同种异体骨髓间充质干细胞移植对家兔股骨头缺血性坏死的治疗作用

Therapeutic effects of allogenic bone marrow mesenchymal stem cell transplantation on ischemic necrosis of the femoral head in rabbits

目的:虽然骨髓间充质干细胞的体外培养和扩增技术已经成熟,但是自体细胞移植仍处于细胞培养和临床治疗相互分离的现状,不便于临床实际应用。为建立骨髓间充质干细胞库,以及为临床应用提供实验证据,实验拟比较同种异体和自体骨髓间充质干细胞移植,对实验性家兔股骨头缺血性坏死的治疗作用。方法:实验于2004-03/2007-04在河北医科大学人体解剖学教研室,白求恩国际和平医院骨科完成。①实验材料:实验所用新西兰大白兔由白求恩国际和平医院实验动物中心提供,雌雄不拘,四五月龄,体质量2.5～3.5kg。实验过程中对动物的处置符合动物伦理学标准。②实验方法:采用全骨髓培养法制备新西兰大白兔骨髓间充质干细胞。采用液氮冷冻法制作兔股骨头缺血性坏死模型。将造模后家兔随机分成自体细胞移植组和同种异体细胞移植组,12只/组,自体细胞移植组钻孔后,植入含有体外培养的自体骨髓间充质干细胞的明胶海绵;同种异体细胞移植组钻孔后,植入含有同种异体骨髓间充质干细胞的明胶海绵。③实验评估:分别于术后2,4,6,8周,进行股骨头标本的X射线和组织学观察。结果:24只新西兰大白兔均进入结果分析。①X射线观察显示,2周时自体细胞移植组与同种异体细胞移植组,股骨头钻孔区边缘密度增加,8周时两组的钻孔区,均出现骨小梁结构。②组织学观察表明,2周时两组的钻孔区内,都有大量的成骨细胞,边缘有较多的骨组织形成;在同种异体细胞移植组的钻孔区内,出现少量淋巴细胞和浆细胞。4周时两组的钻孔区内,充满了新生的骨小梁结构,8周时钻孔区内的骨小梁趋于成熟,两组间骨小梁面积图像分析无明显差异。结论:同种异体和自体骨髓间充质干细胞移植,对实验性家兔股骨头缺血性坏死的治疗效果相似。

AIM： There is an interval between cell culture and clinical treatment by using bone marrow mesenchymal stem cells to treat skeletal diseases. This experiment is to compare the repair effect of treating femoral head ischemia necrosis in rabbits with autografting and allografting of bone marrow mesenchymal stem cells and to supply theory for establishing bone marrow mesenchymai stem cells bank and clinical practice. METHODS： Experiments were conducted from March 2004 to April 2007 in Department of Human Anatomy of Hebei Medical University and Department of Orthopaedics, Bethune International Peace Hospital. （1）New Zealand rabbits of either sex and aged 4 or 5 months weighting 2.5-3.5 kg were offered by medical experimental animal center of Bethune International Peace Hospital. Animal intervention met the animal ethics standard.（2）The bone marrow mesenchymal stem cells were isolated and cultured by cultivating whole bone marrow. Rabbit models of femoral head ischemia necrosis were established by liquid nitrogen frozen method. The rabbit models were divided into two groups randomly, group A as bone marrow mesenchymal stem cells autografting group and group B as one of allografting with 12 in each group. Gelatin sponge containing autologous bone marrow mesenchymal stem cells was transplanted in the rabbit models of the group A. Gelatin sponge containing allograft was transplanted in rabbit models of the group B. （3）Specimens in every group were subjected to radiograph and microscope examination respectively 2, 4, 6, 8 weeks after operation. RESULTS： All the 24 New Zealand rabbits were subjected to result analysis. （1）Radiograph examination： At week 2, density around the drilled holes in groups A and B all increased. At week 8, trabeculae appeared in the drilled holes in both groups. （2）Microscope examination： In both groups, at week 2, there were a large amount of osteoblasts in the drilled hole and some bone tissue formed around the drilled hole. At the same time, some plasma cells and lymphocytes presented in the drilled hole in group B. At week 4, the drilled holes were full of trabeculae. At week 8, the trabeculae were mature and bone marrow appeared in the intertrabecular space. There was no-statistical difference in both groups by analyzing trabeculae area image. CONCLUSION： There are similar repaired effects by treating femoral head ischemia necrosis in rabbits with autografting and allografting of bone marrow mesenchyrnal stem cells.