[骨髓基质细胞中骨形态发生蛋白-2和碱性成纤维细胞生长因子在异位和原位骨形成中的反应]

[Response of bone morphogenetic protein-2 and basic fibroblast growth factor in bone marrow stromal cells in ectopic and in situ bone formation].

作者信息

Wang Lei, Zhang Yan, You Sulan, Tan Luanjun, Huang Yuanliang

机构信息

Dept. of Stomatology, Hongqiao Community Medical Service Center in Changning District of Shanghai, Shanghai 200051, China.

出版信息

Hua Xi Kou Qiang Yi Xue Za Zhi. 2012 Aug;30(4):420-4.

PMID:22934504

Abstract

OBJECTIVE

We ascertained the effect of bone morphogenetic protein-2 (BMP-2) and basic fibroblast growth factor (bFGF) by a series of experiments: Proliferation and differentiation of bone marrow stromal cells (BMSCs) in vitro, ectopic and in situ bone formation and loaded porous calcium phosphate cement (CPC) on the repair of bone defects around dental implants.

METHODS

BMSCs from Beagle dogs were cultured in vitro with basic culture medium containing BMP-2, bFGF, and BMP-2+bFGF. Proliferation and differentiation of BMSCs were quantified using methyl thiazolyl tetrazolium (MTT) and alkaline phosphatase (ALP) test. The CPC seeded with BMSCs and BMP-2, bFGF, combined BMP-2 with bFGF were implanted subcutaneously into nude rats in ectopic bone formation, and were implanted into critical-sized bone defects of Beagle dogs in the in situ bone formation. The bone formation was detected by histology examination and quantified using an image analysis system. Polychrome sequential fluorescent labels and fluorescence histological examinations of undecalcified sections were performed post-operatively.

RESULTS

It was determined that BMP-2+bFGF promoted BMSCs statistically significant proliferation and differentiation compared to either BMP-2 or bFGF in vitro. The CPC with BMP-2+bFGF group yielded more bone than those with either BMP-2 or bFGF in ectopic bone formation test. The percentages of newly ectopic formed bone were higher in the BMP-2+bFGF group (48.79% +/- 11.31%) than those in other groups (BMP-2 group, 30.71% +/- 10.85%; bFGF group, 27.33% +/- 9.67%; and the control group, 10.65% +/- 6.05%). Undecalcified showed that new bone was actively formed in the BMP-2+bFGF group after 12 weeks in the in situ bone formation test. The bone mineralization apposition rate (MAR) was better in the BMP-2+bFGF group than in other groups (P<0.01).

CONCLUSION

BMP-2 combined with bFGF are more effective than one alone in promoting the formation of new bone.

摘要

目的

我们通过一系列实验确定了骨形态发生蛋白-2（BMP-2）和碱性成纤维细胞生长因子（bFGF）对骨髓基质细胞（BMSCs）体外增殖与分化、异位和原位骨形成以及负载多孔磷酸钙骨水泥（CPC）对牙种植体周围骨缺损修复的影响。

方法

将比格犬的BMSCs在含有BMP-2、bFGF以及BMP-2+bFGF的基础培养基中进行体外培养。使用甲基噻唑基四氮唑（MTT）和碱性磷酸酶（ALP）检测对BMSCs的增殖与分化进行定量分析。将接种有BMSCs以及BMP-2、bFGF、BMP-2与bFGF组合的CPC皮下植入裸鼠进行异位骨形成实验，并植入比格犬的临界尺寸骨缺损处进行原位骨形成实验。通过组织学检查检测骨形成情况，并使用图像分析系统进行定量分析。术后进行多色顺序荧光标记和未脱钙切片的荧光组织学检查。

结果

确定在体外，与单独使用BMP-2或bFGF相比，BMP-2+bFGF能促进BMSCs的增殖与分化，差异具有统计学意义。在异位骨形成实验中，含BMP-2+bFGF组的CPC生成的骨量比含BMP-2或bFGF组更多。BMP-2+bFGF组新异位形成骨的百分比（48.79%±11.31%）高于其他组（BMP-2组，30.71%±10.85%；bFGF组，27.33%±9.67%；对照组，10.65%±6.05%）。未脱钙切片显示，在原位骨形成实验12周后，BMP-2+bFGF组有新骨活跃形成。BMP-2+bFGF组的骨矿化沉积率（MAR）优于其他组（P<0.01）。