目的 研究海螵蛸-骨形态发生蛋白(BMP)复合材料对骨缺损大鼠成骨及再血管化的影响. 方法 将海螵蛸去外壳,制成圆柱体(直径约5 mm,高约2mm),浸泡于重组人骨形态发生蛋白2溶液中制作海螵蛸-BMP复合材料.将30只SD大鼠制成颅骨缺损模型,按照随机数字表法分成海螵蛸-BMP组与单纯海螵蛸组,每组15只,分别植入相应材料修复颅骨缺损.移植术后4、6、8周,2组各取5只大鼠,分批放血处死同时行墨汁灌注.ld后取出全部植入物和周围部分颅骨同时进行植入区大体观察,制作切片行HE及Masson染色,通过组织病理学技术、图像采集系统,观察、分析微血管面积和新生骨面积.对数据行两因素析因设计方差分析和LSD检验,并对海螵蛸-BMP组大鼠微血管面积和新生骨面积的相关性行Pearson相关分析. 结果 (1)植入区大体观察可见植入物被纤维结缔组织包膜包裹,移植术后4周包膜质地柔软、较厚;移植术后6、8周包膜质韧、薄,但较致密.植入物移植术后4周时转为胶冻样物质,移植术后6、8周时与软骨组织类似.(2)组织学观察显示,移植术后4周,2组大鼠植入物结构均遭到破坏,并可见中等数量的炎性细胞浸润;海螵蛸-BMP组初级骨小梁、微血管数量明显多于单纯海螵蛸组,破骨细胞数量则少于单纯海螵蛸组.移植术后6周,2组大鼠植入物炎性细胞明显减少,海螵蛸进一步降解,新生微血管更加丰富,植入物周边均形成明显的成熟骨小梁,但海螵蛸-BMP组在降解后的海螵蛸-BMP中也出现了成熟骨小梁.移植术后8周,2组大鼠植入物炎症反应消失,成熟骨小梁显著增多,海螵蛸的网格状结构基本被破坏,单纯海螵蛸组的成熟骨小梁仍分布在海螵蛸周边;而海螵蛸-BMP组中,海螵蛸-BMP中央、周围均可见成熟骨小梁,微血管数量仍多于单纯海螵蛸组.(3)移植术后4~8周,海螵蛸-BMP组微血管面积[(63±4)、(136 ±36)、(347±31) μm2]大于单纯海螵蛸组[(44±7)、(73±4)、(268±42) μm2,P<0.05或P<0.01];新生骨面积[(236±26)、(339 ±42) 、(553 ±40)μm2]大于单纯海螵蛸组[(137±15)、(243 ±21)、(445±29) μm2,P值均小于0.01].(4)海螵蛸-BMP组微血管面积与新生骨面积呈显著正相关(r =0.948,P=0.00l) 结论 海螵蛸-
Objective To study the effects of cuttlefish bone-bone morphogenetic protein (BMP)composite material on osteogenesis and revascularization of bone defect in rats.Methods The cuttlefish bone was formed into cylinder with the dianeter of about 5 mm and height of about 2 mm after the shell was removed,and then it was soaked in the recombinant human BMP 2 to make a cuttlefish bone-BMP (CBB)composite material.Thirty SD rats,with a defect of skull in every rat,were divided into the CBB and pure cuttlefish bone (PCB) groups according to the random numbertable,with 15 rats in each group.The rats in the group CBB and group PCB were transplanted with the corresponding material to repair the skull defect.At post transplantation week (PTW) 4,6,and 8,5 rats from every group were sacrificed by exsanguination,and ink perfusion was performed.One day later,all the transplants and part of the skull surrounding the defect were harvested,and general observation was conducted at the same time.The specimens