Shi Yong, Zhang Xiaoxuan, Liu Rui, Shao Xiaoyan, Zhao Yuanjin, Gu Zhuxiao, Jiang Qing
State Key Laboratory of Pharmaceutical Biotechnology, Division of Sports Medicine and Adult Reconstructive Surgery, Department of Orthopedic Surgery, Nanjing Drum Tower Hospital, The Affiliated Hospital of Nanjing University Medical School, 321 Zhongshan Road, Nanjing, 210008, Jiangsu, People's Republic of China.
Branch of National Clinical Research Center for Orthopedics, Sports Medicine and Rehabilitation, Nanjing, People's Republic of China.
Biomater Res. 2022 Dec 22;26(1):87. doi: 10.1186/s40824-022-00335-w.
Volumetric muscle loss (VML) due to various reasons may cause motor dysfunction and tissue engineering has been proposed for muscle regeneration. However, developing three-dimensional (3D) tissue-engineered scaffolds that can mimic oriented cell growth of muscle tissues are challenging for regeneration medicine. Herein, we propose a novel self-curling 3D oriented scaffold (SCOS) composed of fish derived gelatin methacrylate (GelMA) and fish scales for repairing skeletal muscles.
Fish scales of tilapia were decellularized and decalcified. Then, SCOSs were constructed by ultraviolet-coating methylated fish gelatin on the back of fish scales. C2C12 myoblasts were cultured on SCOSs, and after induction of myogenic differentiation, SCOS/C2C12 transplants were prepared for in vivo experiments.
Decellularized and decalcified fish scales (DDFSs) became soft and retained the original oriented microgroove surface structure that could induce oriented cell growth. SCOSs could self-curl into 3D structures when immersing in culture medium due to different swelling properties of fish GelMA and DDFSs. Cell experiments demonstrated that SCOSs enhanced the oriented growth and myogenic differentiation of C2C12 myoblasts. By integrating SCOSs and myogenic differentiated C2C12 myoblasts, the resultant SCOS/C2C12 transplants promoted de novo muscle regeneration and functional restoration of muscle activity in the mouse model of VML.
Our results suggest that SCOSs loaded with myogenic differentiated C2C12 myoblasts can promote muscle regeneration in mice with skeletal muscle injuries, indicating application prospects of such scaffolds in muscle tissue engineering and other related fields.
由于各种原因导致的体积性肌肉损失(VML)可能会引起运动功能障碍,组织工程已被提出用于肌肉再生。然而,开发能够模拟肌肉组织定向细胞生长的三维(3D)组织工程支架对再生医学来说具有挑战性。在此,我们提出一种由鱼源甲基丙烯酸明胶(GelMA)和鱼鳞组成的新型自卷曲3D定向支架(SCOS),用于修复骨骼肌。
罗非鱼的鱼鳞进行脱细胞和脱钙处理。然后,通过在鱼鳞背面紫外线涂层甲基化鱼明胶构建SCOS。将C2C12成肌细胞接种在SCOS上,诱导成肌分化后,制备SCOS/C2C12移植物用于体内实验。
脱细胞和脱钙的鱼鳞(DDFS)变得柔软,并保留了能够诱导细胞定向生长的原始定向微槽表面结构。由于鱼GelMA和DDFS的膨胀特性不同,SCOS浸入培养基时可自卷曲成3D结构。细胞实验表明,SCOS增强了C2C12成肌细胞的定向生长和成肌分化。通过整合SCOS和经成肌分化的C2C12成肌细胞,所得的SCOS/C2C12移植物促进了VML小鼠模型中的新生肌肉再生和肌肉活动功能恢复。
我们的结果表明,负载经成肌分化的C2C12成肌细胞的SCOS可促进骨骼肌损伤小鼠的肌肉再生,表明这种支架在肌肉组织工程和其他相关领域的应用前景。