壳聚糖-人骨形态发生蛋白-2纳米粒基因载体的构建分析  

Construction and Analysis of One Kind of Chitosan-coated BMP-2 Nanoparticles as Genetic Carrier

作　　者：刘梦桃 张庆鸿[2] 翟浚江[3] 梁星[3] 

机构地区：[1]浙江省杭州哼哈口腔医院修复科,杭州310006 [2]浙江大学医学院附属口腔医院修复科,杭州310006 [3]四川大学华西口腔医院修复科,成都610041

出　　处：《四川大学学报（医学版）》2011年第4期485-489,共5页Journal of Sichuan University(Medical Sciences)

基　　金：浙江省教育厅基金(N20080180;N20110143);浙江省自然科学基金(Y2080340)资助

摘　　要：目的旨在构建一种良好性能的壳聚糖-人骨形态发生蛋白(BMP)-2纳米粒基因载体。方法通过复凝聚法制备的壳聚糖纳米粒与基因重组构建的BMP-2质粒相结合,形成含50、100和200μg/mL质粒BMP-2的壳聚糖纳米粒基因载体(分别命名为C-B50、C-B100和C-B200)。采用包封率测定壳聚糖与BMP-2目的基因的结合情况,场发射扫描电子显微镜观察纳米粒的形态和大小,DNaseⅠ消化实验检测该基因载体对目的基因的保护作用,以及MTT试验检测基因载体对骨髓基质干细胞的生长抑制。结果壳聚糖-BMP-2纳米粒基因载体呈不规则球形,结构紧密,粒径约为(90±20)nm。C-B50、C-B100和C-B200的包封率分别为76.31%±1.58%、84.49%±1.81%和81.69%±1.77%。壳聚糖-BMP-2纳米粒基因载体有效地保护BMP-2目的基因不被DNaseⅠ消化,随着剂量和时间的增加,对细胞的生长抑制率也增加(P<0.05),但最大剂量100 mg/L作用后绝大部分细胞的正常生长没有受到明显影响。结论该壳聚糖-BMP-2纳米粒基因载体有望应用于骨组织工程。Objective To construct an excellent kind of chitosan-coated BMP-2 nanoparticles as genetic carrier.Methods Chitosan-coated BMP-2 nanoparticles as genetic carrier,including C-B50 group,C-B100 group and C-B200 group,were constructed through re-coacervation and gene recombination methods.This carrier was studied by using morphological observation,enveloping test,anti-DNaseⅠ digested test and MTT assay.Results One kind of excellent chitosan-coated BMP-2 nanoparticles as genetic carrier was constructed.Particle diameter was about（90±20） nm.Enveloping ratio of C-B50 group,C-B100 group and C-B200 group was 76.31%＋1.58%,84.49%＋1.81% and 81.69%＋1.77%,respectively.Genetic carrier could prevent objective gene from DNase Ⅰ digestion.The growth inhibiting ratio to bone marrow stem cells was increased as the increase of genetic carrier dose and contact time（P0.05）,but maximal dose of carrier had no obvious toxic effect on cells growth in vitro.Conclsuion The constructed chitosan-coated BMP-2 nanoparticle material was a promising genetic carrier in bone tissue engineering.

关 键 词：壳聚糖 BMP-2 纳米粒 骨髓基质干细胞 基因载体 基因工程 组织工程 

分 类 号：R346[医药卫生—基础医学]