聚乳酸-O-羧甲基壳聚糖纳米粒子的制备及其对培养猪肝细胞的影响  

Preparation of polylactic acid O-carboxymethylated chitosan nanoparticles and their influences on the cultured porcine hepatocytes

作　　者：陈钟[1] 戴新征[1] 杨欣荣[1] 祝文彩[1] 

机构地区：[1]南通大学附属医院肝胆外科南通大学肝胆外科研究所,江苏省南通市226001

出　　处：《世界华人消化杂志》2006年第17期1669-1674,共6页World Chinese Journal of Digestology

基　　金：江苏省卫生厅重大基金资助项目.No.H200215南通大学创新基金资助项目;No.CX0301~~

摘　　要：目的:制备聚乳酸-O-羧甲基壳聚糖纳米粒子并探讨其对培养猪肝细胞形态和功能的影响．方法:以聚乳酸和O-羧甲基壳聚糖为基质材料,采用超声波法制备了聚乳酸-O-羧甲基壳聚糖纳米粒子,用原子力显微镜对其进行了物理表征,用X射线光电子光谱法对其表面的化学结构进行了表征．用聚乳酸-O-羧甲基壳聚糖纳米粒子进行原代猪肝细胞培养,并以普通培养作为对照组．观察培养后1 wk内肝细胞的形态和功能变化．结果:聚乳酸-O-羧甲基壳聚糖纳米粒子制备成功,原子力显微镜下其直径300-500 nm．经过XPS光谱分析显示,C,O,N的比例分别为72．51％,23．69％,3．80％．普通培养的肝细胞,48 h后逐渐失去多边形特征．而与PLA-O- CMC共培养48 h后,肝细胞形成细胞黏附聚集的球形体,大部分肝细胞重建细胞极性,呈现较典型、均匀的多边形形态特征．培养后 24 h,3 d和5 d时,纳米细胞培养组ALB含量高于普通培养组(3．53±0．052,3．48±0．075,3．57 ±0．137g/L vs 3．10±0．179,3．17±0．186,3．10 ±0．219 g／L,均P<0．05);3,5,7 d时,纳米细胞培养组ALT水平明显低于普通组(15．33± 13．05,8．84±8．87,7．00±5．22 nkat／L vs 24．17 ±20.35,16．17±27．49,15．50±11．95 nkat／L, 均P<0．05),而BUN含量虽然有所升高,但是不显著(P>0．05)．结论:聚乳酸-O-羧甲基壳聚糖纳米粒子是肝细胞培养的良好材料．AIM： To prepare polylactic acid-O-carboxymethylated chitosan （PLA-O-CMC） nanoparticles and evaluate their influences on the morphology and functions of cultured porcine hepatocytes. METHODS： PLA-O-CMC nanoparticles were prepared from PLA and O-CMC by ultrasonic method. The nanoparticles were characterized by atomic force microscopy and X-ray photoelectron spectrometry. Isolated porcine hepatocytes were divided into two groups： commonly culturing group and nanomaterial group （co-culturing hepatocytes with PLA-O-CMC nanoparticles）. The morphology and functions of the cultured hepatocytes were observed within 1 wk. RESULTS： PLA-O-CMC nanoparticles were successfully prepared, and the diameter was 300-500 nm under atomic force microscope. X-ray photoelectron spectrometry showed that the proportions of C, O, and N element were 72.51%, 23.69%, and 3.80%, respectively. The hepatocytes lost their features of polygons after common culture for 48 h. However, after co-culture with PLA-O-CMC nanoparticles, the hepatocytes clustered into conglobulations, and most of them restored their cell polarities, shaping as typical polygons. After culture for 1, 3, and 5 d, the content of albumin （ALB） was significantly higher in nanomaterial group than that in commonly culturing group （3.53 ± 0.052, 3.48 ± 0.075, 3.57 ± 0.137 g/L vs 3.10 ± 0.179, 3.17 ± 0.186, 3.10 ± 0.219 g/L, all P 〈 0.05）, while on day 3, 5 and 7, the level of alanine aminotransferase （ALT） was markedly lower （15.33 ± 13.05, 8.84± 8.87, 7.00 ± 5.22 nkat/L vs 24.17 ± 20.35, 16.17 ± 27.49, 15.50 ± 11.95 nkat/L, all P 〈 0.05）. From day 3 to 7, the content of supernant blood urea nitrogen （BUN） was higher in nanomaterial group than that in commonly culturing group, but there was no significant difference （P 〉 0.05）. CONCLUSION： PLA-O-CMC nanoparticles are successfully prepared, and can be used in the culture of hepatocytes.

关 键 词：聚乳酸-O-羧甲基壳聚糖 纳米材料 肝细胞 培养 肝细胞移植 

分 类 号：R575.3[医药卫生—消化系统]