肌腱干细胞定性及其与支架的相互作用研究

发布时间：2018-04-04 05:16

本文选题：肌腱组织工程　切入点：肌腱分化　出处：《浙江大学》2013年博士论文

【摘要】：肌腱损伤是一种非常常见的运动损伤。目前,临床上对肌腱损伤的治疗主要停留在理疗、手术缝合以及自体或异体移植阶段,虽有一定效果,但是疗效有限,即使是自体肌腱移植修复也只能达到正常肌腱力学性能的40%左右,且伴随有大量疤痕组织增生。这主要是因为成体肌腱不具备完全再生能力,所以修复后肌腱的质量远不如正常肌腱,易出现肌腱粘连,肌腱结构和力学性能低下,常重复断裂。因此,寻找新型的、可促进肌腱生理性再生修复的方法具有极其重要的临床意义。近年来,通过利用干细胞复合支架材料的组织工程手段,促使身体自主再生已损伤组织,为提高肌腱等软组织的修复质量带来了全新的希望。肌腱组织工程技术临床转化前需迫切解决的两个关键问题：1)获得足够量具有腱系分化能力的种子细胞；2)具有诱导腱系分化性能的生物支架材料。所以,本论文针对这两个问题,立足于本研究室前期的干细胞与生物支架相关研究成果,进行了成系列的研究,最终目的为运用肌腱干细胞与生物支架构建组织工程肌腱和促进肌腱损伤的再生。本论文研究内容包含：第一部分研究,分离培养了人成体和胚胎来源的肌腱干细胞,并证实其均具有克隆形成能力和多向分化潜能等干细胞特性；第二部分研究进一步探索巢蛋白标志的肌腱干／祖细胞亚群特性及其巢蛋白对肌腱干细胞行为调控的功能,发现巢蛋白对于肌腱干细胞腱系分化成熟和存活能力有至关重要的作用,尤其是参与了胶原纤维的聚合；第三部分研究肌腱干细胞与支架的相互作用,仿造肌腱组织纳米级平行细胞外基质设计出平行纳米纤维支架,评估平行纳米拓扑结构对肌腱干细胞分化的影响及其应用于体内修复的效果,发现平行纳米材料会激发肌腱干细胞向腱系分化,并适合作为肌腱组织工程的支架材料；第四部分在前三部分的基础上进一步研究不同来源的具有不同微结构的脱细胞基质对于肌腱干细胞分化的影响,选择最佳微结构的脱细胞基质复合肌腱干细胞,构建组织工程肌腱,通过动物模型评估其对肌腱损伤修复的再生作用,证实肌腱来源的脱细胞基质是肌腱组织工程的理想支架材料,并与干细胞联合会获得更好的修复效果。本研究的成果和发现为利用干细胞和生物材料修复再生肌腱提供了新的理论依据和新技术手段,为未来肌腱损伤治疗带来新的种子细胞和支架。
[Abstract]:Tendon injury is a common sports injury. At present, the clinical treatment of tendon injury mainly in physical therapy, surgical suture and autologous or allogeneic transplantation, has a certain effect, but the effect is limited, even about autologous tendon graft repair also can only achieve the performance of normal tendon mechanics 40%, with a large number of the scar tissue hyperplasia. This is mainly because the body does not have full tendon regeneration ability, so after the repair of tendon quality than normal tendon, prone to tendon adhesion, low tendon structure and mechanical properties, often repeated fracture. Therefore, the search for new, but it has important clinical significance to promote tendon regeneration methods.
In recent years, by means of the use of stem cells in tissue engineering scaffolds, prompting the body self regeneration has damaged tissue, has brought new hope to improve the quality of repair tendon and soft tissue. The tendon tissue engineering clinical transformation to be solved two key issues: 1) to acquire sufficient seed cells with tendon. Differentiation; 2) with scaffold materials for tendon induced differentiation properties. So, this paper focuses on these two problems, based on the previous research of stem cells and scaffold related research results, the series of research, the ultimate goal is to use the tendon stem cells and biological scaffold for tissue engineering tendon and promote the regeneration of tendon injury.
The content of this paper includes: the first part of the study, the in vitro culture of adult human embryonic stem cells and tendon, and confirmed that they have clonogenic cell characteristics and multipotency stem; the second part of the study to further explore the nestin markers of tendon stem / progenitor cell subsets and its characteristics of nestin on tendon stem cell behavior the regulation function of stem cells for tendon tendon found nestin differentiation maturation and survival ability plays an important role, especially in the polymerization of collagen fibers; the third part of the study of tendon stem cells and support the interaction of counterfeit tendon tissue nano parallel extracellular matrix designed parallel nanofiber scaffold, evaluation of parallel nano topology the structure effect on the stem cell differentiation and its application in tendon repair effect, found that parallel nano materials can stimulate tendon stem cells Cell differentiation to tendon, and suitable as scaffold materials for tendon tissue engineering; the fourth part further study with different sources on the basis of the three parts of cells from different micro structure matrix for cell differentiation of tendon stem, choose the best microstructure of acellular matrix composite tendon stem cells to construct tissue engineered tendon and through animal model to evaluate its role in the regeneration of tendon repair, confirmed acellular tendon derived matrix is an ideal scaffold materials for tendon tissue engineering, and the Federation of stem cells and obtain better repair effect. The results of this study and the findings provide a new theoretical basis and new techniques for using stem regeneration of tendon cells and biological materials for the future treatment of tendon injuries brought new seed cells and scaffold.

【学位授予单位】：浙江大学
【学位级别】：博士
【学位授予年份】：2013
【分类号】：R873

【共引文献】