br Nanomaterials nanoparticles cause cell death when
Nanomaterials/nanoparticles cause cell death when taken up by the human tissues and cellular system and hence cell died causes via oxidative stress led by toxicity of nanoparticles. Size of
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E-mail addresses: [email protected] (M. Fakhar-e-Alam), [email protected]
nanoparticles depends on kinetics of uptake process, localization in WY 14643 and exocytose, physicochemical, surficial properties and on the ability to bond. Kinetics of uptake of nanoparticles also depends upon morphology of cell and its nature [6e9].
Silver oxide nanoparticles are used as antiviral therapeutics against such type of viruses that causes disease. Silver nano-particles are mainly focused to study for possible destruction against bacteria. It is also realized that silver oxide nanoparticles are beneficial against several types of viruses. Synthesized nano-materials can be applied as Nano medicinal and Nano therapeutic drugs due to their anti-cancer, anti-viral properties and other sur-face charge property which might be beneficial for treating cancer and other viral/bacterial diseases. Due to strong antibacterial property it may be used for living organisms, including the human body, in the food, and also through skin or the respiratory system .
Silver nanoparticles go into individuals by oral absorption, inhalation, through damage skin still during hurdle of retina in mature and transmission or endocytosis through the skin of em-bryos. Ag nanoparticles penetrate into female genital tract and commonly female use different hygienic products consisting of
silver (Ag) NPs [11e14]. Thus, nanomaterials provide an opportu-nity to overcome this challenge of drug delivery in the target site . It is expected that nanotechnology will bring a central change in biological applications such as drug delivery and diagnostics in the coming few decades .
In this experimental study, after successful synthesis and char-acterization steps, authors are interested to explore the real cyto-toxicity and photo toxicity mechanism via proper pathway/chain reaction for cell killing effect.
2.1. Materials and methods
In current work co-precipitation method was applied for syn-thesis of silver oxide nanoparticles. This method is simple, direct and straight forward method. The detail for the preparation of sil-ver oxide nanoparticles is discussed in detail in the sample preparation.
2.2. Sample preparation
Silver oxide nanoparticles were prepared via well-developed co-precipitation method using silver nitrate (1.70 g), de-ionized water (10 mL). AgNO3 was mixed with de-ionized water and stirred to have homogeneous solution by magnetic stirrer. Triethylamine (10 mL) included in the solution and stirred for 200 min. Centrifu-gation process of obtained precipitates proceeded at 6500 rpm for 7e10 min. These precipitates are washed with distilled water or ethanol (10 mL). Washed precipitates were dried at 30 C then placed for 24 h in an incubator. Prepared amount of precipitates were placed in an electric oven at 200 C for 4 h than again in furnace at 800 C for 24 h. Furnace can be used to dry particles and also to minimize the size of particles. By controlling the morality ratio of solution, pH, time, temperature and addition of possible seed solution the desired shape and size of silver oxide nano-particles can be achieved. For cancer cell toxicity the size of developed nanoparticles must be within the range of 100 nm which is less than the pore size of Biological model.
2.3. Cell culturing and labelling conditions
In cell culturing process, HepG2 cell line was cultivated in tissue cultured plastic flasks (Nunc Wiesbaden Germany) in Minimum Essential Medium (MEM) with Hanks salts, supplemented with 10% fetal bovine serum (FBS), 2 mL glutamine and with some nones-sential amino acids. Furthermore, for attachment to the substratum properly, the cells were incubated at 37 C for 24 h. The cells were also sub cultured for two or three times in a week. After that, the cells were harvested via trypsin 0.25% once reached to the confluence of 75e85% . HepG2 cells having concentration of 1 105 cells/well were incubated with different concentrations ranging from to 90 mg/mL of silver oxide nanoparticles.
2.4. Cell viability assay
Cell life is determined by using dye (neutral red assay) [5,6]. Culture media was replaced after 24 h with fresh 200 mL MEM as well as 50 mL (50 mg/mL) of neutral red assay. In first four columns of 96 well plates poured concentrated neutral red assay and in eleventh and twelfth column remains empty. The 96-well-plates were placed in an incubator for 3 h and then media was removed and washed 2e3 times each well with 250 mL PBS, and evaluated the neutral red by using a mixture 1:1 of 50 vol % ethanol and 1 vol % acetic acid. The plate was shaken for 60sec and then left for