Ad-BMP-2/BMSCs/DBM对兔股骨头坏死修复的实验研究
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摘要
目的应用聚合酶链式反应(RT-PCR)、电镜扫描(SEM)和X射线能谱分析技术(EDS)评价腺病毒-骨形态发生蛋白-2/骨髓间充质干细胞/脱钙松质骨(Ad-BMP-2/BMSCs/DBM)对兔股骨头坏死的治疗效果,为临床治疗股骨头坏死探索新途径。方法 1采用髓芯减压联合液氮冰冻法制备兔股骨头坏死模型;2将造模成功的兔子随机均等分为A、B、C、D组(n=12),A组不植入材料,为造模组,B、C、D分别植入DBM、DBM/BMSCs、BMP-2/BMSCs/DBM;3术后4、8、12周各组分别处死4只兔子,运用RT-PCR、SEM、EDS检测股骨头BMP-2表达量、表面形貌和钙磷比(Ca/P),观察股骨头的修复情况。结果 RT-PCR结果显示各时间点BMP-2表达量A组 Objective To evaluate the effect of the Ad-BMP-2/BMSCs/DBM for rabbits' femoral head necrosis by the RT-PCR,SEM and EDSand explore the new treatments for femoral head necrosis. Methods 1 Prepare femoral head necrosis models by clinical core decompression combined with liquid nitrogen frozen. 2 After building models,animals were randomly divided into 4 groups( n = 12) as follows: group A was not implanted anything as control group,group B was implanted into DBM,group C was implanted into h BMP-2/DBM,group D was implanted into h BMP-2/BMSCs/DBM. 3 Four rabbits of each group were killed at 4,8,12 weeks after surgery to evaluate the reparation of ONFH through the RT-PCR,SEM and EDS. Results The RT-PCR showed: the expression of BMP-2in group A < group B < group C < group D( P < 0. 05). SEM showed: there was no calcific depositions in group A. Group B and group C had some calcific depositions and low calcific bone. There was new mature bone in group D and there was no obvious difference compared with the normal bone tissue. EDS showed: The Ca/P of group A
引文
[1]Kim S Y,Kim T H.Genetic Studies in osteonecrosis of the femoral head[M]//Osteonecrosis.Springer Berlin Heidelberg,2014:61-9.
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[3]Chamberlain J R,Schwarze U,Wang P R,et al.Gene targeting in stem cells from individuals with osteogenesis imperfecta[J].Science,2004,303(5661):1198-201.
[4]Wang C J,Huang C C,Wang J W,et al.Long-term results of extracorporeal shockwave therapy and core decompression in osteonecrosis of the femoral head with eight-to nine-year follow-up[J].Biomed J,2012,35(6):481-5.
[5]Pittenger M F,Mackay A M,Beck S C,et al.Multilineage potential of adult human mesenchymal stem cells[J].Science,1999,284(5411):143-7.
[6]Bose S,Roy M,Bandyopadhyay A.Recent advances in bone tissue engineering scaffolds[J].Trends Biotechnol,2012,30(10):546-54.
[7]陈佳滨,李强,茹嘉,等.腺病毒介导BMP-2和EGFP基因转染兔骨髓基质干细胞及种植脱钙骨的体外观察[J].中国骨质疏松杂志,2014,20(8):869-74.
[8]李海冰,瞿向阳,李明,等.应用软骨下钻孔结合液氮冷冻制作兔股骨头缺损坏死模型[J].第三军医大学学报,2014,36(19):23.
[9]Lebouvier A,Poignard A,Cavet M,et al.Development of a simple procedure for the treatment of femoral head osteonecrosis with intra-osseous injection of bone marrow mesenchymal stromal cells:study of their biodistribution in the early time points after injection[J].Stem Cell Res Ther,2015,6:68.
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[11]李强,唐际存,王锐英,等.完全脱钙松质骨的生物学特性(英文)[J].中国组织工程研究与临床康复,2009,47:14.
[12]李强,孙正义,严冬雪,等.脱钙骨基质材料的生物学表现:降解性能、孔隙率及其黏附性能特征[J].中国组织工程研究与临床康复,2007,11(31):6121-4.
[13]程谷,李祖兵.骨组织工程中的检测方法及其效果评估展望[J].国际口腔医学杂志,2010,37(5):593-6.
[14]Melling M,Karimian·Teherani D,Mosfler S,et al.3-D morphological characterization of the liver parenehyma by atomic force microscopy and by scanning electron micmacopy[J].Microsc Res Tech,2004,64(1):1-9
[15]颜彦,周海静,聂红兵.医学领域中骨组织元素分析方法[J].口腔生物医学,2012,3(1):41-4.
[2]Coll R,Aguirre M,Hernandez A,et al.Mesenchymal stromal cells for osteonecrosis of the femoral head(ONFH).data from ongoin clinica trial[J].Cytotherapy,2014,16(4):S97-8.
[3]Chamberlain J R,Schwarze U,Wang P R,et al.Gene targeting in stem cells from individuals with osteogenesis imperfecta[J].Science,2004,303(5661):1198-201.
[4]Wang C J,Huang C C,Wang J W,et al.Long-term results of extracorporeal shockwave therapy and core decompression in osteonecrosis of the femoral head with eight-to nine-year follow-up[J].Biomed J,2012,35(6):481-5.
[5]Pittenger M F,Mackay A M,Beck S C,et al.Multilineage potential of adult human mesenchymal stem cells[J].Science,1999,284(5411):143-7.
[6]Bose S,Roy M,Bandyopadhyay A.Recent advances in bone tissue engineering scaffolds[J].Trends Biotechnol,2012,30(10):546-54.
[7]陈佳滨,李强,茹嘉,等.腺病毒介导BMP-2和EGFP基因转染兔骨髓基质干细胞及种植脱钙骨的体外观察[J].中国骨质疏松杂志,2014,20(8):869-74.
[8]李海冰,瞿向阳,李明,等.应用软骨下钻孔结合液氮冷冻制作兔股骨头缺损坏死模型[J].第三军医大学学报,2014,36(19):23.
[9]Lebouvier A,Poignard A,Cavet M,et al.Development of a simple procedure for the treatment of femoral head osteonecrosis with intra-osseous injection of bone marrow mesenchymal stromal cells:study of their biodistribution in the early time points after injection[J].Stem Cell Res Ther,2015,6:68.
[10]Sekiya I,Larson B L,Vuoristo J T,et al.Adipogenic differentiation of human adult stem cells from bone marrow stroma(MSCs)[J].J Bone Miner Res,2004,19(2):256-64.
[11]李强,唐际存,王锐英,等.完全脱钙松质骨的生物学特性(英文)[J].中国组织工程研究与临床康复,2009,47:14.
[12]李强,孙正义,严冬雪,等.脱钙骨基质材料的生物学表现:降解性能、孔隙率及其黏附性能特征[J].中国组织工程研究与临床康复,2007,11(31):6121-4.
[13]程谷,李祖兵.骨组织工程中的检测方法及其效果评估展望[J].国际口腔医学杂志,2010,37(5):593-6.
[14]Melling M,Karimian·Teherani D,Mosfler S,et al.3-D morphological characterization of the liver parenehyma by atomic force microscopy and by scanning electron micmacopy[J].Microsc Res Tech,2004,64(1):1-9
[15]颜彦,周海静,聂红兵.医学领域中骨组织元素分析方法[J].口腔生物医学,2012,3(1):41-4.