Regeneration in denervated toad (bufo viridis) gastrocnemius muscle and the promotion of the process by low energy laser irradiation

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Abstract
Background: It is known that while denervated skeletal muscles have the ability to regenerate, maturation of regenerated myofibres does not take place under these conditions. Denervation also causes elevation of “invasive” and satellite cells, but the role of these cells in the regeneration process after injury to the denervated muscle is still unknown. Low energy lasers have recently been found to modulate and accelerate physiological processes in cells. The aim of the present study was to compare regeneration in denervated and innervated amphibian muscles and to investigate whether this process in denervated muscles can be stimulated by low energy laser irradiation prior to injury in these muscles. Methods: Denervated gastrocnemius muscles of toads were irradiated with He‐Ne laser (6.0 mW, 31.2 J/cm2) 7 days postdenervation (control muscle received red light irradiation at the same wavelength). Nine days after denervation cold injury was performed on the site of irradiation of both groups of muscles. At 14 days postinjury all muscles were removed and processed for histology and histomorphometric analysis of mononucleated cells, myotubes, and young myofibres in the regenerated zone. Results: The volume fraction (percent of total injured zone) of the various histological structures in the injured zones 14 days after cold injury in the denervated (9 days prior to injury) muscles did not differ from innervated injured muscles at the same time interval postinjury. The mononucleated cells and myotubes in the laser irradiated muscles comprised 49 ± 4% and 6 plusmn; 1% of the injured area, respectively, which was significantly lower than their volume fraction (67 plusmn; 2% and 11 plusmn; 2%, respectively) in the control muscles. The young myofibres populated 34 plusmn; 4% of the total injured area in the denervated and laser irradiated muscles which was significantly higher than their volume fraction (12 ± 2%) in control denervated muscles. Conclusions: It is concluded that initial stages of regeneration can also take place in skeletal denervated and injured muscles of amphibians. The kinetics of the regeneration process are identical in denervated and innervated muscles. The process of regeneration in denervated muscles can be markedly enhanced if the muscle is irradiated by low energy laser prior to injury, probably by activation (stimulation of proliferation and/or differentiation) cells in the muscles that are “recruited” and participate in the process of regeneration.