Department of Pediatric Surgery, Second Affiliated Hospital of Harbin Medical University, Harbin, China.
Department of Pediatric Surgery, Second Affiliated Hospital of Harbin Medical University, Harbin, China.
Cytotherapy. 2014 May;16(5):662-73. doi: 10.1016/j.jcyt.2013.07.014. Epub 2013 Oct 3.
After injury, tendons often heal with poor tissue quality and inferior mechanical properties. Tissue engineering using tendon stem cells (TSCs) is a promising approach in the repair of injured tendon. Tenogenic differentiation of TSCs needs an appropriate environment. More recently, the acellular extracellular matrix (ECM) generated from fibroblasts has been used to construct various engineering tissues. In this study, we successfully developed an engineered tendon tissue formed by seeding TSCs in de-cellularized fibroblast-derived matrix (dFM).
Patellar TSCs and dermal fibroblast were isolated and cultured. Using the method of osmotic shock, dFM was obtained from dermal fibroblast. ECM proteins in dFM were examined. TSCs at passage 3 were seeded in dFM for 1 week. Proliferative capacity and characterization of TSCs cultured in dFM were determined by population doubling time, immunofluorescence staining and quantitative reverse transcriptase polymerase chain reaction. Engineered tendon tissue was prepared with dFM and TSCs. Its potentials for neo-tendon formation and promoting tendon healing were investigated.
dFM is suitable for growth and tenogenic differentiation of TSCs in vitro. Neo-tendon tissue was formed with tendon-specific protein expression when TSCs were implanted together with dFM. In a patellar tendon injury model, implantation of engineered tendon tissue significantly improved the histologic and mechanical properties of injured tendon.
The findings obtained from our study provide a basis for potential use of engineered tendon tissue containing dFM and TSCs in tendon repair and regeneration.
受伤后，肌腱常常愈合组织质量差，机械性能差。使用肌腱干细胞（TSCs）的组织工程是修复受损肌腱的一种很有前途的方法。TSCs 的腱向分化需要一个合适的环境。最近，从成纤维细胞中获得的无细胞细胞外基质（ECM）已被用于构建各种工程组织。在这项研究中，我们成功地开发了一种由种子 TSCs 接种在脱细胞化成纤维细胞衍生基质（dFM）中形成的工程化肌腱组织。
分离和培养髌腱 TSCs 和真皮成纤维细胞。使用渗透压休克的方法从真皮成纤维细胞中获得 dFM。检查 dFM 中的 ECM 蛋白。将第 3 代 TSCs 接种于 dFM 中培养 1 周。通过细胞倍增时间、免疫荧光染色和实时定量聚合酶链反应测定 TSCs 在 dFM 中的增殖能力和特征。用 dFM 和 TSCs 制备工程化肌腱组织。研究其促进新腱形成和促进腱愈合的潜力。
dFM 适合 TSCs 的体外生长和腱向分化。当 TSCs 与 dFM 一起植入时，形成了具有腱特异性蛋白表达的新腱组织。在髌腱损伤模型中，植入工程化肌腱组织显著改善了损伤肌腱的组织学和机械性能。
本研究结果为含有 dFM 和 TSCs 的工程化肌腱组织在肌腱修复和再生中的潜在应用提供了依据。
