辛伐他汀诱导内皮祖细胞归巢及促进骨缺损修复的作用

[Effect of simvastatin on inducing endothelial progenitor cells homing and promoting bone defect repair].

作者信息

Song Quansheng, Wang Lingying, Zhu Jinglin, Han Xiaoguang, Li Xu, Yang Yanlin, Sun Yan, Song Chunli

机构信息

Department of Orthopaedics, Third Hospital, Peking University, Beijing, 100191, PR China.

出版信息

Zhongguo Xiu Fu Chong Jian Wai Ke Za Zhi. 2010 Sep;24(9):1103-6.

PMID:20939484

Abstract

OBJECTIVE

To investigate the effect of simvastatin on inducing endothelial progenitor cells (EPCs) homing and promoting bone defect repair, and to explore the mechanism of local implanting simvastatin in promoting bone formation.

METHODS

Simvastatin (50 mg) compounded with polylactic acid (PLA, 200 mg) or only PLA (200 mg) was dissolved in acetone (1 mL) to prepare implanted materials (Simvastatin-PLA material, PLA material). EPCs were harvested from bone marrow of 2 male rabbits and cultured with M199; after identified by immunohistochemistry, the cell suspension of EPCs at the 3rd generation (2 x 10(6) cells/mL) was prepared and transplanted into 12 female rabbits through auricular veins (2 mL). After 3 days, the models of cranial defect with 15 cm diameter were made in the 12 female rabbits. And the defects were repaired with Simvastatin-PLA materials (experimental group, n=6) and PLA materials (control group, n=6), respectively. The bone repair was observed after 8 weeks of operation by gross appearance, X-ray film, and histology; gelatin-ink perfusion and HE staining were used to show the new vessels formation in the defect. Fluorescence in situ hybridization (FISH) was performed to show the EPCs homing at the defect site.

RESULTS

All experimental animals of 2 groups survived to the end of the experiment. After 8 weeks in experimental group, new bone formation was observed in the bone defect by gross and histology, and an irregular, hyperdense shadow by X-ray film; no similar changes were observed in control group. FISH showed that the male EPC containing Y chromosome was found in the wall of new vessels in the defect of experimental group, while no male EPC containing Y chromosome was found in control group. The percentage of new bone formation in defect area was 91.63% +/- 4.07% in experimental group and 59.45% +/- 5.43% in control group, showing significant difference (P < 0.05).

CONCLUSION

Simvastatin can promote bone defect repair, and its mechanism is probably associated with inducing EPCs homing and enhancing vasculogenesis.

摘要

目的

探讨辛伐他汀对诱导内皮祖细胞（EPCs）归巢及促进骨缺损修复的作用，并探究局部植入辛伐他汀促进骨形成的机制。

方法

将辛伐他汀（50 mg）与聚乳酸（PLA，200 mg）复合或仅用PLA（200 mg）溶于丙酮（1 mL）中制备植入材料（辛伐他汀-PLA材料、PLA材料）。从2只雄性兔骨髓中获取EPCs，用M199培养基培养；经免疫组织化学鉴定后，制备第3代EPCs细胞悬液（2×10⁶个细胞/mL），通过耳静脉将其移植到12只雌性兔体内（2 mL）。3天后，在12只雌性兔身上制作直径为15 mm的颅骨缺损模型。分别用辛伐他汀-PLA材料（实验组，n = 6）和PLA材料（对照组，n = 6）修复缺损。术后8周通过大体观察、X线片及组织学观察骨修复情况；采用明胶-墨汁灌注及HE染色显示缺损区新生血管形成。进行荧光原位杂交（FISH）以显示EPCs在缺损部位的归巢情况。

结果

两组所有实验动物均存活至实验结束。实验组术后8周，大体及组织学观察可见骨缺损处有新骨形成，X线片显示为不规则高密度影；对照组未见类似变化。FISH显示实验组缺损区新生血管壁中发现含Y染色体的雄性EPCs，而对照组未发现含Y染色体的雄性EPCs。实验组缺损区新骨形成百分比为91.63%±4.07%，对照组为59.45%±5.43%，差异有统计学意义（P < 0.05）。