Nanodiamonds activate blood platelets and induce thromboembolism.

AIM: Nanodiamonds (NDs) have been evaluated for a wide range of biomedical applications. Thus, thorough investigation of the biocompatibility of NDs has become a research priority. Platelets are highly sensitive and are one of the most abundant cell types found in blood. They have a central role in hemostasis and arterial thrombosis. In this study, we aim to investigate the direct and acute effects of carboxylated NDs on platelet function. METHODS: In this study, pro-coagulant parameters such as platelet aggregability, intracellular Ca(2+) flux, mitochondrial transmembrane potential (ΔΨm), generation of reactive oxygen species, surface exposure of phosphatidylserine, electron microscopy, cell viability assay and in vivo thromboembolism were analyzed in great detail. RESULTS: Carboxylated NDs evoked significant activation of human platelets. When administered intravenously in mice, NDs were found to induce widespread pulmonary thromboembolism, indicating the remarkable thrombogenic potential of this nanomaterial. CONCLUSION: Our findings raise concerns regarding the putative biomedical applications of NDs pertaining to diagnostics and therapeutics, and their toxicity and prothrombotic properties should be critically evaluated.

Amine-modified graphene: thrombo-protective safer alternative to graphene oxide for biomedical applications.

Graphene and its derivatives have attracted significant research interest based on their application potential in different fields including biomedicine. However, recent reports from our laboratory and elsewhere have pointed to serious toxic effects of this nanomaterial on cells and organisms. Graphene oxide (GO) was found to be highly thrombogenic in mouse and evoked strong aggregatory response in human platelets. As platelets play a central role in hemostasis and thrombus formation, thrombotoxicity of GO potentially limits its biomedical applications. Surface chemistry of nanomaterials is a critical determinant of biocompatibility, and thus differentially functionalized nanomaterials exhibit varied cellular toxicity. Amine-modified carbon nanotubes have recently been shown to possess cytoprotective action, which was not exhibited by their relatively toxic carboxylated counterparts. We, therefore, evaluated the effect of amine modification of graphene on platelet reactivity. Remarkably, our results revealed for the first time that amine-modified graphene (G-NH(2)) had absolutely no stimulatory effect on human platelets nor did it induce pulmonary thromboembolism in mice following intravenous administration. Further, it did not evoke lysis of erythrocytes, another major cellular component in blood. These findings contrasted strikingly the observations with GO and reduced GO (RGO). We conclude that G-NH(2) is not endowed with thrombotoxic property unlike other commonly investigated graphene derivatives and is thus potentially safe for in vivo biomedical applications.

Thrombus inducing property of atomically thin graphene oxide sheets.

Graphene oxide (GO), the new two-dimensional carbon nanomaterial, is extensively investigated for potential biomedical applications. Thus, it is pertinent to critically evaluate its untoward effects on physiology of tissue systems including blood platelets, the cells responsible for maintenance of hemostasis and thrombus formation. Here we report for the first time that atomically thin GO sheets elicited strong aggregatory response in platelets through activation of Src kinases and release of calcium from intracellular stores. Compounding this, intravenous administration of GO was found to induce extensive pulmonary thromboembolism in mice. Prothrombotic character of GO was dependent on surface charge distribution as reduced GO (RGO) was significantly less effective in aggregating platelets. Our findings raise a concern on putative biomedical applications of GO in the form of diagnostic and therapeutic tools where its prothrombotic property should be carefully investigated.

Characterization of graphene oxide by flow cytometry and assessment of its cellular toxicity.

Because of its unique physicochemical properties, graphene oxide (GO) has found significant applications in a wide spectrum of biomedical utilities. In the present report, we have presented flow cytometry as an alternative tool for analysis of size distribution and intrinsic fluorescence of GO sheets, its interaction with cells and cytotoxicity evaluations.

Atomic-scale observation of rotational misorientation in suspended few-layer graphene sheets.

Single or few-layer graphene (FLG) sheets offer extraordinary electronic, thermal and mechanical properties and are expected to find a variety of applications. Fully exploiting the properties of graphene will require a method for the production of high-quality graphene sheets (almost pristine graphene) in large quantities. In this regard, we report a two-step method for obtaining a homogenous colloidal suspension of single or FLG sheets up to 0.15 mg ml(-1) in N,N-dimethylformamide solution. The graphene nanostructures are directly imaged using a high-resolution transmission electron microscope (HRTEM) operated at 200 kV with a point resolution of 0.16 nm. We observed rotational misorientation within the flake in the HRTEM images of 2, 4 and 6 layers of graphene sheets, giving rise to Moiré patterns. By filtering in the frequency domain using a Fourier transform, we reconstruct the graphene lattice of each sheet and determine the relative rotation between consecutive graphene layers up, to six separate sheets. Direct evidence is obtained for FLG sheets with packing that is different to the standard AB Bernal packing of bulk graphite. Furthermore, we observed periodic ripples in suspended graphene sheets in our TEM measurements. Electrostatic force microscopy was used to characterize the electric potential distribution on the surface of FLG sheets on SiO2/Si substrates in ambient conditions. The FLG sheets were found to exhibit a conducting nature with small potential variations on the surface.