镶嵌纳米级再生丝素蛋白聚乙烯醇膜的制备与促细胞生长性能

Preparation of Polyvinyl Alcohol Film Inlaid with Silk Fibroin Peptide Nano-scale Particles and Evaluation of Its Function to Promote Cell Growth

从废弃蚕茧及缫丝下脚料中提取纯净丝素蛋白,采用盐溶酶解法制备纳米级丝素蛋白（SFP）,按照不同配比与聚乙烯醇（PVA）采用水相混合后制备出嵌入纳米级丝素蛋白颗粒的聚乙烯醇（SFP in PVA）膜,对膜进行表征并测定其促细胞生长性能。使用60%NaSCN溶解丝素蛋白后,采用α-糜蛋白酶、胰蛋白酶、中性蛋白酶进行酶解筛选试验,通过场发射扫描电镜观察,发现中性蛋白酶酶解得到的SFP粉末尺寸约为80~150 nm,颗粒均匀。红外光谱表征确认了SFP/PVA膜的基团间结合;扫描电镜显示SFP以微细线段状镶嵌在PVA膜中。对镶嵌膜的SFP/PVA进行了10/90、20/80、30/70、40/60四种配比的筛选试验,通过膜的表面观察、水中形态稳定性观测和促CHO细胞生长性能测试（MTT法）,结果显示,SFP/PVA配比为30/70的镶嵌膜表面光滑、水中形态稳定,促进CHO细胞快速黏附和生长性能最优。

Nano-scale particles of silk fibroin peptide（SFP） were prepared from discarded materials of cocoon or filature by dissolving and enzymolysis.Polyvinyl Alcohol films inlaid with silk fibroin peptide nano-scale particles（SFP in PVA） were prepared by blending nano-SFP and PVA in water according to different blending ratios.The films＇ characteristics and their promoting cell growth functions were investigated.Silk fibroin fiber was dissolved in 60%NaSCN solution,and was decomposed with α-Chymotrypsin,Trypsin and Neutral,respectively.The uniformity of size of SFP nano-particles prepared by Neutral was better and appeared about 80-150nm.（SFP in PVA） films were characterized by infrared spectroscopy（IR） measurement which demonstrated the combination of SFP and PVA.Scanning electron microscopy revealed the PVA films already inlaid with SFP micro-segment.The surface and form stability in water of the（SFP in PVA） films with blending ratios of 10/90,20/80,30/70 and 40/60 were observed.And the results showed that SFP/PVA film with the blending ratio of 30/70 has smoother surface and better stability in water.The Chinese hamster ovary（CHO） cells were cultured,and the promoting cell growth function of（SFP in PVA） films was assessed by MTT colorimetric assay.These findings indicate that SFP /PVA（30/70） film has excellent function of promoting cell growth.