Antioxidant thymoquinone and eugenol alleviate TiO2 nanoparticle-induced toxicity in human blood cells in vitro.

Titanium dioxide (TiO2) nanoparticles (NPs) are used extensively in a variety of commercial, industrial, and medical products, due to which human exposure is inevitable. This study aimed to explore the potential of eugenol and thymoquinone (TQ), two well-known antioxidants, in counteracting the NP-induced toxicity in human blood cells in vitro. Fresh lymphocytes and erythrocytes were isolated from volunteer human blood donors and incubated with 50 µg/mL of TiO2 NPs in the presence and absence of 50 µM of TQ and 20 µg/mL of eugenol for 3 h. Results showed that NP-treatment-induced hemolysis, oxidative stress, lactate dehydrogenase (LDH) leakage, and reduced ATPase activity in the erythrocytes. In the lymphocytes treated with NPs alone (50 µg/mL), cytotoxicity in MTT assay and DNA damage in comet assay were observed; in addition, mitochondrial membrane potential collapsed and ADP/ATP ratio increased indicating mitochondrial function impairment. However, in the presence of antioxidants, all these NP-induced changes were mitigated significantly. The results were more significant when both antioxidants eugenol and TQ were given together. Thus, it seems that antioxidants eugenol and TQ can be used as a protective agent against TiO2 NP-induced toxicity.

Coenzyme Q10 protects isolated human blood cells from TiO2 nanoparticles induced oxidative/antioxidative imbalance, hemolysis, cytotoxicity, DNA damage and mitochondrial impairment.

TiO2 NPs have been investigated for their toxic potential and studies have reported their toxicity is due to generation of oxidative stress. In the present study, we investigated the toxicity of TiO2 NPs and explored the potential of well-known antioxidant coenzyme Q10 (CoQ10) in counteracting the NP-induced toxicity in isolated human blood cells. When the isolated blood cells were treated with varying concentrations of TiO2 NPs (25-100 µg/ml), only 50 µg/ml dose induced statistically significant hemolysis in erythrocytes and cytotoxicity in lymphocytes (p < 0.05). None of the concentrations induced any significant increase in platelet aggregation. To investigate the protective effect of CoQ10, we incubated the isolated blood cells with 50 µg/ml of TiO2 NPs in the presence and absence of 25 µM of CoQ10 for 3 h. Hemolysis, oxidative stress, LDH leakage and ATPase enzyme activity were studied in erythrocytes; cytotoxic and DNA damaging potential of NPs were determined in lymphocytes, along with mitochondrial membrane potential (MMP) and ADP/ATP ratio. Hemolysis, generation of oxidative stress, LDH leakage and reduced ATPase activity were observed in the erythrocytes treated with NPs alone (50 µg/ml), the results were statistically significant at p < 0.05. Oxidative stress was evident by increased levels of malonaldehyde, indicating lipid peroxidation and generation of reactive oxygen species including hydrogen peroxide, together with statistically significant decrease in the activities of catalase and superoxide dismutase and reduced glutathione levels. In the lymphocytes treated with NPs alone (50 µg/ml), cytotoxicity in MTT assay and DNA damage in comet assay were observed; in addition, mitochondrial membrane potential collapsed and ADP/ATP ratio increased indicating mitochondrial function impairment. However, in the presence of CoQ10, hemolysis, oxidative stress and LDH leakage in the erythrocytes and lymphocyte cytotoxicity and DNA damage were drastically reduced, enzyme activities, MMP and ADP/ATP ratio were restored towards normal levels. TiO2 NPs induce cytotoxicity, damage DNA in lymphocytes, and induce oxidative/anti-oxidative imbalance in erythrocytes. Antioxidant CoQ10 protects erythrocytes and lymphocytes from toxicity induced by TiO2 NPs.

Influence of zinc levels on the toxic manifestations of lead exposure among the occupationally exposed workers.

Lead and zinc are usually found at the same occupational places and occur as co-contaminants. Effects of lead toxicity are detrimental on human health as it is probable carcinogen and impairs normal growth and development. On the other hand, zinc is an important nutritional element, the deficiency of which causes debilitating effects on growth and development. The purpose of this study was to examine the possible association of blood lead and zinc levels and any influence of zinc over DNA damage, blood cell membrane aberration and oxidative stress among lead and zinc co-exposed workers. Atomic absorption spectroscopy was used for lead and zinc measurement and comet assay for DNA damage assessment. Haematological aberrations were studied using light and electron microscopy (LM and EM) followed by electron density X-ray spectroscopy (EDS) and elemental mapping. Occupational exposure was observed to cause significant elevation in blood lead levels among workers. This elevation in lead levels and associated DNA damage among workers was significantly high in comparison to controls. Further light and electron micrographs of red blood cells revealed significant morphological alterations associated with increased lead ions in workers. It was clear from SEM-based elemental maps and EDS graphs that elevated lead levels were associated with low levels of zinc. The results suggest that lead absorption is highly influenced due to zinc levels in body which has an impact over DNA damage, blood cell aberration and oxidative stress caused by lead exposure. Efforts are going on to understand the role of other trace metals on lead toxicity in order to develop a sustainably nutrition-based therapeutic intervention. Graphical abstract.