Effect of Infrared and Green Photobiomodulation on the Number of MyoD-Positive Cells in the Connective Tissue of the Regenerating Skeletal Muscle Injury Site

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Abstract

BACKGROUND: Laser irradiation promotes accelerated regeneration of striated muscle tissue by enhancing cell proliferation and differentiation. The effectiveness of photobiomodulation depends on multiple factors, including target tissue, exposure duration, wavelength, and irradiation power. MyoD (myogenic differentiation) is a transcription factor that regulates myogenesis. We found no literature data on the effect of infrared and green photobiomodulation of varying durations on the enhancement of functional activity of MyoD-positive (MyoD⁺) cells and the increase in their number at the injury site. Meanwhile, the search for effective methods to restore skeletal muscle fibers after injury remains relevant.

AIM: To analyze the effect of infrared and green-spectrum laser irradiation on the number of MyoD⁺ cells in the connective tissue at the injury site of regenerating skeletal muscle.

METHODS: The study was conducted on 208 male Wistar rats divided into 6 experimental groups: control (group 0, n = 8); incised muscle wound (group 1, n = 40); incised muscle wound followed by short-term (60 s) infrared laser exposure to the wound area (group 2, n = 40); incised wound with prolonged (180 s) infrared laser exposure (group 3, n = 40); incised wound with short-term (60 s) green laser exposure (group 4, n = 40); and incised wound with prolonged (180 s) green laser exposure (group 5, n = 40). Laser irradiation was applied once in continuous mode immediately after muscle injury. Histological sections stained with hematoxylin and by immunohistochemical techniques using MyoD antibodies were examined to count the number of MyoD⁺ cells per 1 mm2 of tissue in the focal area of injured striated skeletal muscle on days 1, 3, 7, 14, and 30 of the observation period.

RESULTS: It was established that photobiomodulation increased the number of nuclei in the connective tissue within the focal area of muscle injury at various time points during the experiment. Moreover, a significant increase in the number of MyoD⁺ cells per 1 mm2 was observed 1 day after short-term photobiomodulation with both green and infrared wavelengths. The most pronounced stimulatory effect on MyoD⁺ cells was noted following short-term green laser irradiation.

CONCLUSION: Green and infrared photobiomodulation promoted an early increase in the number of MyoD⁺ cells in the connective tissue at the site of skeletal muscle injury. The greatest increase in cells undergoing myogenic differentiation was observed in rats following short-term green laser photobiomodulation.

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About the authors

Rostislav V. Takhaviev

South Ural State University

Email: rkenpachi@bk.ru
ORCID iD: 0000-0002-8994-570X
SPIN-code: 9619-9800
Russian Federation, Chelyabinsk

Gennady V. Bryukhin

South Ural State University

Author for correspondence.
Email: bgenvas@mail.ru
ORCID iD: 0000-0002-3898-766X
SPIN-code: 7691-8383

Dr. Sci. (Medicine), Professor

Russian Federation, Chelyabinsk

Elena S. Golovneva

South Ural State University

Email: micron30@mail.ru
ORCID iD: 0000-0002-6343-7563
SPIN-code: 1728-1640

Dr. Sci. (Medicine), Associate Professor

Russian Federation, Chelyabinsk

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2. Fig. 1. MyoD-positive cells (marked with asterisks) in the injury site of rat skeletal muscle on day 14 of the experiment, showing an increased number of MyoD-positive cells: a, after incision injury modeling (group 1); b, after prolonged infrared photobiomodulation of the wound area; c, after prolonged green laser irradiation of the wound area. Hematoxylin staining and immunohistochemistry using MyoD-specific antibodies; magnification ×1000 (objective ×100, eyepiece ×10).

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3. Fig. 2. MyoD-positive cells (marked with asterisks) in the injury site of rat skeletal muscle on day 7 of the experiment, showing high MyoD expression in intrafascicular cells and reduced expression in connective tissue cells surrounding the fibers: a, c, after green photobiomodulation; b, after infrared photobiomodulation. Hematoxylin staining and immunohistochemistry using MyoD-specific antibodies; magnification ×1000 (objective ×100, eyepiece ×10).

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