【作者】 王秋玲；

【导师】 谢德明；

【作者基本信息】 暨南大学 ， 生物医学工程， 2011， 硕士

【摘要】 制备Ⅰ型胶原/纳米羟基磷灰石(nano-HAp)复合物,TEM、IR、XRD表征；盐滤沥法制备分别由PLGA/I型胶原/nano-HAp及PLGA/透明质酸钠组成的一体化双层骨-软骨支架,SEM观察支架表面形貌,MTT法测细胞毒性及与BMSCs复合培养检测该支架生物相容性。用静电纺丝的方法制备不同纳米HAp含量的PLGA/Hap复合电纺膜,分别接种大鼠BMSCs复合培养。14d和21d后,进行碱性磷酸酶、茜素红染色,观察BMSCs成骨分化情况。静电纺丝法制备PLGA/HAp/Zein复合纤维膜,FT-IR、DSC分别检测；采用多层沉积静电纺丝方法,制备HAP、透明质酸分别呈梯度分布的三维支架,植入兔膝关节骨-软骨缺损处,对支架自身的变化及缺损修复效果进行初步的研究。结果表明,一体化双层骨-软骨支架具有良好的微观结构,无细胞毒性,细胞与支架生物相容性良好,但细胞生长缓慢；不同HAp含量的PLGA能电纺成微米/纳米纤维,BMSCs能够良好地粘附、生长和增殖,碱性磷酸酶和茜素红染色表明BMSCs具有向骨细胞分化的趋势；PLGA/Hap/Zein能获得纳米/微米级电纺纤维。通过多层沉积方式,成功制备纳米梯度骨-软骨支架。植入兔骨软骨缺损4W后,该支架生物相容性良好,与宿主骨-软骨整合较好,支架与宿主界面有粘连并有少量的新生组织长入,支架本身暂无溶胀、收缩及分层等现象。更多还原

【Abstract】 Complex of collagen I and nano-HAp was prepared and characterized by TEM, IR and XRD, and the integrated bilayed osteochondral scaffold was manufactured by combining nano-HAp, collagen I, PLGA and sodium hyaluronate(HA) and PLGA with sodium leaching method. The surface topography, biocompatibility and cytoxicity of the scaffold were assessed by SEM, MTT assay when seeded with bone marrow stem cells (BMSCs).Complex of different concentration of nano-HAp and PLGA was prepared by electrospinning. With alkaline phosphatase and alizarin red staining after co-culturing with BMSCs and PLGA/HAp electrospun membrane for 14 and 21 days, the osteogenic differentiation of BMSCs and osteoblasts activity expression were observed.PLGA/HAp/Zein co-electrospun fibers were prepared and evaluated by FT-IR and DSC separately. With multi-layer deposit under electrospinning, PLGA/HAp/Zein and PLGA/HA electrospun to 3-D scaffolds, while the density of HAp and HA in gradient, and was transplanted into osteochondral defects of rabbits. The change of the scaffold and the repair status of defects were also evaluated preliminarily.The results indicated that the integrated bilayed osteochondral scaffold had good micro-structure and biocompatibility for rat BMSCs, BMSCs were able to adhere to the surface of the nano- or microfibrous PLGA/HAp electrospun scaffold with different HAp contents and good proliferation. The results of alkaline phosphatase and alizarin red staining indicated that BMSCs has the tendency of osteogenic differentiation. Nanofibrous gradient osteochondral scaffold was successfully prepared by multi-layed deposit electrospinning. After 4 weeks of scaffold transplation in rabbits articular cartilage in vivo, the results indicated that the scaffold had good biocompatibility, and integrated with osteochondral defects well, neo-tissue adhesion and growth were found in the interface of scaffold and osteochondral defects, as well, the swelling, shrink and delamination of implants were not found for all animals.更多还原