Center for Research in Micro- and Nano-engineering (CRIM) Laboratory, Scuola Superiore Sant'Anna, Pisa, Italy.
Nanomedicine. 2009 Dec;5(4):424-31. doi: 10.1016/j.nano.2009.02.006. Epub 2009 Mar 31.
There are conflicting data concerning the safety and biocompatibility of carbon nanotubes (CNTs). In some reports CNTs have been used for gene delivery without significant toxicity, whereas in others various cytotoxic effects were observed, including induction of intracellular reactive oxygen species (ROS), DNA damage, and apoptosis. Although it is clear that CNT production methods, purity, and functionalization treatments impact on biocompatibility, most of the published reports lack detailed characterization of the CNT samples used. We investigated the effect of various physicochemical features of multiwalled carbon nanotubes (MWCNTs) on toxicity and biocompatibility with cultured human neuroblastoma cells by using MTT, WST-1, Hoechst, and oxidative stress assays. In vitro experiments confirm that after 3 days of incubation with three different types of CNTs dispersed in Pluronic F127 solution, 0.01% cell viability is not affected and apoptosis and ROS are not induced in the SH-SY5Y cells. With prolonged cultures and continued propagation in the presence of MWCNTs, the loss of cell viability was minimal for pure MWCNTs (99% purity), but cell proliferation decreased significantly for 97% purity MWCNTs and acid-treated MWCNTs (97% purity, surface oxidation 8%); no intracellular ROS were detected. When the concentration of CNTs increases, purity and surface oxidation seem to affect cell viability (ED(25) is 48, 34.4, and 18.4 mug/mL, respectively, for 99% purity MWCNTs, 97% purity MWCNTs, and acid-treated 97% purity MWCNTs. Our results indicate that concentrations of 5-10 mug/mL MWCNTs seem ideal for studies on the design and development of artificial MWCNT nanovectors for gene and drug therapy against cancer.
With prolonged cultures, loss of cell viability was minimal for preparations with 99% purity, but significant adverse effects were detected with 97% purity and with acid-treated preparations. A concentrations of 5-10 mug/mL of MWCNTs seems ideal for gene and drug therapy against cancer.
关于碳纳米管（CNTs）的安全性和生物相容性存在相互矛盾的数据。在一些报道中，CNTs 已被用于基因传递而没有明显的毒性，而在其他一些报道中则观察到了各种细胞毒性作用，包括诱导细胞内活性氧（ROS）、DNA 损伤和细胞凋亡。尽管很明显，CNT 的生产方法、纯度和功能化处理会影响生物相容性，但大多数已发表的报告缺乏对所用 CNT 样品的详细描述。我们通过使用 MTT、WST-1、Hoechst 和氧化应激测定法，研究了多壁碳纳米管（MWCNTs）的各种物理化学特性对培养的人神经母细胞瘤细胞毒性和生物相容性的影响。体外实验证实，在 3 天的孵育期内，三种不同类型的 CNT 分散在 Pluronic F127 溶液中，0.01%的细胞活力不受影响，SH-SY5Y 细胞不会发生凋亡和 ROS 诱导。随着培养时间的延长和在 MWCNTs 存在下的持续繁殖，纯 MWCNTs（纯度为 99%）的细胞活力损失最小，但 97%纯度的 MWCNTs 和酸处理的 MWCNTs（纯度为 97%，表面氧化 8%）的细胞增殖显著下降；未检测到细胞内 ROS。当 CNT 浓度增加时，纯度和表面氧化似乎会影响细胞活力（ED（25）分别为 48、34.4 和 18.4μg/mL，用于 99%纯度 MWCNTs、97%纯度 MWCNTs 和酸处理的 97%纯度 MWCNTs）。我们的结果表明，浓度为 5-10μg/mL 的 MWCNTs 似乎是用于设计和开发用于癌症基因和药物治疗的人工 MWCNT 纳米载体的理想选择。
随着培养时间的延长，纯度为 99%的制剂的细胞活力损失最小，但 97%的纯度和酸处理的制剂则检测到明显的不良反应。MWCNTs 的浓度为 5-10μg/mL 似乎是癌症基因和药物治疗的理想选择。
