目的:利用聚乙二醇对单壁碳纳米管(single-walled carbon nanotubes, SWNTs)进行功能化修饰,并利用斑马鱼胚胎评价其被修饰前后的生物毒性,为SWNTs在生物医药方面的应用提供毒性依据。方法:将甲氧基聚乙二醇胺5000(methoxy-polyethylene glycol amine, mPEG-NH2 , Mw = 5 ku) 与羧基化单壁碳纳米管( carboxylated single-walled carbon nanotubes,SWNT—COOH)非共价偶联制备SWNT-mPEG。以紫外-可见分光光度法、傅里叶转换红外光谱和透射电子显微镜考查SWNT—mPEG的组成、浓度、形貌和粒径大小;利用水浴给药对斑马鱼胚胎进行染毒,并分别在受精后24h(24hpf)和受精后96h(96hpf)观察和统计孵化率、畸形率和死亡率。结果:与SWNT—COOH相比,SWNT—mPEG在水中具有更好的分散性;胚胎毒性实验表明,在24hpf和96hpf,SWNT—mPEG各浓度组的胚胎的致畸率和致死率仅略高于空白组,只有80mg/LSWNT—mPEG实验组在96hpf的对斑马鱼胚胎的致死率(11.11%)明显高于同一试验时段的空白组(3.33%)(P〈0.05)。结论:SWNT—COOH和SWNT—mPEG对斑马鱼胚胎均呈现低生物毒性,原因可能与斑马鱼胚胎绒包膜对纳米颗粒的低通透性有关。
Objective:To evaluate the biotoxicity of the as-prepared single-walled carbon nanotube (SWNTs) modified with polyethylene glycol by using zebrafish embyos, and to provide toxicity evidence for biomedical application of single-walled carbon nanotubes. Methods: Preparation of SWNTs modified by methoxypolyethylene glycol amine 5 000 (mPEG-NH2, Mw = 5 ku) was performed by non-covalently associating SWNT-COOH with mPEG-NH2. Components, mor- phology, concentrations, and particle sizes of SWNT-mPEG were determined and analyzed by using UV- Visible spectroscopy, Fourier transform infrared spectroscopy (FT-IR), and transmission electron micros- copy, respectively. Embryos were exposured to graded concetrations of SWNT-mPEG solutions in 96-well cell culture plates. Hatching rate, malformed ratio and mortality were investigated at 24 hpf (24 hours post-fertilization) and 96 hpf, respectively. Results: Compared with SWNT-COOH, SWNT-mPEG was a kind of non-covalent conjugate, and exhibited better dispersity in water. Embryotoxicity assay indicated that malformed ratios and mortalities of all test groups were only slightly greater than that of control group. However, only embryos treated with 80 mg/L SWNT-mPEG had a statistically significant ( P d0. 05) greater mortality (11.11%) than that (3.33%) of control group at 96 hpf. Conclusion: Both SWNT- COOH and SWNT-mPEG have low embryotoxieity to zebrafish embryos, which seems to be due to low permeability of embryos' chorion for nanoparticles.