Uysal A Cagri, Mizuno Hiroshi
Department of Plastic and Reconstructive Surgery, Faculty of Medicine, Baskent University, Ankara, Turkey.
Methods Mol Biol. 2011;702:443-51. doi: 10.1007/978-1-61737-960-4_32.
The goal of primary tendon repair is to increase tensile strength at the time of mobilization. Tendon repair and regeneration using mesenchymal stem cells have been described in several studies; however, the use of adipose derived stem cells (ASCs) for tendon repair has only recently been considered. In order to establish a suitable experimental model for the primary tendon repair using ASCs, this chapter describes the detailed methods for: (1) isolating stem cells from adipose tissue, (2) generation of a primary tendon injury and repair model, (3) evaluating functional restoration by measuring tensile strength, and (4) investigating the mechanisms involved in ASC-mediated tendon healing by histological and immunohistochemical analyses. Topical administration of ASCs to the site of injury accelerates tendon repair, as exhibited by a significant increase in tensile strength, direct differentiation of ASCs toward tenocytes and endothelial cells, and increases in angiogenic growth factors. These findings suggest that ASCs may have a positive effect on primary tendon repair and may be useful for future cell-based therapy.
一期肌腱修复的目标是在活动时提高拉伸强度。多项研究描述了使用间充质干细胞进行肌腱修复和再生;然而,脂肪来源干细胞(ASC)用于肌腱修复的应用只是最近才被考虑。为了建立一个使用ASC进行一期肌腱修复的合适实验模型,本章描述了以下详细方法:(1)从脂肪组织中分离干细胞,(2)建立一期肌腱损伤和修复模型,(3)通过测量拉伸强度评估功能恢复情况,以及(4)通过组织学和免疫组织化学分析研究ASC介导的肌腱愈合所涉及的机制。将ASC局部应用于损伤部位可加速肌腱修复,表现为拉伸强度显著增加、ASC直接向肌腱细胞和内皮细胞分化以及血管生成生长因子增加。这些发现表明,ASC可能对一期肌腱修复有积极作用,并且可能对未来基于细胞的治疗有用。