Mitani Genya, Sato Masato, Lee Jeong I K, Kaneshiro Nagatoshi, Ishihara Miya, Ota Naoshi, Kokubo Mami, Sakai Hideaki, Kikuchi Tetsutaro, Mochida Joji
Department of Orthopaedic Surgery, Surgical Science, Tokai University School of Medicine, Kanagawa, Japan.
BMC Biotechnol. 2009 Mar 6;9:17. doi: 10.1186/1472-6750-9-17.
Although the clinical results of autologous chondrocyte implantation for articular cartilage defects have recently improved as a result of advanced techniques based on tissue engineering procedures, problems with cell handling and scaffold imperfections remain to be solved. A new cell-sheet technique has been developed, and is potentially able to overcome these obstacles. Chondrocyte sheets applicable to cartilage regeneration can be prepared with this cell-sheet technique using temperature-responsive culture dishes. However, for clinical application, it is necessary to evaluate the characteristics of the cells in these sheets and to identify their similarities to naive cartilage.
The expression of SOX 9, collagen type 2, 27, integrin alpha 10, and fibronectin genes in triple-layered chondrocyte sheets was significantly increased in comparison to those in conventional monolayer culture and in a single chondrocyte sheet, implying a nature similar to ordinary cartilage. In addition, immunohistochemistry demonstrated that collagen type II, fibronectin, and integrin alpha 10 were present in the triple-layered chondrocyte sheets.
The results of this study indicate that these chondrocyte sheets with a consistent cartilaginous phenotype and adhesive properties may lead to a new strategy for cartilage regeneration.
尽管近年来基于组织工程程序的先进技术使自体软骨细胞植入治疗关节软骨缺损的临床效果有所改善,但细胞处理问题和支架缺陷仍有待解决。一种新的细胞片技术已经开发出来,并且有可能克服这些障碍。使用温度响应培养皿通过这种细胞片技术可以制备适用于软骨再生的软骨细胞片。然而,对于临床应用,有必要评估这些细胞片中细胞的特性,并确定它们与天然软骨的相似性。
与传统单层培养和单个软骨细胞片中的细胞相比,三层软骨细胞片中SOX 9、Ⅱ型胶原蛋白、整合素α10和纤连蛋白基因的表达显著增加,这意味着其性质与普通软骨相似。此外,免疫组织化学显示三层软骨细胞片中存在Ⅱ型胶原蛋白、纤连蛋白和整合素α10。
本研究结果表明,这些具有一致软骨表型和黏附特性的软骨细胞片可能会为软骨再生带来新策略。