Evaluation Of Electrical Stimulation On Muscle Satellite Cell Development And Differentiation

Background:A variety of pacemaker is gradually used in clinic. The muscles stimulated by the pacemaker, like paralized cricoarytenoid muscle pacemaker, the paralysed detrusor of bladder pacemaker and apnea muscle pacemaker has been applied in clinical trials and in clinic. At present, the effect of electrical stimulation on muscle paralysis of muscles and normal muscle remains unclear. Pacemakers recover the muscle function instead of nerve, while it also changes the muscle cell. Muscle satellite cells as main source of muscle fiber, have unclearly effects on muscle cells regeneration and proliferation undergoing pacer electrical stimulation. In recent years, the many studies found muscle satellite cells are the main source of muscle regeneration undergoing electrical stimulation. The reports about satellite cell undergoing electrical are little.Head and neck muscles are important muscle for sound, breathing, swallowing and head and neck movement. The nerve of muscles of head and neck irreversible damaged, muscles become atrophy. Satellite stem cells as main source of muscle cells, keep important effect on muscle regeneration and repair. So we begin our study on head and neck satellite cells with electrical stimulation. We investigated the changes of satellite activity, proliferation and differentiation undergoing electrical stimulation. These studies will improve our understanding regarding the potential electrophysiologic mechanisms of satellite cell proliferation and differentiation, facilitate the development of novel electrical stimulation system approach for future study, and provide useful information on the optimal electrical stimulation pattern which can be achieved in head neck muscle pacer application.Objective:Using the C2C12cell lines and beagle dogs genioglossus muscle satellite cells in vitro, which undergo fixed voltage, fixed stimulation time, different pulse widths and frequencies, investigate the effects of electrical stimulation on satellite stem cell proliferation and differentiation.Methods:1.In vitro cultured C2C12cell line, fixed voltage (5V), different pulse width (0ms,1ms,2ms,4ms,8ms,20ms six groups), the different frequency (Opps,2pps, lOpps,20pps,40pps,80pps six groups) train stimulation (5S,3s-on,2s-off) stimulated cell lines. The MTT cell viability assay, flow cytometry cycle proliferation assessment, qRT-PCR were applied in this study.2. Canine genioglossus muscle satellite cell, were stimulated with fixed voltage (5V), lms duration, different frequency (Opps,2pps,10pps,40pps,80pps,160group six) train electrical current(5S,3s-on,2s-off) in vitro. The MTT cell viability assay, flow cytometry cycle proliferation assessment, qRT-PCR and Western blotting were used in this study to assess cell proliferation and different.3. Spearmam r coefficient was calculated to evaluate correlations. Data are reported as mean±D. Data were analyzed using SPSS release19.0for Windows(SPSS, Chicago, IL, USA). Kruskal-Wallis ANOVA tests was performed to compare groups subjected to different treatments, followed by multiple pairwise comparisons. There were statistically significant differences in P<0.05.Results:1. With the pulse width pulse increment electrical stimulation will lead to reduced myogenic C2C12cell viability(r=-0.724, P<0.01). Its differentiation ability has improved, especially in pulse width20ms (p=0.044). With the frequency of electrical stimulation increasing, the C2C12cells proliferation(r=0.643, P=0.004) and differentiation (r=0.637, P=0.004; r=0.660, P=0.003)were enhanced. The trend of muscle cell transit from fast to slow type is obvious, especially in80pps frequency (P<0.05).2. As the stimulation frequency increasing, muscle satellite cells viability and proliferation activity had no change, but cell differentiation was enhanced. The transform of muscle cell phenotypes from fast to slow happened undergoing different frequency electrical stimulation, especially160pps frequency (P<0.01).Conclusion:In vitro, with the pulse width pulse electrical stimulation increment will lead to reduced activity of satellite stem cells and enhanced differentiation ability, elicit muscle phenotypes transform. The frequency increment of electrical stimulation enhances satellite stem cells differentiation, fast-to-slow phenotypes transformation.