聚己内酯和β-磷酸三钙复合支架经血小板衍生生长因子BB修饰后的促血管生成作用  

Promoting angiogenesis of platelet-derived growth factor BB modified polycaprolactone and beta-tricalcium phosphate composite scaffolds

作　　者：赵金龙[1] 刘继超[1] 于洋[1] Zhao Jinlong;Liu Jichao;Yu Yang(Department of Orthopedics,The 3201 Hospital Affiliated to Xi’an Jiaotong University School of Medicine,Hanzhong 723000,Shaanxi Province,China)

机构地区：[1]西安交通大学医学院附属三二〇一医院骨科,陕西省汉中市723000

出　　处：《中国组织工程研究》2023年第21期3300-3306,共7页Chinese Journal of Tissue Engineering Research

摘　　要：背景:组织工程技术的发展为骨缺损的修复重建提供了新的思路,但是血管化问题使组织工程骨应用于临床受到了制约。血小板衍生生长因子在促进骨细胞形成的同时也可促进骨组织中血管的形成。目的:观察聚己内酯/β-磷酸三钙/血小板衍生生长因子BB支架对骨髓间充质干细胞成骨分化及人脐静脉内皮细胞增殖、黏附的影响,以及修复骨缺损的效果。方法:①利用3D快速成形机制备聚己内酯/β-磷酸三钙支架(记为PCL/β-TCP支架),将支架浸渍于血小板衍生生长因子BB溶液中制备聚己内酯/β-磷酸三钙/血小板衍生生长因子BB支架(记为PCL/β-TCP/PDGF BB支架)。②体外实验:将骨髓间充质干细胞、人脐静脉内皮细胞分别接种于两种支架上,检测骨髓间充质干细胞的成骨分化情况,检测人脐静脉内皮细胞的增殖、黏附与成血管基因表达。③体内实验:取21只成年大鼠,建立双侧胫骨缺损模型,实验组植入PCL/β-TCP/PDGF BB支架,对照组植入PCL/β-TCP支架,空白组不植入支架,每组7只。术后12周,进行Micro-CT扫描、骨组织形态学观察及成骨与成血管基因检测。结果与结论:①体外实验:与PCL/β-TCP支架相比,PCL/β-TCP/PDGF BB支架可促进骨髓间充干细胞的成骨分化,促进人脐静脉内皮细胞的增殖与黏附,促进人脐静脉内皮细胞血管内皮生长因子、CD31 mRNA的表达。②体内实验:Micro-CT扫描显示,空白组可见明显的骨缺损,对照组、实验组均可见大量新生的骨组织,实验组修复效果更明显。苏木精-伊红染色、Masson和番红固绿染色显示,空白组没有明显骨组织形成;对照组可见大量成熟度较高的骨基质及相对较少的不成熟软骨组织;实验组可见大量的骨样组织及成熟度较高的软骨组织,大部分髓腔再通。RT-PCR检测显示,实验组骨形态发生蛋白2、碱性成纤维细胞生长因子、碱性磷酸酶、骨钙素、血管内皮生长�BACKGROUND:The development of tissue engineering technology provides new ideas for the repair and reconstruction of bone defects,but the problem of vascularization restricts the clinical application of tissue engineered bone.Platelet-derived growth factor can promote the formation of blood vessels in bone tissue while promoting the formation of bone cells.OBJECTIVE:To observe the effects of polycaprolactone/β-tricalcium phosphate/platelet-derived growth factor BB(PCL/β-TCP/PDGF BB)scaffold on osteogenic differentiation of bone marrow mesenchymal stem cells and proliferation and adhesion of human umbilical vein endothelial cells in vitro,as well as the effect of its application in animal bone defects.METHODS:(1)Polycaprolactone/β-tricalcium phosphate scaffolds were prepared by 3D rapid prototyping machine,and the scaffolds were immersed in platelet-derived growth factor BB solution to prepare PCL/β-TCP/PDGF BB scaffold.(2)In vitro experiment:Bone marrow mesenchymal stem cells and human umbilical vein endothelial cells were inoculated on two kinds of scaffolds,respectively,to detect the osteogenic differentiation of bone marrow mesenchymal stem cells,and to detect the proliferation,adhesion and angiogenic gene expression of human umbilical vein endothelial cells.(3)In vivo experiment:21 adult rats were taken to establish a bilateral tibial defect model.The experimental group(n=7)was implanted with PCL/β-TCP/PDGF BB scaffolds;the control group(n=7)was implanted with PCL/β-TCP scaffolds;the blank group(n=7)was not implanted with scaffolds.At 12 weeks after operation,micro-CT scanning,bone histomorphological observation,and gene detection of osteogenesis and angiogenesis were performed.RESULTS AND CONCLUSION:(1)In vitro experiment:Compared with PCL/β-TCP scaffold,PCL/β-TCP/PDGF BB scaffold could promote the osteogenic differentiation of bone marrow mesenchymal stem cells,promote the proliferation and adhesion of human umbilical vein endothelial cells,and promote vascular endothelial growth factor and CD31 mRNA

关 键 词：骨缺损 聚己内酯 Β-磷酸三钙 血小板衍生生长因子(PDGF)BB 血管形成 组织工程 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R683