重组合异种骨复合浓缩自体骨髓移植修复股骨头缺损性坏死

Implantation of concentrated autologous bone-marrow cells of reconstituted bone xonograft in repair of bone defect of femoral head

目的:将复合重组合异种骨的自体浓缩骨髓移植到按Trapdoor法制作的股骨头缺损内,以观察其对股骨头坏死后骨缺损的修复和重建能力。方法:实验于2005-05/12在安徽医科大学附属安徽省立医院中心实验室完成。将成年健康新西兰兔24只48髋按Trapdoor法制作双侧股骨头缺损模型并随机分为6组:自体骨+自体骨髓组;自体骨组;重组合异种骨+自体骨髓组;重组合异种骨组;空白对照组;正常对照组。其中自体骨组和异种骨组实验动物右侧同时植入自体骨髓和载体,左侧仅植入载体。自体骨为取自自体髂骨的皮质松质混合骨;重组合异种骨由天津金世公司提供;自体浓缩骨髓取自体,用肝素化的注射器在股骨大转子处穿刺抽取骨髓2～3mL,低温离心机以1500r/min离心,获浓缩骨髓0.5mL备用。在制作缺损模型后即按分组植入不同复合物,空白对照组自然愈合。于植入后4,8,12周通过X射线片、CT、组织学等方法观察其骨缺损修复能力。结果:实验兔24只均进入结果分析。①X射线检查结果:术后4周,异种骨组与自体骨组关节面光滑,缺损区可见少量絮状影,自体骨组较重组合异种骨组稍明显,而此二组右侧较左侧为好。自然愈合组见扩大的低密度影。术后12周,异种骨组与自体骨组关节面光滑,缺损区可见接近正常骨小梁结构影,右侧较左侧为佳。自然愈合组可见硬化和囊变影。②CT检查结果:术后12周,异种骨组与自体骨组CT表现相近,关节面光整,缺损区被接近正常骨组织影替代,右侧优于左侧。自然愈合组仍为低密度影,伴硬化和囊变影。③组织学观察结果:术后4周,大体观察,自体骨组与异种骨组关节面光滑。镜下观察,自体骨组右侧较左侧炎症反应轻,新骨生成稍多。异种骨组可见骨小梁样结构,右侧无明显坏死和组织细胞,左侧可见局灶坏死和少量组织细胞。空白对照组见脂肪性骨髓,骨坏死及炎性细胞。术后12周,大体观察,自体骨组与异种骨组关节面光滑。镜下观察,自体骨组右侧骨小梁丰富,形粗,排列整齐,其周围可见骨母细胞及新生的血管;左侧骨小梁稀疏,形细。异种骨组右侧新生骨骨小梁排列整齐,近似板层骨;左侧新生骨骨小梁排列稍乱。自然愈合组缺损区域周围有脂性骨髓组织,中央有纤维组织增生,碎片样软骨组织,缺损区无成熟骨组织。结论:复合重组合异种骨的浓缩自体骨髓修复股骨头缺损具有优良的骨传导性和骨诱导性及良好的机械支撑。

AIM： Concentrated autologous bone-marrow cells of reconstituted bone xonograft was implantated to the defect of femoral head with Trapdoor method, so as to observe its effect on repair and reconstruction ability of bone defect after necrosis of femoral head. METHODS： This experiment was conducted at the Central Laboratory of Anhui Provincial Hospital of Anhui Medical University from May to December 2005. Totally hips of 24 adult healthy New Zealand rabbits were made into femoral head defect models of two sides according to Trapdoor method and randomly divided into 6 groups： autologous bone ＋ autologous bone marrow group; autogolous bone group; xenograft bone ＋ autologous bone marrow group; xenograft bone group; blank control groUp; normal control group. Autogolous bone marrow and vector were implanted into the right side of the animals in the autologous bone marrow group and xenograft bone group, only vector was implanted into the left side of the animals . Autologous bone was the mixture of os integumentale and trabocular bone obtained from ilium; xenograft was made by Jinshi Company, Tianjin; Concentrated autologous bone-marrow was harvested from the tested rabbits. 2 to 3 mL bone marrow was extracted from the femoral bone with injector treated by heparin. Centrifugation was performed at 1 500 r/min, 0.5 mL concentrated bone marrow was obtained. After defect model making, different compound was implanted in each group according to grouping, and rabbits in the blank control group healed by themselves. Bone defect repair ability was observed through X-ray, CT and histological methods at 4,8 and 12 weeks after transplantation. RESULTS：All the 24 rabbits entered the stage of result analysis. ① Result of X-ray detection： At postoperative 4 weeks, articular surface of xenograft bone group and autologous bone group , few flocculent shadow was seen in the defect area . The shadow was more obvious in the autogolous group than xenograft group, and the right side was better than the left side in each group. Expanded low-density shadow was seen in the natural healing group. At postoperative 12 weeks, articular surface of xenograft bone group and autologous group was smooth, structure shadow of bone trabecula close to normal one was seen in the natural healing group. The fight side was better than the left side. Sclerosis and capsule change were found in the natural healing group. ②CT examination： At postopera： five 12 weeks, there was no difference in CT examination between xenograft bone group and autogolous bone group. Articular surface was smooth. Defect area was replaced by bone tissue close to the normal one, and the right side was better than the left side. There was still low-density shadow with sclerosis and capsule change in the natural healing group. ③Histological observation： At postoperative 4 weeks, in gross observation, articular surface of autologous bone group and xenograft bone group was smooth. The inflammation was less severe on the right side than on the left side in the autologous bone group. Bone trabecula-like structure was found in the xenograft group, there was no obvious necrosis and tissue cells on the right side. Local necrosis and few tissue cells appeared on the left side. Fatty bone marrow, bone necrosis and inflammatory cells were found in the blank control group. At postoperative 12 weeks, in the gross observation, articular surface was smooth in the autagolous bone group and xenograft bone group. It was observed under microscope that the bone trabecula was abundant, thick and well arranged on the right side in the autagolous group; there were also osteoblast and newborn blood vessels around the bone trabecula on the fight side. The bone trabecula was sparse and thin on the left side. In the xenograft group, well-arranged new born bone trabocula , which was close to lamellar bone , was found on the right side ; Disordered new born bone trabecula was found on the left side. In the natural healing group, there was fatty bone marrow tissue around th.e defected area, proliferated fibrous tissue, fragment-like cartilage tissue in the center, but no mature bone tissue was found in the defected area. CONCLUSION： Concentrated autologous bone marrow cells with reconstituted bone xenograft possess excellent osteeconductivity, and osteoinducibility as well as good mechanical supporting in repairing of bone defect of femoral head.