Evidence of a non-apoptotic mode of cell death in microglial BV-2 cells exposed to different concentrations of zinc oxide nanoparticles.

Zinc oxide nanoparticles (ZnO NPs) possess huge application potential. However, the toxicity of ZnO NPs is a great cause of concern. Indeed, ZnO NPs have been found to cause neurotoxicity. As microglial dysfunctions have been linked to the neurotoxic potential of NPs, the physico-chemical properties of ZnO NPs were determined and their cytotoxic effects were characterised on murine microglial BV-2 cells. In-house prepared and meticulously characterised ZnO NPs exhibited narrow size distribution with an average size of around 20 nm and a zeta potential at physiological pH around 24 mV. ZnO NPs did not exhibit aggregation in the cell culture medium. When microglial BV-2 cells were exposed for 6 and 24 h to ZnO NPs (5, 10, 20, 40, and 80 µg/mL), several cell damages were observed. Cellular accumulation of NPs in microglial BV-2 cells was associated with cell growth inhibition and cell death induction, measured by the trypan blue exclusion and MTT assays. Mitochondrial dysfunction and lysosomal alteration were associated with increased plasma membrane permeability measured by staining with DiOC6(3), acridine orange, and propidium iodide, respectively. In addition, an accumulation of reactive oxygen species (ROS) was detected after staining with dihydroethidium and dihydrorhodamine 123. No apoptotic features were present: no cells with condensed and/or fragmented nuclei (Hoechst staining) characteristic of apoptotic cells, absence of subG1 cells, absence of caspase-3 cleavage, and PARP fragmentation. With ZnO NPs (80 µg/mL), with the annexin V/propidium iodide (PI) assay, few apoptotic cells (annexin V+/PI- cells) were detected whereas (annexin V+/PI+ cells) evocating necrotic cells were mainly identified. No modification of the cells in the different phases of the cell cycle was found. Altogether, our data show that ZnO NPs induce a non-apoptotic mode of cell death associated with an accumulation of ROS, mitochondrial, and lysosomal dysfunction and plasma membrane damages in microglial BV-2 cells.Graphical abstract.

Engineered Nanoparticles with Antimicrobial Property.

BACKGROUND: The urge for the development and manufacture of new and effective antimicrobial agents is particularly demanding especially in the present scenario of emerging multiple drug resistant microorganisms. A promising initiative would be to converge nanotechnology to develop novel strategies for antimicrobial treatment. These distinct nano scale properties confer impressive antimicrobial capabilities to nanomaterials that could be exploited. Nanotechnology particularly modulates the physicochemical properties of organic and inorganic nanoparticles, rendering them suitable for various applications related to antimicrobial therapy compared to their bulk counterparts. However, a major issue associated with such usage of nanomaterials is the safety concern on heath care system. Hence, a thorough put knowledge on biocompatible nanostructures intended for antimicrobial therapy is needed. METHODS: A systematic review of the existing scientific literature is being attempted here which includes the properties and applications of a few nano structured materials for antimicrobial therapy and also the mechanism of action of nanomaterials as antimicrobial agents. Silver (Ag), Graphene, Quantum dots (QDs), Zinc oxide (ZnO) and chitosan nanoparticles are taken as representatives of metals, semiconductors, metal oxides and organic nanoparticles that have found several applications in antimicrobial therapy are reviewed in detail. RESULTS AND CONCLUSION: An ideal anti microbial should selectively kill or inhibit the growth of microbes but cause little or no adverse effect to the host. Each of the engineered nanomaterials reviewed here has its own advantages and disadvantages. Nanomaterials in general directly disrupt the microbial cell membrane, interact with DNA and proteins or they could indirectly initiate the production of reactive oxygen species (ROS) that damage microbial cell components and viruses. Some like silver nanoparticles have broad spectrum antibacterial activity while others like cadmium containing QDs shows both antibacterial as well as antiprotozoal activity. Nano material formulations can be used directly or as surface coatings or as effective carriers for delivering antibiotics. Polycationic nature of Chitosan NPs helps in conjugation and stabilization of metallic nanoparticles which will enhance their effective usage in antimicrobial therapy.

Engineered Zinc Oxide Nanoparticles; Biological Interactions at the Organ Level.

Zinc oxide nanoparticles (ZnO NPs) are one of the widely used nanoparticles with spectrum of application, in the areas like daily care products, sensors, antibacterial agents, and biomedical sector. With extensive application the risk of exposure at occupational and consumer level also increases. Huge amount of data are available on the biointeraction of ZnO NPs. Though the toxicity of ZnO NPs is attributed to particle dissolution inside the cellular compartments and their ability to generate the reactive oxygen species, the ambiguity prevails over the exact mechanism of toxicity. The in vivo studies on different animal models and humans suggest different level of toxicity in these organisms. However the synthetic route, physiochemical properties of the nanoparticle, mode of exposure and nature of the test system often influences these studies. Hence the study results vary and sometimes contradict on one another. The current review focuses on the interaction of ZnO NPs with different organ systems. It also points to the factors to be considered while undertaking such studies in order to ensure reliability of the results.

Assessing the systemic toxicity in rabbits after sub acute exposure to ocular irritant chemicals.

Eye is a highly vascularised organ. There are chances that a foreign substance can enter the systemic circulation through the eye and cause oxidative stress and evoke immune response. Here the eyes of rabbits were exposed, for a period of 7 days, to 5 known ocular irritants: Cetyl pyridinium chloride (CPC), sodium salicylate (SS), imidazole (IMI), acetaminophen (ACT) and nicotinamide (NIC). The eyes were scored according to the draize scoring. Blood collected from the treated rabbit were analyzed for haematological and biochemical parameters. After sacrifice, histological analysis of the eye and analysis of pro-inflammatory biomarkers (IL-1α, IL-1ß, IL-8 and TNF-α) in the cornea using ELISA was carried out. Spleen was collected and the proliferation capacities of spleenocytes were analyzed. Liver and brain were collected and assessed for oxidative stress. The eye irritation potential of the chemicals was evident from the redness and swelling of the conjunctiva and cornea. Histopathological analysis and ELISA assay showed signs of inflammation in the eye. However, the haematological and biochemical parameters showed no change. Spleenocyte proliferations showed only slight alterations which were not significant. Also oxidative stress in the brain and liver were negligible. In conclusion, chemicals which cause ocular irritation and inflammation did not show any systemic side-effects in the present scenario.