生物交联剂京尼平对静电纺明胶纳米纤维膜改性的影响

Genipin effects on chemical modification of electrospun gelatin nanofibrous membrane

背景:生物高分子纳米纤维膜极不稳定,易水解,所以需要进行交联改性。而以往所采用的交联剂具有一定的细胞毒性,降低了材料的生物相容性。目的:用生物交联剂京尼平对静电纺明胶纤维膜进行交联处理,观察交联产物的理化性能和生物相容性。设计、时间及地点:观察性实验,于2008-03/10在东华大学生物材料与组织工程研究实验室完成。材料:将交联剂京尼平按质量比为0.0%,2.5%,5.0%,7.5%,10%加入明胶溶液中共混,通过静电纺制备纳米纤维膜。方法:扫描电镜样品经表面喷金后在10kV加速电压下观察纤维表面的形貌。在万能材料测试机测试其拉伸力学性能。采用MTT法测试猪动脉血管内皮细胞在纳米纤维膜上的黏附与增殖情况。主要观察指标:共混静电纺明胶纳米纤维的形态结构、力学性能、生物相容性。结果:通过扫描电镜观测发现京尼平共混交联的明胶纳米纤维尺寸略有增大,当京尼平含量为5.0%时,纤维直径最大,增大了约200nm;力学测试显示材料的力学性能在添加京尼平之后有了明显提高,当京尼平含量为5.0%时,应力达到了(2.45±0.09)MPa,应变达到了(3.85±0.57)%;生物相容性实验表明猪动脉血管内皮细胞在经京尼平处理过的明胶纳米纤维膜上能有效地黏附与增殖。结论:含有京尼平的明胶纳米纤维膜各项理化性能都有了显著的提高,与猪动脉血管内皮细胞复合具有良好的生物相容性。

BACKGROUND： Biopolymer nanofibrous mats were instable, and hydrolyzed easily. Thus, crosslinking was needed. While crosslinging agents used before had cell cytotoxicity, and the biocompatibility were decreased. OBJECTIVE： Electrospin gelatin fibrous scaffolds were crosslinked with genipin, a natural crosslinking agent, then to investigate the physicochemical properties and biocompatibility of crosslinged products. DESIGN, TIME AND SETFING： An experiment based on observation was conducted at the Institute of Biomaterial and Tissue Engineering, Donghua University from March to October 2008. MATERIALS： Genipin （mass ratior of 0.0%, 2.5%, 5.0%, 7.5%, 10%） was mixed with gelatin to prepare nanofibrous mats by electrospinning technique. METHODS： The properties of electrospun mats were investigated by the scanning electron microscopy following gold sputtering under 10 kV accelerating voltage. Tension mechanical property was tested on a testing machine. Adhesion and proliferation of porcine artery endothelial cells on the nanofibrous mats were examined by MTT. MAIN OUTCOME MEASURES： Morphous structure, mechanical test, biocompatibility of electrospun gelatin fibers. RESULTS： Scanning electron microscope images indicated the diameter of fibers increased after crosslinked by genipin. While genipin reached to 5.0%, the diameter of fibers achieved maximum, and increased by 200 nm. The mechanical test showed that tensile strength enhanced after adding genipin. While genipin reached to 5.0%, the stress reached to （2.45±0.09） Mpa, and the strain reached to （3.85±0.57）%. Biocompatibility test suggested that porcine artery endothelial cells could effectively adhere and proliferate on the gelatin nanofibrous mats following genipin treatment, CONCLUSION： The physicochemical properties of gelatin/genipin fibers can be enhanced obviously and their biocompatibility is good with porcine artery endothelial cells.