Regenerated keratin proteins as potential biomaterial for drug delivery
Regenerated keratin proteins as potential biomaterial for drug delivery
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DOI：
10.1002/pat.3168
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发表时间：
2013-11-01
影响因子：
3.4
通讯作者：
Bellosta, S.
中科院分区：
文献类型：
--
作者：
Cilurzo, F.;Selmin, F.;Bellosta, S.
本研究旨在通过研究再生角蛋白（RKs）的可加工性和细胞毒性，初步评估其在微颗粒给药系统设计中的可靠性。以羊毛废料为原料，采用亚硫酸盐裂解法提取RKs。以4.5%w/w的RK溶液喷雾干燥，在压力下用蒸汽对微粒进行灭菌。采用扫描电镜、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳、傅里叶变换红外光谱等方法对RKs及其微粒进行了表征。通过乳酸脱氢酶对人单核细胞系Tamm-Horsfall糖蛋白-1的释放测定体外细胞毒性。获得了粒径分布窄且单峰的r基微粒子（类似于6 μ m）。它们有一个表面光滑的葡萄干状结构。灭菌后的微颗粒形态和RK分子量均保存完好。酰胺I带的曲线拟合表明，微颗粒中的RK普遍存在于无序/-螺旋二级结构中，这使得蛋白质可溶于水。为了促进薄片二级结构的结晶，从而提高不溶于水性，将r基微颗粒浸泡在pH3.5的醋酸水溶液中过夜。RK在任何浓度（在1ml细胞培养基中从1到1000 μ g无菌微粒）或时间点（24-72h）均未诱导任何明显的细胞毒性。这些初步数据表明，以废羊毛为原料生产RK生物相容性微颗粒是可行的。版权所有：John Wiley & Sons， Ltd。
This work aims to preliminarily evaluate the reliability of regenerated keratins (RKs) in the design of microparticulate drug delivery systems by studying their processability and cytotoxicity. RKs were extracted by sulfitolysis from wool waste. A 4.5%w/w RK solution was spray-dried, and microparticles were sterilized by steam vapor under pressure. Scanning electron microscope, sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and Fourier transform infrared spectroscopy were used to characterize RKs and microparticles thereof. The in vitro cytotoxicity was determined by assessing the release of lactate dehydrogenase in the human monocytic cell line Tamm-Horsfall glycoprotein-1. RK-based microparticles with a narrow and unimodal particle size distribution (similar to 6 mu m) were obtained. They had a raisin-like structure with a smooth surface. Both microparticle morphology and RK molecular weight were well-preserved after sterilization. The curve fitting of the amide I bands showed that RK in the microparticles was prevalently present in the disordered/-helix secondary structures which made the protein soluble in water. To promote crystallization in the -sheet secondary structure and, therefore, water insolubility, RK-based microparticles were immersed in an aqueous solution of acetic acid at pH3.5 overnight. RK did not induce any appreciable cellular cytotoxicity at any of the concentrations (from 1 up to 1000 mu g sterile microparticles in 1ml cell culture medium) or time-points (24-72h) tested. These preliminary data suggest the feasibility of producing RK biocompatible microparticles using waste wool as starting material. Copyright (c) 2013 John Wiley & Sons, Ltd.