Ando Wataru, Tateishi Kosuke, Hart David A, Katakai Daisuke, Tanaka Yoshinari, Nakata Ken, Hashimoto Jun, Fujie Hiromichi, Shino Konsei, Yoshikawa Hideki, Nakamura Norimasa
Department of Orthopaedics, Osaka University Graduate School of Medicine, 2-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
Biomaterials. 2007 Dec;28(36):5462-70. doi: 10.1016/j.biomaterials.2007.08.030. Epub 2007 Sep 14.
The objective was to in vitro generate a mesenchymal stem cell (MSC)-based tissue-engineered construct (TEC) to facilitate in vivo repair in a porcine chondral defect model. Porcine synovial MSCs were cultured in monolayer at high density and were subsequently detached from the substratum. The cell/matrix complex spontaneously contracted to develop a basic TEC. Immunohistochemical analysis showed that the basic TEC contained collagen I and III, fibronectin, and vitronectin. The basic TEC exhibited stable adhesion to the surface of a porcine cartilage matrix in an explant culture system. The TEC cultured in chondrogenic media exhibited elevated expression of glycosaminoglycan and chondrogenic marker genes. The TEC were implanted in vivo into chondral defects in the medial femoral condyle of 4-month-old pigs, followed by sacrifice after 6 months. Implantation of a TEC into chondral defects initiated repair with a chondrogenic-like tissue, as well as secure biological integration to the adjacent cartilage. Histologically, the repair tissue stained positively with Safranin O and for collagen II. Biomechanical evaluation revealed that repair tissue exhibited mechanical properties similar to those of normal porcine cartilage in static compression and friction tests. This technology is a unique and promising method for stem cell-based cartilage repair.
目的是在体外生成一种基于间充质干细胞(MSC)的组织工程构建体(TEC),以促进猪软骨缺损模型的体内修复。将猪滑膜间充质干细胞高密度单层培养,随后从基质上脱离。细胞/基质复合物自发收缩以形成基本的TEC。免疫组织化学分析表明,基本的TEC含有I型和III型胶原蛋白、纤连蛋白和玻连蛋白。在器官培养系统中,基本的TEC对猪软骨基质表面表现出稳定的粘附。在软骨形成培养基中培养的TEC表现出糖胺聚糖和软骨形成标记基因表达升高。将TEC体内植入4月龄猪股骨内侧髁的软骨缺损处,6个月后处死。将TEC植入软骨缺损处可启动软骨样组织修复,并与相邻软骨实现牢固的生物整合。组织学上,修复组织对番红O和II型胶原蛋白染色呈阳性。生物力学评估显示,在静态压缩和摩擦试验中,修复组织表现出与正常猪软骨相似的力学性能。这项技术是一种独特且有前景的基于干细胞的软骨修复方法。