Department of Chemical Engineering, National Cheng Kung University, Tainan 70101, Taiwan.
Department of Cell Biology and Anatomy, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
Mater Sci Eng C Mater Biol Appl. 2020 Jul;112:110964. doi: 10.1016/j.msec.2020.110964. Epub 2020 Apr 13.
The adipose stem cell is a potential candidate for the autologous chondrocytes repairing approach because of the abundance of fat in the animal body and its versatile differentiation capability. In this study, rat adipose stem cells (rASCs) were seeded into anti-oxidative N-acetylcysteine (NAC) grafted polyurethane (PU) scaffold and then combined with short dynamic compressive stimulation (24 h) to induce rASCs chondrogenesis differentiation in vitro. The inner pore surface of the PU scaffold was first modified via alginate and type I collagen to promote rASCs adherence. The modified layers crosslinked by genipin showed outstanding stability after ultrasonic treatment, indicating the modified layers were stable and can keep the cells adhesion well during dynamic compressive stimulation. After inner pore surface modification and 10 mM NAC grafting, the PU scaffold-A-C-G (graft 10 mM NAC) has shown the best proliferation efficiency with homogeneous cell distribution after 72hr static culture. After short term dynamic compressive stimulation, significant gene expression in chondrogenic markers, Sox-9, and Aggrecan, were noted in both PU scaffold-A-C-G and PU scaffold-A-C-G (graft 10 mM NAC). Considering the cell proliferation efficiency and gene expression, the anti-oxidative NAC grafted PU scaffold combined with short term dynamic compressive stimulation could be useful for cell culturing in stem cell therapy.
脂肪干细胞因其在动物体内丰富的脂肪含量和多样的分化能力,是自体软骨细胞修复方法的潜在候选者。在本研究中,将大鼠脂肪干细胞(rASCs)接种到抗氧化 N-乙酰半胱氨酸(NAC)接枝的聚氨酯(PU)支架上,然后结合短期动态压缩刺激(24 h),在体外诱导 rASCs 软骨分化。首先通过海藻酸钠和 I 型胶原对 PU 支架的内孔表面进行修饰,以促进 rASCs 的黏附。经京尼平交联的修饰层经超声处理后表现出出色的稳定性,表明修饰层稳定,在动态压缩刺激过程中能保持细胞良好的黏附性。内孔表面修饰和接枝 10 mM NAC 后,PU 支架-A-C-G(接枝 10 mM NAC)在 72 小时静态培养后显示出最佳的增殖效率和均匀的细胞分布。短期动态压缩刺激后,PU 支架-A-C-G 和 PU 支架-A-C-G(接枝 10 mM NAC)中软骨标志物 Sox-9 和 Aggrecan 的基因表达均显著增加。考虑到细胞增殖效率和基因表达,抗氧化 NAC 接枝的 PU 支架结合短期动态压缩刺激可用于干细胞治疗中的细胞培养。