PLA-O-CMC纳米粒子培养的猪肝细胞移植对急性肝衰竭大鼠肝再生作用的初步研究

目的:探讨胶原凝胶包埋聚乳酸-O-羧甲基壳聚糖(PLA-O-CMC)纳米粒子粘附培养猪肝细胞移植对急性肝衰竭(ALF)大鼠肝的再生作用。方法:D-氨基半乳糖腹腔内注射制作大鼠ALF模型;48h后分别将5mlⅠ型胶原凝胶包埋的PLA-O-CMC纳米粒子粘附培养24h猪的肝细胞、5mlⅠ型胶原凝胶固定培养24h的猪肝细胞、5ml培养24h的猪肝细胞悬液(均含5.0×107个)移植到各组ALF大鼠腹腔内,并以5mlRPMI1640腹腔内注射作为阴性对照。观察大鼠14天存活率和受体肝脏的病理变化。结果:移植后14天ALF大鼠的存活率:单纯肝细胞移植组(Ⅰ组)为56.25%,胶原肝细胞移植组(Ⅱ组)为62.5%,纳米胶原肝细胞移植组(Ⅲ组)为75%,阴性对照组(Ⅳ组)为18.75%,各移植组14天存活率高于对照组(P<0.05),各移植组间无统计学意义(P>0.05)。ALF大鼠肝脏再生于ALF后5～7天最为显著;各组肝脏病理变化以Ⅲ组恢复最好,双核细胞最多,肝再生最快。结论:应用PLA-O-CMC纳米粒子和Ⅰ型胶原联合培养猪肝细胞移植于ALF大鼠腹腔内能促进肝脏的再生。

The preliminary study of liver regeneration effects of porcine hepatocytes transplantation by using PLA-O-CMC nano-particles on treating rats with acute liver failure

Objective:To evaluate the liver regeneration effects of intraperitoneal transplantation of polylactic acid-O-carboxymethylated chitosan(PLA-O-CMC)nanoparticles-attached porcine hepatocytes embedded in TypeⅠcollagen gel on acute liver failure(ALF)in rats induced by D-galactosamine(D-Gal).Methods:The isolated porcine hepocytes were co-cultured with PLA-O-CMC nanoparticles and Type Ⅰcollagen gel for 24h.5ml chitosan nanoparticles-attached porcine hepatocytes(5.0×107cells)embedded in Type Ⅰcollagen gel were transplanted into the abdominal cavity of SD rats at 48h after D-gal introperitonear injection(1.2g/kg)(Group Ⅲ).5ml pig hepatocytes(5.0×107cells)embedded in Type Ⅰcollagen gel(Group Ⅱ)and 5ml pig free hepatocyte suspension(5.0×107cells)(GroupⅠ)were transplanted into abdominal cavity of SD rats.5ml RPMI 1640 injection(Group Ⅳ)was used as the negative group.The 14th survival rate and the pathological change of ALF rat livers were observed.Results:There was significant difference in the survival rate at 14th day among Group I(56.25%),Group II(62.5%),Group III(75%),Group IV(18.75%)(P<0.05).The survival rate of Group III was the highestp in the four groups,but there wase no significant difference in GroupⅠ,GroupⅡand Group Ⅲ.The liver pathological damage were quickly restored in GroupⅠ,GroupⅡand Group Ⅲ as compared with Group Ⅳ.The liver regeneration mainly appeared about 5-7d after hepatocyte transplantation and it in Group Ⅲ was the most apperent,follwed by Group Ⅱ and GroupⅠ,with the slowest one in Group Ⅳ(The number of double nuclear liver cells was the hightest in Group III).Conclusion:Intraperitoneal transplantation of PLA-O-CME nanoparticle-attached porcine hepatocytes embedded in collagen gel can improve the survival rates of ALF rats induced by D-Gal and can promote regeneration of the damaged recipient liver.