Particle gun-mediated bone morphogenetic protein-2 gene transfection for treatment of chronic bone defects

doi:10.3969/j.issn.2095-4344.2014.11.007

Abstract

Abstract:

BACKGROUND: Both in vitro and in vivo studies have confirmed that, bone morphogenetic protein (BMP) regulates the differentiation of osteoblasts and chondroblasts, induces heterotopic bone formation, promotes fracture healing, and controls the morphology of skeleton in mammals.

OBJECTIVE: To treat chronic bone defects using particle gun containing BMP2 gene eukaryotic expression plasmid via local injection.

METHODS: A total of 72 healthy New Zealand white rabbits were applied to establish chronic bone defect model in the rabbit radius. According to the length of bone defect, the rabbits were divided into three groups: 1.5 cm group, 2.0 cm group, 2.5 cm group. Each group was further randomly assigned into two subgroups: treatment group (BMP-2 gene transfection) and control group (naturally healing). X-ray examinations were performed at 1, 3, 8 and 9 weeks after transfection, and soft tissue between the bone defects was harvested to detect BMP-2 using western blot analysis; and radius specimens were taken for gross observation at the same time points, to evaluate the healing.

RESULTS AND CONCLUSION: (1) Gross specimen observation: bone callus formation in treatment group was generally more than that in control group. (2) Lane-Sandhu X-ray score in treatment group was significantly higher than that in control group at 1, 3, 8, 9 weeks after transfection (P < 0.05). (3) BMP-2 concentration in treatment group was significantly higher than that in control group at each time point (P < 0.05). The local transfer of particle gun-mediated BMP-2 gene is an effective therapy of chronic bone defect.

中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

全文链接：

Key words: fractures, bone, bone morphogenetic proteins, osteotomy, biolistics, fracture healing

CLC Number:

R318

Zhu Xiao-meng, Wang Chong, Song Xing-hua, Zhan Yu-lin, Li Wen-ju. Particle gun-mediated bone morphogenetic protein-2 gene transfection for treatment of chronic bone defects[J]. Chinese Journal of Tissue Engineering Research, 2014, 18(11): 1680-1686.

Figures/Tables 2

2.1 大体标本观察基因转染后1周：骨缺损区被不规则、断续的暗褐色物质填充，质地略硬，对照组骨缺损间填充物稍低于治疗组。多数骨缺损表面凹凸不平，较易与周围组织剥离，骨髓腔均已闭，大部分对照组骨折断端有异常活动。基因转染后3周：机化组织填充于各组骨缺损区，多数1.5 cm组断端间连接有粗细不一的纤维性骨桥。2.0 cm治疗组断端间有少量纤细骨组织连接，断端间骨痂均呈向心性延伸生长。2.5 cm组骨痂较少、向心性延伸。断端无异常活动。治疗组整体愈合情况优于对照组。其中1.5 cm组最为突出。基因转染后8周：1.5 cm治疗组断端间可见大量不规则连续骨痂形成，呈向心性分布，但骨痂中央区相对薄弱。2.0 cm组断端间均有骨痂，亦呈向心性生长趋势，对照组缺损部位形成弧形凹槽，治疗组两端骨痂间多形成较细的弓形骨桥。2.5 cm治疗组断端骨痂亦呈向心性生长，形成下凹槽，缺损长度有所缩短，偶可见弓形骨桥；而对照组骨缺损断端仅可见少量骨痂形成，有骨不连倾向。基因转染后9周：大体情况如基因转染后8周时，治疗组修复部位直径普遍大于对照组。 2.2 骨折修复后X射线检查结果基因转染后1周：治疗组与对照组比较差异有显著性意义(1.5 cm组：t=4.547，P < 0.05；2.0 cm组：t=2.237， P < 0.05；2.5 cm组：t=3.169，P < 0.05)，见图2。基因转染后3周：治疗组优于对照组(1.5 cm组：t=6.436，P < 0.05；2.0 cm组：t=4.452，P < 0.05；2.5 cm组：t=6.185，P < 0.05)，见图3。治疗后8周：治疗组优于对照组(1.5 cm组：t=5.612，P < 0.05；2.0 cm组：t=5.571，P < 0.05；2.5 cm组：t=5.476，P < 0.05)，见图4。治疗后9周：治疗组优于对照组(1.5 cm组：t=5.854，P < 0.05；2.0 cm组：t=11.362，P < 0.05；2.5 cm组：t=13.308，P < 0.05)，见图5。治疗组与对照组在转染后1，3，8，9周拍摄X射线平片的Lane-Sandhu X射线评分结果组内比较：各骨缺损长度，各时间点治疗组与对照组X射线评分差异有显著性意义(P <0.05)：治疗组优于对照组(表2)。 2.3 所取组织标本中骨形态发生蛋白2的western-blot检测分别在基因转染后处死以上各组实验动物3只，取骨缺损间软组织行Western blot检测，样本骨形态发生蛋白2浓度定量结果显示，各骨缺损长度，各时间点治疗组骨形态发生蛋白2浓度优于对照组，差异有显著性意义(P < 0.05，表3)。"

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