组织工程半月板的支架仿生设计、生长进程调控和功能构筑研究

Due to inferior potential of mini-invisive accessibility and autotransplantation of seed cells, defective structure and configuration of scaffolds, the current tissue engineering meniscus (TEM) own inferior mechanical strenth, defective regulation of tissue formation, poor structure and function. Thus it could not meet the clinical needs. In our study, Polylactone scaffold with highly bionic mechanical property and structure, double network hydrogel with ultra high strength are prepared, and then combined both the former features to fabricate the biophysic scaffold. Meanwhile, peripheral blood mesenchymal stem cells (PB-MSCs) are actived, acquired mini-invisively and seperated as seed cells. In vitro, with regulation of 3D space, surface modification, growth factor controlled by microcapsules in different time and zone and biomechanical stimulation, PB-MSCs are cultured with the above scaffolds to construct the TEM. Based on the gross observation, histology, biomechanics and gene/proteomics chip test, the greatest TEM protocol is explored. The above suprerior tissue engineered meniscus then are transplanted partial or total for menicus repair and reconstruction. Besides, the new TEM might be explored for its mechanism in micro-environment, gene and protein exprssion. This study would provide promising stratagem for clinal treatment of both partial and total meniscectomy.

现有组织工程半月板（TEM）方案，因支架材料和构型缺陷，种子细胞微创获得和自体移植潜能不足和生长调控不合理，最终构建的TEM的力学强度、组织结构和功能均满足不了临床需求。本研究拟制备力学强度和三维结构均高度仿真的聚内酯支架，并制备双重网络增强型超高强度水凝胶支架，以及聚内酯/水凝胶双相支架。用动员后可微创抽血分离、有自体移植潜能的外周血间充质干细胞为种子细胞，研究多种TEM构建方案。用体外试验观察支架的三维空间、材料界面、生长因子区域差异、生长因子时间差异和应力刺激对多种TEM方案生长进程的调控效果。通过大体观察、组织学、生物力学、影像学、基因芯片和蛋白芯片评估，筛选出最优体外TEM方案。在体内验证中，分别行半月板部分切除的TEM修复和半月板全切除后的TEM移植替代，并对体外和体内过程中功能基因和功能蛋白的参与机制进行研究，为临床治疗半月板部分切除和全切除提供新的方案。

现有组织工程半月板（TEM）方案，因支架材料和构型缺陷，种子细胞微创获得和自体移植潜能不足和生长调控不合理，最终构建的TEM的力学强度、组织结构和功能均满足不了临床需求。本研究拟制备力学强度和三维结构均高度仿真的聚内酯支架，并制备双重网络增强型超高强度水凝胶支架，以及聚内酯/水凝胶双相支架。用动员后可微创抽血分离、有自体移植潜能的外周血间充质干细胞为种子细胞，研究多种TEM构建方案。用体外试验观察支架的三维空间、材料界面、生长因子区域差异、生长因子时间差异和应力刺激对多种TEM方案生长进程的调控效果。通过大体观察、组织学、生物力学、影像学、基因芯片和蛋白芯片评估，筛选出最优体外TEM方案。在体内验证中，分别行半月板部分切除的TEM修复和半月板全切除后的TEM移植替代，并对体外和体内过程中功能基因和功能蛋白的参与机制进行研究，为临床治疗半月板部分切除和全切除提供新的方案。. 我们全面完成了原项目申请书中的研究工作，分离、鉴定了多种PB-MSCs，精准获取了新西兰兔和比格犬半月板的相关参数并进行三维重建和3D打印，筛选制备了多种组织工程半月板构建方案，初步阐明了支架三维空间结构、材料界面、生化刺激及应力刺激对构建与天然半月板相似的异质性TEM的影响，同时，我们还自制了微型卡具和等距垂直切割组织的手术刀，以精准获得兔半月板不同区域的力学性能。另外，我们制备并改善了具有超声响应释放微球特性的可注射水凝胶，以用于TEM的异质性构建。