Osteopontin调控间充质干细胞移植预防硬膜外纤维化及其机制研究

Fibrosis post surgery/lesion is a hot area of global research, epidural fibrosis (EF) is a focus question in the area of spinal surgery. As previous reported, mesenchymal stem cells (MSCs) transplantation can inhibit scar hyperplasia and prevent keloid formation. However, the efficiency of MSCs transplantation needs improving. Based on our previous report, osteopotin(OPN) is significantly correlated with EF formation post laminectomy. Meanwhile, OPN was reported to regulate MSCs’ differentiation and migration. In the present project, in order to find out the optimal time for MSC transplantation, imaging scans and histological analysis were combined to study the expression tendency of OPN in EF formation process. Both isolation and culture of MSCs will be performed. The optimal transplantation route and cell dose will be decided based on the experiment in vivo. Lentivirus OPN shRNA will be created. With the combination of MSCs transplantation, OPN shRNA will be locally injected into EF modelling. The safety and efficacy of MSCs transplantation on preventing EF formation will be evaluated. Further research on studying the mechanism of OPN regulation MSCs transplantation efficiency will be performed. The results of the present project will give a hint on improving stem cell transplantation efficiency, and lay the translational foundation of MSCs transplantation on preventing EF.

术后/病变区域的纤维化预防是国内外研究热点，硬膜外纤维化（EF）已成为脊柱外科领域亟待解决的焦点问题。近期研究证实间充质干细胞（MSCs）移植可有效抑制肥厚瘢痕增生和预防瘢痕疙瘩形成，然而，MSCs移植效率有待进一步提高。申请人近期研究证明骨桥蛋白（OPN）表达与EF形成有密切联系。近期有研究表明OPN参与调控间充质干细胞（MSCs）分化与迁移。本项目将通过影像学扫描与组织学染色联合分析OPN在EF形成过程中表达趋势，确定MSCs移植的最佳时机。体外分离、培养大鼠MSCs，以动物在体实验确定MSCs的最佳移植途径和剂量。制备慢病毒OPN-shRNA，通过采用MSCs移植联合OPN-shRNA原位注射的方法，开展高效MSCs移植预防EF的研究，评估该方法安全性和疗效性，探索OPN调控MSCs移植效率的机制，提出BMSCs移植效率提升的新思路，为MSCs移植预防EF临床转化奠定动物实验基础。

术后/病变区域的纤维化预防是国内外研究热点，硬膜外纤维化（EF）已成为脊柱外科领域亟待解决的焦点问题。近期研究证实间充质干细胞（MSCs）移植可有效抑制肥厚瘢痕增生和预防瘢痕疙瘩形成，然而，MSCs移植效率有待进一步提高。申请人前期研究证明骨桥蛋白（OPN）表达与EF形成有密切联系。前期有研究表明OPN参与调控间充质干细胞（MSCs）分化与迁移。本项目通过影像学扫描与组织学染色联合分析OPN在EF形成过程中表达趋势，再次证实了OPN调控EF的形成，并伴随着EF的形成而表达上调。体外分离、培养大鼠MSCs，以动物在体实验确定MSCs预防EF的移植途径为原位移植，外周移植途径难以达到满意的MSCs聚集效果。移植剂量对比方面，4×106与5×106预防效果无显著差异，相较其它组别的剂量，效果具有显著差异。同时，在动物实验水平，OPN-shRNA体现出了更高效的EF预防效果，并且无明显不良反应，通过调控OPN联合MSCs移植预防EF具有较好的安全性和可行性，未来仍需进一步临床研究探索和证实。基于本项目的开展，项目申请人开展了一系列中国与俄罗斯的医学领域的科研合作，在干细胞、人工智能、肿瘤防控和转移癌预测等方面均取得了成果，并获批了国家自然科学基金中俄合作专项（82011530050），同时在天津市科学技术局的指导下，建立了天津市中俄恶性肿瘤骨转移癌联合研究中心。为未来进一步拓展中俄合作方医学领域的合作奠定了基础。