Patel Anjali, Vendrell-Gonzalez Sara, Haas Gabriel, Marcinczyk Madison, Ziemkiewicz Natalia, Talovic Muhamed, Fisher Jonathan S, Garg Koyal
Department of Biomedical Engineering, Parks College of Engineering, Aviation, and Technology, Saint Louis University, St. Louis, Missouri.
Department of Biology, College of Arts and Sciences, Saint Louis University, St. Louis, Missouri.
Biores Open Access. 2019 Jul 30;8(1):129-138. doi: 10.1089/biores.2019.0016. eCollection 2019.
Skeletal muscle has a remarkable regenerative capacity in response to mild injury. However, when muscle is severely injured, muscle regeneration is impaired due to the loss of muscle-resident stem cells, known as satellite cells. Fibrotic tissue, primarily comprising collagen I (COL), is deposited with this critical loss of muscle. In recent studies, supplementation of laminin (LM)-111 has been shown to improve skeletal muscle regeneration in several models of disease and injury. Additionally, electrical stimulation (E-stim) has been investigated as a possible rehabilitation therapy to improve muscle's functional recovery. This study investigated the role of E-stim and substrate in regulating myogenic response. C2C12 myoblasts were allowed to differentiate into myotubes on COL- and LM-coated polydimethylsiloxane molds. The myotubes were subjected to E-stim and compared with nonstimulated controls. While E-stim resulted in increased myogenic activity, irrespective of substrate, LM supported increased proliferation and uniform distribution of C2C12 myoblasts. In addition, C2C12 myoblasts cultured on LM showed higher Sirtuin 1, mammalian target of rapamycin, desmin, nitric oxide, and vascular endothelial growth factor expression. Taken together, these results suggest that an LM substrate is more conducive to myoblast growth and differentiation in response to E-stim .
骨骼肌在受到轻度损伤时具有显著的再生能力。然而,当肌肉受到严重损伤时,由于肌肉驻留干细胞(即卫星细胞)的丧失,肌肉再生会受到损害。主要由I型胶原蛋白(COL)组成的纤维化组织会随着肌肉的这种关键损失而沉积。在最近的研究中,已表明补充层粘连蛋白(LM)-111可在多种疾病和损伤模型中改善骨骼肌再生。此外,电刺激(E-stim)已作为一种可能的康复疗法进行研究,以改善肌肉的功能恢复。本研究调查了E-stim和底物在调节成肌反应中的作用。使C2C12成肌细胞在COL和LM包被的聚二甲基硅氧烷模具上分化为肌管。对肌管进行E-stim,并与未刺激的对照进行比较。虽然无论底物如何,E-stim都会导致成肌活性增加,但LM支持C2C12成肌细胞的增殖增加和均匀分布。此外,在LM上培养的C2C12成肌细胞显示出更高的沉默调节蛋白1、雷帕霉素哺乳动物靶蛋白、结蛋白、一氧化氮和血管内皮生长因子表达。综上所述,这些结果表明,LM底物更有利于成肌细胞在E-stim作用下的生长和分化。