摘要
目的:以矿化诱导的骨髓基质细胞作为种子细胞,应用陶瓷化骨粉/水凝胶复合材料修复大鼠颅骨极量骨缺损,观察骨髓基质细胞修复自体颅骨缺损效果。方法:实验于2002-05/08在解放军第四军医大学口腔医学院口腔组织病理教研室完成。选取健康SD大鼠8只,随机分为重组人骨形态蛋白-2/转化生长因子-β组、陶瓷化骨粉/水凝胶复合物对照组,4只/组。重组人骨形态蛋白-2/转化生长因子-β组将矿化诱导的大鼠自体骨髓基质细胞与水凝胶、陶瓷化骨粉混合,向其中加入重组人骨形态蛋白-210μg、转化生长因子-β50ng,修复自体颅骨8mm直径缺损。陶瓷化骨粉/水凝胶复合物对照组以单独水凝胶/陶瓷化骨粉修复颅骨8mm直径缺损。术后6周取材,应用苏木精-伊红染色、改良Mallory三色法以及彩色图像分析观察骨缺损修复情况。结果:实验纳入8只大鼠,全部进入结果分析。①两组术后6周大体标本观察:两组大鼠的植入物牢固嵌在骨缺损处,未见脱落、移位,表面被纤维被膜包绕,被膜内可见少量毛细血管,两组植入物无明显差别。②两组术后6周骨形成情况比较:苏木精-伊红染色:重组人骨形态蛋白-2/转化生长因子-β组有大量网格状的骨形成,仍有部分水凝胶残留,周围是条索状软骨样组织和纤维组织。陶瓷化骨粉/水凝胶复合物对照组水凝胶部分吸收,陶瓷化骨颗粒和水凝胶粒周围只有少量的骨样或软骨样基质形成,大部分为纤维结缔组织。改良Mallory’s三色法:重组人骨形态蛋白-2/转化生长因子-β组的创缘断端附近的网状骨为砖红色的成熟骨,植入物的中心部可见蓝色的新生骨,以及散在的黄色毛细血管。而陶瓷化骨粉/水凝胶复合物对照组只见少量不规则红色的骨样基质,以及黄色毛细血管。③两组术后6周骨基质及成熟骨基质面积值的比较:重组人骨形态蛋白-2/转化生长因子-β组骨基质面积、成熟骨基质面积明显均多于陶瓷化骨粉/水凝胶复合物对照组[(8207.22±46.31),(375.05±25.83);(6376.73±78.35),(134.44±14.62)μm2;P均<0.05]。结论:陶瓷化骨粉/水凝胶与骨髓基质细胞在重组人骨形成蛋白-2、转化生长因子-β等生长因子联合作用下能够成功构建组织工程骨,修复颅骨极量缺损明显好于单独应用陶瓷化骨粉/水凝胶。
AIM： To repair calvarial defects of rats with ceramic bovine bone particle/ hydrogel hybrid by taking the mineralization induced mesenchymal stem cells （MSCs） as the seed cells, and observe the effect of MSCs in repairing autogenous calvarial defects.
METHODS： The experiment was carried in the Department of Histopathology, Stomatology Medical College, Fourth Military Medical University of Chinese PEA from May to August 2002. Eight healthy SD rats were randomly divided into recombinant human bone morphogenetic protein-2/transforming growth factor-beta group （rhBMP-2/TGF-β group, n =4） and ceramic bovine bone particle/hydrogel hybrid group （control group, n=4）. In the rhBMP-2/TGF-β group, the mineralization induced MSCs were mixed with the ceramic bovine bone particle/hydrogel powder, and 10 μg rhBMP-2 and 50 ng TGF-β were added, and the 8-mm autogenous calvarial defect was repaired, In the control group, the 8-tmn autogenous calvarial defect was repaired only by ceramic bovine bone particle/hydrogel hybrid. The materials were taken after 6 weeks, and the osteogenesis was observed with hematoxylin and eosin （HE） staining, modified Mallory＇s trichrome staining, and color image analysis.
RESULTS： All the 8 rats were involved in the analysis of results. ① Gross observation of the samples after 6 weeks in the two groups： In both groups, the implants were fixed at the defect, no abscission and displacement was observed, the surface was wrapped with fibrous capsule, and a few blood capillaries were observed in capsule, the implants in the two groups had no obvious difference. ② Comparison of bone formation after 6 weeks between the two groups： The HE staining showed that there were plenty of net-like bone formations in the rhBMP-2/TGF-β group, there was still some remain of hydrogel, strip-shape cartilage-like tissue and fibrous tissue were observed in the surrounding; While part of the hydrogel in the control group was absorbed, only a bit of osteoid or cartilage-like matrix formed around the ceramic bone particles and hydrogel particles, and most were fibrous connective tissue. The modified Mallory＇s trichrome staining indicated that the net-like bone around the wounded edge was brick-red mature bone in the rhBMP-2/TGF-β group, and the blue new bone and scattered yellow blood capillaries could be observed in the center of the implants; While only a bit of irregular red osteoid matrix and yellow blood capillaries could be observed in the control group. ③ Comparison of the areas of bone matrix and mature bone matrix after 6 weeks between the two groups： The areas of bone matrix and mature bone matrix in the areas of bone matrix and mature bone matrix were obviously more than those in the control group ]（8 207.22±46.31）, （375.05±25.83）; （6 376.73±78.35）, （134.44±14.62） μm^2,P 〈 0.05].
CONCLUSION： Under the combined effects of rhBMP-2, TGF-β and other growth factors, MSCs-ceramic bovine bone particle/hydrogel hybrid can successfully construct tissue engineering bone, and it is better in repairing calvarial defect than the application of ceramic bovine bone particle/hydrogel alone.
出处
《中国临床康复》
CSCD
北大核心
2005年第46期39-41,T0002,共4页
Chinese Journal of Clinical Rehabilitation
