摘要
目的 探讨甲基丙烯酸缩水甘油酯右旋糖酐 (dex GMA)重组人骨形态发生蛋白(rhBMP 2 )凝胶微球 (rhBMP2 dex HM )对成骨细胞的生物学作用。方法 将单纯rhBMP 2 (A组 )、空白dex GMA凝胶微球dex HM (B组 )和rhBMP2 dex HM (C组 )加入成骨细胞培养液中 ,用细胞计数法、噻唑蓝比色法 (MTT法 )、流式细胞仪观察细胞增殖情况 ,并检测成骨细胞上清液中骨钙素 (BGP)含量。结果 培养 1～ 2d后 ,3组细胞计数、吸光度 (A)值差异无统计学意义 (P >0 .0 5 ) ;4～ 6d时A、C组细胞计数和A值开始高于B组 ,但A、C组间差异无统计学意义 (P >0 .0 5 ) ;培养 6～ 8d后 ,C组明显高于其他组 ,差异有统计学意义 (P <0 .0 1)。14d后 ,A、B、C 3组细胞数分别为 (2 2 .97± 0 .2 3 )、(13 .89± 0 .5 7)和 (3 2 .46± 0 .67)× 10 4个细胞 /ml,差异有统计学意义(P <0 .0 1)。流式细胞仪检测结果显示 ,培养 2d后 ,A组的G2 /M +S期百分数最高 ;6～ 8d后 ,C组的G2 /M +S期百分数最高。成骨细胞上清液中BGP含量 ,C微球组最高 ,其次为A组。结论 rhBMP 2 dex HM可以较长时间持续释放活性rhBMP 2 ,作为rhBMP 2的缓释载体 ,可以明显促进成骨细胞增殖和分化。
Objective To investigate the effect of recombinant human bone morphogenetic protein-2 sustained release hydrogel microspheres (reBMP2-dex-HM) on the proliferation and differentiation of osteoblasts.Methods The secondary culture osteoblasts were divided into three groups according to the different ingredients being added into the DMEM culture medium:rhBMP-2 group (A),dex-HM group (B) and rhBMP2-dex-HM group (C).The proliferation of the cultured osteoblasts was measured by using cell counting method,MTT method and flow cytometry.The contents of bone BGP secreted by osteoblasts were also measured by RIA method.Results The in vitro cellular study showed no significant difference in the cell number and cell viability on the second day after plate culture,but 2 days later the cell number and cell viability in group A were significantly increased as compared with those in other groups (P<0.05).Six to 8 days after plate culture,the cell number and cell viability in group C were increased as compared with other two groups (P<0.01).After 14 days,the cell number in groups A,B and C was ( 22.97± 0.23),( 13.89± 0.57) and ( 32.46± 0.67)×104 cells/ml,with the difference being significant among them (P<0.01).The flow cytometry revealed that the G_2/M+S percentage in group C were the highest 4-8 days after plate culture (P< 0.05).Among the three groups,the content of BGP in group C was the highest.Conclusion The rhBMP2 -dex-HM can promote the proliferation and differentiation of osteoblasts through a long period of controlled release of rhBMP-2.Hydrogel microspheres may be an ideal controlled release carrier for growth factors.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2005年第2期151-153,共3页
Chinese Journal of Experimental Surgery
基金
国家高技术研究发展 863计划资助项目(2 0 0 2AA2 0 50 4 1 )
