微构建论文：管状仿生血管支架制备及诱导VSMCs取向生长研究.docVIP

微构建论文：管状仿生血管支架的制备及诱导VSMCs取向生长研究 【中文摘要】在人工血管工程化中,种子细胞(VSMC、EC、Fibroblast)若不能形成其天然的三维微堆砌结构,就不能发挥正常的功能。除了使用有应力的生物反应器,制备管状仿生支架是诱导种植细胞产生天然微堆砌结构和功能的另一条重要途径。静电纺丝和熔融纺丝能直接、连续地制备聚合物纳米/微米纤维,配合缠绕接收技术,可制备具有周向微槽结构的仿生血管支架。本实验研究了不同电纺工艺(纺丝液浓度、电压、推进速率、接收距离、缠绕转速等)和不同熔纺工艺(熔纺温度、平移速度、缠绕转速等)对仿生管状支架微结构的影响。用静电纺丝结合熔融纺丝并利用溶剂/非溶剂粘结的方式制得了具有两层管壁、外壁具有周向排列微槽的管状仿生血管支架。上述支架用胶原蛋白固定改性后,在其上种植人脐带静脉平滑肌细胞(VSMCs)。用扫描电镜和荧光显微镜观察了纤维不同缠绕角度对VSMCs定向诱导能力的影响。结果表明,该仿生支架具有良好的生物相容性,其表面周向微槽结构对VSMCs的取向排列有明显的诱导作用。这对制备临床用血管替代物具有重要指导意义。在细胞体外评价的基础上,本项目还进行了兔子体内移植试验,以研究仿生支架原位诱导血管再生的可能性。术后1周、1月分别取出管状支架,发现其外层已与兔子自身组织紧密结合在一起(剥开发现支架外层呈粉红色),内腔则有厚约0.4 mm的血栓生成。组织学观察发现,支架外表面附着了周向排列的VSMCs和成纤维细胞。 【英文摘要】In vascular tissue engineering, if the vacular cells, e.g., smooth muscle cells (SMCs), could not regenerate their native 3D micro architectures or organization structures, their functions would disappear. Except for applying bioreactor introducing extra stress during vascular culture, fabricating biomimetic and tubular scaffold is another approach to induce the SMCs circumferencial alignment and elongation. Electrospinning and melt-spinning are two convenient ways that can produce nanoscale or microscale fibers continuously. Combining suitable receiving/wraping technology, tubular scaffold with circumferencial microchannel on outside and interconnected nano/micro pores within the wall would be prepared easily.In this study, the influence of the electrospinning parameters (solution concentration, voltage, the feed rate of syringe pump, the distance between syringe and collecting mandril, wraping speed) and the melt-spinning parameters (polymer melting temp., the right-and left removing and rotating speed of collecting mandril) on fibers抦orphology and weave structure were investigated. Novel biodegradable tubular scaffolds with two layers and circumferential nano/micro channels on outside surface and interconnected nano/micro pores within the walls were fabricated by electrospinning, melting spinning, and solvent/nonsolvent bonding tech