基于纤维丝组装的组织工程心肌体外构建关键工艺研究

发布时间：2018-03-29 10:07

本文选题：心肌组织工程　切入点：纤维束　出处：《清华大学》2012年硕士论文

【摘要】：本论文在分析天然心肌组成与结构特点的基础上，依照组织工程的仿生设计原则与“自下而上”组元化构建思路，提出了以含心肌细胞水凝胶纤维丝有序组装构建组织工程心肌纤维束的工艺路线。论文系统地研究了与组织工程心肌纤维束的设计、成形、应用相关的科学与技术问题，主要工作包括：从材料和结构两个方面进行了纤维束结构体的设计与分析；设计开发了水凝胶纤维丝的成形工艺和装置，完成了工艺原理分析和工艺参数研究；对构建的纤维束结构体，对其进行了力学性能的表征与评价；将工艺应用于环状组织工程心肌纤维束的构建，并进行初步的生物学评价。论文取得的主要结果包括：（1）模仿天然心肌组织的组成、结构及其内部微环境，设计了组织工程心肌纤维束，由含心肌细胞水凝胶纤维丝有序组装构建而成；纤维丝采用“海藻酸钠-纤维蛋白-Matrigel”仿生材料体系，平均直径为150~250μm，以满足氧在水凝胶材料中的扩散距离限制；纤维丝之间的间隙间接构成了微通道网络，通过FLUENT软件和理论计算对微通道灌流流场和供氧状态进行了分析与评价，得出了能在整个结构体内部营造富氧环境的有益结论；（2）设计开发了基于海藻酸钠的搅拌缠绕湿法成丝工艺，实现了纤维丝的连续稳定成形，并利用磁转子搅拌实现了纤维丝的缠绕有序收集，成功制备了纤细均匀的纤维丝和细胞复合的纤维丝，并以此为组装单元实现了多种纤维束结构体的构建；（3）深入研究了工艺参数对纤维丝成形质量的影响关系，根据实验结果确定了成形参数方案，同时对纤维丝组成的纤维束结构体的力学性能进行了研究，为满足后续的组织工程体外培养和训练的要求提供了依据；（4）完成了环状组织工程心肌纤维束的成形构建、体外培养和力学拉伸训练，对其进行了生物学评价，实验结果表明：纤维丝的材料体系具有良好的生物相容性；结合含钙培养环境和力学拉伸训练，纤维束结构能长时间稳定保持，局部的心肌细胞实现了定向排布与连接，形成了具有方向性的纤维状类组织形态。
[Abstract]:On the basis of analyzing the composition and structural characteristics of natural myocardium, according to the bionic design principle of tissue engineering and the idea of "bottom-up" component construction, In this paper, the process of constructing tissue engineered myocardial fiber bundle by orderly assembly of hydrogel fiber filament containing cardiomyocytes is presented. The scientific and technical problems related to the design, forming and application of tissue engineering myocardial fiber bundle are systematically studied in this paper. The main work includes: the design and analysis of fiber bundle structure from two aspects of material and structure, the design and development of forming process and equipment of hydrogel fiber filament, the process principle analysis and process parameter research; The mechanical properties of the fiber bundle were characterized and evaluated, and the process was applied to the construction of annular tissue engineering myocardial fiber bundle, and the preliminary biological evaluation was carried out. The main results obtained in this paper are as follows: (1) mimicking the composition, structure and internal microenvironment of natural myocardial tissue, a tissue engineered myocardial fiber bundle was designed and constructed from the ordered assembly of hydrogel filament containing cardiomyocytes. The fiber is made of "alginate-fibrin-Matrigel" biomimetic material system, with an average diameter of 150 ~ 250 渭 m, to satisfy the limit of oxygen diffusion distance in hydrogel materials, and the gap between fibers indirectly forms a microchannel network. The flow field and oxygen supply state of microchannel were analyzed and evaluated by FLUENT software and theoretical calculation. It is concluded that the oxygen-enriched environment can be created in the whole structure. (2) A wet spinning process based on sodium alginate is designed and developed to realize the continuous and stable forming of the filament. The winding and orderly collection of the filament was realized by magnetic rotor stirring, and the fine and uniform filament and the cell-composite filament were successfully prepared. The influence of process parameters on fiber forming quality is studied deeply, and the forming parameter scheme is determined according to the experimental results. At the same time, the mechanical properties of the fiber bundle structure were studied, which provided the basis for the in vitro culture and training of the subsequent tissue engineering, and completed the forming and construction of the annular tissue engineering myocardial fiber bundle. The biological evaluation of in vitro culture and mechanical stretching training was carried out. The experimental results showed that the material system of fiber had good biocompatibility, combined with calcium containing culture environment and mechanical stretching training, The fibrous bundle structure can be kept stable for a long time, and the local cardiac myocytes can be arranged and connected in a directional way, forming a directional fibrous tissue shape.
【学位授予单位】：清华大学
【学位级别】：硕士
【学位授予年份】：2012
【分类号】：R318.11

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