Human Peripheral Blood Lymphocyte Cells Research Articles

Evaluation of the genotoxicity, cytotoxicity, and bioactivity of calcium silicate-based cements

BackgroundAs calcium silicate-based cements (CSCs) have found success in various vital pulp therapy applications, several new CSC products have emerged. This study aimed to assess the genotoxicity, cytotoxicity, and bioactivity of four CSCs by comparing the newly introduced materials Bio MTA+ and MTA Cem with previously studied materials, Biodentine and NeoMTA.MethodsGenotoxicity was evaluated using the micronucleus (MN) assay in human peripheral blood lymphocyte cells, measuring MN frequency and nuclear division index (NDI). Cytotoxicity was assessed in human dental pulp stem cells through the Water-Soluble Tetrazolium Salt-1 (WST-1) colorimetric assay. Bioactivity was determined by ELISA, measuring the levels of angiogenic and odontogenic markers (BMP-2, FGF-2, VEGF, and ALP). Statistical analyses included ANOVA, Dunnet and Sidak tests, and Wald chi-square test. (p < .05).ResultsThe MN frequency in the groups was significantly lower than that in the positive control group (tetraconazole) (p < .05). NDI values decreased with increasing concentration (p < .05). Bio MTA+ and NeoMTA showed decreased cell viability at all concentrations in 7-day cultures (p < .01). All materials increased BMP-2, FGF-2, and VEGF levels, with Biodentine and NeoMTA showing the highest levels of BMP-2 and FGF-2 on day 7. Biodentine displayed the highest VEGF levels on day 7. Biodentine and NeoMTA groups exhibited significantly higher ALP activity than the Bio MTA+ and MTA Cem groups by day 7.ConclusionBio MTA+ and MTA Cem demonstrated no genotoxic or cytotoxic effects. Moreover, this study revealed bioactive potentials of Bio MTA+ and MTA Cem by enhancing the expression of angiogenic and osteogenic growth factors.

In Vitro and In Vivo Genotoxicity Analysis of Gold Nanoparticles

Gold nanoparticles have been well recognized for biomedical applications, especially cancer therapeutics. As a preliminary step, it is necessary to investigate the genotoxicity of gold nanoparticles under in vitro and in vivo conditions. In vitro, chromosomal aberration study results showed that gold nanoparticles of 5 mg/ml concentration did not increase the percentage of chromosomal aberration in human peripheral blood lymphocyte cells compared to the negative control. Gold nanoparticles were evaluated erythrocyte micronucleus formation by in vivo micronucleus assay, and results showed no apparent micronucleus formation in mice after 42h exposure of gold nanoparticles. Gold nanoparticles of 600 mg/kg (body weight) were non-clastogenic in swiss albino mice.

Cytotoxic and mutagenic effects of green silver nanoparticles in cancer and normal cells: a brief review

Applications of silver nanoparticles (AgNPs) have revolutionized the medicinal industry. Due to small size (1–100 nm) these nanoparticles are able to cross cell and nuclear membrane and induce cyto/genotoxicity. Green synthesized (plant based) AgNPs (GSAgNPs) have emerged as alternative antimicrobial agent to chemically synthesized nanoparticles assuming their methods of synthesis will be environment friendly and non toxic. In this review we report that the GSAgNPs were found to be cytotoxic and genotoxic to different cancer cell lines. They affected cell proliferation caused apoptosis and cell death of human cancer cells but did not cause such damages to normal human peripheral blood lymphocyte cells. This property can be utilized in cancer therapeutics to overcome the major drawbacks of chemotherapeutics with surface modifications of the nanoparticles by adding functionalized AgNPs.

Minocycline induces apoptosis in acute lymphoblastic leukemia Jurkat cells

Acute lymphoblastic leukemia (ALL) is still an incurable hematologic malignant disorder. Therefore, novel therapeutic approaches are urgently required. In this study, we tested the pro-apoptotic properties of minocycline (Mino) in human peripheral blood lymphocyte cells (hPBLCs) and Jurkat cells (clone E6-1). hPBLCs and Jurkat cells were exposed to increasing concentrations of minocycline (Mino; 10–200 μM). We found that Mino induces apoptosis in Jurkat cells through an H2O2-mediated signaling pathway. Indeed, H2O2 serves as a signaling molecule, thus triggering a molecular cascade involving the oxidation of the DJ-1 cysteine106-thiol residue into DJ-1 cysteine106-sulfonic acid; activation of transcription factors NF-kappaB, P53 and c-JUN; up-regulation of pro-apoptotic proteins BAX/PUMA and proteins involved in the maintenance of mitochondrial function PINK1/PARKIN; and loss of mitochondrial membrane potential and activation of CASPASE-3/AIF, which are responsible for nuclear fragmentation, demonstrated by fluorescence microscopy, flow cytometry and Western blot analysis. Interestingly, Mino induced none of the aforementioned oxidative stress and/or apoptosis markers in hPBLCs. We report for the first time that minocycline, a second-generation, semi-synthetic tetracycline derivative, selectively induces apoptosis in Jurkat cells through an oxidative stress-mediated mechanism. Additionally, since Mino has well-established clinical pharmacokinetic and pharmacodynamic properties, we propose that this tetracycline is a promising anti-leukemic drug against ALL.

In vitro evaluation of the genotoxicity of CeO2 nanoparticles in human peripheral blood lymphocytes using cytokinesis-block micronucleus test, comet assay, and gamma H2AX.

Engineered nanoparticles (ENPs) are used in a wide range of applications because of their unique properties. Cerium dioxide nanoparticles (CeO2 NPs) are one of the important ENPs, and they can cause negative health effects, such as genotoxicity, in humans and other living organisms. The aim of this work was to analyze the genotoxic effects of short-term (3-24 h) CeO2 NPs exposure to cultured human blood lymphocytes. Three genotoxicity systems "cytokinesis-block micronucleus test, comet assay, and gamma H2AX test" were used to show the genotoxic potential of CeO2 NPs (particle size <25 nm, concentrations: 6, 12, and 18 µg/mL). Hydrogen peroxide was selected as the positive-control genotoxic agent. Our results indicate that CeO2 NPs have genotoxic potential on human peripheral blood lymphocytes cells even at 3-24 h exposure under in vitro conditions.

Synergistic activity of sorafenib and betulinic acid against clonogenic activity of non-small cell lung cancer cells.

The highly selective multi‐targeted agent sorafenib is an inhibitor of a number of intracellular signaling kinases with anti‐proliferative, anti‐angiogenic and pro‐apoptotic effects in various types of tumors, including human non‐small cell lung cancer (NSCLC). Betulin displays a broad spectrum of biological and pharmacological properties, including anticancer and chemopreventive activity. Combination of drugs with different targets is a logical approach to overcome multilevel cross‐stimulation among key signaling pathways in NSCLC progression. NSCLC cell lines, A549, H358 and A427, with different KRAS mutations, and normal human peripheral blood lymphocyte cells, were treated with sorafenib and betulinic acid alone and in combination. We examined the effect of different combined treatments on viability (MTS test), proliferation and apoptotic susceptibility based on flow cytometry, alterations in signaling pathways by western blotting and colony‐forming ability. The combination of sorafenib with betulinic acid had a strong effect on the induction of apoptosis of different NSCLC cell lines. In addition, this combination was not toxic for human peripheral blood lymphocytes. Combination treatment changed the expression of proteins involved in the mitochondrial apoptosis pathway and induced apoptotic death by caspase activation. Importantly, combination treatment with low drug concentrations tremendously reduced the colony‐forming ability of A549, H358 and A427 cells, as compared to both compounds alone. In this study, we showed that combination therapy with low concentrations of sorafenib and betulinic acid had the capacity to induce high levels of cell death and abolish clonogenic activity in some NSCLC cell lines regardless of KRAS mutations.

Metal chelator TPEN selectively induces apoptosis in K562 cells through reactive oxygen species signaling mechanism: implications for chronic myeloid leukemia.

Chronic myeloid leukemia (CML) is a hematologic disorder characterized by the constitutive expression of BCR-ABL tyrosine kinase. Although successful implementation of tyrosine kinase inhibitors for the treatment of CML remain a traditional choice for molecularly targeted therapy, some patients present primary or secondary resistance to such therapy. Therefore, alternative therapeutic strategies are required to treat resistant CML cells. Accordingly, new anti-proliferative and/or pro-apoptotic compounds would be needed for clinical treatment. In the present investigation, we demonstrate that TPEN (e.g. 3μM), a lipid-soluble metal chelator, induces apoptosis in K562 cells via a molecular cascade involving H2O2≫JNK, NF-κB>c-JUN, P73>PUMA, BAX>loss of ΔΨm>CASPASE-3>nuclei/DNA fragmentation. Fragmentation of the nuclei and DNA are indicative of cell death by apoptosis. Remarkably, the antioxidant N-acetyl-cysteine, and inhibitors of the transcription factors CASPASE 3 and (JNK) kinase, decreased oxidative stress (OS) and cell death in these cells. This is evidenced by fluorescence microscopy, flow cytometry and immunocytochemistry for OS markers (e.g. generation of H2O2 and DJ 1 oxidation) and nuclear expression of apoptotic markers (e.g. activated caspase 3 and JNK kinase). In addition, TPEN causes no detectable damage in human peripheral blood lymphocyte cells (hPBLCs). We conclude that TPEN selectively induces apoptosis in K562 cells via an OS-mechanism. Our findings may provide insight into more effective CML anticancer therapies.

Total velvet-antler polypeptide extract from Cervus nippon Temminck induces cell proliferation and activation of the PI3K–Akt signalling pathway in human peripheral blood lymphocytes

Velvet antler (VA) has been used for centuries as a traditional medicine to treat a wide range of health problems. VA was first recorded in the Chinese medical classics Shen Nong Ben Cao Jing 2000 years ago. VA has been extensively used to strengthen bones and muscles, promote blood flow and increase immune function. In the present study, we used VA (Cervus nippon Temminck) protein extract (VAPE) to induce cell proliferation and differentiation in human peripheral blood lymphocyte cell line 1301 cells. The results showed that 100 µg/mL of VAPE could significantly induce 1301-cell proliferation (151.3%), increase the population of S-phase cells and increase the expression of CDK4. In addition, VAPE could effectively increase the expression levels of PI3K and Akt and activate PI3K–Akt signal pathway. These findings have provided novel insights into the further understanding of the molecular and signalling pathway mechanisms of VAPE in promoting proliferation of human peripheral blood lymphocyte cells.

Genotoxic and cytotoxic effects of formic acid on human lymphocytes in vitro

Objective: Formic acid is an ubiquitous chemical constituent in the environment, being produced by sources as diverse as vegetation, ants, soil, vehicles, biomass burning, and photochemical reactions. The present work is focused on in vitro analysis of cytotoxic and genotoxic effects of formic acid, using cytogenetic tests such as the cytokinesis-block micronucleus assay (CBMN) and chromosomal aberration analysis, in human lymphocytes. Method: This study was carried out using blood samples from healthy, non-smoking adults aged 18–22 years, of whom 10 were male and 6 were famale. Different concentrations (0.07, 0.1, 0.2, 0.3, 0.4, 0.5, 0.8 mM) of formic acid was added to the lymphocyte culture test for chromosomal aberration (CA) analysis. Mitomycin-C (0.3 mg/ml) was used as the positive control. Human peripheral blood lymphocyte cells were treated with 20, 40, 60, 80 mM concentrations of formic acid for 48 h. for the CBMN test. Mitomycin-C (0.5 mg/ml) was added to the Lymphocyte culture as a positive control. The present research was carried out to assess the cytotoxic and genotoxic effects of formic acid on human peripheral lymphocytes in vitro using the cytokinesis-block micronucleus assay (CBMN), as well as chromosomal aberration (CA) analysis.

Recent microwave research studies of Gazi Biophysics &amp; Gazi Non-Ionizing Radiation Protection Center

Recent microwave research studies of Gazi Biophysics &amp; Gazi Non-Ionizing Radiation Protection Center

Genotoxicity of thimerosal in cultured human lymphocytes with and without metabolic activation sister chromatid exchange analysis proliferation index and mitotic index

Thimerosal is an antiseptic containing 49.5% of ethyl mercury that has been used for years as a preservative in many infant vaccines and in flu vaccines. Thimerosal is an organic mercurial compound used as a preservative in biomedical preparations. In this study, we evaluated the genotoxic effect of thimerosal in cultured human peripheral blood lymphocytes using sister chromatid exchange analysis in culture conditions with and without S9 metabolic activation. This study is the first report investigating the genotoxic effects of thimerosal in cultured human peripheral blood lymphocyte cells using sister chromatid exchange analysis. An analysis of variance test (ANOVA) was performed to evaluate the results. Significant induction of sister chromatid exchanges was seen at concentrations between 0.2 and 0.6 microg/ml of thimerosal compared with negative control. A significant decrease (p<0.001) in mitotic index (MI) and proliferation index (PRI) as well as an increase in SCE frequency (p<0.001) was observed compared with control cultures. Our results indicate the genotoxic and cytotoxic effect of TH in cultured human peripheral blood lymphocytes at tested doses in cultures with/without S9 fraction.

Inhibition of B(a)P induced strand breaks in presence of curcumin

Incidence of cancer at different sites may be related to oxidative damage to host genome by genotoxicants. These oxidative actions may be modified by phytochemicals present in foods. The non-nutritive dietary constituents which possess antimutagenic property appear to be promising chemopreventive agents. This study reports the protective effect of curcumin on B(a)P induced DNA damage in human peripheral blood lymphocyte cells. The study group consisted of 10 male smokers, 10 non-smokers and 10 non-smoking females aged between 25 and 45. The DNA damage was assessed using comet assay. In all the groups curcumin showed a dose-dependent inhibitory effect. The effect appeared to be sex dependent. There was no correlation between DNA damage and GST-Mu levels and levels of micronutrients namely Vitamins A, E and beta carotene. The results of this study are in line with our earlier observations on turmeric/curcumin as a potential chemopreventer.

Antiproliferative and cytotoxic effect of a novel organotin compound on mammalian cells both in vitro and in vivo

Organotin compounds are organometallic compounds showing various toxicological properties. Several organotin compounds also showed an antineoplastic effect. However, their relative mutagenic potential is not well established. In this study Et 2SnCl 2·L [L= N-(2-pyridylmethylene)-4-toluidine] (OTC) has been subjected to investigation for its cytotoxic effect in mouse bone marrow cells (BMCs) and human peripheral blood lymphocyte cells (HPBLs). The SnN bond in OTC is 2.46 Å which is greater than 2.39 Å and therefore, a better formation of tin–DNA complex can be expected. The present data indicate that OTC induced significant delay in cell kinetics and sister chromatid exchanges (SCEs) in both BMCs and HPBLs, whereas, induction of chromosome aberrations was found only in HPBLs. The presence of buthionine sulfoximine (BSO) modulated cellular sensitivity towards OTC in both cell systems. It may be inferred that the OTC could bind on DNA more easily owing to its structural advantage and this may explain the induction of DNA damage and the delay in cell proliferation. Since the cytotoxic effect of OTC is more in glutathione depleted cells, the concentration of OTC may be reduced to get an antitumour effect in GSH-depleted cells and thus minimizes its toxic side effect.