Salehi Majid, Bagher Zohreh, Kamrava Seyed Kamran, Ehterami Arian, Alizadeh Rafieh, Farhadi Mohammad, Falah Masoumeh, Komeili Ali
Department of Tissue Engineering, School of Medicine, Shahroud University of Medical Sciences, Shahroud, Iran.
Tissue Engineering and Stem Cells Research Center, Shahroud University of Medical Sciences, Shahroud, Iran.
J Cell Physiol. 2019 Sep;234(9):15357-15368. doi: 10.1002/jcp.28183. Epub 2019 Jan 31.
Regeneration and functional recovery after peripheral nerve damage still remain a significant clinical problem. In this study, alginate/chitosan (alg/chit) hydrogel was used for the transplantation of olfactory ectomesenchymal stem cells (OE-MSCs) to promote peripheral nerve regeneration. The OE-MSCs were isolated from olfactory mucosa biopsies and evaluated by different cell surface markers and differentiation capacity. After creating sciatic nerve injury in a rat model, OE-MSCs were transplanted to the injured area with alg/chit hydrogel which was prepared and well-characterized. The prepared hydrogel had the porosity of 91.3 ± 1.27%, the swelling ratio of 379% after 240 min, weight loss percentages of 80 ± 5.56% after 14 days, and good blood compatibility. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, 4',6-diamidino-2-phenylindole, and LIVE/DEAD staining were done to assay the behavior of OE-MSCs on alg/chit hydrogel and the results confirmed that the hydrogel can provide a suitable substrate for cell survival. For functional analysis, alg/chit hydrogel with and without OE- MSCs was injected into a 3-mm sciatic nerve defect of Wistar rats. The results of the sciatic functional index, hot plate latency, electrophysiological assessment, weight-loss percentage of wet gastrocnemius muscle, and histopathological examination using hematoxylin-eosin and Luxol fast blue staining showed that utilizing alg/chit hydrogel with OE-MSCs to the sciatic nerve defect enhance regeneration compared to the control group and hydrogel without cells.
周围神经损伤后的再生和功能恢复仍然是一个重大的临床问题。在本研究中,藻酸盐/壳聚糖(alg/chit)水凝胶被用于移植嗅外胚间充质干细胞(OE-MSCs)以促进周围神经再生。OE-MSCs从嗅黏膜活检组织中分离出来,并通过不同的细胞表面标志物和分化能力进行评估。在大鼠模型中造成坐骨神经损伤后,将OE-MSCs与制备好且特性明确的alg/chit水凝胶一起移植到损伤部位。制备的水凝胶孔隙率为91.3±1.27%,240分钟后的溶胀率为379%,14天后的失重百分比为80±5.56%,并且具有良好的血液相容性。进行了3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐试验、4',6-二脒基-2-苯基吲哚试验和活/死染色,以检测OE-MSCs在alg/chit水凝胶上的行为,结果证实该水凝胶可为细胞存活提供合适的基质。为了进行功能分析,将含有和不含有OE-MSCs的alg/chit水凝胶注入Wistar大鼠3毫米的坐骨神经缺损处。坐骨神经功能指数、热板潜伏期、电生理评估、腓肠肌湿重减轻百分比以及苏木精-伊红染色和卢戈氏碘蓝染色的组织病理学检查结果表明,与对照组和不含细胞的水凝胶相比,将含有OE-MSCs的alg/chit水凝胶应用于坐骨神经缺损处可增强再生。