Fattori Ana Carolina M, Brassolatti Patrícia, Feitosa Karina A, Pedrino Matheus, Correia Ricardo de O, Albuquerque Yulli R, Rodolpho Joice M de A, Luna Genoveva L F, Cancino-Bernardi Juliana, Zucolotto Valtencir, Speglich Carlos, Rossi Karina N Z P, Anibal Fernanda de F
Laboratório de Inflamação e Doenças Infecciosas, Departamento de Morfologia e Patologia, Universidade Federal de São Carlos, São Carlos, SP, Brazil,
Laboratório de Inflamação e Doenças Infecciosas, Departamento de Morfologia e Patologia, Universidade Federal de São Carlos, São Carlos, SP, Brazil.
Cell Physiol Biochem. 2023 Mar 22;57(2):63-81. doi: 10.33594/000000616.
BACKGROUND/AIMS: Titanium dioxide nanoparticles (TiO NPs) are extensively applied in the industry due to their photocatalytic potential, low cost, and considerably low toxicity. However, new unrelated physicochemical properties and the wide use of nanoparticles brought concern about their toxic effects. Thereby, we evaluated the cytotoxicity of a TiO NP composed of anatase and functionalized with sodium carboxylate ligands in a murine fibroblast cell line (LA-9).
Scanning Electron Microscopy (SEM), Dynamic Light Scattering (DLS), and ATR-FTIR spectroscopy were applied to determine nanoparticle physicochemical properties. The cell viability (MTT assay) and clonogenic survival were analyzed in fibroblasts exposed to TiO NP (50, 150, and 250 µg/mL) after 24h. Moreover, oxidative stress, proinflammatory state, and apoptosis were evaluated after 24h.
TiO NP characterization showed an increased hydrodynamic size (3.57 to 7.62 nm) due to solvent composition and a heterogeneity dispersion in water and cell culture media. Also, we observed a zeta potential increased from -20 to -11 mV in function of protein adsorption. TiO NP reduced fibroblast cell viability and induced ROS production at the highest concentrations (150 and 250 µg/mL). Moreover, TiO NP reduced the fibroblasts clonogenic survival at the highest concentration (250 µg/mL) on the 7th day after the 24h exposure. Nevertheless, TiO NP did not affect the fibroblast proinflammatory cytokines (IL-6 and TNF) secretion at any condition. Early and late apoptotic fibroblast cells were detected only at 150 µg/mL TiO NP after 24h.
Probably, TiO NP photocatalytic activity unbalanced ROS production which induced apoptosis and consequently reduced cell viability and metabolic activity at higher concentrations.
背景/目的:二氧化钛纳米颗粒(TiO NPs)因其光催化潜力、低成本和相当低的毒性而在工业中得到广泛应用。然而,新的不相关物理化学性质以及纳米颗粒的广泛使用引发了对其毒性作用的担忧。因此,我们评估了由锐钛矿组成并用羧酸钠配体功能化的TiO NP在小鼠成纤维细胞系(LA-9)中的细胞毒性。
应用扫描电子显微镜(SEM)、动态光散射(DLS)和衰减全反射傅里叶变换红外光谱(ATR-FTIR)来确定纳米颗粒的物理化学性质。在24小时后,对暴露于TiO NP(50、150和250μg/mL)的成纤维细胞进行细胞活力(MTT法)和克隆形成存活率分析。此外,在24小时后评估氧化应激、促炎状态和细胞凋亡。
TiO NP表征显示,由于溶剂组成,流体动力学尺寸增加(从3.57到7.62nm),并且在水和细胞培养基中存在异质性分散。此外,我们观察到随着蛋白质吸附,zeta电位从-20mV增加到-11mV。TiO NP在最高浓度(150和250μg/mL)下降低了成纤维细胞活力并诱导了ROS产生。此外,在暴露24小时后的第7天,TiO NP在最高浓度(250μg/mL)下降低了成纤维细胞的克隆形成存活率。然而,在任何条件下,TiO NP均未影响成纤维细胞促炎细胞因子(IL-6和TNF)的分泌。仅在24小时后150μg/mL的TiO NP处理下检测到早期和晚期凋亡的成纤维细胞。
可能是TiO NP的光催化活性使ROS产生失衡,从而诱导细胞凋亡,进而在较高浓度下降低细胞活力和代谢活性。