He Yan-jin, Lin Ting-ting, Zhu Li-min, Luo Hao, Chang Jin, Zhang Hong, Zhang Hong-mei
Tianjin Medical University Eye Center, Tianjin 300070, China.
Zhonghua Yan Ke Za Zhi. 2010 Sep;46(9):795-801.
To prepare and observe the properties of Folate Receptor-mediated VCR-loaded nanoparticles, which is abbreviated as FA-PLGA (VCR)-NP and to study the inhibitory effect of FA-PLGA (VCR)-NP in ACC-2 cells in vitro and in ACC in BALB/c-nu mice.
The mAified W/O/W extraction-evaporation technique was chosen to prepare FA-PLGA (VCR)-NP. Tumor cells were divided into three groups: VCR, PLGA (VCR)-NP and FA-PLGA (VCR)-NP. Seven doses (0.05, 0.25, 0.50, 1.00, 5.00, 10.00, and 30.00 mg/L) of VCR were tested in the cell culture mAel. After 1, 2, 3, 4 and 5 days, the cell growth inhibition ratio was evaluated by MTT colorimetry. Nude mice mAel of orbital ACC was built by injecting ACC cell suspension and divided into four groups: VCR, PLGA (VCR)-NP, FA-PLGA (VCR)-NP, and control group. After 1 day, 7 days and 14 days, the inhibition ratio of gross tumor volume was observed. Residual concentrations of VCR in tumors were evaluated by HPLC. The feature of histopathology was observed by electron microscopy. The effect of empty nanoparticles on ACC-2 cells was compared with normal control group using t-test to analyze. On account of different drugs, concentration, time and the interaction of them, multivariate analysis of variance was used to analyze their relationship. Inhibition rate and residual volume of drug concentrations were compared using one-factor analysis of variance and LSD method.
FA-PLGA (VCR)-NP were smooth and spherical with a mean particle size of 249.2 nm. The drug loading efficiency was 4.53%. The release of VCR from PLGA nanoparticles can persist for 14 d. After blank particles PLGA-NP and ACC-2 cells were co-cultured for 5 days, cell viability had remained at more than 80 percent (t = 1.952 ∼ 3.285, P = 0.081 ∼ 0.190). The inhibitory effect of FA-PLGA (VCR)-NP was more effective than VCR alone after a period of time (F = 4.798 ∼ 563.479, P = 0.000 ∼ 0.006). The effects of the treatment were both in dose-dependent and time-dependent manner. Targeting particles could attach to tumor surface, via folate receptor. FA was competitive inhibitor of this recolonization. The volume inhibition ratios of FA-PLGA (VCR)-NP and PLGA (VCR)-NP were significant higher than VCR (P = 0.016, P = 0.029). The inhibition ratio of FA-PLGA (VCR)-NP was higher than that of PLGA (VCR)-NP, but there was no statistical difference (P = 0.376). There was significant different between residual concentrations of VCR on the 1(st), 7(th) and 14(th) days. TEM pictures showed a mass of electron-dense microspheres in tumor cells on the 14(th) day. Tumor necrosis was obvious, while surrounding tissues were normal.
FA-PLGA (VCR)-NP are stable and have high drug entrapment efficiency and high effect of growth inhibition in vitro. It can be proposed as a potentially controlled and targeted delivery system for the treatment of ACC.
制备并观察叶酸受体介导的载长春新碱纳米粒（FA-PLGA(VCR)-NP）的性质，研究其对体外培养的ACC-2细胞及BALB/c-nu小鼠ACC的抑制作用。
采用改良的W/O/W萃取-蒸发技术制备FA-PLGA(VCR)-NP。将肿瘤细胞分为三组：长春新碱组、PLGA(VCR)-NP组和FA-PLGA(VCR)-NP组。在细胞培养体系中测试七剂（0.05、0.25、0.50、1.00、5.00、10.00和30.00mg/L）长春新碱。1、2、3、4和5天后，采用MTT比色法评估细胞生长抑制率。通过注射ACC细胞悬液建立眼眶ACC裸鼠模型，并分为四组：长春新碱组、PLGA(VCR)-NP组、FA-PLGA(VCR)-NP组和对照组。1天、7天和14天后，观察肿瘤总体积抑制率。采用高效液相色谱法评估肿瘤中长春新碱的残留浓度。通过电子显微镜观察组织病理学特征。使用t检验分析空纳米粒对ACC-2细胞的影响与正常对照组的差异。由于不同药物、浓度、时间及其相互作用，采用多因素方差分析来分析它们之间的关系。使用单因素方差分析和LSD法比较抑制率和药物浓度残留量。
FA-PLGA(VCR)-NP呈光滑球形，平均粒径为249.2nm。载药效率为4.53%。长春新碱从PLGA纳米粒中的释放可持续14天。空白颗粒PLGA-NP与ACC-2细胞共培养5天后，细胞活力保持在80%以上（t=1.952～3.285，P=0.081～0.190）。一段时间后，FA-PLGA(VCR)-NP的抑制作用比单独使用长春新碱更有效（F=4.798～563.479，P=0.000～0.006）。治疗效果呈剂量依赖性和时间依赖性。靶向颗粒可通过叶酸受体附着于肿瘤表面。叶酸是这种再定植的竞争性抑制剂。FA-PLGA(VCR)-NP和PLGA(VCR)-NP的体积抑制率显著高于长春新碱组（P=0.016，P=0.029）。FA-PLGA(VCR)-NP的抑制率高于PLGA(VCR)-NP，但无统计学差异（P=0.376）。第1、7和14天长春新碱残留浓度有显著差异。透射电镜照片显示第14天肿瘤细胞中有大量电子致密微球。肿瘤坏死明显，而周围组织正常。
FA-PLGA(VCR)-NP稳定，具有较高的载药效率和体外生长抑制效果。可作为一种潜在的可控靶向递送系统用于治疗ACC。
