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

›› 2019, Vol. 10 ›› Issue (2): 73-82.

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

陈飞扬,卜寿山,庄海,龚春玲,张继生

南京医科大学第一附属医院口腔科

收稿日期:2019-03-01 修回日期:2019-03-13 出版日期:2019-06-25 发布日期:2019-07-01
通讯作者: 卜寿山 E-mail:bushsh@vip.sina.com
基金资助:
复合3-D仿生基质骨的构建及其修复骨缺损的研究;BMSCs复合3-D仿生基质骨的构建及其修复颌骨缺损的研究

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

Received:2019-03-01 Revised:2019-03-13 Online:2019-06-25 Published:2019-07-01

摘要/Abstract

摘要： 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21 d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。

关键词: 静电纺丝, 丝素蛋白, 壳聚糖, 人骨髓间充质干细胞, 增殖, 成骨

Abstract: 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.

Key words: electrospun, silk fibroin, chitosan, ,hBMSCs, proliferation, osteogenic differentiation

陈飞扬卜寿山庄海龚春玲张继生. 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[J]. , 2019, 10(2): 73-82.

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