The characteristics and mechanisms of uptake of PLGA nanoparticles in rabbit conjunctival epithelial cell layers
The characteristics and mechanisms of uptake of PLGA nanoparticles in rabbit conjunctival epithelial cell layers
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DOI：
10.1023/b:pham.0000022411.47059.76
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发表时间：
2004-04-01
影响因子：
3.7
通讯作者：
Lee, VHL
中科院分区：
文献类型：
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作者：
Qaddoumi, MG;Ueda, H;Lee, VHL
目的.目的探讨生物可降解聚乳酸-羟基乙酸共聚物（PLGA）纳米粒在原代培养的兔结膜上皮细胞（RCECs）中的摄取特性及机制。使用含有6-香豆素（作为荧光标记物）的聚（D，L-丙交酯-共-乙交酯）纳米颗粒（PLGA 50：50，直径100 nm）。使用各种粒度（100 nm，800 nm和10 μ m）的大小的影响进行了研究。研究了细胞松弛素D、诺考达唑和代谢抑制剂对纳米颗粒摄取的影响。纳米颗粒的能力，以提高包封的蛋白质，结合到得克萨斯红（TR-BSA）的摄取进行了评估。在37 degreesC的纳米粒子的最大吸收发生在2小时，100 nm的颗粒在RCEC中的吸收最高，与800 nm和10 μ m的颗粒相比。纳米颗粒摄取在0.1 - 4 mg/ml浓度范围内是饱和的。通过共聚焦显微镜证实了纳米颗粒摄取，并且通过无香豆素纳米颗粒（相似大小）、较低的孵育温度以及代谢抑制剂和细胞松弛素D的存在显著抑制了纳米颗粒摄取。RCEC对包封TR-BSA的摄取在4 h时比游离BSA高28%。我们的研究结果表明，原代培养的兔结膜上皮细胞中的PLGA纳米颗粒摄取发生最有可能通过吸附型内吞作用。
Purpose. To delineate the characteristics and mechanisms of uptake of biodegradable poly(D,L-lactide-co-glycolide) (PLGA) nanoparticles in primary cultured rabbit conjunctival epithelial cells (RCECs).Methods. Poly(D, L-lactide-co-glycolide) nanoparticles (PLGA 50: 50, 100 nm in diameter) containing 6-coumarin (as a fluorescent marker) were used. The effect of size was studied using various particle sizes ( 100 nm, 800 nm, and 10 mum). The effect of cytochalasin D, nocodazole, and metabolic inhibitors on nanoparticle uptake was investigated. The capability of nanoparticles to enhance the uptake of an encapsulated protein, BSA bound to Texas red (TR-BSA), was evaluated.Results. Maximal uptake of nanoparticles at 37 degreesC occurred at 2 h, and 100-nm particles had the highest uptake in RCECs in comparison with 800-nm and 10-mum particles. Nanoparticle uptake was saturable over the 0.1 - 4 mg/ml concentration range. Nanoparticle uptake was confirmed by confocal microscopy and was inhibited significantly by coumarin-free nanoparticles ( of similar size), by lower incubation temperature, and by the presence of metabolic inhibitors and cytochalasin D. The uptake of encapsulated TR-BSA in RCECs at 4 h was 28% higher than free BSA application.Conclusion. Our findings suggest that PLGA nanoparticle uptake in primary cultured rabbit conjunctival epithelial cells occurs most likely by adsorptive-type endocytosis.