hBMP2修饰的β-TCP/胶原支架对MC3T3-E1细胞分化的影响  

The evaluation of β tricalcium phosphate/collagen scaffold modified by human bone morphogenetic proteins 2 plasmid on the differentiation of MC3T3-E

作　　者：阮蔷 赵刚[1] 肖月[1] 孙佳宁[1] 

机构地区：[1]佳木斯大学口腔医学院,黑龙江佳木斯154003

出　　处：《黑龙江医药科学》2016年第5期5-7,4,共4页Heilongjiang Medicine and Pharmacy

基　　金：黑龙江省自然科学基金项目;编号:H201487;佳木斯大学研究生创新项目;编号:LZR2015_015

摘　　要：目的:构建含h BMP2(Human Bone morphogenetic proteins2,h BMP2)质粒DNA的β-TCP/胶原(β-Tricalcium Phosphate/Collagenβ-TCP/胶原)支架材料,并研究其对MC3T3-E1细胞分化的影响。方法:制备纳米级多孔β-TCP/胶原支架并负载含BMP2目的基因的质粒DNA形成基因修饰的支架材料。建立MC3T3-E1细胞株与复合支架的体外培养体系。将其分为支架组和平皿组,再根据质粒的不同浓度每组再分为4个小组。复合培养后取样通过扫描电镜和光镜进行细胞形态学观察;1、3、7、14d ALP活性检测测量细胞碱性磷酸酶的活性,1、3、7、14d茜素红染色观察,实时荧光定量PCR检测细胞周期1、3、7、14d观察Runx2、OCN、ALP、OPN因子,检测结果处理并进行统计学分析。结果:含h BMP2为目的基因的质粒DNA修饰纳米β-TCP/I型胶原溶液复合材料上细胞黏附数、分化数量及材料表面分布都高于单纯含h BMP2为目的基因的质粒DNA,具有统计学意义(P<0.05),通过用碱性磷酸酶活性ALP、茜素红染色检测结果显示支架组对MC3T3-E1细胞成骨能力的影响优于平皿组,具有统计学意义(P<0.05)。结论:负载h BMP2基因修饰的β-TCP/胶原支架材料具有良好的生物相容性、优良的骨诱导性和骨传导性,是一种理想的骨缺损修复材料。Objective： To build containing h BMP2（ Human Bone morphogenetic proteins2,h BMP2） plasmid DNA β-TCP（ β-Tricalcium Phosphate/Collagen β-TCP）/collagen scaffold material,and to study its effect on the differentiation ability of MC3T3-E1 cells. Methods： The nanoscale porous β-TCP/collagen/collagen scaffold loaded by BMP2 gene plasmid DNA genetic modification of scaffold materials was prepared. MC3T3-E1 cells lines and composite scaffolds in vitro culture system was established. It can be divided into stent group and dish group. According to different concentration of plasmid,each group could be also divided into four groups. The sample cell morphology was selected after compound training by scanning electron microscope and light microscope for further observation. 1,3,7,14 days activity detection measurement cells the activity of alkaline Phosph-atase1,3,7,14 days observe the alizarin red staining. Test results were processed and statistically analyzed. Results：For the purpose gene plasmid DNA containing h BMP2 modified nano beta TCP/solutions of type I collagen composite cells adhesion number,quantity and distribution of material surface is higher than that of pure containing h BMP-2 for the purpose gene plasmid DNA（ P〈0. 05）. By using alkaline phosphatase activity of ALP,and Alizarin red staining,the test results show that the stent group of cells of MC3T3-E1 osteogenesis ability than the influence of the dish group. Conclusion： We conclude that porous β-TCP/collagen scaffold loaded with BMP2 DNA is an appropriate candidate for bone defect repair material.

关 键 词：BMP β-TCP/胶原 分化 骨缺损 

分 类 号：Q254[生物学—细胞生物学]