装载肝素PLGA纳米粒的制备及体外细胞相容性研究

摘要

目的 采用双次乳化法制备装载有肝素的PLGA纳米粒,并评价其体外缓释性能和细胞相容性.方法 ①使用双次乳化法制备PLGA-肝素纳米粒(PLGA-Hep NPs)；②对PLGA-Hep纳米粒进行理化分析和体外缓释效果评价,主要指标有:纳米粒径分析、表面形态观察,测定药物载药量和绘制体外缓释曲线等；③采用细胞增殖实验评价PLGA-Hep纳米粒的细胞毒性.结果 ①所制备的PLGA-Hep纳米粒呈球形,纳米粒的粒径、Zeta电位和肝素载药量与初始肝素投入量相关,当肝素投入量为100 mg时,粒径平均大小为(184.8±3.0)nm,Zeta电位为(-20.24±0.83)mV,1mg PLGA-Hep纳米粒装载(48.7±2.3)μg肝素；②体外缓释试验提示:突释阶段肝素释放率在24 h内达(26.6±2.8)％,缓释阶段纳米粒可稳定释放,在14 d时释放达(54.9±1.9)％；③细胞增殖实验提示PLGA- Hep纳米粒对细胞体外生长无不良影响,细胞相容性好.结论 采用双次乳化法制备的PLGA-Hep纳米粒具有良好的缓释效应和良好的细胞相容性,显示了PLGA纳米粒在药物缓释领域的广泛应用前景.%Objective To prepare heparin-loaded PLGA nanoparticles by double emulsion method,and to study the properties of controlled release of heparin and biocompatibility in vitro. Methods Double emulsion method was used to prepare PL-GA-heparin nanoparticles (PLGA-Hep NPs). The physical and chemical properties and release effect of PLGA-Hep NPs in vitro were evaluated. The main outcome measures included size distribution,SEM of nanoparticles,and drug content. The in vitro release curve was drawn. The cytotoxicity of PLGA-Hep NPs was evaluated by using cell proliferation assay. Results The PLGA-Hep NPs were spherical,and the mean diameters and Zeta potential of the spheres and the amount of heparin loaded were related to the amount of heparin used initially. When the amount of heparin used initially was 100 mg,the mean diameters of the spheres were(184. 8 ± 3. 0)nm,Zeta potential was( -20. 24 ±0. 83)mV,and(48. 7± 2. 3)jzg of heparin was loaded per 1 mg PLGA-Hep NPs- In vitro release test showed that heparin release was(26. 6 ± 2. 8)% at 24th h at the burst release phase,and up to(54. 9± 1. 9)% at 14th day longer in the slower release phase. Cell proliferation assay revealed that the PLGA-Hep NPs did not damage the cell growth in vitro, indicating good compatibility. Conclusion The PLGA-heparin nanoparticles prepared by double emulsion method have a good release effect and good biocompatibility in vitro , showing the broad prospect of PLGA nanoparticles in the field of drug delivery.