MC3T3-E1细胞在四种生物膜上粘附增殖能力的比较研究

›› 2014, Vol. 5 ›› Issue (3): 130-134.

MC3T3-E1细胞在四种生物膜上粘附增殖能力的比较研究

戴沄¹,陈汉帮¹,李娜²,陈刚³,夏阳⁴,章非敏⁵

1. 江苏省口腔医院
2. 南京医科大学口腔研究所
3. 南京医科大学口腔医学研究所?附属口腔医院修复科
4. 南京医科大学口腔医学研究所附属口腔医院
5. 南京医科大学口腔医学研究所附属口腔医院修复科

收稿日期:2014-08-14 修回日期:2014-09-15 出版日期:2014-09-25 发布日期:2014-09-24
通讯作者: 章非敏 E-mail:fmzhang@njmu.edu.cn
基金资助:
江苏高校优势学科建设工程项目;江苏省医学领军人才与创新团队

Comparison of four kinds of biological membranes and their influence on adhesion and proliferation of MC3T3-E1 cells

Received:2014-08-14 Revised:2014-09-15 Online:2014-09-25 Published:2014-09-24

摘要/Abstract

摘要： 目的：研究四种生物膜的表面形态及生物相容性，比较它们对MC3T3-E1细胞初期粘附增殖能力的影响，及成膜材料与制备方法之间的交互作用。方法：以聚己内酯[poly（ε-caprolactone），PCL]与聚乳酸-羟基乙酸[poly（lactic-co-glycolic acid），PLGA]作为膜材料，浇铸及电纺两种方法分别制备生物膜，使用场发射扫描电子显微镜观察四种膜表面结构，CCK-8法检测膜上MC3T3-E1细胞1、3、7 d增殖量，激光共聚焦显微镜评价细胞在材料表面的粘附情况，实时荧光定量PCR反应检测粘附相关基因。结果：扫描电镜观察到浇铸膜表面具有孔隙结构，电纺膜表面电纺丝交织成网；CCK-8检测中3 d、7 d时PLGA膜上细胞增殖量大于PCL膜（P<0.05），与制备方法无关（P＞0.05）；激光共聚焦显微镜观察到除PCL电纺膜表面细胞铺展不佳外，其余膜上细胞生长良好；实时荧光定量PCR检测发现PCL浇铸膜上细胞整合素基因表达量高于其他组（P<0.05），且不同材料与制备方法间存在交互作用。结论：PLGA膜有利于MC3T3-E1细胞增殖，而PCL浇铸膜适合细胞粘附。因此，相对单一材料或成膜方式，PLGA与PCL及浇铸、电纺法的混合膜可成为以后发展的方向。

Abstract: Objective：To study the surface characteristics and biocompatibility of four kinds of biological membranes in order to compare their applications of cell adhesion and proliferation，as well as interaction between materials and membrane types. Methods：By casting or electrospinning, we fabricated poly(ε-caprolactone)(PCL) membranes and poly(lactic-co-glycolic acid)(PLGA) membranes. Field emission scanning electron microscope (FESEM) was used to detect the surface microstructure of the membranes. After Cell line MC3T3-E1 was seeding to all membranes, we tested cell proliferation of 1,3,7days by Cell counting Kit-8 (CCK-8) assay, while estimating cell adhesion by confocal laser scanning microscope (CLSM) and real-time quantitative polymerase chain reaction (RT-qPCR). Results：FESEM pictures showed pores on cast membranes, while interwoven fibers were observed on electrospun membrane surfaces. CCK-8 tests showed MC3T3-E1cells prefer two PLGA groups than PCL groups（P<0.05）, no matter how the membranes were fabricated（P＞0.05）. Despite PCL electrospun membranes, cells were observed spreading well on other membranes by CLSM. RT-qPCR tests showed there are interactions between materials and membrane types, and PCL cast membrane group showed highest integrin gene expression（P<0.05）. Conclusions：PLGA membranes contributed to MC3T3-E1 cell proliferation, while PCL cast membranes were suitable for cellular adhesion. Other than singly using one kind of material or fabrication method, combination use of PLGA, PCL by casting or electrospinning will become the trend to fabricate biological membranes.

戴沄陈汉帮李娜陈刚夏阳章非敏. MC3T3-E1细胞在四种生物膜上粘附增殖能力的比较研究[J]. , 2014, 5(3): 130-134.

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