BMP-2和地塞米松对成人成骨细胞增殖和分化的影响  

Effects of BMP-2 and dexamethasone on proliferation and differentiation of adult osteoblasts in vitro

作　　者：温学红[1] 郭若霖[2] 程瑞芳[3] 张福江[2] 谈志龙[2] 叶伟胜[2] 王学谦[2] 

机构地区：[1]天津中医学院第二附属医院检验科,天津300150 [2]天津医院骨研所,天津300211 [3]天津口腔医院,天津300078

出　　处：《临床检验杂志》2005年第1期34-37,i001,共5页Chinese Journal of Clinical Laboratory Science

基　　金：国家自然科学基金 (青年 ) (No .3 0 2 0 0 13 4)

摘　　要：目的　观察不同浓度骨形态发生蛋白 2 (bonemorphogeneticprotein 2 ,BMP 2 )和地塞米松 (dexamathasone,DEX)对体外培养的人成骨细胞增殖、分化的影响。方法　成人髂骨松质骨胰蛋白酶消化后获得成骨细胞进行培养 ,用倒置显微镜进行活体观察并做碱性磷酸酶 (ALP)、骨钙素 (OCN)、Ⅰ型及Ⅲ型胶原染色观察成骨细胞生物学特征。培养的细胞加入不同浓度BMP 2(1、10、10 0、10 0 0ng/ml)和DEX(10 8、10 7、10 6mol/L)作用 4 8h后 ,用MTT法检测细胞增殖 ,同时测定细胞裂解液中ALP活性和OCN含量。结果　体外培养的成人成骨细胞可合成碱性磷酸酶、骨钙素、Ⅰ型胶原 ,并形成钙化结节。 3个浓度的DEX在 4 8h后均能刺激成骨细胞的增殖 ,增加成骨细胞ALP活性和OCN含量 ,4个浓度BMP 2在 4 8h后对成骨细胞无增殖作用 ,但都能增加ALP活性和OCN含量。结论　地塞米松促进体外培养的成人成骨细胞增殖和分化 ;短期作用下 ,BMP 2对成骨细胞的增殖无明显作用 。Objective To investigate the effects of bone morphogenetic protein-2 (BMP-2) and dexamethasone (DEX) on proliferation and differentiation of adult osteoblasts in vitro.Methods Adult osteoblasts were isolated from iliac trabecula and cultured in vitro.The biological characteristics were observed by phase-contrast microscope and investigated by alkaline phosphatase (ALP),osteocalcin (OCN),collagen Ⅰ and Ⅲ staining.The cells were subjected to the treatment with different concentrations of BMP-2 (1,10,100 and 1 000 ng/ml) and DEX (10 -8,10 -7,10 -6 mol/L).The proliferation was measured with methylthiazol tetrazolium (MTT) method.Alkaline phosphatase activity and osteocalcin were measured to evaluate the differentiation of osteoblasts after 48h.Results ALP,OCN and collagen Ⅰ were synthesized by the osteoblasts.but collagen Ⅲ staining was negative.Compared with the control group,the OD value of MTT,ALP activity and OCN level were increased with different concentrations of DEX.However,ALP and OCN were significantly increased in different concentrations of BMP-2,but OD value of MTT did not significantly increase.Conclusions DEX has significant effect on proliferation and differentiation of adult osteoblasts in vitro.After short-term treatment BMP-2 also has differentiating effect on adult osteoblasts,but no significant effect on proliferation.

关 键 词：BMP-2 人成骨细胞 增殖 地塞米松 DEX 分化 ALP 活体观察 体外培养 细胞裂解液 

分 类 号：R318[医药卫生—生物医学工程] R68[医药卫生—基础医学]