Effects of photobiomodulation therapy in chondrocyte response by in vitro experiments and experimental model of osteoarthritis in the knee of rats.

The purpose of this study is to evaluate the effects of photobiomodulation (PBM) therapy in chondrocyte response by in vitro experiments and cartilage repair using an experimental model of osteoarthritis (OA) in the knee of rats. The in vitro experiment was performed with chondrocyte cells, and they were divided into two groups: non-irradiated and irradiated with PBM (808 nm; 0.8 J or 1.4 J). Then, cell proliferation was evaluated after 1, 3, and 5 days. The experimental model of osteoarthritis (OA) was performed in the knee of 64 Wistar rats, and they were assorted into control group (CG), PBM (808 nm; 1.4 J). The results of in vitro showed that PBM 1.4 J increased cell proliferation, on days 1 and 5. However, after 3 days was demonstrated a significant increase in cell proliferation in PBM 0.8 J. The in vivo experiment results demonstrated, on histological analysis, that PBM presented less intense signs of tissue degradation with an initial surface discontinuity at the superficial zone and disorganization of the chondrocytes in the cartilage region when compared to CG, after 4 and 8 weeks. These findings were confirmed by immunohistochemistry and qRT-PCR analysis which showed that PBM increased IL-4, IL-10, COL-2, Aggrecan, and TGF-ß which are anabolic factors and acts on extracellular matrix. Also, PBM reduces the IL1-ß, an inflammatory marker that operates as a catabolic factor on articular cartilage. In conclusion, these results suggest that PBM may have led to a return to tissue homeostasis, promoting chondroprotective effects and stimulating the components of the articular tissue.

Bone substitutes and photobiomodulation in bone regeneration: A systematic review in animal experimental studies.

In general, bone fractures are able of healing by itself. However, in critical situations such as large bone defects, poor blood supply or even infections, the biological capacity of repair can be impaired, resulting in a delay of the consolidation process or even in non-union fractures. Thus, technologies able of improving the process of bone regeneration are of high demand. In this context, ceramic biomaterials-based bone substitutes and photobiomodulation (PBM) have been emerging as promising alternatives. Thus, the present study performed a systematic review targeting to analyze studies in the literature which investigated the effects of the association of ceramic based bone substitutes and PBM in the process of bone healing using animal models of bone defects. The search was conducted from March and April of 2019 in PubMed, Web of Science and Scopus databases. After the eligibility analyses, 16 studies were included in this review. The results showed that the most common material used was hydroxyapatite (HA) followed by Biosilicate associated with infrared PBM. Furthermore, 75% of the studies demonstrated positive effects to stimulate bone regeneration from association of ceramic biomaterials and PBM. All studies used low-level laser therapy (LLLT) device and the most studies used LLLT infrared. The evidence synthesis was moderate for all experimental studies for the variable histological analysis demonstrating the efficacy of techniques on the process of bone repair stimulation. In conclusion, this review demonstrates that the association of ceramic biomaterials and PBM presented positive effects for bone repair in experimental models of bone defects.

Biphasic Dose/Response of Photobiomodulation Therapy on Culture of Human Fibroblasts.

Objective: The objective of this study was to evaluate the effects of application of different fluences and energies of laser in the 24-, 48-, and 72-h periods in fibroblasts originating from human skin (HFF-1). Methods: The cell used as a template for cell proliferation was HFF-1. For the photobiomodulation (PBM) application, a 660 nm laser with a power of 40 mW and energies of 0.84, 1.40, 5.88, and 6.72 J was used. Five experimental groups were studied: one control group (CG) with simulated PBM and four groups that received PBM in different doses. The changes observed after laser irradiation were evaluated by cell viability (trypan blue) and proliferation [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)] tests. Intergroup comparisons were performed using two-way analysis of variance and the Tukey post hoc test (software GraphPad Prism 7.0). Results: In the trypan blue test, the total number of cells was significantly different between the irradiated groups and the CG at all times studied. The total number of cells increased in laser group (LG)1 (0.84 J) and LG2 (1.40 J) and decreased in LG4 (6.72 J). The mitochondrial activity increased significantly in LG1 and LG2 at 48 and 72 h and decreased in LG3 (5.88 J) and LG4 (6.72 J) compared with CG. Conclusions: The results indicate that the lower doses (0.45 and 0.75 J/cm2) of PBM induce the highest mitochondrial activity and cellular viability.

Effect of photobiomodulation associated with cell therapy in the process of cutaneous regeneration in third degree burns in rats.

Due to the complexity involved in the healing process of full thickness burns, the literature looks for alternatives to optimize tissue reconstruction. The objective of this study was to explore the action of photobiomodulation therapy associated with MSCs in the healing process of third degree burns. A total of 96 male Wistar rats were used, distributed in four groups with 24 animals each: Control Group, Laser Group, Cell Therapy Group, and Laser Group and Cell Therapy. The burn was performed with aluminum plate (150 °C). We performed analysis of wound contraction, histology, immunohistochemistry, birefringence analysis, and immunoenzymatic assay to evaluate tissue quality. Our results demonstrate that the association of the techniques is able to accelerate the repair process, modulating the inflammatory process, presenting a cutaneous tissue with better quality. Thus, we conclude that the use of photobiomodulation therapy associated with cell therapy is a promising treatment in the repair of total thickness burns.

Photobiomodulation therapy and transcutaneous electrical nerve stimulation on chronic neck pain patients: Study protocol clinical trial (SPIRIT Compliant).

INTRODUCTION: Chronic neck pain is a common musculoskeletal disorder that is associated with functional disability and decreased of quality of life. Electrophysical agents are commonly used to relieve pain, however the effects of combined use of these agents are little studied. The objective is to investigate the efficacy of photobiomodulation and electrical stimulation to relieve pain, both in isolation and combined. MATERIALS AND METHODS: This a 4-arm randomized placebo-controlled trial with patient and evaluator blinded. This study will be performed in Department of Physical Therapy at Federal University of São Carlos, São Carlos/SP, Brazil. One hundred and forty-four patients with chronic neck pain will be randomized into 4 groups: active photobiomodulation therapy with active electrical stimulation, active photobiomodulation therapy, active electrical stimulation, or placebo treatment. They will receive 10 sessions of treatment. PRIMARY OUTCOME: pain intensity (measured by pain numerical rating scale) posttreatment. SECONDARY OUTCOMES: pain during movement, neck disability, range of motion, pressure pain threshold, temporal summation, conditioned pain modulation, depressive symptoms, pain catastrophizing, quality of life, analgesic intake, and global perceived effect at posttreatment (10 sessions). Pain intensity and global perceived effect will also be measured after 6 weeks randomization. DISCUSSION: The findings of this study might clarify the importance of using the photobiomodulation therapy and transcutaneous electrical nerve stimulation for patients with chronic neck pain. TRIAL REGISTRATION: NCT04020861. https://clinicaltrials.gov/ct2/show/NCT04020861?term=NCT04020861&draw=2&rank=1.

Photobiomodulation effect on the proliferation of adipose tissue mesenchymal stem cells.

The use of mesenchymal stem cells (MSCs) in tissue engineering has been extensively investigated. The greater the proliferation of this cellular group, the greater the regenerative and healing capacity of the tissue to which they belong. In this context, photobiomodulation (PBM) is an efficient technique in proliferation of distinct cell types. However, its parameters and mode of action are still unclear and require further investigation. This study aimed to evaluate the PBM action with different energies in MSCs of adipose tissue (hASCs). We used hASCs, seeded in 24-well plates, with 3 × 104 cells per well, in culture media. We used a total of four experimental groups, one with hASCs and simulated PBM and three other groups, which received PBM irradiation at 24, 48, and 72 h, with a 660-nm laser and power of 40 mW and energy of 0.56, 1.96, and 5.04 J. We performed analyses of MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromidefor) and trypan blue to evaluate cell proliferation and viability, 1 h after PBM irradiation. Software Graph PadPrism 7.0 was used. Intergroup comparisons were performed with ANOVA two-way and we used the Tukey post hoc test. Mitochondrial activity evaluated by MTT revealed the statistical difference in the first 24 h for group with more high energy when compared to control group; and in the 72 h for two irradiated groups when compared to the control group. The trypan blue test showed significant differences at the end of the experiment for two irradiated groups LG1 (4.52 × 104 ± 0.2) and LG2 (4.85 × 104 ± 0.8), when compared to the control group (1.87 × 104 ± 0.7). Both tests failed to be statistically different at the end of the experiment for groups LG1 and LG2 and observed a reduction in cellular mitochondrial growth and activity for group LG3. We conclude that PBM with energy close to 0.56 and 1.96 J promote proliferation of hASCs, and higher energy, such as 5.04 J, can be harmful.

Photobiomodulation on critical bone defects of rat calvaria: a systematic review.

Bone defects following trauma represent a high impact on the quality of life of millions of people around the world. The aim of this study was to review photobiomodulation (PBM) action in the treatment of bone critical defects in rat calvaria, related to evaluation of the current protocols applied. One hundred and forty-seven articles related to the subject were found by searching the main databases (Pubmed, Lilacs, Web of Science, and Scopus) considering the period of publication until the year 2017, and only 14 corresponded the inclusion criteria established for this systematic review. The main parameters of the PBM were expressed in Table 1. In addition, it was possible to observe the use of two different wavelengths (red and infrared), which are considered therapeutic. Most of the evaluated articles presented positive results that describe a greater amount of neoformed bone, an increase in collagen synthesis, and a contribution to microvascular reestablishment. However, two studies report no effect on the repair process when the PBM was used. In addition, we observed considerable variations between the values of power, fluence, and total energy, which make it difficult to compare the results presented between the selected studies. It was possible to conclude that the infrared laser was more effective in positively stimulating the bone repair process of critical defects. Furthermore, a discrepancy was found in the parameter values used, which made it difficult to choose the best protocol for the treatment of this type of lesion.

Evaluation of the low-level laser therapy application parameters for skin burn treatment in experimental model: a systematic review.

Burn is defined as a traumatic injury of thermal origin, which affects the organic tissue. Low-level laser therapy (LLLT) has gained great prominence as a treatment in this type of injury; however, the application parameters are still controversial in the literature. The aims of this study were to review the literature studies that use LLLT as a treatment in burns conducted in an experimental model, discuss the main parameters used, and highlight the benefits found in order to choose an appropriate therapeutic window to be applied in this type of injury. The selection of the studies related to the theme was carried out in the main databases (PubMed, Cochrane Library, LILACS, Web of Science, and Scopus in the period from 2001 to 2017). Subsequently, the articles were then chosen that fell within the inclusion criteria previously established. In the end, 22 were evaluated, and the main parameters were presented. The analyzed studies presented both LLLT use in continuous and pulsed mode. Differences between the parameters used (power, fluence, and total energy) were observed. In addition, the protocols are distinct as to the type of injury and the number of treatment sessions. Among the results obtained by the authors are the improvements in the local microcirculation and cellular proliferation; however, a study reported no effects with LLLT as a treatment. LLLT is effective in accelerating the healing process. However, there is immense difficulty in establishing the most adequate protocol, due to the great discrepancy found in the applied dosimetry values.

Eficácia da terapia a laser de baixa intensidade no controle da dor neuropática em camundongos / Eficacia de la terapia láser de baja potencia en el control del dolor neuropático en ratones / Low intensity laser therapy effectiveness in controlling neuropathic pain in mice

RESUMO A terapia a laser de baixa intensidade (LLLT) vem sendo amplamente discutida na literatura como forma alternativa de tratamento para diversos tipos de dor, com destaque para a neuropática. Essa terapia sobressai pelo fato de não ser invasiva, raramente causar efeitos colaterais e ser de baixo custo. Em contrapartida, para sua eficácia, é necessário o detalhamento dos parâmetros, que ainda são muito discrepantes na literatura. Assim, este trabalho tem como objetivo investigar o efeito da LLLT, na faixa do infravermelho, com fluência de 30J/cm², no controle da dor neuropática em modelo animal. Foram utilizados 24 camundongos da cepa suíço albino, machos, pesando 25­30 gramas, divididos em três grupos: Grupo Placebo (GP), Grupo Laser (GL30) e Grupo Sham (GS). A indução da neuropatia foi feita através do modelo de constrição crônica do nervo isquiático (CCI), e o tratamento da LLLT realizou-se da seguinte maneira: GP com o laser com fluência de 0J/cm2, GL30 tratado com fluência de 30J/cm2, e GS com simulação de cirurgia sem intervenção. Executaram-se as irradiações 3 vezes por semana, durante 90 dias, no ponto de compressão do nervo, utilizando-se a técnica de contato. A fins de avaliação, foram utilizados o teste da placa quente, para hiperalgesia térmica, e o Teste de Randall-Selitto para hiperalgesia mecânica. Nos resultados do GP, observamos que não houve melhora significativa nos dias após a cirurgia em nenhum dos testes realizados e, no GL30, observou-se uma melhora expressiva em ambos os testes a partir do 30º dia de tratamento para o teste de Placa Quente e a partir do 45º para o Randall-Selitto, em que os camundongos apresentaram restauração total da sensibilidade. Concluímos, pois, que a utilização de LLLT com fluência de 30J/cm2 no tratamento da dor neuropática em modelo animal é eficaz.

RESUMEN La terapia láser de baja potencia (LLLT) ha sido ampliamente discutida en la literatura como forma alternativa de tratamiento para muchos tipos de dolor, especialmente para el neuropático. Esa terapia se destaca por el hecho de no ser invasiva, raramente causar efectos secundarios y ser de bajo costo. En cambio, para su eficacia, es necesario que se presenten los detalles de los parámetros, que todavía son muy discordantes en la literatura. Así, este trabajo tiene como objetivo investigar el efecto de la LLLT, en el rango del infrarrojo, con fluidez de 30 J/cm², en el control del dolor neuropático en modelos animales. Se utilizaron 24 ratones de la cepa suizo-albino, machos, con peso de 2530 gramos, divididos en tres grupos: Grupo Placebo (GP), Grupo Láser (GL30) y Grupo Sham (GS). Se llevó a cabo la inducción de la neuropatía por medio del modelo de constricción crónica del nervio isquiático y se realizó el tratamiento de la LLLT como sigue: GP con el láser con fluidez de 0 J/cm2, GL30 con fluidez de 30 J/cm2, y GS con simulación de cirugía sin intervención. Se realizaron las irradiaciones tres veces por semana, durante 90 días, en el punto de compresión del nervio, utilizándose la técnica de contacto. A efectos de evaluación, se utilizaron la prueba de placa caliente para hiperalgesia térmica y la prueba de Randall y Selitto para hiperalgesia mecánica. En los resultados del GP, observamos que no hubo mejora significativa en los días después de la cirugía en ninguna de las pruebas realizadas y, en el GP30, se verificó mejora significativa en ambas pruebas: desde el 30.º día de tratamiento para la prueba de placa caliente y desde el 45.º día para la prueba de Randall y Selitto, en que los ratones presentaron restauración total de la sensibilidad. Concluimos, por lo tanto, que el uso de la LLLT con fluidez de 30 J/cm2 en el tratamiento del dolor neuropático en modelos animales es efectivo.

ABSTRACT Low-level laser therapy (LLLT) has been widely discussed in the literature as an alternative form of treatment for several types of pain, especially neuropathic pain. This kind of therapy stands out for not being invasive, rarely causing side effects and being cost effective. However, for its effectiveness, it is necessary more detailed parameters, which are still very discrepant in the literature. Thus, this study aims to investigate the effect of LLLT, in the infrared range, with 30J/cm² fluency, on the control of neuropathic pain in animal models. A total of 24 male Swiss albino mice, weighing 25.30 grams, were divided into three groups: the Control Group (CG), the Laser Group (L30G) and the Sham Group (SG). The induction of neuropathy was held through the model of chronic constriction of the sciatic nerve (CCI), and the LLLT treatment was conducted as follows: The CG was treated with 0 J/cm2 fluency, whereas the L30G was treated with 30 J/cm2 fluency, and the SG with simulation of surgery without intervention. The irradiations were performed 3 times a week, for 90 days, at the nerve's point of compression, using the contact technique. For the evaluation, the Hot Plate Test was used for thermal hyperalgesia, and the Randall-Selitto test was used for mechanical hyperalgesia. In the CG's results, we observed no significant improvement in the days after surgery in any of the tests conducted, and, in the GL30, a significant improvement in both tests was observed: from the 30th day of treatment for the Hot Plate Test, and from the 45th for the Randall-Selitto Test, in which the mice showed total restoration of sensitivity. We thus conclude that the use of LLLT with 30 J/cm2 fluency in the treatment of neuropathic pain in animal models is effective.

Effect of photobiomodulation therapy (808 nm) in the control of neuropathic pain in mice.

Neuropathic pain can be defined as the pain initiated or caused by a primary lesion or dysfunction of the central or peripheral nervous system. Photobiomodulation therapy (PBM) stands out among the physical therapy resources used for analgesia. However, application parameters, especially the energy density, remain controversial in the literature. Therefore, this study aimed to investigate the PBM effect, in different energy densities to control neuropathic pain in mice. Fifty (50) mice were induced to neuropathy by chronic constriction surgery of the sciatic nerve (CCI), treated with PBM (808 nm), and divided into five groups: GP (PBM simulation), GS (sham), GL10, GL20, GL40 (energy density of 10, 20, and 40 J/cm2, respectively). The evaluations were carried out using the hot plate test and Randall and Selitto test, before and after the CCI surgery, every 15 days during the 90 days experiment. ß-Endorphin blood dosage was also tested. For both the hot plate and Randall and Selitto tests, the GL20 and GL40 groups presented reduction of the nociceptive threshold from the 30th day of treatment, the GL10 group only after day 75, and the GP group did not show any improvement throughout the experiment. The ß-endorphin dosage was higher for all groups when compared to the GP group. However, only the GL20 group and GL40 presented a significant increase. This study demonstrates that PBM in higher energy density (20, 40 J/cm2) is more effective in the control of neuropathic pain.

Use of low level laser therapy to control neuropathic pain: A systematic review.

Neuropathic pain can be defined as pain initiated or caused by a primary lesion or dysfunction in the central or peripheral nervous system. The low level laser therapy (LLLT) has gained great prominence as a treatment in this type of pain; however, the application parameters are still controversial in the literature. This study aimed to review the literature on the use of LLLT in neuropathic pain with the goal of establishing a "therapeutic window" for the effective use of this treatment. We analyzed 14 articles, 10 in experimental animals and 4 in humans. The results are presented in three tables, the first being for comparison of the studies' application parameters, the second showing the average and median parameters experimental studies and third showing the clinical studies embodiment. The experimental studies revealed better results for LLLT and infrared laser powers above 70mW. Clinical studies are inconclusive as to the application parameters, due to the discrepancy; however all demonstrate the effectiveness of LLLT. According to the data presented, it was concluded that LLLT has positive effects on the control of analgesia for neuropathic pain, but further studies with high scientific rigor are needed in order to define treatment protocols that optimize the action LLLT in neuropathic pain.