Development of a novel technology using mesenchymal stem cells from synovium for bone and cartilage regeneration.

关节软骨具有抗粘连特性，移植组织与邻近软骨基质的整合一直是软骨修复领域关注的问题。我们开发了一种新型的无支架三维合成组织(3DBT)，它由滑膜细胞及其合成的细胞外基质组成。3DST富含黏附分子，与软骨基质高度黏附，具有高效的软骨分化潜能。在本研究中，我们探讨了3DST修复部分厚度软骨损伤的可行性。猪滑膜细胞在抗坏血酸培养下形成了细胞和合成基质的单层硬质复合体。当从底物分离时，这种单层复合体主动收缩以形成3D组织。在4月龄猪股骨内侧髁处造成直径6.5 mm、深度2 mm的部分厚度缺损。3DSTs植入缺损处，无缝合。在对照组中，缺损处为空。动物被处死并进行组织学评估。3个月时，对照组未修复，而植入组骨缺损处已被修复的组织填充，与邻近软骨顺利结合。6个月后，修复组织保持良好的组织整合，藏红素O染色阳性。3DST与软骨基质的粘附性很强，因此与邻近组织的整合良好。3DST在体内也显示出很强的软骨分化潜能。此外，由于无支架结构，3DST最低限度地包含外部材料，如聚合物和动物材料。因此，与传统的基于支架的细胞疗法相比，3DST植入有望获得长期的安全性。综上所述，3DST技术可以为肌肉骨骼组织再生提供各种治疗干预。

Articular cartilage has anti-adhesive properties and the integration of implanted tissue to the adjacent cartilage matrix has been a concern in cartilage repair. We have developed a novel scaffold-free 3D synthetic tissue (3DBT) composed from synovial cells and their synthesized extracellular matrix. The 3DST is abundant in adhesion molecules, is highly adhesive to cartilage matrix, and has efficient chondrogenic differentiation potential. In the present study, we investigated the feasibility of the 3DST to the repair of partial-thickness chondral injury.Porcine synovial cells developed a stiff monolayer complex of the cells and the synthesized matrix when cultured with ascorbic acid. This monolayer complex actively contracts to develop a 3D-tissue when detached from the substratum. A 6.5mm diameter, 2mm deep partial-thickness defect was created at porcine medial femoral condyle of four-month-old pig. The 3DSTs were implanted in the defect without suture. In the control groups, the defect was left empty. The animals were sacrificed and evaluated histologically. At 3 months, the defect was not repaired in the control group, while the defect was filled with the repaired tissue with smooth integration to the adjacent cartilage in the implantation group. At 6 months, the repaired tissue maintained good tissue integration with positive Safranin O stainingThe 3DST is highly adhesive to the cartilage matrix and thus securely attains good integration to the adjacent tissue. The 3DST also exhibits strong chondrogenic differentiation potential in vivo. Furthermore, due to scaffold-free constitution, the 3DST minimally contains extrinsic materials such as polymers and animal materials. Thus, the 3DST implantation could potentially expect long-term safety than that of conventional scaffold-based cell therapies. Taken together, the 3DST technology could provide various therapeutic interventions for musculoskeletal tissue regeneration.