红细胞膜包载粉防己碱PLGA纳米粒的制备与体外评价

Preparation and in vitro evaluation of erythrocyte membrane-camouflaged tetrandrine-loaded PLGA nanoparticles

目的制备红细胞膜包载粉防己碱PLGA纳米粒(RPTNs),并进行体外评价。方法采用乳化溶剂挥发法制备粉防己碱PLGA纳米粒(PTNs);单因素考察了PLGA的质量浓度、有机相体积、稳定剂浓度;优化了红细胞膜囊泡(RVs)和RPTNs的制备方法;对纳米粒的粒径、PDI、电位、包封率、形态和稳定性进行了表征;透析法考察了PTNs和RPTNs的体外释放特性;CCK-8法考察了该载药系统的体外细胞毒性。结果最佳制备工艺为:PLGA质量浓度20 g·L-1,稳定剂质量浓度10 g·L-1,油水体积比1∶20,RVs的制备采用超声合并挤压法;制备的PTNs形状圆整,粒径为(147.9±0.25) nm,PDI<0.15,包封率为(84.1±0.41)%,在30 d内保持稳定;RPTNs具有清晰的壳-核双层结构,粒径为(164.1±1.65) nm,PDI<0.2,在15 d内保持稳定;体外释放实验显示PTNs和RPTNs在120 h内对粉防止碱(tetrandrine,TET)均有缓释作用,且RPTNs平均累计释放量减少了5.33%;体外细胞毒性实验表明在正常细胞和肿瘤细胞中,RPTNs的IC50值均高于游离TET组和PTNs组,且对293T细胞的IC50值高于对MCF-7的IC50值。结论采用该制备方法得到的纳米粒包封率高、稳定性好,能显著延缓药物的释放,降低体外细胞毒性,增强对肿瘤细胞的抑制作用。

Objective To prepare the erythrocyte membrane-camouflaged tetrandrine-loaded PLGA nanoparticles(RPTNs) and investigate their characterization,stability,release and cytotoxicity in vitro.Methods Methods The tetrandrine-loaded PLGA nanoparticles(PTNs) were prepared by emulsification method.Three factors including PLGA concentration,oil phase volume,stabilizer concentration were investigated.Then the preparation methods of erythrocyte membrane vesicles(RVs) and RPTNs were optimized,and the particle size,polydispersity index(PDI),potential,encapsulation efficiency,morphology and stability of the nanoparticles were characterized.The release characteristic of PTNs and RPTNs were investigated by dialysis method.Furthermore,the CCK-8 kit assay was used to investigate the cytotoxicity of RPTNs.Results Optimal preparation technology was as follows:PLGA concentration was 20 g·L-1,stabilizer concentration was 10 g·L-1,oil-water phase ratio was 1∶20 and RVs was prepared by ultrasonic combined extrusion method.The PTNs had the spherical structure and kept their basic structure within 30 days.The average particle size was(147.9±0.25) nm,PDI was less than 0.15 and entrapment efficiency was(84.1±0.41)%.RPTNs showed shell-core double-layer structure and kept their basic structure within 15 days.The average size was(164.1±1.65)nm,PDI was less than 0.2.In vitro release test showed that both PTNs and RPTNs showed obvious sustained release effect within 120 h,and the release effect of RPTNs was reduced by 5.33%.in vitro cytotoxicity experiments showed that the IC50 of RPTNs was higher than that of the PTNs and free TET both in normal cells and tumor cells.And the IC50 of RPTNs in 293 T cells was significantly higher than that in MCF-7 cells.Conclusion The RPTNs prepared by the optimal method have a high encapsulation efficiency,good stability,sustained releasecharacteristic.They can significantly reduce the cytotoxicity of the free drug and passively enhance the inhibitory effect in tumor cells.