β-磷酸三钙（β-TCP）基多孔骨修复体的功能化构建及研究
发布时间：2018-08-25 17:03
【摘要】：由于炎症、外伤与肿瘤等引起的骨缺损是临床常见的疾病。传统的自体/异体骨移植由于来源短缺等原因,在临床的应用受到很大限制。随着材料科学的不断进步和发展,人工合成骨修复体成为解决临床使用骨替代物的最佳方法。在人工合成骨修复体的材料中,β-磷酸三钙(β-TCP)由于与人体骨骼相似无机成份和可生物降解性成为较为理想的材料。在结构方面,骨修复要求修复体材料具有多孔的结构便于细胞和血管的长入。为进一步提高β-TCP修复体材料的成骨诱导能力,促进骨组织再生修复,本研究基于3D打印制备的β-TCP多孔修复体,采用物理包被、负载和细胞工程技术方法对修复体进行功能化改性。主要研究内容和结果如下:(一)β-TCP多孔骨修复体的制备及生物安全性评价利用3D打印技术构建孔结构精确可控的β-TCP多孔修复体,对其进行体内和体外的生物安全性评价。研究发现,在体外,β-TCP多孔修复体有利于骨髓间充质干细胞(mBMSCs)的粘附与增殖;在体内,β-TCP多孔修复体生物相容性好,不刺激结缔组织增生,降解产物不引起主要脏器的毒性反应,具有良好的生物相容性和生物安全性。(二)Ⅰ型胶原纳米纤维功能化β-TCP多孔修复体通过物理包被的方法在β-TCP多孔修复体表面构建重组Ⅰ型胶原纳米纤维。研究发现β-TCP多孔修复体表面的Ⅰ型胶原纳米纤维能促进mBMSCs的增殖与成骨分化,同时,刺激mBMSCs基质小泡的分泌,促进基质矿化。(三)Ⅰ型胶原纳米纤维负载生长因子PIGF功能化β-TCP多孔修复体采用Ⅰ型胶原纳米纤维负载生长因子PIGF修饰β-TCP多孔修复体表面,实现PIGF的有效缓释。研究发现PIGF的负载能促进mBMSC细胞增值及成血管相关生长因子的分泌,更重要的是,PIGF还能刺激mBMSC细胞成骨相关基因的表达,诱导其成骨分化。因此,该修复体具有同时促成血管及促成骨的双重诱导调控作用。(四)转基因pigf-tk-CHO细胞功能化β-TCP多孔修复体构建基于四环素诱导表达系统(Tet-on induce expression system)的pLVX-Tet-On-Tight-myc-PIGF-PGK-Puro质粒和基于更昔洛韦诱导自杀系统(GCV/HSV-tk suicide gene system)的pIRES2-ZsGreen1-TK质粒,同时转染到CHO细胞中。经过转基因CHO修饰的β-TCP多孔修复体,可实现诱导表达PIGF生长因子和细胞凋亡。这种“智能双安全调控表达系统”骨修复体提高了转基因治疗骨修复的生物安全性和有效性,是一种理想的基于基因工程对骨修复体功能化方法。
[Abstract]:Bone defect caused by inflammation, trauma and tumor is a common clinical disease. Traditional autologous / allogeneic bone transplantation is limited in clinical application due to shortage of sources. With the progress and development of material science, artificial bone prosthesis has become the best method to solve the clinical use of bone substitutes. Among the synthetic bone restoration materials, 尾 -TCP (尾 -TCP) is an ideal material due to its inorganic composition and biodegradability similar to human bone. In terms of structure, bone repair requires porous structures to facilitate the growth of cells and blood vessels. In order to further improve the osteogenic induction ability of 尾 -TCP restorative material and promote the regeneration of bone tissue, a 尾 -TCP porous prosthesis based on 3D printing was prepared with a physical coating. The functional modification of the prosthesis was carried out by loading and cell engineering techniques. The main contents and results are as follows: (1) preparation and biosafety evaluation of 尾 -TCP porous bone prosthesis. 3D printing technique was used to construct 尾 -TCP porous prosthesis with precise and controllable pore structure and to evaluate its biosafety in vivo and in vitro. It was found that 尾 -TCP porous repair was beneficial to the adhesion and proliferation of bone marrow mesenchymal stem cells (mBMSCs) in vitro, and that 尾 -TCP porous repair had good biocompatibility, did not stimulate connective tissue proliferation, and the degradation products did not cause toxic reactions in main organs. It has good biocompatibility and safety. (2) Recombinant type 鈪，
本文编号：2203540
[Abstract]:Bone defect caused by inflammation, trauma and tumor is a common clinical disease. Traditional autologous / allogeneic bone transplantation is limited in clinical application due to shortage of sources. With the progress and development of material science, artificial bone prosthesis has become the best method to solve the clinical use of bone substitutes. Among the synthetic bone restoration materials, 尾 -TCP (尾 -TCP) is an ideal material due to its inorganic composition and biodegradability similar to human bone. In terms of structure, bone repair requires porous structures to facilitate the growth of cells and blood vessels. In order to further improve the osteogenic induction ability of 尾 -TCP restorative material and promote the regeneration of bone tissue, a 尾 -TCP porous prosthesis based on 3D printing was prepared with a physical coating. The functional modification of the prosthesis was carried out by loading and cell engineering techniques. The main contents and results are as follows: (1) preparation and biosafety evaluation of 尾 -TCP porous bone prosthesis. 3D printing technique was used to construct 尾 -TCP porous prosthesis with precise and controllable pore structure and to evaluate its biosafety in vivo and in vitro. It was found that 尾 -TCP porous repair was beneficial to the adhesion and proliferation of bone marrow mesenchymal stem cells (mBMSCs) in vitro, and that 尾 -TCP porous repair had good biocompatibility, did not stimulate connective tissue proliferation, and the degradation products did not cause toxic reactions in main organs. It has good biocompatibility and safety. (2) Recombinant type 鈪，
本文编号：2203540
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