Bodewein Lambert, Schmelter Frank, Di Fiore Stefano, Hollert Henner, Fischer Rainer, Fenske Martina
Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Project Group Translational Medicine and Pharmacology, Frankfurt & Aachen, Germany; Institute of Environmental Research (Biology V), RWTH Aachen University, Germany.
Fraunhofer Institute for Molecular Biology and Applied Ecology IME, Molecular Biology Division, Aachen, Germany.
Toxicol Appl Pharmacol. 2016 Aug 15;305:83-92. doi: 10.1016/j.taap.2016.06.008. Epub 2016 Jun 8.
Dendrimers are an emerging class of polymeric nanoparticles with beneficial biomedical applications like early diagnostics, in vitro gene transfection or controlled drug delivery. However, the potential toxic impact of exposure on human health or the environment is often inadequately defined. Thus, polyamidoamine (PAMAM) dendrimers of generations G3.0, 3.5, 4.0, 4.5 and 5.0 and polypropylenimine (PPI) dendrimers G3.0, 4.0 and 5.0 were tested in zebrafish embryos for 96h and human cancer cell lines for 24h, to assess and compare developmental in vivo toxicity with cytotoxicity. The zebrafish embryo toxicity of cationic PAMAM and PPI dendrimers increased over time, with EC50 values ranging from 0.16 to just below 1.7μM at 24 and 48hpf. The predominant effects were mortality, plus reduced heartbeat and blood circulation for PPI dendrimers. Apoptosis in the embryos increased in line with the general toxicity concentration-dependently. Hatch and dechorionation of the embryos increased the toxicity, suggesting a protective role of the chorion. Lower generation dendrimers were more toxic in the embryos whereas the toxicity in the HepG2 and DU145 cell lines increased with increasing generation of cationic PAMAMs and PPI dendrimers. HepG2 were less sensitive than DU145 cells, with IC50 values≥402μM (PAMAMs) and ≤240μM (PPIs) for HepG2 and ≤13.24μM (PAMAMs) and ≤12.84μM (PPIs) for DU145. Neither in fish embryos nor cells toxicity thresholds were determinable for anionic PAMAM G3.5 and G4.5. The study demonstrated that the cytotoxicity underestimated the in-vivo toxicity of the dendrimers in the fish embryos.
树枝状大分子是一类新兴的聚合物纳米颗粒，在早期诊断、体外基因转染或可控药物递送等生物医学应用方面具有优势。然而，其暴露对人类健康或环境的潜在毒性影响往往定义不充分。因此，对第G3.0、3.5、4.0、4.5和5.0代的聚酰胺胺（PAMAM）树枝状大分子以及第G3.0、4.0和5.0代的聚丙撑亚胺（PPI）树枝状大分子在斑马鱼胚胎中进行了96小时的测试，并在人类癌细胞系中进行了24小时的测试，以评估和比较体内发育毒性与细胞毒性。阳离子PAMAM和PPI树枝状大分子对斑马鱼胚胎的毒性随时间增加，在24和48小时胚胎期时，半数有效浓度（EC50）值范围为0.16至略低于1.7μM。主要影响是死亡率，以及PPI树枝状大分子导致的心跳和血液循环减少。胚胎中的凋亡随着一般毒性浓度的增加而呈浓度依赖性增加。胚胎的孵化和去绒毛膜增加了毒性，表明绒毛膜具有保护作用。较低代的树枝状大分子在胚胎中毒性更大，而在HepG2和DU145细胞系中，阳离子PAMAM和PPI树枝状大分子的毒性随着代数增加而增加。HepG2细胞比DU145细胞敏感性低，HepG2的半数抑制浓度（IC50）值≥402μM（PAMAM）和≤240μM（PPI），DU145的IC50值≤13.24μM（PAMAM）和≤12.84μM（PPI）。对于阴离子PAMAM G3.5和G4.5，在鱼类胚胎和细胞中均无法确定毒性阈值。该研究表明，细胞毒性低估了树枝状大分子在鱼类胚胎中的体内毒性。
