适宜电刺激对成骨细胞平面培养下细胞增殖分化及BMP-2表达的实验研究

Proliferation,differentiation and BMP-2 expression of osteoblasts cultured in the plane under suitable electrical stimulation

目的:观察适宜电流刺激成骨细胞在平面培养下成骨细胞增殖及分化,明确是否具有促进增殖、分化作用,并观察电流刺激下BMP-2表达的变化。方法:将来自家兔幼仔颅骨提取的成骨细胞进行平面培养,2代之后进行分类培养,实验组进行电流刺激,并给予相同的刺激时间和刺激强度,对照组未进行电流刺激。之后在不同时间点进行细胞增殖、分化检测及BMP-2蛋白表达分析。结果:实验组8d内光镜下可见大量成骨细胞增殖,成骨细胞呈多形性改变,6～8d细胞内出现少量钙化点,而对照组骨细胞增殖缓慢。实验组分化明显,碱性磷酸酶及钙结节染色阳性,碱性磷酸酶定量分析显示逐渐增加。实验组经免疫组化分析显示BMP-2量逐渐增加。结论:电刺激可促进成骨细胞的增殖分化而达到实现细胞数目的短时间增加,细胞内部经过免疫组化染色后显示BMP-2表达增加。

Objective ：To observe proliferation and differentiation of osteoblasts cultured in the plane on appropriate elec- trical stimulation ,to specify whether it promote the proliferation, and observe expression of BMP-2 on electrical stimulation. Methods ： Osteoblasts were extracted from the skull of rabbit offspring and cultured. Ceils after the 2nd generations were cultured. In experimental group, cells had electrical stimulation, and same stimulation time and intensity were given. In control group cells had not electrical stimulation. The proliferation and differentiation were detected at different time, and BMP-2 pro- tein expression was analyzed. Results ：The cell morphology of experimental group in 8 days under the light microscope was ob- served and showed a lot of proliferation of osteoblasts, pleomorphic changes,in 6 to 8 days a small amount of Calcified spots was also observed ; while in the control group, proliferation was slower. Differentiation of the experimental group was signifi- cantly,alkaline phosphatase staining and calcium nodules were positive,quantitative analysis of alkaline phosphatase in- creaseed significantly. Experimental group showed that BMP-2 was gradually increased by immunohistochemistry analysis. Conclusion：Electrical stimulation can promote the proliferation and differentiation of osteoblasts and achieved the increasement the number of cells in short-term,intracellular staining by immunohistochemistry showed the inereasement in expression of BMP-2.