强力学仿生细胞外基质纳米纤维支架介导DCN shRNA长效转染ASCs的肌腱缺损修复研究

项目名称： 强力学仿生细胞外基质纳米纤维支架介导DCN shRNA长效转染ASCs的肌腱缺损修复研究

项目编号： No.81201408

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学四处

项目作者： 银国利

作者单位： 浙江医学高等专科学校

项目金额： 23万元

中文摘要： 肌腱缺损是临床常见疾病，组织工程肌腱已成为临床肌腱缺损修复的理想选择。但目前组织工程肌腱尚存在以下不足：种子细胞来源受限、缺乏降解速率与细胞增殖相匹配的强力学生物支架、修复后肌腱因缺少大直径胶原纤维而导致力学性能低下等根本性问题。为此，我们拟将核心蛋白聚糖（Decorin）shRNA质粒包裹于壳聚糖纳米微囊中，以高压静电纺丝技术混纺壳聚糖纳米微囊-丝素蛋白制备强力学三维纳米纤维支架，并将脂肪干细胞（ASCs）种植于该支架，在持续拉伸力刺激下构建组织工程肌腱；然后将组织工程肌腱植入肌腱缺损动物模型进行肌腱缺损修复，以期在肌腱再生修复过程中，该支架不仅能提供相匹配的力学强度，为ASCs向肌腱细胞分化提供仿生细胞外基质的生长微环境，而且随着支架材料逐步降解，释放出的壳聚糖纳米微囊携带DCN shRNA进入新生肌腱细胞，抑制decorin表达，促进大直径胶原纤维形成，从而实现肌腱的功能性再生修复。

中文关键词： 肌腱组织工程；韧带组织工程；丝素蛋白；壳聚糖；无疤痕

英文摘要： Tendon defect is a common clinical disease. And tissue engineered tendon has become the ideal choice for the regeneration and repair of the damaged tendon. However, tendon tissue engineering still has many problems to be solved: limited source of seed cells, lack of scaffolds whose degradation rate matching the cell proliferation, healed tendon with the absence of large diameter collagen fibers and low mechanical properties, etc. In this project, we intent to solve these problems by the following researches: preparation of chitosan nanocapsules with the plasmid vector of Decorin shRNA, preparation of three dimensional fibrous scaffolds with the chitosan nanocapsules and silk fibroin by electrospinning, and construction of tissue engineered tendon by planting the adiopse stem cells on the scaffold and stimulating under sustained tensile force, regeneration and repair of tendon defects by transplantation of tissue engineered tendon into the rabbit Achilles tendon defect. Through these researches, we hope this three dimensional fibrous scaffold could not only provide enough mechanical strength during the tendon repairing, but also provide a extracellular matrix-like microenvironment to ASCs. And with the degradation of the scaffold, chitosan nanocapsules with Decorin shRNA vector will be released and transected int

英文关键词： Tendon tissue engineering；Ligament tissue engineering；Silk fibroin；Chitosan；Scar-free