Comparative toxicological-hygienic assessment, structural-morphological, physicochemical characteristics, and virucidal properties of new nanopowder materials TiO2 and TiO2@Ag

O. Yavorovsky, V. M. Riabovol, T. O. Zinchenko, M. Zahornyi, A. Ragulya, N. I. Tyschenko, O. Povnitsa, L. Artiukh, S. Zahorodnia, D. Ostapiv
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Abstract

 In order to address safety concerns related to the acquisition and utilization of TiO2 and TiO2@Ag nanomaterials, as well as to investigate their disinfectant and biological effects, the structural-morphological, morpho­metry, toxicological, cytotoxic, and virucidal properties of these specified nanomaterials have been studied through experiments conducted on laboratory animals and in vitro. It has been demonstrated that the TiO2@Ag nanocomposite exhibited distinct physicochemical characteristics: it consisted of TiO2 nanoparticles ranging in size from 13 nm to 20 nm and Ag nanoparticles ranging from 35 nm to 40 nm with 4.0 wt% of silver localized on the surface of titanium dioxide. The purity of the modification of synthesized nano-TiO2 and nano-TiO2@Ag has been confirmed. Acute intraperitoneal administration of nanopowders revealed LD50 values of 4783.30 mg/kg for nano-TiO2 and 724.44 mg/kg for nano-TiO2@Ag. A slight accumulation was observed upon repeated (28-fold) intragastric administration of nano-TiO2. The cumulative dose administered, which equated to 15.9 multiples of the LD50 (76040 mg/kg), did not result in animal mortality but led to retardation in body weight gain. TiO2 and TiO2@Ag nanopowders do not irritate the skin, induce mild conjunctival irritation, and may exhibit a weak sensitizing effect. Nano-TiO2 and nano-TiO2@Ag powders accumulate in the tissues of internal organs and cause damage to the liver, kidneys, and lungs of laboratory animals upon intraperitoneal administration. The most characteristic morphological signs of the toxic effect of nano-TiO2 on liver tissue were observed at a level of 67.7% (cytoplasmic vacuolization in hepatocytes), while in the case of nano-TiO2@Ag initial necrotic changes were at a level of 70.0% (hepatocytes with pyknotic nuclei). Immunoassay analysis has demonstrated that TiO2@Ag and TiO2 nanomaterials at concentrations of 30 µg/ml can enhance the functional activity of peripheral blood mononuclear cells in vitro by increasing the production of cytokines IL-1, IL-6, TNF-α, and IL-4 in donors (p<0.05). This indicates the potential for chronic inflammation and allergic reactions among synthesis operators. In the study of the impact of nanomaterials on murine germ cells, it has been established that they affect the activity of mitochondrial enzymes and exert a damaging effect on mitochondrial membranes and overall cell integrity. Estimated approximate safe exposure levels in the workplace air are 0.3 mg/m3 for nano-TiO2 and 0.2 mg/m3 for nano-TiO2@Ag. Nano-TiO2@Ag and nano-TiO2 at a concentration of 100 µg/ml exhibit pronounced extracellular virucidal activity against human adenovirus serotype 2. The TiO2@Ag nanocomposite has a less damaging effect on Нер-2 cells compared to nano-TiO2.
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新型纳米粉体材料 TiO2 和 TiO2@Ag 的毒理学-卫生学评估、结构形态、理化特性和杀病毒性能比较
为了解决与获取和利用 TiO2 和 TiO2@Ag 纳米材料有关的安全问题,以及研究其消毒和生物效应,我们通过在实验动物和体外进行的实验,研究了这些特定纳米材料的结构形态、形态计量、毒理学、细胞毒性和杀病毒特性。结果表明,TiO2@Ag 纳米复合材料具有明显的理化特征:它由尺寸为 13 纳米至 20 纳米的 TiO2 纳米粒子和尺寸为 35 纳米至 40 纳米的 Ag 纳米粒子组成,其中 4.0 wt%的银被定位于二氧化钛表面。合成的纳米二氧化钛和纳米二氧化钛@银的改性纯度已得到证实。急性腹腔给药显示,纳米二氧化钛的半数致死剂量为 4783.30 毫克/千克,纳米二氧化钛@银的半数致死剂量为 724.44 毫克/千克。在重复(28 倍)胃内给药纳米二氧化钛后,观察到轻微的蓄积。累积给药剂量相当于半数致死剂量(76040 毫克/千克)的 15.9 倍,不会导致动物死亡,但会导致体重增加缓慢。纳米二氧化钛和 TiO2@Ag 粉末不会刺激皮肤,对眼结膜有轻微刺激,并可能表现出微弱的致敏作用。纳米二氧化钛和纳米二氧化钛@银粉在内脏器官组织中蓄积,腹腔给药会对实验动物的肝、肾和肺造成损害。纳米二氧化钛对肝脏组织毒性作用的最典型形态学迹象为 67.7%(肝细胞胞浆空泡化),而纳米二氧化钛@Ag 的初始坏死变化为 70.0%(肝细胞核坏死)。免疫测定分析表明,浓度为 30 微克/毫升的 TiO2@Ag 和 TiO2 纳米材料可提高供体外周血单核细胞的功能活性,增加细胞因子 IL-1、IL-6、TNF-α 和 IL-4 的产生(p<0.05)。这表明合成操作者有可能出现慢性炎症和过敏反应。在研究纳米材料对小鼠生殖细胞的影响时,已确定纳米材料会影响线粒体酶的活性,并对线粒体膜和整体细胞完整性产生破坏作用。据估计,工作场所空气中纳米二氧化钛的安全接触水平约为 0.3 毫克/立方米,纳米二氧化钛@银的安全接触水平约为 0.2 毫克/立方米。浓度为 100 µg/ml 的纳米二氧化钛@银和纳米二氧化钛对人类腺病毒血清 2 型具有明显的细胞外杀病毒活性。与纳米二氧化钛相比,纳米二氧化钛@银复合材料对Нер-2细胞的破坏作用较小。
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