Abstract：
BACKGROUND: Based on recent advances in tissue engineering and stem cell therapy in nervous system diseases treatments, this study aimed to investigate sciatic nerve regeneration using human endometrial stem cells (hEnSCs) encapsulated fibrin gel containing chitosan nanoparticle loaded by insulin (Ins-CPs). Stem cells and also Insulin (Ins), which is a strong signaling molecule in peripheral nerve regeneration, play an important role in neural tissue engineering.
METHODS: The fibrin hydrogel scaffold containing insulin loaded chitosan particles was synthesized and characterized. Release profiles of insulin from hydrogel was determined through UV-visible spectroscopy. Also, human endometrial stem cells encapsulated in hydrogel and its cell biocompatibility were assigned. Furthermore, the sciatic nerve crush injury was carried out and prepared fibrin gel was injected at the crush injury site by an 18-gage needle. Eight and twelve weeks later, the recovery of motor and sensory function and histopathological evaluation were assessed.
RESULTS: The in vitro experiments showed that the insulin can promote hEnSCs proliferation within a certain concentration range. Animals\' treatment confirmed that developed fibrin gel containing Ins-CPs and hEnSCs significantly improves motor function and sensory recovery. Hematoxylin and Eosin (H&E) images provided from cross-sectional and, longitudinal-sections of the harvested regenerative nerve showed that regenerative nerve fibers had been formed and accompanied with new blood vessels in the fibrin/insulin/hEnSCs group.
CONCLUSIONS: Our results demonstrated that the prepared hydrogel scaffolds containing insulin nanoparticles and hEnSCs could be considered as a potential biomaterial aimed at regeneration of sciatic nerves.
METHODS: The fibrin hydrogel scaffold containing insulin loaded chitosan particles was synthesized and characterized. Release profiles of insulin from hydrogel was determined through UV-visible spectroscopy. Also, human endometrial stem cells encapsulated in hydrogel and its cell biocompatibility were assigned. Furthermore, the sciatic nerve crush injury was carried out and prepared fibrin gel was injected at the crush injury site by an 18-gage needle. Eight and twelve weeks later, the recovery of motor and sensory function and histopathological evaluation were assessed.
RESULTS: The in vitro experiments showed that the insulin can promote hEnSCs proliferation within a certain concentration range. Animals\' treatment confirmed that developed fibrin gel containing Ins-CPs and hEnSCs significantly improves motor function and sensory recovery. Hematoxylin and Eosin (H&E) images provided from cross-sectional and, longitudinal-sections of the harvested regenerative nerve showed that regenerative nerve fibers had been formed and accompanied with new blood vessels in the fibrin/insulin/hEnSCs group.
CONCLUSIONS: Our results demonstrated that the prepared hydrogel scaffolds containing insulin nanoparticles and hEnSCs could be considered as a potential biomaterial aimed at regeneration of sciatic nerves.
摘要：
背景：基于组织工程和干细胞疗法在神经系统疾病治疗中的最新进展，本研究旨在研究使用人子宫内膜干细胞（hEnSCs）包裹的纤维蛋白凝胶（含胰岛素负载的壳聚糖纳米颗粒（Ins-CPs））的坐骨神经再生。干细胞和胰岛素（Ins），它是周围神经再生中的一种强信号分子，在神经组织工程中发挥着重要作用。
方法：合成并表征了含有负载胰岛素的壳聚糖颗粒的纤维蛋白水凝胶支架。通过UV-可见光谱法测定胰岛素从水凝胶中的释放曲线。此外，我们对封装在水凝胶中的人子宫内膜干细胞及其细胞生物相容性进行了鉴定。此外，进行坐骨神经挤压损伤，并在挤压损伤部位用18针注射制备的纤维蛋白凝胶。八周和十二周后，评估了运动和感觉功能的恢复以及组织病理学评估。
结果：体外实验表明，胰岛素在一定浓度范围内可促进hEnSCs的增殖。动物治疗证实，含有Ins-CPs和hEnSC的开发的纤维蛋白凝胶显着改善运动功能和感觉恢复。苏木精和伊红(H&E)图像从横截面和,采集的再生神经纵切面显示,在纤维蛋白/胰岛素/hEnSC组中,再生神经纤维已经形成,并伴有新血管.
结论：我们的结果表明，制备的含有胰岛素纳米颗粒和hEnSC的水凝胶支架可以被认为是一种潜在的旨在再生坐骨神经的生物材料。
方法：合成并表征了含有负载胰岛素的壳聚糖颗粒的纤维蛋白水凝胶支架。通过UV-可见光谱法测定胰岛素从水凝胶中的释放曲线。此外，我们对封装在水凝胶中的人子宫内膜干细胞及其细胞生物相容性进行了鉴定。此外，进行坐骨神经挤压损伤，并在挤压损伤部位用18针注射制备的纤维蛋白凝胶。八周和十二周后，评估了运动和感觉功能的恢复以及组织病理学评估。
结果：体外实验表明，胰岛素在一定浓度范围内可促进hEnSCs的增殖。动物治疗证实，含有Ins-CPs和hEnSC的开发的纤维蛋白凝胶显着改善运动功能和感觉恢复。苏木精和伊红(H&E)图像从横截面和,采集的再生神经纵切面显示,在纤维蛋白/胰岛素/hEnSC组中,再生神经纤维已经形成,并伴有新血管.
结论：我们的结果表明，制备的含有胰岛素纳米颗粒和hEnSC的水凝胶支架可以被认为是一种潜在的旨在再生坐骨神经的生物材料。
