Energy-dependent effect trial of photobiomodulation on blood pressure in hypertensive rats.

The main purpose of this work was to construct an energy-dependent response curve of photobiomodulation on arterial pressure in hypertension animal model. To reach this objective, we have used a two-kidney one clip (2K-1C) rat model. Animals received acute laser light irradiation (660 nm) on abdominal region using different energy (0.6, 1.8, 3.6, 7.2, 13.8, 28.2, 55.8, and 111.6 J), the direct arterial pressure was measured by femoral cannulation, and systolic arterial pressure (SAP), diastolic arterial pressure (DAP), heart rate (HR), and time of effect were obtained. Our results indicated that 660 nm laser light presents an energy-dependent hypotensive effect, and 28.2 J energy irradiation reached the maximum hypotensive effect, inducing a decreased SAP, DAP, and HR (decrease in SAP: - 19.23 ± 1.82 mmHg, n = 11; DAP: - 9.57 ± 2.23 mmHg, n = 11; HR: - 39.15 ± 5.10 bpm, n = 11; and time of hypotensive effect: 3068.00 ± 719.00 s, n = 11). The higher energy irradiation evaluated (111.6 J) did not induce a hypotensive effect and induced an increase in HR (21.69 ± 7.89 bpm, n = 7). Taken together, our results indicate that red laser energy irradiation from 7.2 to 55.8 J is the effective therapeutic window to reduce SAP, DAP, MAP, and HR and induce a long-lasting hypotensive effect in rats, with effect loss at higher energy irradiation (111.6 J).

Photobiomodulation can improve ovarian activity in polycystic ovary syndrome-induced rats.

Follicular cystic ovary disease is a common reproductive disorder in women and females of domestic animals, characterized by anovulation and the persistence of follicle is a common cause of reproductive failure in mammalian. Polycystic ovary syndrome (PCOS) is characterized by hyperandrogenism (HA), chronic anovulation and polycystic ovaries, and it is a common reproductive endocrine disease with clinical manifestations including hirsutism, acne, infertility and obesity that can affect 5-20% of women in their reproductive age. Photobiomodulation (PBM) has been investigated and used in clinical practice, related to biomodulatory influences on cellular functions in animals and humans, both in vivo and in vitro. In this study, we include endocrine and reproductive features in a rat model for PCOS and the effects of PBM on ovarian activities. Forty-five adult female Wistar rats PCOS-induced by a single dose of the estradiol valerate (EV) were used in the study. After the EV injection for PCO induction, rats were divided into 9 groups (n = 5/group) named C30, C45 and C60 (Control group), S30, S45 and S60 (PCO group) and L30, L45 and L60 (PCO/Laser group). The rats were irradiated with laser 3 times/week. The results shown that EV PCO-induced rats had increased body mass, reduced ovary mass, and reduced GSI. The plasma levels of P4 and T were increased, and the LH plasma level was decreased by PBM stimulation. The number of ovarian follicles and corpus luteum were increased, and the number of ovarian cysts was decreased by PBM stimulation. Thus, reproductive and endocrine characteristics were modulated by PBM.

Comparison of two different laser photobiomodulation protocols on the viability of random skin flap in rats.

To identify the best low level laser photobiomodulation application site at the same irradiation time to increase the viability of the skin flap in rats. Eighteen male rats (Rattus norvegicus: var. Albinus, Rodentia Mammalia) were randomly distributed into three groups (n = 6). Group I (GI) was submitted to simulated laser photobiomodulation; group II (GII) was submitted to laser photobiomodulation at three points in the flap cranial base, and group III (GIII) was submitted to laser photobiomodulation at 12 points distributed along the flap. All groups were irradiated with an Indium, Galium, Aluminum, and Phosphorus diode laser (InGaAlP), 660 nm, with 50 mW power, irradiated for a total time of 240 s in continuous emission mode. The treatment started immediately after performing the cranial base random skin flap (10 × 4 cm2 dimension) and reapplied every 24 h, with a total of five applications. The animals were euthanized after the evaluation of the percentage of necrosis area, and the material was collected for histological analysis on the seventh postoperative day. GII animals presented a statistically significant decrease for the necrosis area when compared to the other groups, and a statistically significant increase in the quantification of collagen when compared to the control. We did not observe a statistical difference between the TGFß and FGF expression in the different groups evaluated. The application of laser photobiomodulation at three points of the flap cranial base was more effective than at 12 points regarding the reduction of necrosis area.

Effect of photobiomodulation (670 nm) associated with vitamin A on the inflammatory phase of wound healing.

Wound healing is a complex biological process with specific phases. Photobiomodulation (PBM) decreases the inflammatory infiltrate, stimulating fibroblast proliferation and angiogenesis, and therefore, is indicated for wound healing. Vitamin A is used to reverse the inhibitory effects on wound healing and accelerate the healthy granulation tissue. The study aimed to evaluate the effect of topical vitamin A and PBM (GaAlAs) in inflammatory phase of cutaneous wounds. Forty Wistar male rats were separated into four groups: (1) control (CG); (2) laser group (LG) GaAlAs, 670 nm, 30 mW, energy per point of 0.9 J, radiating by 1 point in 30 s; (3) vitamin A group (VitAG); and (4) laser group plus vitamin A (LG + VitAG). Wounds were surgically made by a punch biopsy with 10 mm of diameter on the back of the animals and all treatments were started according to the experiment. The treatments were administered for four consecutive days and biopsy was performed on day 4. We performed both H&E and immunohistochemistry analysis. The results were compared between groups by one-way analysis of variance ANOVA test with post hoc Tukey (p < 0.05). Inflammatory infiltrate increased significantly in LG compared to CG and VitAG (p < 0.05). Regarding angiogenesis, VEGF expression was increased significantly in LG and LG + VitAG groups, p < 0.01. The results indicate that proposed treatments were effective on the healing process improved by LG and LG + VitAG. We show that laser plus vitamin A enhances healing by reducing the wound area and may have potential application for clinical management of cutaneous wounds.

Hypotensive acute effect of photobiomodulation therapy on hypertensive rats.

The purpose of this study was to evaluate the acute effect of photobiomodulation therapy (PBM) on arterial pressure in hypertensive and normotensive rats with application in an abdominal region. Normotensive (2K) and hypertensive (2K-1C) wistar rats were treated with PBM. Systolic arterial pressure (SAP), diastolic arterial pressure (DAP), mean arterial pressure (MAP) and heart rate (HR) were measured before, during and after PBM application. The nitric oxide (NO) serum concentration was measured before and after PBM application. Vascular reactivity study was performed in isolated thoracic aortas. Aluminum gallium arsenide (GaAlAs) diode laser was used, at 660nm wavelength and 100mW optical output. The PBM application induced a decrease of SAP in 2K-1C rats. In 2K rats, the PBM application had no effect on SAP, DAP and MAP. Moreover, the magnitude of hypotensive effect was higher in 2K-1C than in 2K rats. The PBM application induced a decrease of HR in 2K-1C and 2K, with higher effect in 2K-1C rats. In 2K-1C, the hypotensive effect induced by PBM was longer than that obtained in 2K rats. PBM application induced an elevation of NO concentration in serum from 2K-1C and 2K rats, with higher effect in 2K-1C. In isolated aortic rings PBM effect is dependent of NO release, and is not dependent of nitric oxide synthase (NOS) activation. Our results indicate that the abdominal acute application of PBM at 660nm is able to induce a long lasting hypotensive effect in hypertensive rats and vasodilation by a NO dependent mechanism.

The effects of laser irradiation on osteoblast and osteosarcoma cell proliferation and differentiation in vitro.

OBJECTIVE: The aim of this study was to investigate the effects of 670-nm, 780-nm, and 830-nm laser irradiation on cell proliferation of normal primary osteoblast (MC3T3) and malignant osteosarcoma (MG63) cell lines in vitro. BACKGROUND: Some studies have shown that laser phototherapy is able to stimulate the osteogenesis of bone tissue, increasing osteoblast proliferation and accelerating fracture consolidation. It has been suggested that laser light may have a biostimulatory effect on tumor cells. However, the mechanism by which the laser acts on cells is not fully understood. MATERIALS AND METHODS: Neonatal, murine, calvarial, osteoblastic, and human osteosarcoma cell lines were studied. A single laser irradiation was performed at three different wavelengths, at the energies of 0.5, 1, 5, and 10 J/cm(2). Twenty-four hours after laser irradiation, cell proliferation and alkaline phosphatase assays were assessed. RESULTS: Osteoblast proliferation increased significantly after 830-nm laser irradiation (at 10 J/cm(2)) but decreased after 780-nm laser irradiation (at 1, 5, and 10 J/cm(2)). Osteosarcoma cell proliferation increased significantly after 670-nm (at 5 J/cm(2)) and 780-nm laser irradiation (at 1, 5, and 10 J/cm(2)), but not after 830-nm laser irradiation. Alkaline phosphatase (ALP) activity in the osteoblast line was increased after 830-nm laser irradiation at 10 J/cm(2), whereas ALP activity in the osteosarcoma line was not altered, regardless of laser wavelength or intensity. CONCLUSION: Based on the conditions of this study, we conclude that each cell line responds differently to specific wavelength and dose combinations. Further investigations are required to investigate the physiological mechanisms responsible for the contrasting outcomes obtained when using laser irradiation on cultured normal and malignant bone cells.

Influência de diferentes comprimentos de onda da laserterapia de baixa intensidade na regeneração tendínea do rato após tenotomia / Influence of different low-intensity laser therapy wavelengths for rat tendon regeneration following tenotomy

OBJETIVO: Comparar a influência da terapia laser baixa intensidade (TLBI) na organização das fibras de colágeno, na dose de 3J/cm² em dois comprimentos de onda (lambda) (670nm e 904nm) e na associação de ambos, durante o reparo do tendão calcâneo de ratos. MÉTODO Foram utilizadas 37 ratas da raça Wistar divididas em 5 grupos, submetidas a tenotomia total por meio de cisalhamento transversal do tendão na porção média entre a inserção calcânea e a junção miotendínea, com exceção do grupo GP (n= 6) que não houve lesão. O grupo GA (n= 8) foi submetido à irradiação laser lambda= 904nm, diodo de GaAs, emissão pulsada, duração do pulso de 180ns, freqüência de pulso de 2000Hz e potência de pico de 15W e dose de 3J/cm². O grupo GB (n= 8) foi submetido à irradiação laser lambda= 670nm, diodo de AlGaInP, emissão contínua, potência 30mW e dose de 3J/cm². O grupo GAB (n= 8) recebeu irradiação de ambos os lasers 670nm e 904nm associados. O grupo GCL (n= 7) foi submetido a um tratamento placebo, assim como um grupo padrão. Após um total de 12 sessões de laserterapia, os animais foram submetidos à eutanásia no 14º dia pós-operatório. Após processamento histológico padrão, foi realizada a quantificação do retardo óptico pela birrefringência. RESULTADOS: A análise estatística mostra que todos os grupos irradiados apresentaram valores de retardo óptico maiores quando comparados ao grupo placebo (p= 0,00001), o que indica que apresentaram melhor organização das fibras colágenas ao longo do eixo longitudinal do tendão, evidenciando melhor qualidade no reparo. O grupo GAB foi o que apresentou melhores resultados (p= 0,00001) quando comparado aos demais grupos, excetuando-se o GP, que é o referencial de retardo óptico para esta faixa etária.

OBJECTIVE: The aim of this study was to compare the effect of the low level laser therapy in the organization of the collagen fibers with 3 J/cm² of dose in two wavelengths (670nm and 904nm) and in the combination of both on the tissue repair of the calcaneus tendon. METHODS: We used 37 female Wistar rats divided in 5 groups which had been submitted to the total tenotomy. Group GA (n= 8) was submitted to the laser irradiation in lambda= 904nm, GaAS diode, pulsated emission, peak power 15W and dose of 3J/cm², group GB (n= 8) was submitted to laser irradiation lambda= 670nm AlGaInP diode, continuous emission, power 30 mW and dose 3J/cm², group GAB (n= 8) was submitted to association of both of lasers and group GCL (n= 7) which the calcaneum tendon was submitted to placebo treatment. The group GP (n= 6) was a standard group without lesion and treatment. The animals received 12 sessions of lasertherapy and submitted to euthanasia on 14th day after surgery. The tendons were submitted to histological procedure and were analyzed to evaluate the optic retardation (in nm) for the birefringence measurement. RESULTS: The statistic analysis showed that the irradiated groups have higher values of optic retardation statistically significant than placebo group (p= 0,00001). It indicates that all the treated groups show better organization of collagen fibers in the longitudinal axis of the tendon, evidencing better quality in the tissue repair. The group GAB was that shown best results (p= 0,00001) when compared to another groups.

Adaptação enzimática da LDH em ratos submetidos a treinamento aeróbio em esteira e laser de baixa intensidade / Adaptation of LDH enzyme in rats undergoing aerobic treadmill training and low intensity laser therapy

OBJETIVO: Verificar a atividade e padrão eletroforético da LDH em resposta ao exercício em esteira em ratos submetidos à fotoestimulação por laser de baixa intensidade (780 etam). MÉTODO: Foram utilizados nesse estudo 54 ratos machos jovens (30dias), Winstar, com peso inicial médio de 112 ± 4,7g, os quais foram divididos em quatro grupos: dois permaneceram em repouso, GRC (grupo repouso controle, n= 9) e GRL (grupo repouso laser, n= 10), sendo o segundo irradiado por laser, e outros dois foram submetidos ao treinamento aeróbio e testes de esforço crescente em esteira (em degraus descontínuos) visando à determinação do limiar de anaerobiose (LA) durante 5 semanas, GEC (grupo exercício controle, n= 16) e GEL (grupo exercício laser, n= 19), sendo o último também irradiado por laser. O laser foi aplicado no quadríceps, glúteo máximo, sóleo e tibial anterior (TA), bilateralmente, imediatamente após cada sessão de treinamento, usando: 3,8J/cm², 15mW, 10s, modo contínuo, durante 5 semanas. Amostras de sóleo, TA e coração foram removidas 48 horas após a última sessão de exercício para análise eletroforética e espectrofotométrica. A estatística foi realizada através do teste ANOVA e post-hoc de TUKEY. O nível de significância foi considerado (p< 0,05). RESULTADOS: Foram observadas reduções na atividade da LDH causadas pelo laser (de 19 por cento a 30 por cento), exercício (de 41 por cento a 66 por cento) e as ações combinadas (de 47 por cento a 66 por cento) (p< 0,01). A eletroforese mostrou uma predominância da subunidade B para o sóleo e coração, e uma predominância da subunidade A no TA. CONCLUSÕES: Estes dados sugerem uma melhor resposta adaptativa dos animais ao treinamento aeróbio associado à terapia laser, no que se refere à otimização da via oxidativa.