Nishiwaki Hitoshi, Fujita Mitsugu, Yamauchi Makoto, Isogai Noritaka, Tabata Yasuhiko, Kusuhara Hirohisa
Department of Plastic and Reconstructive Surgery, Faculty of Medicine, Kindai University, Osaka-sayama, Japan.
Cells Tissues Organs. 2017;204(5-6):251-260. doi: 10.1159/000479790. Epub 2017 Oct 4.
Cartilage tissue is characterized by its poor regenerative properties, and the clinical performance of cartilage grafts to replace cartilage defects has been unsatisfactory. Recently, cartilage regeneration with mature chondrocytes and stem cells has been developed and applied in clinical settings. However, there are challenges with the use of mature chondrocytes and stem cells for tissue regeneration, including the high costs associated with the standard stem cell isolation methods and the decreased cell viability due to cell manipulation. Previous studies demonstrated that cartilage can be regenerated from chondrocyte clusters that contain stem cells. Based upon some of the existing techniques, the goal of this study was to develop a novel and practical method to induce cartilage regeneration. A microslicer device was developed to process cartilage tissues into micron-size cartilage (microcartilage) in a minimally invasive manner. We evaluated microcartilage sizes and demonstrated 100-400 µm as optimal for generating a high cell yield with collagenase digestion. In addition, autologous intrafascial implantation of the composites of microcartilage and an absorbable scaffold with a slow-release system of basic fibroblast growth factor (bFGF) was carried out to induce cartilage regeneration. Our results demonstrated that the extent of bFGF diffusion depends on the size of microcartilage, and that cartilage regeneration was induced most effectively with 100 µm of microcartilage via SOX5 upregulation. These findings suggest that cartilage regeneration is possible with microcartilage as a source of cells without ex vivo cell expansion.
软骨组织的特点是再生能力差，用于替代软骨缺损的软骨移植临床效果并不理想。近年来，利用成熟软骨细胞和干细胞进行软骨再生的技术已得到发展并应用于临床。然而，使用成熟软骨细胞和干细胞进行组织再生存在一些挑战，包括标准干细胞分离方法成本高以及细胞操作导致细胞活力下降。先前的研究表明，含有干细胞的软骨细胞簇可以再生软骨。基于一些现有技术，本研究的目标是开发一种新颖且实用的方法来诱导软骨再生。我们开发了一种切片机装置，以微创方式将软骨组织加工成微米级软骨（微软骨）。我们评估了微软骨的大小，并证明100 - 400 µm是通过胶原酶消化获得高细胞产量的最佳尺寸。此外，进行了微软骨与具有碱性成纤维细胞生长因子（bFGF）缓释系统的可吸收支架复合材料的自体筋膜内植入，以诱导软骨再生。我们的结果表明，bFGF的扩散程度取决于微软骨的大小，并且通过上调SOX5，100 µm的微软骨最有效地诱导了软骨再生。这些发现表明，以微软骨作为细胞来源无需体外细胞扩增即可实现软骨再生。
