Balaram Das

Toxicity of cobalt oxide nanoparticles to normal cells; an in vitro and in vivo study

The aim of this study was to find out the intracellular signaling transduction pathways involved in cobalt oxide nanoparticles (CoO NPs) mediated oxidative stress in vitro and in vivo system. Cobalt oxide nanoparticles released excess Co++ ions which could activated the NADPH oxidase and helps in generating the reactive oxygen species (ROS). Our results showed that CoO NPs elicited a significant (p<0.05) amount of ROS in lymphocytes. In vitro pretreatment with N-acetylene cystine had a protective role on lymphocytes death induced by CoO NPs. In vitro and in vivo results showed the elevated level of TNF-α after CoO NPs treatment. This TNF-α phosphorylated the p38 mitogen-activated protein kinase followed by activation of caspase 8 and caspase 3 which could induce cell death. This study showed that CoO NPs induced oxidative stress and activated the signaling pathway of TNF-α-caspase-8-p38-caspase-3 to primary immune cells. This study suggested that bare CoO NPs are a toxic for primary human immune cells that deals directly with human health. Surface modification or surface functionalization may open the gateway for further use of CoO NPs in different industrial use or in biomedical sciences.

Chitosan-modified cobalt oxide nanoparticles stimulate TNF-α-mediated apoptosis in human leukemic cells

The objective of this study was to develop chitosan-based delivery of cobalt oxide nanoparticles to human leukemic cells and investigate their specific induction of apoptosis. The physicochemical properties of the chitosan-coated cobalt oxide nanoparticles were characterized using transmission electron microscopy, dynamic light scattering, X-ray diffraction, and Fourier transform infrared spectroscopy. The solubility of chitosan-coated cobalt oxide nanoparticles was higher at acidic pH, which helps to release more cobalt ions into the medium. Chitosan-coated cobalt oxide nanoparticles showed good compatibility with normal cells. However, our results showed that exposure of leukemic cells (Jurkat cells) to chitosan-coated cobalt oxide nanoparticles caused an increase in reactive oxygen species generation that was abolished by pretreatment of cells with the reactive oxygen species scavenger N-acetyl-l-cysteine. The apoptosis of Jurkat cells was confirmed by flow-cytometric analysis. Induction of TNF-α secretion was observed from stimulation of Jurkat cells with chitosan-coated cobalt oxide nanoparticles. We also tested the role of TNF-α in the induction of Jurkat cell death in the presence of TNF-α and caspase inhibitors. Treatment of leukemic cells with a blocker had a greater effect on cancer cell viability. From our findings, oxidative stress and caspase activation are involved in cancer cell death induced by chitosan-coated cobalt oxide nanoparticles.Graphical abstract

Cobalt oxide nanoparticles induced oxidative stress linked to activation of TNF-α/caspase-8/p38-MAPK signaling in human leukemia cells.

The purpose of this study was to determine the intracellular signaling transduction pathways involved in oxidative stress induced by nanoparticles in cancer cells. Activation of reactive oxygen species (ROS) has some therapeutic benefits in arresting the growth of cancer cells. Cobalt oxide nanoparticles (CoO NPs) are an interesting compound for oxidative cancer therapy. Our results showed that CoO NPs elicited a significant (P <0.05) amount of ROS in cancer cells. Co‐treatment with N‐aceyltine cystine (an inhibitor of ROS) had a protective role in cancer cell death induced by CoO NPs. In cultured cells, the elevated level of tumor necrosis factor‐alpha (TNF‐α) was noted after CoO NPs treatment. This TNF‐α persuaded activation of caspase‐8 followed by phosphorylation of p38 mitogen‐activated protein kinase and induced cell death. This study showed that CoO NPs induced oxidative stress and activated the signaling pathway of TNF‐α‐Caspase‐8‐p38‐Caspase‐3 to cancer cells. Copyright

Self-assembled betulinic acid protects doxorubicin induced apoptosis followed by reduction of ROS–TNF-α–caspase-3 activity

Doxorubicin (DOX) is a well-known drug used to treat a wide range of solid tumor and hematological malignancies, but the use of this drug is now restricted owing to its severe side effects, including normal cellular toxicity. This study was conducted to evaluate the potency of self-assembled betulinic acid (SA-BA) against DOX induced chemotherapeutic toxicity in human peripheral blood lymphocytes (PBLs). The isolated betulinic acid from the bark of Ziziphus jujuba tree was purified by column chromatography and characterized by FT-IR, XRD, (1)H NMR and self-assembly property was investigated by SEM imaging. DOX treatment produced significant reduction of viability of PBLs mainly by lowering cellular anti-oxidant pool and elevating the reactive oxygen species level. Pre-treatment with SA-BA followed by DOX exposure for 24h protected the PBLs from DOX induced oxidative stress. Potent anti-apoptotic role of SA-BA was also confirmed by FACS analysis and western blot assay. Severe inflammation is one of the major concerns in DOX treatment. We found that pre-treatment with SA-BA on PBLs significantly protected the PBLs from DOX induced inflammation. Thus, our finding confirms that SA-BA can be used to ameliorate the cytotoxic effects of DOX, which can be a helpful strategy during DOX mediated chemotherapy in cancer patients.

Bio-fabricated silver nanoparticles preferentially targets Gram positive depending on cell surface charge.

Recently bio-inspired experimental processes for synthesis of nanoparticles are receiving significant attention in nanobiotechnology. Silver nanoparticles (Ag NPs) have been used very frequently in recent times to the wounds, burns and bacterial infections caused by drug-resistant microorganisms. Though, the antibacterial effects of Ag NPs on some multi drug-resistant bacteria specially against Gram positive bacteria has been established, but further investigation is needed to elicit its effectiveness against Gram negatives and to identify the probable mechanism of action. Thus, the present study was conducted to synthesize Ag NPs using Andrographis paniculata leaf extract and to investigate its antibacterial efficacy. After synthesis process the biosynthesized nanoparticles were purified and characterized with the help of various physical measurement techniques which raveled their purity, stability and small size range. The antimicrobial activity of Ag NPs was determined against both Gram-positive Enterococcus faecalis and Gram-negative Proteus vulgaris. Results showed comparatively higher antibacterial efficacy of Ag NPs against Gram positive Enterococcus faecalis strains. It was found that greater difference in zeta potential values between Gram positive bacteria and Ag NPs triggers better internalization of the particles. Thus the cell surface charge played vital role in cell killing which was confirmed by surface zeta potential study. Finally it may be concluded that green synthesized Ag NPs using Andrographis paniculata leaf extract can be very useful against both multi drug resistant Gram-positive and Gram-negative bacteria.

Green one step morphosynthesis of silver nanoparticles and their antibacterial and anticancerous activities

Bile salts carry out a vital bioactive responsibility among the various physiologically important molecules. The use of bile acids and their conjugates in nanoscience is a novel idea, which opens up fascinating prospects and gives rise to various versatile properties. Sodium cholate, a biosurfactant which is environmentally safe, has been used to reduce and stabilize silver nitrate solution. The silver nanostructures thus produced are crystalline and anisotropic in nature and show different types of particle shapes and sizes depending on the reaction conditions including star and wire shapes which have not been reported earlier, by simply varying the reaction conditions. The silver precursor concentration and the ratio of sodium cholate to silver precursor were the key factors which controlled the morphosynthesis. The formation of the silver NPs was monitored using UV-vis spectrophotometry. The transmission electron microscopy (TEM) technique was used to study the morphology of the synthesized NPs. Fourier transform infrared (FT-IR) spectroscopy was used to identify the interaction of sodium cholate with the synthesized NP. The possible mechanistic role of sodium cholate in the reduction and stabilization of silver nanoparticles (Ag NPs) has also been discussed. The obtained Ag NPs exhibit antibacterial and anticancerous activities. The present study also explores the cytotoxic role of Ag NPs in KG-1A (human acute myeloid leukemia) and K562 (human chronic myeloid leukemia) cell lines.

Preparation and characterization of ferromagnetic nickel oxide nanoparticles from three different precursors: application in drug delivery

Three varieties of nickel oxide nanoparticles [NiO(I), NiO(Br) and NiO(Cl)] have been prepared from three simple mononuclear nickel(II) Schiff-base complexes using a pyrolytic technique. The synthesized nanoparticles are characterized by FT-IR, UV-Vis, XRPD, DLS, SEM, TEM and EDX methods. All the techniques suggest the production of highly pure nickel oxides. The magnetic measurements reveal a small hysteresis loop at room temperature, confirming the super-paramagnetic (weak ferromagnetic) nature of the synthesized NiO nanoparticles. We have applied these nanoparticles for drug delivery. For this purpose, erythromycin, the well known broad spectrum antibiotic is conjugated with the NiO nanoparticles to develop NiO(I)-Ery, NiO(Br)-Ery and NiO(Cl)-Ery. These conjugated nanoparticles successfully deliver erythromycin towards both Gram positive and Gram negative bacteria and show effective antimicrobial activity against erythromycin resistant Staphylococcus aureus and Escherichia coli as model microbial species, evidenced from the Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) values. The order of efficiency toward drug delivery is NiO(I)-Ery > NiO(Br)-Ery > NiO(Cl)-Ery. Thus these conjugates can be applied to overcome the drug resistant properties of bacteria which will be a beneficial strategy in anti-bacterial therapy.

Theoretical and experimental study of folic acid conjugated silver nanoparticles through electrostatic interaction for enhance antibacterial activity

In this paper, folic acid conjugated silver nanoparticles (Ag NPs) are developed for enhancing antibacterial activity. Here triethylamine is used as a capping agent as well as reducing agent during the synthesis of silver nanoparticles. Folic acid is conjugated on the surface of the functionalized silver nanoparticles through electrostatic interaction. The folic acid conjugated silver nanoparticles are characterized in terms of size and morphology by transmission electron microscopy (TEM) and field emission scanning electron microscopy (FESEM) respectively. The phase formation and surface functional groups of nanoparticles are analyzed by X-ray diffraction (XRD) and Fourier transform infrared (FTIR) spectroscopy respectively. Minimum inhibitory concentration study, minimum bactericidal concentration, growth pattern analysis and fluorescence carbon dot tagged nanoparticles uptake study reveal that the folic acid conjugated silver nanoparticles show good prospects against both Gram-negative (Escherichia coli) and Gram-positive (Staphylococcus aureus) bacteria.

Double Mutation in the pfmdr1 Gene Is Associated with Emergence of Chloroquine-Resistant Plasmodium falciparum Malaria in Eastern India

ABSTRACT Malaria is a major public health problem in tropical and subtropical countries, including India. This study elucidates the cause of chloroquine treatment failure (for Plasmodium falciparum infection) before the introduction of artemisinin combination therapy. One hundred twenty-six patients were randomized to chloroquine treatment, and the therapeutic efficacy was monitored from days 1 to 28. An in vitro susceptibility test was performed with all isolates. Parasitic DNA was isolated, followed by PCR and restriction digestion of different codons of the pfcrt gene (codons 72 to 76) and the pfmdr1 gene (N86Y, Y184F, S1034C, N1042D, and D1246Y). Finally, sequencing was done to confirm the mutations. Forty-three (34.13%) early treatment failure cases and 16 (12.69%) late treatment failure cases were observed after chloroquine treatment. In vitro chloroquine resistance was found in 103 isolates (81.75%). Twenty-six (60.47%) early treatment failure cases and 6 (37.5%) late treatment failure cases were associated with the CVMNK-YYSNY allele (the underlined amino acids are those that were mutated). Moreover, the CVIEK-YYSNY allele was found in 8 early treatment failure (18.60%) and 2 late treatment failure (12.5%) cases. The presence of the wild-type pfcrt (CVMNK) and pfmdr1 (YYSNY) double mutant allele in chloroquine-nonresponsive cases was quite uncommon. In vivo chloroquine treatment failure and in vitro chloroquine resistance were strongly correlated with the CVMNK-YYSNY and CVIEK-YYSNY haplotypes (P < 0.01).

One-pot synthesis of multifunctional nanoscale metal-organic frameworks as an effective antibacterial agent against multidrug-resistant Staphylococcus aureus

Drug-resistant bacteria are an increasingly serious threat to global public health. In particular, infections from multidrug-resistant (MDR) Gram-positive bacteria (i.e. Staphylococcus aureus) are growing global health concerns. In this work, we report the first use of nanoscale metal-organic frameworks (NMOFs) coencapsulating an antibiotic (vancomycin) and targeting ligand (folic acid) in one pot to enhance therapeutic efficacy against MDR S. aureus. Zeolitic imidazolate framework (ZIF-8) NMOFs, which have globular morphologies coencapsulating vancomycin and folic acid, are characterized by transmission electron microscopy, field-emission scanning electron microscopy, powder x-ray diffraction, ulltraviolet-visible spectroscopy, and dynamic light-scattering techniques. We determined that the presence of folic acid on the surface of the NMOFs is significant in the sense of effective uptake by MDR S. aureus through endocytosis. The functionalized NMOFs transport vancomycin across the cell wall of MDR S. aureus and enhance antibacterial activity, which has been confirmed from studies of the minimum inhibitory concentration, minimum bactericidal concentration, cytotoxicity of bacterial cells, and generation of reactive oxygen species. This work shows that functionalized NMOFs hold great promise for effective treatment of MDR S. aureus.