. Photodynamic therapy may be an option for treatment of periodontal disease in renal-transplanted patients and its effectiveness is similar to conventional therapy.

HIV/Aids patients present a change of microbiota associated with host immunodeficiency. Photodynamic therapy (PDT) showed as a promising and viable alternative in reducing microbiota. Present study evaluate effectiveness of photodynamic therapy in periodontal disease of AIDS patients with highly activity antiretroviral therapy (HAART) failure, measuring the clinical periodontal parameters and periodontal microbiota. Twelve patients with HARRT resistance (R group) divided into two groups (control and PDT) and 12 patients with no HAART resistance (NR group) divided into two groups (control and PDT). The results show the difference in baseline of CD4 cells count, NR group 640.0 ± 176.2 cells/mm³ R group and 333.3 ± 205.8 cells / mm³ (p<0.05), and in 8.3% detectable viral load in NR group and 75% detectable (p <0.001) in R group. As clinical periodontal parameters (PD and CAL), PDT was more effective than the control group only in the NR group (p <0.05%), moreover, there was no difference in the evaluation of clinical periodontal parameters between the both R groups (p>0.05%). Microbiological evaluation in R group presents a general reduction in the Aa at 3 and 6 months. Furthermore, demonstrated a reduction of Pg in all groups at 6 months and in R group at 3 months. The impact assessment of photodynamic therapy in patients with different levels of immunosuppression determined that the combination of mechanical periodontal treatment with photodynamic therapy in patients with HAART failure did not cause additional benefits. Therefore, PDT in this study could not been indicated in HAART resistance patients.

Bone neoformation is essential in the osteointegration of implants and has been correlated with the repair capacity of tissues, the blood supply and the function of the cells involved. Laser therapy accelerates the mechanical imbrication of peri-implant tissue by increasing osteoblastic activity and inducing ATP, osteopontin and the expression of sialoproteins. Objective: The aim of the present study was to assess peri-implant bone repair using the tibia of dogs that received dental implants and laser irradiation (AsGaAl 830nm – 40mW, CW, f~0.3mm) through Energy Dispersive X-ray Fluorescence (EDXRF). Methodology: Two groups were established: G1 (Control, n=20; two dental implants were made in the tibia of each animal; 10 animals); G2 (Experimental, n=20, two dental implants were made in the tibia each animal + Laser therapy; 10 animals). G2 was irradiated every 48 hours for two weeks, with a total of seven sessions. The first irradiation was conducted during the surgery, at which time a point in the surgical alveolus was irradiated prior to the placement of the implant and four new spatial positions were created to the North, South, East and West (NSEW) of the implant. The subsequent sessions involved irradiation at these four points and at one infra-implant point (in the direction of the implant apex). Each point received 4J/cm2 and a total dose of 20J/cm2 per session (treatment dose=140J/cm2). The specimens were removed 15 and 30 days after the operation for the EDXRF test. The Mann- Whitney statistical test was used to assess the results. Results: The increase in the calcium concentration in the periimplant region of the irradiated specimens (G2) was statistically significant (p < 0.05), when compared with the control group (G1). Conclusion: The results of the present study show that irradiation with the AsGaAl laser promoted an acceleration in bone repair in the peri-implant region.