静电纺SF/CS复合纤维支架对hBMSCs体外细胞增殖及成骨分化的影响

Effect of electrospun SF/CS composite fiber scaffold on cell proliferation and osteogenic differentiation of hBMSCs in vitro

目的:用静电纺丝法制备丝素(SF)/壳聚糖(CS)纳米纤维膜支架,评价其性能及对人骨髓间充质干细胞(hBMSCs)增殖和成骨分化的影响。方法:将SF、CS以质量比1∶0和1∶1共同溶于三氟乙酸和二氯甲烷混合溶剂中,采用静电纺丝法制备SF电纺膜支架和SF/CS电纺膜支架。通过扫描电子显微镜(SEM)、红外光谱(FTIR)、热重分析(TG/DTG)等对电纺膜的结构性能进行表征。hBMSCs分别接种于SF、SF/CS支架表面(对照组直接接种于培养皿),进行成骨诱导培养,CCK-8实验研究hBMSCs生长增殖情况,能谱仪(EDS)、茜素红染色(ARS)检测各组hBMSCs成骨矿化能力。结果:SEM和FTIR结果显示,与SF支架比较,SF/CS支架纤维直径更为均一,构象更为稳定;而TG/DTG结果显示,SF组热稳定性更佳;与对照组比较,SF组和SF/CS组hBMSCs增殖能力无显著差异(P>0.05);培养21d后,元素分析表明SF/CS组钙元素含量更高;与对照组及SF组相比,SF/CS组hBMSCs钙化结节明显增多且染色较深。结论:静电纺SF/CS纳米纤维支架生物相容性佳,对hBMSCs成骨分化有促进作用。

Objective: Using electrospinning to preparesilk fibroin/chitosan(SF/CS) nanofiber membrane scaffolds, and then evaluating its properties and effects on proliferation and osteogenic differentiation of human bone marrow mesenchymal stem cells(hBMSCs). Methods: The regenerated silk fibroin(SF) and chitosan(CS) were dissolved in the mixed solvent system of trifluoroacetic acid and dichloromethane by mass ratio (1∶0, 1∶1). SF and SF/CS nanofiber scaffolds then were prepared by electrospinning. The structure and properties of the electrospun films were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), and thermal gravity/differential thermal gravity analysis (TG/DTG). Cells in the experimental group were inoculated on the surface of SF and SF/CS membrane respectively. Cells in the control group were directly inoculated in culture dish. hBMSCs were used in each group to induce osteogenesis. CCK-8 was used to study the growth and proliferation of cells. Energy dispersive spectrometer (EDS) and alizarin red staining (ARS) were used to detect the ability of osteogenesis and mineralization. Results: Compared with SF scaffolds, SF/CS scaffolds had more uniform fiber diameter (SEM) and more stable conformation (FTIR);TG/DTG results showed that SF scaffolds had more thermal stability. CCK-8 showed that compared with the control group, there was no significant difference in proliferation of hBMSCs between SF and SF/CS groups when co-cultured for 5 and 7 days( P >0.05). After 21 days of culture, elemental analysis indicated that the SF/CS group had higher calcium content. Compared with the control group and SF group, calcified nodules of hBMSCs in SF/CS group were significantly increased and staining was deep. Conclusions: Electrospinning SF/CS nanofibers scaffolds have good biocompatibility and can promote osteogenic differentiation of hBMSCs.