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| 骨形态生发蛋白2基因修饰脂肪干细胞对去卵巢骨质疏松性大鼠骨缺损的修复作用 |
| The repairing effect of BMP 2 gene-modified adipose stem cells on the bone defects in ovariectomized osteoporotic rats |
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| DOI：10.3969/j.issn.1006.7108.2012.08.006 |
| 中文关键词: 骨形态生发蛋白2 脂肪干细胞 骨质疏松 成骨 |
| 英文关键词:BMP-2 ADSCs Osteoporosis Osteogenesis |
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| 中文摘要: |
| 目的 构建人骨形态生发蛋白2( hBMP-2 )基因真核表达载体，转染脂肪干细胞（ADSCs)后，将其转入去卵巢骨质疏松性大鼠骨缺损模型体内，观察其成骨、修复骨缺损能力。方法①选择雌性未孕Wistar大鼠40只，行双侧卵巢切除，术后定期检测骨密度、骨形态计量学指标及血、尿生化指标，以监测去卵巢骨质疏松性大鼠模型的成功建立；②通过噬菌斑原位杂交筛选人混合细胞cDNA文库获得人骨形态生发蛋白2基因，与真核表达载体pcDNA3.1-连接，构建重组质粒pcDNA3.1 - hBMP-2，
利用脂质体LipofectamineTM2000介导hBMP-2基因转染第4代脂肪干细胞，并经G418筛选；③将40 只模型兔随机分成4组后，于股骨中下段钻孔造成骨缺损，后分别将hBMP2+pcDNA3. 1、ADSCs+pcDNA3. 1、pcDNA3. 1 - hBMP -2+ ADSCs注入前3组兔的骨缺损处，最后1组为对照组；④定期抽静脉血检测Ca+、P及碱性磷酸酶（ALP)等生化指标，X线观察成骨情况；8周后处死动物取骨缺损区组织，HE染色观察成骨情况及炎症反应。结果术后实验组的检测指标较对照组有明显不同，pcDNA3. 1 - hBMP -2+ ADSCs组的血清Ca+、P及ALP水平明显升高，X线检查可看到有明显的成骨、骨缺损得到修复，HE染色可见到大量的成骨细胞生成，较对照组有明显的差异，有统计学意义（P<0. 05 )。结论hBMP-2基因修饰的脂肪干细胞对去卵巢大鼠骨质疏松性骨缺损有很好的修复作用，为绝经后所致的骨质疏松性骨缺损提示了新的治疗思路。 |
| 英文摘要: |
| Objective We built a eukaryotic expression vector with hBMP-2 gene,then we transfected it into ADSCs. Finally we transptanted the cells to the bone defect model of ovariectomized rats to observe the osteogenesis and the capacity of bone defect repair. Methods 1 ) Forty female Wistar rats were undergone bilateral ovariectomy. Bone minerat density,bone histomorphometry indexes,and blood and urine biochemical indicators were monitored to establish the ovariectomized rat model. 2 ) Germinal plaque in situ hybridization was used to screen the hBMP\2 gene in the human hybrid cell cDNA library. The gene was connected to the eukaryotic expression vector pcDNA3. 1. hBMP-2 gene was transfected into the 4th generation of fat stem cells using tiposome Lipofectamine TM2000，and it was selected with G418. 3 ) Forty rabbits randomly divided into 4 groups. The distal femurs of all the rabbits were drilled to make bone defects. Then hBMP2 + pcDNA3. 1,ADSCs + pcDNA3. 1,or pcDNA3. 1 - hBMP - 2 + ADSCs were injected into the bone defects of the rabbits in the first three groups，respectively. The last group was control group. 4 ) Serum Ca，P，ALP，and other biochemical indicators were detected regularly. Bone formation was observed using X-ray. The animals were sacrificed 8 weeks later. Bone formation and inflammation were observed using HE staining.Results Indicators in the experimental group as significantly different comparing to those in the control group postoperatively. Ca,P,and ALP levels in the serum of the pcDNA 3. 1 - hBM P - 2 + ADSC s group increased significantly. Osteogenesis was seen in the X- ray examination. The bone defects were fixed. Proliferation of osteoblasts was seen in HE staining. All of the differences were statistically significant comparing to the control group(P?0.05). Conclusion hBMP-2 gene-modified fat stem cells have good effect on bone defect repairing in ovariectomized osteoporotic rats. This provides a new idea for treatment of bone defects induced by postmenopausal osteoporosis. |
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