载肝细胞生长因子聚乳酸－O-羧甲基壳聚糖纳米粒子制备、表征及其对培养大鼠肝细胞活力的影响
摘要
BACKGROUND: Hepatocyte growth factor (HGF) has short half-life period and no target, it is difficult to culture a large amount of mature hepatocytes in vitro and maintain their viability, which restricts the hepatocytes transplantation and bioartificial liver application. OBJECTIVE: To prepare HGF loaded polylactic acid-O-carboxymethylated chitosan (PLA-O-CMC) copolyer nanoparticles, to characterize them, and to explore their degradation in vitro, together with release property. The effect of the nanoparticles on the viability of cultured rat hepatocytes is also studied. DESIGN, TIME AND SETTING: Control observations were carried out in the Hepatobiliary Surgery Institute of Nantong University and the Key Laboratory of Neural Regeneration of Jiangsu Province between July 2006 and January 2008. MATERIALS: Ten SD rats were used in this study, HGF was offered by Pepro Tech Company (England), PLA was offered by Sigma Company (USA), and O-CMC was offered by Shanghai Weikang Biological Products Co., Ltd. METHODS: HGF loaded PLA-O-CMC nanoparticles were prepared by ultrasound combine magnetic stirring method. Primary rat hepatocytes cultured were divided into two groups: culture group (co-culturing hepatocyten with HGF loaded nanoparticles) and control group (common culture without HGF). The viability of the cultured hepatocytes was detected by Cell Counting Kit-8 within one week. MAIN OUTCOME MEASURES: The nanoparticles were characterized by electron microscopy, laser particle size analyzer and Zeta potential analyzer. The morphological change and weight loss of the nanoparticles during the degradation were studied. Ph change of degradation medium was also observed as well. RESULTS: The characterion HGF loaded PLA-O-CMC nanoparticles showed that the mean size was 140 nm, polydispersity was 0.108, HGF-loadtng level was 0.126 65%, HGF-embedding ratio was 76.32%, and the Zeta potential was 32.8 eV. The equation of HGF release from the nanoparticles in early 24 hours wa