Single (SWCNTs) and multi-wall carbon nanotubes (MWCNTs) have been demonstrated to affect several cell functions, even if in the literature no Site URL List 1|]# agreement exists on which of them is more toxic. Both SWCNTs and MWCNTs have been reported to induce oxidative stress, release of pro-inflammatory Inhibitors,Modulators,Libraries cytokines, decrease in cell viability in time and dose dependent manner in human keratinocyte cells [4-6] and impair phagocitic function in alveolar macrophages [7]. Moreover, MWCNTs induced apoptosis and necrosis in skin fibroblasts [8] and cytotoxicity and upregulation of TNF-�� in rat peritoneal macrophages [9]. In the toxicity evaluation of CNTs with respect to human health, the response of human blood cells to CNTs is essential if they are to be used for biosensors, drug delivery or bio-imaging in health and disease.
In fact, they would be among the first exposed cell types upon intravenous administration Inhibitors,Modulators,Libraries and, to our knowledge, just an experimental work on human blood cells has been reported Inhibitors,Modulators,Libraries by Bottini and co-workers. Inhibitors,Modulators,Libraries They examined the toxicity of pristine and oxidized MWCNTs on human T cells and Inhibitors,Modulators,Libraries Jurkat Inhibitors,Modulators,Libraries T leukaemia cells. They found that the oxidized MWCNTs were more toxic than the pristine ones, inducing loss of cell viability at doses of 400 ��g/ml [10].In this study, the ability of SWCNTs to induce cytotoxicity in human peripheral blood lymphocytes (HPBL) was assessed.
Cell growth, viability, metabolic activity and apoptosis were monitored in proliferating HPBL treated with increasing doses of SWCNTs (5 to 50 ��g/ml) for variable times on the basis of biological target investigated.
Primary DNA damage was evaluated as marker of cytotoxicity and genotoxicity Inhibitors,Modulators,Libraries in un-stimulated HPBL treated for 6 h with 1, 5 and 10 ��g/ml SWCNTs final concentrations.2.?Material and Methods2.1. ReagentsRPMI-1640 medium and Foetal Bovine Serum (FBS) were from Biowhittaker (Verviers, Belgium); L-glutamine Dacomitinib and phytohemagglutinin (PHA) were from Gibco (Milan, Italy); trypan blue and Tris were from BDH (Poole, England); resazurin, resorufin, Inhibitors,Modulators,Libraries triton X-100, p-iodonitrotetrazolium violet, L-lactic acid, phenazine methosulphate, NAD, dithiothreitol (DTT), HEPES, CHAPS and Methyl Methanesulfonate (MMS) were from SIGMA (St.
Louis, MO, USA); Lymphoprep? was from Axis-Shield (Oslo, Norway); GSK-3 dimethyl sulphoxide (DMSO), Tofacitinib Citrate side effects NaOH, EDTA and Na2EDTA were from Baker (Deventer, the Netherlands); NaCl, sodium acetate and ethanol were from Carlo Erba (Italy); normal-melting point agarose, low-melting point agarose, ethidium bromide, protein assay kit were from BIORAD laboratories (GmbH, Munich, Germany); carbobenzoxy-Asp-Glu-Val-Asp-7-amino-4-trifluoromethyl selleck chem ARQ197 coumarin (Ac- DEVD-AFC) was from Alexis biochemicals (San Diego, CA).2.2. Nanotubes characteristics and preparationSWCNTs were obtained in highly purified form (>90%) from HeJi, Inc. (Hong Kong, China).