hBMP2修饰的β-TCP/胶原支架对MC3T3-E1细胞成骨能力的影响

The effects of β tricalcium phosphate/collagen scaffold loaded with human bone morphogenetic protein 2 plasmid on the osteogenesis ability of MC3T3-E1 cells

目的:构建含hBMP2(human bone morphogenetic proteins 2)质粒DNA的β-TCP/胶原(β-tricalcium phosphate/collagen)支架材料,并研究其对MC3T3-E1细胞成骨能力的影响。方法:制备纳米级多孔β-TCP/胶原支架并负载含hBMP2目的 DNA基因及对照质粒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hBMP2组(Z)对照质粒组(Z0),平皿hBMP2组(M)和对照质粒组(M0)。复合培养后取样通过扫描电镜观察支架表面形态,成骨诱导1、3、7、14 d检测不同浓度BMP2支架组和非支架组细胞碱性磷酸酶(ALP)的活性,并在成骨诱导的不同时间点采用实时荧光定量PCR的方法检测Runx2、OCN、ALP、OPN等成骨相关标志基因表达。检测结果进行统计学分析。结果:含hBMP2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表面呈多孔样结构;支架组和平皿组中加入hBMP2质粒DNA都能提高ALP的活性以及成骨相关标志基因的表达;支架组对MC3T3-E1细胞成骨促进能力优于平皿组。结论:负载hBMP2基因修饰的β-TCP/胶原支架材料具有良好的骨诱导性。

Objective： To study the effects of beta tricalcium phosphate（ β-TCP）/collagen scaffold loaded with human bone morphoge- netic protein 2（hBMP2） plasmid on the osteogenesis ability of MC3T3-E1 cells. Methods： hBMP2 DNA plasmid-modified β-TCP/collagen scaffold and the naked plasmid（control） were constructed. MC3T3-E1 cells were respectively in vitro cultured onto the β-TCP/collagen scaffold with hBMP2 （Z） and with control plasmid（ Z0 ）, on peace dish with the saffold and hBMP2 （M） and with the control plasmid（M0）. The surface morphology of the samples was observed by SEM. Osteogenesis of the cells was examined by alkaline phosphatase activity（ALP） test, real-time fluorescent quantitative PCR for the detection of Runx2, OCN, ALP and OPN mRNA expression. Data were statistically analyzed. Results： The composite sample surface of plasmid DNA containing hBMP2 modified β-TCP/collagen was por- ous ; group Z and M showed highter ALP activity and higher mRNA expression of Runx2, OCN, ALP and OPN than group Z0 and M0 ; so did group Z than group M. Conclusion： Porous β-TCP/collagen scaffold loaded with BMP2 DNA is potential for osteoinduction.