P3HB4HB-(GE-PVA)纤维膜载人骨髓间充质干细胞的生物相容性研究

Biocompatibility of Human Bone Marrow Mesenchymal Stem Cells with Coaxial Electrospinning Poly(3-hydroxybutyrate-co-4-hydroxybutyrate)(gelatin-polyvinyl alcohol) in vitro

目的:探讨人骨髓间充质干细胞(h BMSCs)在同轴电纺3-羟基丁酸-4-羟基丁酸共聚酯(P3HB4HB)-明胶(GE)-聚乙烯醇(PVA)纤维膜上的生物相容性。方法:将第5代h BMSCs细胞接种在同轴电纺P3HB4HB-(GE-PVA)纤维膜上作为实验组,单纯培养的h BMSCs细胞为对照组;采用MTS比色法观察接种h BMSCs细胞后1、2及4 h时两组细胞吸光度(OD)值,并测定接种h BMSCs细胞后的第1、3、5及7天时两组的细胞增殖情况;采用吖啶橙(AO)荧光染色及扫描电镜(SEM)观察细胞-材料复合物培养第5天时纤维膜的外观及结构,同时观察h BMSCs细胞的生长情况。结果:随培养时间的增加,两组接种的h BMSCs细胞OD值呈上升趋势,实验组培养4 h时OD值高于对照组(P<0.05);细胞增殖实验发现,2组h BMSCs细胞的OD值随培养时间延长而升高,实验组h BMSCs细胞的OD值高于对照组(P<0.05);同轴电纺P3HB4HB-(GE-PVA)纤维膜外观呈白色膜状、质地均匀,SEM见纤维直径均匀一致,接种到纤维膜生长5 d的h BMSCs细胞形态舒展、细胞间相互连接,AO染色可见完整的细胞膜,大量细胞黏附于纤维膜表面。结论:同轴电纺P3HB4HB-(GE-PVA)纤维膜与h BMSCs有良好的生物相容性。

Objective： To investigate the biocompatibility of human bone marrow mesenchymal stem cells with coaxial electrospinning poly membrane in vitro. Methods： Human bone marrow mesenchymal stem cells at passage 5 were seeded upon the coaxial electrospinning P3HB4HB-（ GE-PVA） membrane as experimental group and upon conventional cultured h BMSCs cells as control group. MTS assay was adopted to observe the cell absorbance at 1,2 and 4 hours and testing the seeded h BMSCs cells proliferation condition at 1,3,5,7 days of both groups. The out appearance and structure of cell and material compound cultured for 5 days were observed by using acridine orange fluorescent staining and scanning electron microscope,and observing growth condition of h BMSCs cells. Result： With the increasing of cultivating time,cell absorbance rate of both groups were increasing,the OD value of experimental group was higher than the control group at 4 h（ P 〈0. 05）; cell proliferation experiment revealed that OD value of both groups h BMSCs cells increased as the growing time of cultivation,the h BMSCs cells OD value of experimental group increased more than control group（ P〈 0. 05）; coaxial electrospinning P3HB4HB-（ GE-PVA） membrane presented in a white membrane shape with even texture,diameter of SEM fiber showed uniformity; h BMSCs cells seeded on fiber membrane for 5 d presented in a form of stretching and inter-cellular connecting; AO staining showed complete cell membrane with substantial amount of cells adhere to the fiber membrane surface. Conclusion： Electrospinning P3HB4HB-（ GE-PVA） membrane shows a good biocompatibility with human bone marrow mesenchymal stem cells.