RP技术制备PDLLA/TCP人工骨载体与LD-PTH成骨效应相容性的体外实验  

Biocompatibility of RP manufactured PDLLA/TCP tissue engineered bone scaffolds in low dose PTH-induced bone anabolism in vitro

作　　者：马兴[1] 胡蕴玉[1] 颜永年[2] 熊卓[2] 吕荣[1] 王军[1] 徐新智[1] 白建萍[1] 

机构地区：[1]第四军医大学西京医院全军骨科研究所,陕西西安710033 [2]清华大学机械系激光快速成形中心,北京100084

出　　处：《第四军医大学学报》2004年第12期1081-1084,共4页Journal of the Fourth Military Medical University

基　　金：军队"十五"重大课题 (0 1Z0 79) ;国家博士后科学基金项目(2 0 0 30 3352 3)

摘　　要：目的 :应用快速成形技术 (rapidprototyping ,RP)制备聚消旋乳酸 /磷酸三钙 (PDLLA/TCP)复合人工骨载体 ,探讨其在小剂量甲状旁腺激素 (LD PTH)成骨效应中的细胞生物相容性 ,研发仿生活性人工骨 .方法 :采用RP技术制备PDLLA/TCP(7∶3)复合人工骨载体 ,进行超微结构观察 ;制备PDLLA/TCP材料浸提液以及人骨髓基质细胞 (marrowstromalcells,MSCs)进行分组实验 .A组为常规培养对照组 ;B组为材料浸提液培养组 ;C组为LD PTH间歇用药 +常规培养组 :1 0nmol/LhPTH(1 34)× 6h +常规培养× 4 2h ;D组为LD PTH间歇用药 +浸提液培养组 :1 0nmol/LhPTH(1 34)× 6h +浸提液培养× 4 2h .各组均以 4 8h为 1个作用周期× 8周期 .应用细胞倍增 (NCD)、改良钙 钴法、LuciferYellow(LY)分子探针划痕标记荧光传输技术 (SLDT)及VanKossa钙化结节染色分别检测细胞增殖、活细胞间的间隙连接通讯功能 (GJIC)及体外矿化骨基质形成能力 .结果 :采用RP制备的PDLLA/TCP复合人工骨载体具有规则的空间网状支架结构、相互贯通的孔隙以及材料表面微孔特征 .该材料浸提液对MSCs增殖 (P >0 .0 5 )、LD PTH诱导下GJIC功能及体外矿化骨基质形成能力均无不利影响 .结论 :应用RP技术制备的PDLLA/TCP人工骨载体不仅具有良好的空间超微结构 ,AIM: To investigate the cell biocompatibility of tissue engineered bone scaffolds in low dose parathyroid hormone (LD PTH) induced bone formation activities in vitro . METHODS: Tridimensional PDLLA/TCP scaffolds (with component ratio of 7∶3) were synthesized by RP and observed by scanning electron microscopy. Human bone marrow stromal cells (MSCs) were primarily cultured and divided into four groups according to the different mode of exposure. The four groups were: Group A, control culture group; Group B: PDLLA/TCP immersed medium culture group; Group C: LD PTH [10 nmol/L hPTH (1-34)] intermittent exposure group; and Group D: LD PTH+PDLLA/TCP immersed medium culture group. A 48 h incubation cycle was repeated 8 times in 4 groups and the cell proliferation, gap junctional intercellular communication (GJIC) and bone like formation activities in vitro were detected by number of cell doubling (NCD), Lucifer Yellow (LY) scrape loading dye transfer (SLDT) and Von Kossa staining, respectively. RESULTS: The PDLLA/TCP scaffolds established by RP technology had good biomaterial properties such as satisfactory microstructure and porosities. The PDLLA/TCP immersed culture medium had no significant negative effects on cell growth ( P >0.05) and functional differentiation in the presence of LD PTH. CONCLUSION: PDLLA/TCP composite scaffolds established by RP have ideal microstructure, porosities and cell biocompatibilities in LD PTH stimulated bone formation activities.

关 键 词：载体 PTH 骨髓基质细胞 间隙连接蛋白 

分 类 号：Q813.11[生物学—生物工程] R684[医药卫生—骨科学]