Muthuraman Pandurangan

Cytotoxicity of zinc oxide nanoparticles on antioxidant enzyme activities and mRNA expression in the cocultured C2C12 and 3T3-L1 cells.

The present study was aimed to investigate the dose-dependent effect of zinc oxide (ZnO) nanoparticles on antioxidant enzyme activities and messenger RNA (mRNA) expression in the cocultured C2C12 and 3T3-L1 cells. Coculturing experiments are 3D and more reliable compared to mono-culture (2D) experiment. Even though, there are several studies on ZnO nanoparticle-mediated cytotoxicity, but there are no studies on the effect of ZnO nanoparticle on antioxidant enzyme activities and mRNA expression in the cocultured C2C12 and 3T3-L1 cells. A cytotoxicity assay was carried out to determine the effect of ZnO nanoparticles on the C2C12 and 3T3-L1 cell viability. At higher concentration of ZnO nanoparticles, C2C12 and 3T3-L1 cells almost die. ZnO nanoparticles increased reactive oxygen species (ROS) and lipid peroxidation and reduced glutathione (GSH) levels in a dose-dependent manner in the C2C12 and 3T3-L1 cells. In addition, ZnO nanoparticles increased antioxidant enzyme activities and their mRNA expression in the C2C12 and 3T3-L1 cells. In conclusion, the present study showed that ZnO nanoparticles increased oxidative stress, antioxidant enzyme activities, and their mRNA expression in the cocultured C2C12 and 3T3-L1 cells.

Time and Concentration-Dependent Therapeutic Potential of Silver Nanoparticles in Cervical Carcinoma Cells.

Silver nanoparticles (AgNPs) have well-known anti-bacterial properties and have been widely used in daily life as various medical and general products. There is limited information available on the cytotoxicity of AgNPs. Therefore, the present study aimed to investigate the cytotoxicity of AgNPs in HeLa cells. Cytotoxicity and apoptosis have been observed in the AgNPs treated in the HeLa cells. Sulphorhodamine-B assay (SRB assay) showed the cytotoxic effect in the AgNP-treated HeLa cells. Inverted microscope, fluorescence microscope, and confocal laser scanning microscope (CLSM) analyses showed the apoptosis-induced morphological changes such as rounding in shape, nuclear fragmentation, cytoplasm reduction, loss of adhesion, and reduced cell volume. Necrosis and apoptosis were observed in the AgNP-treated HeLa cells by DNA fragmentation study. Mitochondria-derived reactive oxygen species (ROS) have increased in AgNP-treated HeLa cells. Up-regulation of messenger RNA (mRNA) expression of p53, bax, and caspase 3 were found in AgNP-treated HeLa cells. Caspase 3 enzyme activity was found to increase in AgNP-treated HeLa cells. The AgNPs showed the right cytotoxic effect in cervical carcinoma cells. Our results suggest that metal-based nanoparticles might be a potential candidate for the treatment of cervical cancer.

Anticancer studies of synthesized ZnO nanoparticles against human cervical carcinoma cells

A metal oxide nanoparticle has been widely investigated for its potential use in the biomedical application. The present study investigates the cytotoxicity of ZnO nanoparticle in human cervical carcinoma cells. Cell viability was determined, and it showed the possible cytotoxic effect of ZnO nanoparticles. The characteristic apoptotic features such as rounding and loss of adherence were observed in the treated cells. Fluorescence and Confocal Laser Scanning Microscope (CLSM) studies have showed reduced nuclear volume and condensed cytoplasm. The mRNA expression of apoptotic gene p53 and caspase 3 was up-regulated following ZnO nanoparticle exposure, which confirms the occurrence of apoptosis at the transcriptional level. Reactive oxygen species (ROS) was increased in a dose-dependent manner, and initiate lipid peroxidation of the liposomal membrane, which in turn regulate several signaling pathways and influencing the cytokinetic movements of cells. ZnO nanoparticles showed a dynamic cytotoxic effect in cervical carcinoma cells. ZnO nanoparticle might induce the apoptosis through increased intracellular ROS level. Moreover, up-regulated apoptotic gene expression confirms the occurrence of apoptosis. Taking all these data together, it may be concluded that ZnO nanoparticle may exert cytotoxicity on HeLa cell through the apoptotic pathway, implies the probable utility of ZnO nanoparticle in the cancer treatment and therapy.

In Vitro Therapeutic Potential of Tio2 Nanoparticles Against Human Cervical Carcinoma Cells

Cellular and physiological responses to the degradation products of titanium implants are key indicators to determine the quality of biocompatibility of implant devices. The present study investigated titanium dioxide (TiO2) nanoparticle-induced cytotoxicity, apoptotic morphological modification, and apoptotic-related gene expressions in the human cervical carcinoma cells. TiO2 nanoparticle-induced cytotoxicity on cancer cells was determined by the sulphorhodamine-B assay. Apoptotic morphological modification such as nuclear fragmentation, rounding, cytoplasm shrinkage, loss of adhesion, and reduced cell volume were observed by an inverted, fluorescence, and confocal laser scanning microscope (CLSM). The DNA fragmentation study showed the occurrence of necrosis and apoptosis in nanoparticle-treated cells. The qPCR study showed the increased p53 and bax mRNA expression in the nanoparticle-treated cells compared to control. In addition, caspase 3 activity was increased in nanoparticle-treated cells, which indicates the increased auto-catalysis. Taking all these data together, it may suggest that TiO2 nanoparticle could inhibit the growth of HeLa cells.

Probing the impact of quercetin-7-O-glucoside on influenza virus replication influence

BACKGROUND Influenza virus is still at large and seriously affects social welfare and health. Dianthus superbus is a well-known medicinal plant widely used in Mongolian and Chinese traditional medicine for anti-inflammatory purposes. PURPOSE To investigate the influence of this novel herbal medicinal product over virus infection and virus-induced symptoms METHOD Quercetin-7-O-glucoside was isolated by bioassay (anti-influenza)-guided fractionation. The structural elucidation was made with 1H-NMR and 13C-NMR. Influenza A/Vic/3/75 (H3N2), A/PR/8/34 (H1N1), B/Maryland/1/59 and B/Lee/40 viruses were used for the evaluation of the antiviral activity. Virus-induced reactive oxygen species and autophagy formation levels were studied. The antiviral mechanism was elucidated via time-dependent, pre-, post-incubation assay methods. The viral RNA replication inhibition of Q7G was analyzed using quantitative RT-PCR method. The blocking of polymerase basic protein subunits of influenza viral RNA polymerase by Q7G was detected by in silico molecular docking assays using AutoDock Vina program with m(7)GTP. Additionally, Q7G was tested against M-MuLV RNA polymerase. RESULTS Q7G was not cytotoxic (CC50>100µg/ml) in MDCK cells and it showed 3.1µg/ml, 6.61µg/ml, 8.19µg/ml and 5.17µg/ml IC50 values against influenza A/PR/8/34, A/Vic/3/75, B/Lee/40 and B/Maryland/1/59 virus strains, respectively. Treatment of Q7G highly reduced ROS and autophagy formation induced by influenza virus infection. Q7G did not reduce NA activity and did not directly interact with the virus particles. Since viral RNA synthesis was blocked by treatment of Q7G. We targeted viral RNA polymerase for further probing. Interestingly, the binding energy of Q7G on viral PB2 protein was -9.1kcal/mol and was higher than m(7)GTP recorded as -7.5kcal/mol. It also was observe to block M-MuLV RNA polymerase. CONCLUSION Isolated compound Q7G showed strong inhibition activity against influenza A and B viruses. It also reduced virus-induced ROS and autophagy formation. Q7G does not directly bind to the virus particles and did not affect NA activity. These results indicated that Q7G inhibits viral RNA polymerase, and that it occupies the binding site of m(7)GTP on viral PB2 protein.

In vitro anticancer potential of BaCO3 nanoparticles synthesized via green route

Green synthesis of nanoparticles is a growing research area because of their potential applications in nanomedicine. Barium carbonate nanoparticles (BaCO3 NPs) were synthesized using an aqueous extract of Mangifera indica seed as a reducing agent. These particles were characterized by Fourier transform infrared spectroscopy (FT-IR), X-ray diffraction (XRD), Transmission electron microscopy (TEM), selected area electron diffraction (SAED), Energy-dispersive-X-ray (EDX) and X-ray photoelectron spectroscopy (XPS) analysis. HR-TEM images are confirmed that green synthesized BaCO3 NPs have spherical, triangular and uneven shapes. EDX analysis confirmed the presence of Ba, C and O. The peaks at 2θ of 19.45, 23.90, 24.29, 27.72, 33.71, 34.08, 34.60, 41.98, 42.95, 44.18, 44.85, and 46.78 corresponding to (110), (111), (021), (002), (200), (112), (130), (221), (041), (202), (132) and (113) showed that BaCO3 NPs average size was ~18.3 nm. SAED pattern confirmed that BaCO3 NPs are crystalline nature. BaCO3 NPs significantly inhibited cervical carcinoma cells, as evidenced by cytotoxicity assay. Immunofluorescence and fluorescence assays showed that BaCO3 NPs increased the expression and activity of caspase-3, an autocatalytic enzyme that promotes apoptosis. According to the results, green synthesis route has great potential for easy, rapid, inexpensive, eco-friendly and efficient development of novel multifunctional nanoparticles for the treatment of cancer.

Investigation of role of aspartame on apoptosis process in HeLa cells

Aspartame is an artificial sweetener used as an alternate for sugar in several foods and beverages. The study reports that consumption of aspartame containing product could lead to cancer. However, the effect of aspartame on apoptosis process in cancer is not yet understood clearly. HeLa cells were exposed to different concentrations (0.01–0.05 mg/ml) of aspartame for 48 h. Cytotoxicity of aspartame on cancer cells was determined by SRB assay. The result indicates no significant changes on cell viability. Aspartame suppresses apoptosis process in cancer cells by down-regulation of mRNA expression of tumor suppressor gene p53, and pro-apoptotic gene bax. It up-regulates anti-apoptotic gene bcl-2 mRNA expression. In addition, Ki 67 and PCNA mRNA, and protein expressions were determined. Taking all these together, we conclude that aspartame may be a potent substance to slow-down the apoptosis process in HeLa cells. Further works are ongoing to understand the biochemical and molecular mechanism of aspartame in cancer cells.

Green Synthesis: In-vitro Anticancer Activity of Silver Nanoparticles on Human Cervical Cancer Cells

The development of an eco-friendly process for the synthesis of nanoparticles is an important and emerging area in the field of nanotechnology. Silver nanoparticles (AgNPs) were synthesized using Saccharina japonica extract and their cytotoxicity to cervical carcinoma cells was examined. The AgNPs were characterized by ultraviolet–visible absorption spectroscopy, Fourier transform infrared spectroscopy, high resolution-transmission electron microscopy, energy dispersive X-ray spectroscopy, X-ray diffraction, and selected area electron diffraction. The biosynthesized AgNPs were spherical in shape, and HeLa cells exposed to these NPs showed morphological changes, including rounding and loss of adherence. Fluorescence and confocal laser scanning microscopy revealed the characteristic apoptotic features such as reduction in nuclear volume and cytoplasm condensation.

A novel approach for in vitro meat production

The present review describes the possibility of in vitro meat production with the help of advanced co-culturing methods. In vitro meat production method could be a possible alternative for the conventional meat production. Originally, the research on in vitro meat production was initiated by the National Aeronautics and Space Administration (NASA) for space voyages. The required key qualities for accepting in vitro meat for consumption would be good efficiency ratio, increased protein synthesis rate in skeletal muscles, and mimicking the conventional meat qualities. In vitro culturing of meat is possible with the use of skeletal muscle tissue engineering, stem cell, cell co-culture, and tissue culture methods. Co-culture of myoblast and fibroblast is believed as one of the major techniques for in vitro meat production. In our lab, we have co-cultured myoblast and fibroblast. We believe that a billion pounds of in vitro meat could be produced from one animal for consumption. However, we require a great deal of research on in vitro meat production.

Application of cell co-culture system to study fat and muscle cells

Animal cell culture is a highly complex process, in which cells are grown under specific conditions. The growth and development of these cells is a highly unnatural process in vitro condition. Cells are removed from animal tissues and artificially cultured in various culture vessels. Vitamins, minerals, and serum growth factors are supplied to maintain cell viability. Obtaining result homogeneity of in vitro and in vivo experiments is rare, because their structure and function are different. Living tissues have highly ordered complex architecture and are three-dimensional (3D) in structure. The interaction between adjacent cell types is quite distinct from the in vitro cell culture, which is usually two-dimensional (2D). Co-culture systems are studied to analyze the interactions between the two different cell types. The muscle and fat co-culture system is useful in addressing several questions related to muscle modeling, muscle degeneration, apoptosis, and muscle regeneration. Co-culture of C2C12 and 3T3-L1 cells could be a useful diagnostic tool to understand the muscle and fat formation in animals. Even though, co-culture systems have certain limitations, they provide a more realistic 3D view and information than the individual cell culture system. It is suggested that co-culture systems are useful in evaluating the intercellular communication and composition of two different cell types.