聚酰胺-胺对永生化的人角膜上皮细胞的毒性

Cytotoxicity of poly(amidoamine) on immortalized human corneal epithelial cell lines

目的考察一种新型药物载体——聚酰胺-胺[poly(amidoamine),PAMAM]对体外培养的角膜上皮细胞的毒性作用,为其在眼部药物载体的应用提供理论依据。方法建立应用于PAMAM毒性研究的角膜上皮细胞模型,通过细胞毒性分析测定不同浓度(10g·L-1、1g·L-1、0.1g·L-1、0.01g·L-1、0.001g·L-1)、不同代数(G3.0、G3.5、G4.0、G4.5、G5.0)的PAM-AM在共同孵育5min、15min、60min对角膜上皮细胞存活率的影响并计算其IC50值。结果整代PAMAM(G3.0、G4.0、G5.0)和半代PAMAM(G3.5、G4.5)均随着代数的增大,细胞毒性增加,且在一定浓度范围内细胞毒性随着药物浓度增大、药物接触的时间延长而增加。半代PAMAM细胞毒性均低于整代。G3.0、G3.5、G4.5PAMAM在1g·L-1及以下浓度毒性反应均为合格,G4PAMAM在0.1g·L-1浓度及以下毒性评级大部分为合格,而G5PAMAM只在0.01g·L-1和0.001g·L-1下部分毒性评级合格。结论低代或半代PAMAM可能更适合作为眼部药物给药的载体,也可考虑通过结构修饰等手段降低高代PAMAM毒性。

Objective To investigate the cytotoxicity of a potential and novel ocular drug carrier-poly（amidoamine） （PAMAM） on the immortalized human corneal epithelial cells （HCEC）,and provide a theoretical basis in the field of ophthalmic toxicology.Methods HCEC were cultured and then the cells were exposed to a series of PAMAM solution （G3.0,G3.5,G4.0,G4.5 and G5.0） at various concentrations （10 g·L-1,1 g·L-1,0.1 g·L-1,0.01 g·L-1 and 0.001 g·L-1） for a period of 5minutes,15minutes and 60minutes.Toxicity of PAMAM on cultured immortalized HCEC was assessed by WST-1 assay and the IC50 values were also calculated.Results The toxicity data obtained with WST-l test suggested that the low concentrations of PAMAM showed a low toxicity on HCEC compared with high concentrations.And its cytotoxicity also increased with generation growth and exposure time.The corneal epithelial cell toxicity of half generation PAMAM was lesser than whole generation.PAMAM dendrimers （G3.0,G3.5 and G4.5） exhibited toxicity only at concentration above 1 g·L-1,but G4 PAMAM dendrimers showed cytotoxicity toward HCEC at concentration above 0.1 g·L-1 and G5 PAMAM were significantly cytotoxic at most all concentrations examined.Conclusion The low or half generation PAMAM will be more suitable for the ophthalmic drugs carrier and the cytotoxicity of high generation of PAMAM can be considered to decrease by chemically modification.