含缓释生物活性因子的组织工程骨异位成骨

Ectopic Osteogenesis of Tissue Engineered Bone with Slow-released Bioactive Factors

目的研究含重组人骨形态发生蛋白/转移生长因子-β(recombinanthumanbonemorphogeneticprotein/transforminggrowthfactor-β,rhBMP/TGF-β)和WO-1两种生物活性因子的缓释体构建组织工程骨的异位成骨能力。方法以猪部分脱钙骨为支架材料,用真空负压吸附法将rhBMP/TGF-β和WO-1分别涂层在支架材料上,然后再用聚乳酸(polylacticacid,PLA)涂层,制成缓释体。设计单纯材料组(A组),单纯材料复合成骨细胞(B组),含rhBMP/TGF-β为缓释体1(C组)、C组复合成骨细胞(D组)、含WO-1为缓释体2(E组)、E组复合成骨细胞(F组)6组,分别植入36只新西兰兔双后肢,每组6只。术后2、4、6、8周进行组织学、组织化学及生物化学检测,并进行统计学处理。结果在观察时限内,A组无成骨现象,B、D、F组成骨评分显著优于C、E组(P<0.05),C、E组的成骨活动明显延迟,D、F组成骨的质和量较B组提前2周,较C、E组提前4周。C、E组间差异无显著性。新增钙含量B、D、F组显著优于A、C、E组(P<0.05);A、C、E组钙含量递减幅度逐渐减少。结论含生物活性因子1、2的两种缓释体异位诱导成骨出现时间较复合成骨细胞后的成骨活性显著延迟,单纯材料无异位成骨能力。

To study the ectopic osteogenesis of tissue engineered bone with recombinant human bone morphogenetic protein/transforming growth factor-β(rhBMP /TGF-β)or WO-1slow-released factors.Methods Partial demineralized freeze-dried bone (PDFDB)of pig was used as scaffold material.rhBMP /TGF-βor WO-1were pre-coated on the surface of material by means of vacuum negative pressure absorption,and then coated with polylactic acid (PLA)to make slow-released material.There were six group:PDFDB material(group A);PDFDB combined with osteoblasts(group B);PDFDB material with rh BMP /TGF-βslow-released system(group C);PDFDB material combined with rhBMP /TGF-βslow-released system osteoblasts(group D);PDFDB with WO-1slow-released system(group E);PDFDB material combined with WO-1slow-released system and osteoblasts(group F)were implanted in bilateral lower limbs of36Newzealand rabbits respectively(6rabbits in each groups).Histological,histochemical and bio chemical analysis were detected2,4,6,8weeks after operation.Results Within the observation periods,no osteogenesis was observed in group A.The osteogenesis in group B,D,F were superior to that of group C and E(P<0.05).The osteogenetic activity in group C and E was delayed.The quantity and quality of osteogenesis in group D and F were2 weeks ahead of time compared with group B,and4weeks to that of group C and E.The newborn calcification content was superior to that of group A,C,and E(P<0.05).Conclusions The osteogenesis of PDFDB materials with BMP /TGF-βor WO-1is slower than that of which combined with osteoblasts.Simple material PDFDB has no ectopic osteogenesis.