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采用改良Stober法一步合成氨基修饰的阳离子型介孔硅纳米粒(CMSN),结合薄膜水化法和高速离心法制备介孔硅脂质囊纳米粒(LP-CMSN)。所得LP-CMSN呈圆整球形、粒径分布均一、无团聚、具有明显的核-壳结构,粒径为(114.5±3.7)nm,ζ电位为(9.3±2.5)mV,多分散系数(PDI)为0.17±0.05。细胞水平上,LP-CMSN在0~100μg/ml范围内对Hela和A549细胞均无明显细胞毒性。以LP-CMSN为载体,选择多柔比星为模型药,采用饱和溶液吸附法制得了载药量为(15.58±3.25)%的载药纳米粒,有效改善了CMSN中药物易突释缺点。选择异硫氰酸荧光素(FITC)为荧光探针,合成了标记率为0.52%的荧光纳米粒。通过流式细胞仪和激光共聚焦显微镜考察LP-CMSN在Hela细胞的摄取机制和入胞途径。结果显示,CMSN通过网格蛋白介导的内吞途径摄取进入细胞,而LP-CMSN主要经过膜融合途径进入细胞,最终经溶酶体逃逸后释放药物发挥药效。因此,LP-CMSN具有良好的生物相容性和缓控释特性,并能通过无损害的膜融合途径摄取进入细胞,有望成为新型纳米递药载体。
The modified Stober method was used to synthesize amino-modified cationic mesoporous silica nanoparticles (CMSN) in one step. The mesoporous silica liposome nanoparticles (LP-CMSN) were prepared by membrane hydration and high-speed centrifugation. The obtained LP-CMSN was spherical and uniform in size distribution with no agglomeration, with obvious core-shell structure with particle size of (114.5 ± 3.7) nm, zeta potential of (9.3 ± 2.5) mV and polydispersity index ) Was 0.17 ± 0.05. At the cellular level, LP-CMSN showed no significant cytotoxicity on Hela and A549 cells in the range of 0 ~ 100μg / ml. Using LP-CMSN as carrier and selecting doxorubicin as model drug, drug-loaded nanoparticles with drug loading of (15.58 ± 3.25)% were prepared by saturated solution adsorption method, which effectively improved the drug-susceptibility of CMSN. Fluorescein isothiocyanate (FITC) was chosen as fluorescent probe to synthesize fluorescent nanoparticles with the labeling rate of 0.52%. The uptake mechanism and entry pathway of LP-CMSN in Hela cells were investigated by flow cytometry and laser confocal microscopy. The results showed that CMSN uptake into the cells via the clathrin-mediated endocytosis pathway, whereas LP-CMSN enters the cell mainly through the membrane fusion pathway and finally releases the drug through the lysosome escape to exert its efficacy. Therefore, LP-CMSN has good biocompatibility and controlled-release properties, and can be taken into the cells through non-damaging membrane fusion pathway, which is expected to become a new type of nano drug delivery carrier.