Systemic Toxicity and Teratogenicity of Copper Oxide Nanoparticles and Copper Sulfate.

Acceleration in development of metallic nanoparticles for their utility in medical and technological applications due to their unique physicochemical properties has concurrently raised a matter of concern due to their potential toxicity. Of the enormous metallic nanostructures, copper oxide nanoparticles (CuONPs) having optical and electrochemical properties are scrutinized for theranostic applications. Therefore, their safety profile is of a major concern in optimizing a safe dose for its clinical utility. Considering the potency of CuONPs in epitomizing toxicity, we report a dose and time dependent acute, systemic and transgenerational toxicity profile of CuONPs in comparison to the bulk copper as copper sulfate (CuSO4). Acute toxic dose (LD50(14)) of CuONPs (400 mg/kg · b · wt) was found to be three fold higher that of CuSO4(100 mg/kg · b · wt). Comparative steady state evaluation showed that CuONPs (≥5 mg/kg · b · wt.) induce greater dose and time dependent oxidative stress by increase in protein carbonylation and decreased glutathione levels in comparison to the bulk CuSO4. Furthermore, CuONPs were found to disrupt blood brain barrier (BBB) and sneak in to the brain which was quantified by atomic absorption spectroscopy (AAS) and also coax toxicity in liver, kidney and spleen, ascertained by histopathological findings (at ≥5 mg/kg · b · wt.). Considering transgenerational toxicity, CuONPs in comparison to CuSO4 severely affected sperm count and morphology in male animals, though not much teratological effects were observed, except certain extent of embryo resorption. The present study highlights a complete toxicity profile of CuONPs, giving forethought for considering them for clinical applications.

Assessment of genotoxic effect of maleic acid and EDTA: a comparative in vitro experimental study.

OBJECTIVES: This study aims to evaluate and compare the genotoxic and apoptotic effect of aqueous solutions of ethylenediaminetetraacetic acid (EDTA) with that of maleic acid (MA) using Chinese hamster lung fibroblast (V79) cells growing in vitro. MATERIALS AND METHODS: Exponentially growing V79 cells were treated with various concentrations of EDTA or MA alone for 30 min, and genotoxic effect was analyzed by micronucleus as well as comet assays and the type of cell death by apoptotic cell measurements using microscopic and flow cytometric methods. For all the experiments, H2O2 was used as a positive control. RESULTS: Treatment of V79 cells with H2O2 resulted in significantly (P < 0.001) increased micronuclei and levels of DNA damage, whereas, EDTA/MA alone treated cells did not show significant increase of MN frequencies and comet parameters even at their higher concentrations when compared with that of untreated control. V79 cells treated with EDTA/MA for 30 min showed a nonsignificant increase in the percentage of apoptotic and necrotic cells at their lower concentrations (0.025 and 0.05 % for EDTA and MA, respectively). However, at higher concentrations, i.e., >IC50 (0.1 and 0.5 %) for EDTA and MA resulted in increased number of apoptotic and necrotic cells when compared with the untreated group. CONCLUSIONS: This study clearly demonstrates that MA and EDTA are not potentially genotoxic agents and MA induced lesser apoptotic/necrotic death than that of EDTA at their clinically relevant doses. CLINICAL RELEVANCE: MA may have a better clinical acceptability with comparable smear layer removal ability. Hence, the results presented here might be an additional supporting evidence for the use of MA in endodontic practice.

Protective effect of zingerone, a dietary compound against radiation induced genetic damage and apoptosis in human lymphocytes.

Zingerone a dietary compound was investigated for its ability to protect against radiation induced genotoxicity and apoptosis in human lymphocytes growing in vitro. The radiation antagonistic potential of zingerone was assessed by alkaline comet, cytokinesis-block micronucleus, apoptosis and reactive oxygen species inhibition assays. Treatment of lymphocytes with zingerone (10µg/ml) prior exposure to 2Gy gamma radiation resulted in a significant reduction of frequency of micronuclei as compared to the control set of cells evaluated by cytokinesis blocked micronucleus assay. Similarly, treatment of lymphocytes with zingerone prior to radiation exposure showed significant decrease in the DNA damage as assessed by comet parameters, such as percent tail DNA and Olive tail moment. Further, treatment with zingerone (10µg/ml) before irradiation significantly decreased the percentage of apoptotic cells analyzed microscopically method and by DNA ladder assay. Similarly, the radiation induced reactive oxygen species levels were significantly (P<0.01) inhibited by zingerone. Our study demonstrates the protective effect of zingerone against radiation induced DNA damage and antiapoptotic effect in human lymphocytes, which may be partly attributed to scavenging of radiation induced free radicals and also by the inhibition of radiation induced oxidative stress.

Cytotoxic, genotoxic and oxidative stress induced by 1,4-naphthoquinone in B16F1 melanoma tumor cells.

Quinones have diverse pharmacological properties including antibacterial, antifungal, antiviral, anti-inflammatory, antipyretic and anticancer activity. The cytotoxic potential of 1,4-naphthoquinone (NQ14) was studied against B16F1 melanoma cells grown in vitro. NQ14 treatment resulted in a concentration-dependent cytotoxicity as indicated by MTT assay and lactate dehydrogenase leakage assay. Depletion in cellular glutathione levels after 1h incubation with NQ14 correlated with the corresponding increase in reactive oxygen species generation as determined by 2',7'-dicholorofluorescein diacetate assay suggests the role of oxidative stress in cell death. The frequency of micronucleated binucleate cells increased with increasing doses of NQ14 with a corresponding decrease in the cytokinesis block proliferation index indicating the drug induced genotoxicity and cell division delay. Further, a dose-dependent decrease in the clonogenic cell survival indicated the potential of NQ14 to inhibit cell proliferation contributing to cell death. The cell death after NQ14 treatment may be attributed to apoptosis as seen in DNA ladder pattern along with necrosis as indicated in flow cytometric analysis of Annexin V/PI stained cells. Results of the present study demonstrate the cytotoxic and genotoxic potential of NQ14 by the induction of oxidative stress mediated mechanisms leading to tumor cell kill.