Project description:Graphene-based substrate has efficient neuronal differentiation of hMSCs. Stimulatory effects of graphene on hMSCs neurogenesis can be enhanced by ELF-EMF exposure and it is mediated by enhancement of cell adhesion accompanied by intracellular signal pathway. We used microarrays to assess up-regulated genes in human gene expression profiles during neurogenesis induced by ELF-EMF exposure on graphene. 3 groups (HMSCs grown in neuronal medium on glass, graphene, graphene under ELF-EMF exposure) were selected for RNA extraction and hybridization on Affymetrix microarrays. We sought to obtain total human gene expression. To that end, we hand-selected up-regulated gene involved in neurogenesis, extracellular matrix, and cell migration.
Project description:Graphene-based substrate has efficient neuronal differentiation of hMSCs. Stimulatory effects of graphene on hMSCs neurogenesis can be enhanced by ELF-EMF exposure and it is mediated by enhancement of cell adhesion accompanied by intracellular signal pathway. We used microarrays to assess up-regulated genes in human gene expression profiles during neurogenesis induced by ELF-EMF exposure on graphene.
Project description:Occupational and consumer exposure to GRMs will increase but their impact on human health is still largely unknown. We sought to perform a transcriptome comparison of the bioresponses of MDM and THP-1 macrophages exposed to 5 or 20 µg/ml graphene oxide (GO) or graphene nanoplatelets (GNP) for 6 and 24 h to capture early and more persistent acute responses. Cristalline silica (DQ) was included as immunotoxic reference material.
Project description:The genome-wide transcriptome analysis highlight the increased biocompatibility on immune cells of graphene functionalized with amino groups (NH2) compared with graphene oxide (GO); reducing the cell metabolism disfunction. Moreover, GONH2 was found to polarizes T-cell and monocyte activation toward a T helper-1/M1 immune response.
Project description:This study investigates transcriptomic differences in the lung and liver after pulmonary exposure to two Graphene based materials with similar physical properties, but different surface chemistry. Female C57BL/6 mouse were exposed by a single intratracheal instillation of 0, 18, 54 or 162 μg/mouse of graphene oxide (GO) or reduced graphene oxide (rGO). Pulmonary and hepatic transcriptional changes were compared to identify commonly and uniquely perturbed functions and pathways by GO and rGO. These changes were then related to previously analyzed endpoints. GO exposure induced more differentially expressed genes, affected more functions, and perturbed more pathways compared to rGO, both in the lung and liver.
Project description:The present study was conducted in the frame of the EU-funded Graphene Flagship project. We previously evaluated the impact of graphene oxide (GO) on the gut microbiome in adult zebrafish by performing 16S rRNA gene sequencing in wild-type versus AhR-deficient zebrafish. Here, we performed single-cell RNA-sequencing (10x Genomics) on whole (dissociated) germ-free (GF) zebrafish embryos exposed at 5 dpf to GO plus the microbial metabolite butyrate to gain insight into the impact on specific cell populations in GF zebrafish.
Project description:Denitrification, a crucial biochemical pathway prevalent among haloarchaea in hypersaline ecosystems, has garnered considerable attention in recent years due to its ecological implications. Nevertheless, the underlying molecular mechanisms and genetic regulation governing this respiration/detoxification process in haloarchaea remain largely unexplored. In this study, RNA-sequencing was used to compare the transcriptomes of the haloarchaeon Haloferax mediterranei under oxic and denitrifying conditions, shedding light on the intricate metabolic alterations occurring within the cell such as the accurate control of the metal homeostasis. Furthermore, the investigation identifies several genes encoding transcriptional regulators and potential accessory proteins with putative roles in denitrification. Among these are bacterioopsin transcriptional activators, proteins harbouring a domain of unknown function (DUF2249), and a cyanoglobin. Additionally, the study delves into the genetic regulation of denitrification, finding a regulatory motif within promoter regions that activates numerous denitrification-related genes. This research serves as a starting point for future molecular biology studies in haloarchaea, offering a promising avenue to unravel the intricate mechanisms governing haloarchaeal denitrification, a pathway of paramount ecological importance.
Project description:The present study was conducted in the frame of the EU-funded Graphene Flagship project. The aim is to evaluate the impact of graphene oxide (GO) on the (innate) immune system using zebrafish as a model. We previously performed single-cell RNA-sequencing of germ-free zebrafish embryos exposed to GO plus the microbial metabolite butyrate (BA). Here, we performed a follow up experiment using germ-free lck-GFP transgenic fish in which the zebrafish were exposed to GO plus BA at 5 dpf. The embryos were then dissociated and subsequently sorted on lck and submitted for single-cell RNA-sequencing using 10x Genomics.
Project description:Graphene has been selected as a candidate for synthetic feeder-free culture substrate guiding human/mouse multipotent stem cell lineage specification, and culturing pluripotent stem cells in a number of studies. However, conventional graphene is not an ideal biomaterial to maintain the pluripotency of human pluripotent stem cells (hPSC) including hESCs/hiPSCs due to its intrinsic hydrophobicity and relatively flat surface topography. Here, we applied morphology-controlled nanocrystalline graphene (NG) coating onto the culture substrates via diffusion-associated synthesis (DAS) process and cultivated hPSCs. It is found that enhanced hydrophilicity and controlled surface roughness of DAS-NG enabled tight focal adhesion of hPSCs onto the DAS-NG coated culture substrate and retained pluripotency for over 2 weeks. It is also found hPSCs grown on DAS-NG shared comparable global gene expression profile with hPSCs grown on mouse embryonic fibroblast (MEF). Importantly, the similarities in cell adhesion gene expression between hPSCs grown on DAS-NG and hPSCs on MEF suggest DAS-NG may provide comparable physical cues with MEF for sustaining pluripotency. Taken together, our findings show a new reliable method for culturing hPSCs in feeder-free condition using DAS-graphene. Human iPSCs and human ESC H9 were seeded onto DAS-NG coated glass, ITO or QU, CVD-grown graphene coated glass, uncoated glass and mitomycin-C treated CF1. Human pluripotent stem cells seeded on each culture substrate were cultured in human embryonic stem cell medium composed of knockout DMEM (GIBCO-10829) supplemented with 20% knockout serum replacement (GIBCO-10828), 1mM L-glutamine, 1% penicillin/streptomycin (PAA-P11-010), 1% MEM-non essential amino acid (PAA-M11-003), 0.1mM beta-mercaptoethanol, and 5ng/ml human basic fibroblast growth factor.
Project description:Mesenchymal stromal cells from adipose tissue (AD-MSCs) exhibit favourable clinical traits for autologous transplantation and can develop a ‘Schwann-like’ phenotype (sAD-MSCs) to improve peripheral nerve regeneration, where severe injuries yield insufficient recovery. However, sAD-MSCs regress without biochemical stimulation and detach from conduits under unfavourable transplant conditions, negating their paracrine effects. Graphene-derived materials support AD-MSC attachment, regulating cell adhesion and function through physiochemistry and topography. We report graphene oxide (GO) as a suitable substrate for human sAD-MSCs incubation towards severe peripheral nerve injuries, through evaluating transcriptome changes, neurotrophic factor expression over a 7-day period, and cell viability in apoptotic conditions. Transcriptome changes from GO incubation across four patients were minor compared to biological variance.