EIF-GT/PCL纳米纤维电纺膜体外诱导hADSCs向表皮细胞表型转化的相关研究

In Vitro Study of Differentiation of hADSCs into Epidermal Cell Linage Induced by EIF-GT/PCL Electrospun Nanofibrous Membrane

目的探讨表皮诱导因子复合体(Epidermal inducing factors,EIF)-明胶(Gelatin,GT)/聚己内酯(Polycaprolactone,PCL)纳米纤维电纺膜,体外诱导人脂肪干细胞(Human adipose derived stem cells,hADSCs)向表皮细胞表型转化的可行性,以及相关的生物学特性。方法采用混合静电纺丝法制备EIF-GT/PCL纳米纤维电纺膜(实验组)和GT/PCL纳米纤维电纺膜(对照组),进行形态学观察和力学测试;通过酶联免疫吸附测定法绘制EIF-GT/PCL纳米纤维电纺膜的药物释放曲线;将hADSCs接种于两种电纺膜上进行体外培养,扫描电镜观察细胞-纳米纤维电纺膜黏附情况,CCK-8法检测细胞增殖情况,免疫荧光法检测CK10及CD105的表达情况。结果实验组和对照组具有相似的纳米级结构,形态、密度均匀,力学特性佳;实验组能够平稳释放EIF达15 d以上;hADSCs在实验组上黏附更好,增殖更快。培养7 d后,实验组上hADSCs表达CK10,间充质干细胞标志物CD105表达减弱。结论 EIF-GT/PCL纳米纤维电纺膜具有良好的生物学特性,可诱导hADSCs向表皮细胞表型转化。

Objective To study the biological characteristics of epidermal inducing factors(EIF)-gelatin(GT)/polycaprolactone(PCL)electrospun nanofibrous membrane and its ability of inducing hADSCs into epidermal cell linage.Methods EIF-GT/PCL electrospun nanofibrous membrane(experimental group)was produced by blending electrospun technology,and the GT/PCL electrospun nanofibrous membrane was produced as control group.Their morphology and mechanical characteristics were observed.The accumulated EIF releaserate-time curve was studied using ELISA.The hADSCs were seeded onto the membrane of the two groups and were cultured in vitro.Scanning electron microscope(SEM)was used to observe the cell-membrane adhesion.The hADSCs proliferation on was measured by CCK-8 and their differentiation into epidermal cell linage was confirmed by immunofluorescence of CK10(an early marker of epidermal differentiation)and CD105(a marker of mesenchymal stem cell).Results The experimental group and control group have similar nano-scale structure with uniform shape and density as well as good mechanical properties.Experimental group was able to load and stably release the EIF for more than 15 days.After seeded onto experimental group for 7 days,the hADSCs proliferated well and expressed CK10,while the CD105 expression was down regulated.Conclusion Experimental group has good biological characteristics and can induce hADSCs into the epidermal cell linage.