RESUMO
Photobiomodulation therapy (PBM) has shown positive effects when applied locally to modulate the inflammatory process and facilitate muscle repair. However, the available literature on the mechanisms of action of vascular photobiomodulation (VPBM), a non-invasive method of vascular irradiation, specifically in the context of local muscle repair, is limited. Thus, this study aimed to assess the impact of vascular photobiomodulation (VPBM) using a low-level laser (LLL) on the inflammatory response and the process of skeletal muscle repair whether administered prior to or following cryoinjury-induced acute muscle damage in the tibialis anterior (TA) muscles. Wistar rats (n = 85) were organized into the following experimental groups: (1) Control (n = 5); (2) Non-Injury + VPBM (n = 20); (3) Injured (n = 20); (4) Pre-VPBM + Injury (n = 20); (5) Injury + Post-VPBM (n = 20). VPBM was administered over the vein/artery at the base of the animals' tails (wavelength: 780 nm; power: 40 mW; application area: 0.04 cm2; energy density: 80 J/cm2). Euthanasia of the animals was carried out at 1, 2, 5, and 7 days after inducing the injuries. Tibialis anterior (TA) muscles were collected for both qualitative and quantitative histological analysis using H&E staining and for assessing protein expression of TNF-α, MCP-1, IL-1ß, and IL-6 via ELISA. Blood samples were collected and analyzed using an automatic hematological analyzer and a leukocyte differential counter. Data were subjected to statistical analysis (ANOVA/Tukey). The results revealed that applying VPBM prior to injury led to an increase in circulating neutrophils (granulocytes) after 1 day and a subsequent increase in monocytes after 2 and 5 days, compared to the Non-Injury + VPBM and Injured groups. Notably, an increase in erythrocytes and hemoglobin concentration was observed in the Non-Injury + VPBM group on days 1 and 2 in comparison to the Injured group. In terms of histological aspects, only the Prior VPBM + Injured group exhibited a reduction in the number of inflammatory cells after 1, 5, and 7 days, along with an increase in blood vessels at 5 days. Both the Prior VPBM + Injured and Injured + VPBM after groups displayed a decrease in myonecrosis at 1, 2, and 7 days, an increase in newly-formed and immature fibers after 5 and 7 days, and neovascularization after 1, 2, and 7 days. Regarding protein expression, there was an increase in MCP-1 after 1 and 5 days, TNF-α, IL-6, and IL-1ß after 1, 2, and 5 days in the Injured + VPBM after group when compared to the other experimental groups. The Prior VPBM + Injured group exhibited increased MCP-1 production after 2 days, in comparison to the Non-Injury + VPBM and Control groups. Notably, on day 7, the Injured group continued to show elevated MCP-1 protein expression when compared to the VPBM groups. In conclusion, VPBM effectively modulated hematological parameters, circulating leukocytes, the protein expression of the chemokine MCP-1, and the proinflammatory cytokines TNF-α and IL-1ß, ultimately influencing the inflammatory process. This modulation resulted in a reduction of myonecrosis, restoration of tissue architecture, increased formation of newly and immature muscle fibers, and enhanced neovascularization, with more pronounced effects when VPBM was applied prior to the muscle injury.
Assuntos
Terapia com Luz de Baixa Intensidade , Músculo Esquelético , Ratos Wistar , Animais , Ratos , Músculo Esquelético/efeitos da radiação , Músculo Esquelético/metabolismo , Masculino , Biomarcadores/metabolismo , Inflamação/metabolismo , Inflamação/patologia , Interleucina-6/metabolismo , Interleucina-6/sangue , Interleucina-1beta/metabolismo , Interleucina-1beta/sangue , Modelos Animais de Doenças , Fator de Necrose Tumoral alfa/metabolismo , Fator de Necrose Tumoral alfa/sangue , Cicatrização/efeitos da radiação , Quimiocina CCL2/metabolismoRESUMO
OBJECTIVES: Spinal cord injury (SCI) causes the discontinuity of the spinal canal, leading to functional and sensorial losses in areas below the injury, which are often irreversible. Photobiomodulation (PBM) can enhance the neuromuscular repair process, especially in cases of peripheral nerve injuries. However, there is little knowledge regarding the effects of this therapeutic modality on recovery following a SCI, especially the noninvasive systemic form denominated vascular PBM (VPBM). To analyze the effects of VPBM in the immediate, acute and intermediate phases following a compression-induced SCI on morphological aspects of neuromuscular tissue repair, functional recovery and the protein expression of brain-derived neurotrophic factor (BDNF). METHODS: Wistar rats were divided into five groups: control, SCI, SCI + VPBM-Im (immediate administration of VPBM), SCI + VPBM-2h (VPBM administered 2 h after injury) and SCI + VPBM-14d (VPBM administered 14 days after injury). VPBM was administered in the region of the caudal vein/artery with low-level laser (AsGaAl, 780 nm, 80 J/cm², 40 mW for 80 s, totaling an energy of 3.2 J over a single point) for 14 consecutive days. During the analysis periods (1, 3, 7, 14, 21, 28 and 35 days after injury), functioning was evaluated using the Basso-Beattie-Bresnahan (BBB) index. At the end of each experimental period, blood samples were collected for the determination of the concentration of circulating BDNF using ELISA. Muscle tissue and nerve tissue samples were also extracted for morphological and histological analyses using H&E staining. RESULTS: SCI + VPBM-Im and SCI + VPBM-2 h led to the recovery of motor function beginning on the 7th day after injury (p < 0.05), an increase in the cross-sectional area (CSA) of the muscle fibers in the second week (p < 0.05) and an increase in muscle fiber diameter beginning on Day 14 (p < 0.05). Early irradiation had a greater effect on the reduction in the size of the cavity, with stabilization of the cavity found on Day 7 (p < 0.05). Considering the circulating BDNF levels, no changes was found during the experimental periods. CONCLUSION: The present results showed that VPBM was capable of modulating morphological and functional recovery following SCI, especially when administered early. The positive effects on functional recovery were demonstrated by the BBB index; the reestablishment of the structure of the muscle and nerve tissue was demonstrated by the preservation of CSA and diameter of muscle fiber and reduction in the area of the injury (cavity size) respectively. Thus, noninvasive VPBM may be an important component of treatment for spinal cord injuries.
Assuntos
Fator Neurotrófico Derivado do Encéfalo , Traumatismos da Medula Espinal , Ratos , Animais , Fator Neurotrófico Derivado do Encéfalo/metabolismo , Fator Neurotrófico Derivado do Encéfalo/uso terapêutico , Ratos Sprague-Dawley , Ratos Wistar , Traumatismos da Medula Espinal/radioterapia , Traumatismos da Medula Espinal/patologia , Atividade Motora/fisiologia , Recuperação de Função Fisiológica/fisiologiaRESUMO
Peripheral nerve injury (PNI) is associated with considerable functional impairment. Photobiomodulation (PBM) has demonstrated positive effects regarding neuromuscular repair after PNI when applied locally to the nerve or injured muscle. However, the effects of systemic PBM with transcutaneous application over an important artery, which is also denominated vascular PBM (VPBM), remain unclear. The aim of the study was to compare the effects of VPBM with low-level laser (LLL) and light-emitting diode (LED) on gait, sensitivity and muscle morphology following a PNI. PNI was induced on Wistar rats using the sciatic nerve crushing technique. VPBM was performed over the rat's artery tail region with LED (850 nm, 40 mW, 3.2 J) and LLL (780 nm, 40 mW, 3.2 J). Gait functionality, mechanical (nociceptive) sensitivity, and morphology of the tibialis anterior muscle were evaluated at 7, 14, and 21 days after injury. An improvement in functional gait was shown in the VPBM-LLL group in all periods. Motor sensitivity was found after 14 days in the VPBM-LLL group. The left/right (L/R) muscle mass ratio revealed a reduction in muscle atrophy in the VPBM-LLL group at 7 days. Muscle fiber diameter increased in the VPBM-LED group at 14 days and increases in the cross-section area were found in the VPBM-LED and VPBM-LLL groups at 7 days. VPBM with both light sources (LED and LLL) positively modulated functioning and neuromuscular recovery following sciatic nerve injury in rats, with more pronounced results when using LLL.
Assuntos
Terapia com Luz de Baixa Intensidade , Traumatismos dos Nervos Periféricos , Ratos , Animais , Ratos Wistar , Traumatismos dos Nervos Periféricos/radioterapia , Terapia com Luz de Baixa Intensidade/métodos , Nervo Isquiático , LasersRESUMO
The local photobiomodulation (LPBM) has demonstrated positive effects during compensatory hypertrophy (CH) in skeletal muscle as a response to an overload. The aim was to compare the effects of the transcutaneous vascular photobiomodulation (VPBM) and the LPBM on muscle fiber size, gait functionality, and on mechanical sensitivity during the CH model in rats. VPBM was administered over the rat's main tail vein and LPBM was applied over the plantar muscle region. VPBM induced an increase in muscle fiber diameter and cross-sectional area (CSA) after 7 days. At 14 days, an increase in the fiber diameter was found in both irradiated groups. The VPBM and LPBM promoted the reestablishment of normal gait evaluated by the sciatic functional index after 14 days. No changes were found in the mechanical (nociceptive) sensitivity in VPBM and LPBM groups in comparison to the CH group but there was an increase in the nociceptive sensitivity in the CH groups in comparison to the control after 7 and 14 days. In conclusion, both PBM, vascular and local, were able to improve the muscle size and gait during the CH process with more pronounced effects when irradiation was performed systemically (VPBM).
Assuntos
Terapia com Luz de Baixa Intensidade , Ratos , Animais , Ratos Wistar , Fibras Musculares Esqueléticas , Músculo Esquelético , Marcha , HipertrofiaRESUMO
Peripheral nerve injury (PNI) can lead to sensory and/or motor impairment. As a treatment photobiomodulation (PBM) has demonstrated positive effects in terms of the maintenance of muscle activation and trophism. Wistar rats were divided into five groups: control, injury, injury + PBMn (irradiation over injured nerve), injury + PBMm (irradiation over affected muscle) and injury + PBMnm (irradiation over nerve and muscle). The left sciatic nerve was submitted to a crushing injury. Treatment was administered with low-level laser (780 nm, 0.04 cm2 , 1 W cm-2 , 3.2 J) over the injured nerve and/or the tibialis anterior muscle. The effects of PBM were favorable on muscle morphology and gene expression of calcineurin, myogenin and acetylcholine receptors. PBM led to an acceleration on muscle repair process, and effects were more evident in 2 weeks after PNI. Thus, PBM is indicated for the area over both the injured nerve and the affected muscle.
Assuntos
Terapia com Luz de Baixa Intensidade , Traumatismos dos Nervos Periféricos/terapia , Animais , Ratos , Ratos Wistar , Nervo Isquiático/lesões , Nervo Isquiático/efeitos da radiaçãoRESUMO
Moderate levels of a proinflammatory macrophages phenotype are indispensable and play an important role in the skeletal muscle repair process since this response depends on their secreted products concentration to influence and modulate muscle inflammation as well as the differentiation of myoblasts. This study investigated the effects of photobiomodulation (PBM) on undifferentiated and differentiation-induced C2C12 myoblasts cultivated in different concentrations of M1 phenotype macrophage-conditioned media of J774 cells (MCM1) also submitted to PBM using the same irradiation parameters. Irradiation was performed once with low-level laser (780 nm, 70 mW, 1 J) and was evaluated cell viability, proliferation and differentiation, nitric oxide (NO) synthesis and IL-6 and TNF-α protein levels 24 and 48 h after C2C12 irradiation. PBM treatment in undifferentiated myoblasts exhibited lower IL-6 levels in the presence of nonirradiated MCM1 at both concentrations. Myoblasts in proliferation condition cultivated with irradiated MCM1 showed lower IL-6 and TNF-α levels after 48 h in the presence of both concentrations evaluated. PBM induced a decrease in the synthesis of NO on undifferentiated and differentiation-induced myoblasts. PBM was able to reduce the level of proinflammatory protein and markers, which are important to allow the differentiation of myoblasts during the muscle repair process.