电纺胶原和蛛丝纳米纤维膜的制备、表征及生物安全性比较

Comparison of preparation, characterizations and biological safety between electrospun collagen and spider silk nanofibers

目的:制备胶原和蜘蛛丝两种纳米纤维膜,进行理化性能表征和生物相容性比较,以期将蜘蛛丝纳米纤维用于组织工程支架材料。方法:实验于2006-07/2007-08在上海市东华大学生物科学与技术研究所完成。将胶原和蜘蛛丝分别以80g/L溶于六氟异丙醇,采用高15kV压静电纺制成纳米纤维膜,真空干燥后对其理化性能进行表征。扫描电镜观察超微结构,并进行水接触角测量和水解稳定性测量;并采用MTT实验比较猪大动脉内皮细胞在两种纤维膜表面的黏附、生长和增殖等情况。结果:①静电纺胶原和蛛丝纳米纤维膜均具有良好三维多孔结构,但蛛丝纤维直径更均匀。蛛丝膜具有较大的水接触角,在水解稳定性测试中质量损失较少。②MTT实验表明,种植6h后血管内皮细胞在胶原和蛛丝膜上都能黏附,但蛛丝膜上细胞增殖速度较快,2d后超过胶原膜,7d后蛛丝膜上细胞多于胶原膜表面40%以上。结论:胶原和蛛丝都能促进血管内皮细胞黏附、生长。蛛丝膜具有较强的疏水性和水解稳定性,在体外培养过程中蛛丝更有利于细胞增殖,有望作为血管组织工程支架材料。

AIM： To prepare the electrospun collagen and spider silk nanofibers as the scaffolds for tissue engineering cell carrier, and compare their characteristics and biocompatibility. METHODS： The experiment was conducted in the Institute of Biological Science and Biotechnology, Donghua University from July 2006 to August 2007. The collagen and spider silk each 80 g/L were dissolved in hexafluoro isopropanol, respectively. The prepared solutions underwent electrospun to make nanofibrous membrane at 15 kV. The physical and chemical properties were investigated after vacuum drying, and the ultrastructure was observed under scanning electron microscope for water contact angle and hydrolytic stability detection. The attachment, growth and proliferation of porcine aortic endothelial cells （ECs） on both materials were examined by MTT assay. RESULTS： （1）Both the electrospun collagen and spider silk nanofibrous membrane had porous three-dimensional structures, but the spider silk nanofibrous membrane exhibited more uniform diameter distribution, higher value of contact angle and less mass loss than collagen material. （2）MTT assay showed that ECs could adhere on collagen and spider silk materials, but the cells on spider silk nanofibrous membrane proliferated faster than on collagen on day 2, and the number of cells was 40% more than that on collagen material after 7 days culture. CONCLUSION： Collagen and spider silk nanofibers can improve the attachment and growth of ECs, but spider silk material shows better hydrophobicity and hydrolytic stability than collagen one, and can promote cell proliferation. Spider silk is a promising biomaterial as the scaffolds for tissue engineering.