Narayanan Naagarajan, Lengemann Paul, Kim Kun Ho, Kuang Liangju, Sobreira Tiago, Hedrick Victoria, Aryal Uma K, Kuang Shihuan, Deng Meng
Department of Agricultural and Biological Engineering, Purdue University, West Lafayette, Indiana, USA.
Bindley Bioscience Center, Purdue University, West Lafayette, Indiana, USA.
J Biomed Mater Res A. 2021 Mar;109(3):289-299. doi: 10.1002/jbm.a.37022. Epub 2020 Jun 26.
Nerve cells secrete neurotrophic factors that play a critical role in neuronal survival, proliferation, and regeneration. However, their role in regulating myoblast behavior and skeletal muscle repair remains largely unexplored. In the present study, we investigated the effects of PC12 secreted signaling factors in modulating C2C12 myoblast behavior under physiologically relevant conditions. We showed that PC12 conditioned media modulated myoblast proliferation and differentiation in both 2D culture and 3D aligned electrospun fiber scaffold system in a dose-dependent manner. We further developed a biomimetic, tunable hydrogel consisting of hyaluronic acid, chondroitin sulfate, and polyethylene glycol as a 3D matrix encapsulating PC12 cells. The hydrogel-encapsulated PC12 cells promoted survival and proliferation of myoblasts in co-culture. Further proteomics analysis identified a total of 2,088 proteins from the secretome of the encapsulated PC12 cells and revealed the biological role and overlapping functions of nerve-secreted proteins for skeletal muscle regeneration, potentially through regulating myoblast behavior, nerve function, and angiogenesis. These experiments provide insights into the nerve-muscle interactions and pave the way for developing advanced biomaterials strategies incorporating nerve cell secretome for accelerated skeletal muscle regeneration.
神经细胞分泌神经营养因子，这些因子在神经元的存活、增殖和再生中发挥着关键作用。然而，它们在调节成肌细胞行为和骨骼肌修复中的作用在很大程度上仍未得到探索。在本研究中，我们研究了PC12分泌的信号因子在生理相关条件下对C2C12成肌细胞行为的调节作用。我们发现，在二维培养和三维排列的电纺纤维支架系统中，PC12条件培养基均以剂量依赖的方式调节成肌细胞的增殖和分化。我们进一步开发了一种由透明质酸、硫酸软骨素和聚乙二醇组成的仿生、可调节水凝胶，作为包裹PC12细胞的三维基质。水凝胶包裹的PC12细胞在共培养中促进了成肌细胞的存活和增殖。进一步的蛋白质组学分析从包裹的PC12细胞的分泌组中总共鉴定出2088种蛋白质，并揭示了神经分泌蛋白对骨骼肌再生的生物学作用和重叠功能，可能是通过调节成肌细胞行为、神经功能和血管生成来实现的。这些实验为神经-肌肉相互作用提供了见解，并为开发结合神经细胞分泌组的先进生物材料策略以加速骨骼肌再生铺平了道路。
