Suresh K. Verma

Mechanistic insight to ROS and Apoptosis regulated cytotoxicity inferred by Green synthesized CuO nanoparticles from Calotropis gigantea to Embryonic Zebrafish

With the rapid development of nanotechnology, much has been anticipated with copper oxide nanoparticles (CuO NP) due to their extensive industrial and commercial application. However, it has raised concern over the environmental safety and human health effects. In this study, CuO nanoparticles were synthesized using the green method with floral extract of Calotropis gigantea and characterized by standard physiochemical techniques like DLS, Zeta potential determination, UV- Visible Spectroscopy, XRD, FTIR, FESEM, and TEM. Mechanistic cytotoxicity studies were performed using experimental and computational assays including morphological analysis, hatching, and viability rate analysis along with ROS and apoptosis analysis. Physiochemical characterization of CuO NP determined the size and zeta potential of synthesized nanoparticles to be 30 ± 09 nm to 40 ± 2 nm and −38 mV ± 12 mV respectively. Cytotoxicity evaluation with Zebrafish revealed malfunctioned organ development with differential viability and hatching rate at 48 hpf and 72 hpf with LC50 of 175 ± 10 mg/l. Computational analysis depicted the influential role of CuO nanoparticles on zebrafish embryo’s he1a, sod1 and p53 functional expression through hydrophobic and hydrogen bond interaction with amino acid residues. Study demonstrated valuable information of cytotoxic impact which can be influential in further studies of their eco-toxicological effects.

Altered physiochemical properties in industrially synthesized ZnO nanoparticles regulate oxidative stress; induce in vivo cytotoxicity in embryonic zebrafish by apoptosis

This study investigates the in vivo cytotoxicity of ZnO nanoparticles synthesized at industrial scale with embryonic Zebrafish. Industrial synthesis of ZnO nanoparticles was mimicked at lab scale by high energy ball milling technique by milling bulk ZnO particles for 15 h. Synthesized 7 h and 10 h ZnO nanoparticles showed significant alteration of size, zeta potential and optical properties in comparison to Bulk ZnO. Mortality and hatching rate in Zebrafish embryos were influenced by these alterations. Size and charge dependent effect of ZnO nanoparticles exposure on physiology and development of Zebrafish embryos were evident by malfunctioned organ development and abnormal heartbeat rate. Similar dependency on quenching of ROS due to influential hydrogen bond interaction with glycine residue of Sod1 oxidative stress protein and increased apoptosis were observed in cells. The study revealed the mechanism of cytotoxicity in exposed embryonic Zebrafish as an effect of accumulation and internalization inside cells instigating to generation of hypoxic condition and interference with the normal adaptive stress regulation signaling pathways leading towards enhanced apoptosis. The study revealed hidden size and charge dependent in vivo cytotoxicity mechanism of ZnO nanoparticles in Zebrafish embryos insight of the environmental and clinical importance of attention on industrially synthesized ZnO nanoparticles.

Mechanistic Insight into Size-Dependent Enhanced Cytotoxicity of Industrial Antibacterial Titanium Oxide Nanoparticles on Colon Cells Because of Reactive Oxygen Species Quenching and Neutral Lipid Alteration

This study evaluates the impact of industrially prepared TiO2 nanoparticles on the biological system by using an in vitro model of colon cancer cell lines (HCT116). Industrial synthesis of titanium oxide nanoparticles was mimicked on the lab scale by the high-energy ball milling method by milling bulk titanium oxide particles for 5, 10, and 15 h in an ambient environment. The physiochemical characterization by field emission scanning electron microscopy, dynamic light scattering, and UV–visible spectroscopy revealed alteration in the size and surface charge with respect to increase in the milling time. The size was found to be reduced to 82 ± 14, 66 ± 12, and 42 ± 10 nm in 5, 10, and 15 h milled nano TiO2 from 105 ± 12 nm of bulk TiO2, whereas the zeta potential increased along with the milling time in all biological media. Cytotoxicity and genotoxicity assays performed with HCT116 cell lines by MTT assay, oxidative stress, intracellular lipid analysis, apoptosis, and cell cycle estimation depicted cytotoxicity as a consequence of reactive oxygen species quenching and lipid accumulation, inducing significant apoptosis and genotoxic cytotoxicity. In silico analysis depicted the role of Sod1, Sod2, p53, and VLDR proteins–TiO2 hydrogen bond interaction having a key role in determining the cytotoxicity. The particles exhibited significant antibacterial activities against Escherichia coli and Salmonella typhimurium.

Mechanistic insight into the rapid one-step facile biofabrication of antibacterial silver nanoparticles from bacterial release and their biogenicity and concentration-dependent in vitro cytotoxicity to colon cells

Progress in the research and development of green synthesis of silver nanoparticles and their applications has reached new heights in the last decade. In this study, one-step rapid facile biosynthesis of silver nanoparticles is reported, and in vitro cytotoxicity of these nanoparticles has been investigated in an HCT116 cell line. Biogenic silver nanoparticles were synthesized from the culture supernatant of Gram-positive (B. thuringiensis and S. aureus) and Gram-negative bacteria (E. coli and S. typhimurium) using UV light, termed as BTAgNP, SAAgNP, ECAgNP, and STAgNP, respectively. The synthesized silver nanoparticles were characterised by standard characterisation methods such as field emission scanning electron microscopy (FESEM), dynamic light scattering (DLS), UV-visible spectroscopy, and Fourier transform infrared spectroscopy (FTIR). An in silico investigation was performed to elucidate the mechanism of their synthesis. Uniformly distributed ECAgNP, SAAgNP, STAgNP, and BTAgNP with stable zeta potentials were synthesized with the sizes 22.6 ± 5.2 nm, 21.2 ± 4.8 nm, 23.3 ± 6.8 nm, and 29.3 ± 5.2 nm, respectively. The synthesized silver nanoparticles were found to exhibit significant antibacterial activity against their source bacteria. An in vitro assessment revealed their biogenicity and concentration-dependent cytotoxicity and genotoxicity in colon cell lines with the occurrence of morphological deformities, oxidative stress, apoptosis, and cell cycle arrest. The study provided an insight into the biogenic differences in the biological effects of silver nanoparticles.

Molecular aspect of silver nanoparticles regulated embryonic development in Zebrafish ( Danio rerio) by Oct-4 expression

With the enhancement of commercial manifestation of silver nanoparticles, concerned has risen on their accumulation in aquatic system and consequent effects on fish development and metabolism. In this study, experiments were conducted to assess the impacts of silver nanoparticles on early life cycles of fish considering Zebrafish (Danio rerio) as experimental model. Silver nanoparticles were synthesized through chemical reduction method and characterized through UV-visible spectroscopy, dynamic light scattering (DLS), and HR-TEM. Different sub lethal doses of nanosilver were applied (13.6, 21.6, 42.4, 64, and 128 μgL-1) to post-fertilization phases of Zebrafish embryos and their interaction effects were monitored up to six days period. No significant morphological variations were observed at 13.6, 21.6, 42.4 μgL-1 dose of silver nanoparticles, whereas 64 and 128 μgL-1 exposure dose exhibited bending in myotome, deformity in tail region, somites, notochord and swelling in anterior and posterior region of embryos and larva. Hatching performances analysis elicited highest hatching success in 13.6 and 21.6 μgL-1 doses of silver nanoparticles followed by positive and negative control, whereas exposure dose of 64 and 128 μgL-1 exhibited comparatively lower success. Western blot analysis were conducted on developing hatchlings with Oct4 antibody and at 13.6 and 21.6 μgL-1dose,it showed over expression elucidating stimulatory role of nanosilver in these mentioned doses. In silico analysis depicted a firm interaction of nanosilver with Oct4 revealing their key role in growth stimulation of developing embryos. The study demonstrates the function of nanosilver as a growth promoter rather only as a toxicant in fish metabolic system.

In vivo assessment of impact of titanium oxide nanoparticle on zebrafish embryo

Technologies and innovations have attended a new height with recent development in nanotechnology in last few decades. With these developments there has a great raise in demand of metal oxides like TiO2, ZnO having versatile physical, chemical and biological application. However the great rise has raised concern over the effect of these nanoparticles in biological system. In this study, we have assessed the impact of titanium oxide nanoparticles synthesized by high energy ball milling (HEBM) by milling bulk TiO2 particles for 15h. The synthesized particles were characterized with XRD, UV-Visible spectroscopy and DLS for their physiochemical properties. Biological impact of these nanoparticles was then studied on zebrafish embryo as invivo model. Mortality and hatching rate were calculated for 48hpf and 96hpf treatment. To determine the mechanism of mortality effect, Reactive oxygen species (ROS) was determined with the help of flow cytometry. 15h nanoparticles were found to have a LC50 of ( ) for zebrafis...

Nanotoxicity of Rare Earth Metal Oxide Anchored Graphene Nanohybrid: A Facile Synthesis and In Vitro Cellular Response Studies

Graphene, a single sp2 bonded carbon, is now a burgeoning interest with various fascinating properties in a large number of biomedical applications. Consequently, the impact of graphene-based functional nanohybrid and its potential risk to human health have raised considerable public concerns. In this present study, we have synthesized cerium oxide (CeO2) anchored reduced graphene oxide (RGO) nanohybrid and a detailed study on its nanotoxicity profile has also been scrutinized. To confirm the efficient synthesis of nano-CeO2/RGO nanohybrid, the systematic characterization has been carried out using FTIR, Raman and UV-Vis spectroscopic analysis. The successful imprint of CeO2 nanoparticles (NPs) on RGO nanosheet was also evident from the scanning electron microscopy (SEM) and transmission electron microscopy (TEM) micrographs. A dose-dependent in vitro nanotoxicity of the nanohybrid has been assessed by using monocyte macrophage cells-Raw264.7 and colon cancer cells-HCT116 as compared with RGO and CeO2. The results conferred that as compared with single nanostructures (RGO or CeO2), nanohybrid showed excellent biocompatibility and no such prominence morphological alteration of the cell structure. Moreover, after exposure of different nanomaterials to HCT116 cells, the possible cellular interaction was investigated through reactive oxygen species (ROS) measurements using flow cytometry analysis dicholoro-dihydro-fluorescen dia-acetate (DCF) assay. These results conveyed that nanohybrid adapts an oxidative stress mechanism upon cellular interaction where it utilizes the scavenging property of CeO2, which induces the cell proliferation. Overall, the nano-CeO2/RGO nanohybrid exhibits a prolonged biocompatibility and cell viability, which is highly desired for biomedical applications.

Molecular insight to size and dose-dependent cellular toxicity exhibited by a green synthesized bioceramic nanohybrid with macrophages for dental applications

Improvising bioceramics for enhancing their biocompatibility and physical properties has been a focus area for the dental industry. To further explore this area, this study reports a novel green synthesis and molecular in vitro biocompatibility of calcium aluminosilicate-chitosan nanohybrid (CAS-CH). The nanohybrids were synthesized by using a high energy ball milling (HEBM) technique and then characterized for their physiochemical properties using standard techniques including scanning electron microscopy (SEM) and dynamic light scattering (DLS). In vitro cytotoxicity evaluation of a synthesized nanohybrid was made with a RAW264.7 cell line using cell viability assays, such as, MTT, cellular morphology analysis, induction of oxidative stress, and apoptosis. CAS-CH nanohybrids were synthesized at three milling time points: 1H, 2H, and 3H. With increasing milling time, we found a reduction in sizes of particles and increased zeta potential. Viability of cells was found to be decreased with an increase in concentration. Moreover, toxic effects like ROS generation and apoptosis were reduced with increasing milling time. Computational and experimental analysis elucidated the mechanism of toxicity as a consequence of influential functionality of Sod1 and p53 proteins due to interaction and internalization of the nanohybrids with amino acid residues via hydrogen bonds and hydrophobic interactions. The detailed study depicted a novel way of synthesizing biocompatible bioceramic nanohybrids with a mechanistic insight of its cytotoxicity profile.

In vivo molecular toxicity profile of dental bioceramics in embryonic Zebrafish (Danio rerio)

The investigation of the biocompatibility of potential and commercially available dental material is a major challenge in dental science. This study demonstrates that the zebrafish model is a novel in vivo model for investigating the biocompatibility of dental materials. Two commercially available dental materials, mineral trioxide aggregate (MTA) and Biodentine, were assessed for their biocompatibility. The biocompatibility analysis was performed in embryonic zebrafish with the help of standard toxicity assays measuring essential parameters such as survivability and hatching. The mechanistic and comparative analysis of toxicity was performed by oxidative stress analysis by measuring ROS induction and apoptosis in zebrafish exposed to dental materials at different concentrations. The molecular investigation at the protein level was done by a computational approach using in silico molecular docking and pathway analysis. The toxicity analysis showed a significant reduction in hatching and survivability rates along with morphological malformations with an increase in the concentration of exposed materials. ROS and apoptosis assay results revealed a greater biocompatibility of Biodentine as compared to that of MTA which was concentration-dependent. In silico analysis showed the significant role of the tricalcium silicate-protein ( Sod1, tp53, RUNX2B) interaction in an exhibition of toxicity. The study provides a new vision and standard in dental material sciences for assessing the biocompatibility of potential novel and commercially available dental materials.

Molecular investigation to RNA and protein based interaction induced in vivo biocompatibility of phytofabricated AuNP with embryonic zebrafish

Abstract Implication of gold nanoparticles in industrial and day-to-day life products at extensive scale has raised concern about their toxicity to environment and human health. Moreover, quest of new technologies for production of biocompatible nanoparticles increased. This study explores the molecular toxicology of AuNP with enlightenment of their green synthesis using medicinal plant extract as reducing and stabilizing agent. Synthesized CAuNP were characterized for their physiochemical properties by standard techniques like FESEM, TEM, DLS, UV–Vis spectroscopy and FTIR. GCMS analysis revealed the involvement of –OH compounds for CAuNP synthesis. Determined size and zeta potential of CAuNP was found to be 21 ± 08 nm and –24 ± 11 mV with SPR peak at 554 nm. LC50 of CAuNP with zebrafish embryos was 69 ± 12 µg/ml compared to 52 ± 06 µg/ml of AuNP. Gold nanoparticles were found to exhibit concentration dependent morphological abnormalities with acute effect at cellular and molecular level. Experimental and computational analysis depicted the nanotoxicity of gold nanoparticles as a consequence of oxidative stress generation leading to apoptosis due to their influential interaction with Sod1, He1a and tp53 mRNA and proteins. The investigation deciphered the nanotoxicity of gold nanoparticles and suggested the implication of new green methodology for their future productions.