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Protein spot arrays on graphene oxide coatings for efficient single-cell capture.

ABSTRACT: Biomedical applications such as cell screening or cell-cell interaction studies require placement and adhesion of cells on surfaces with controlled numbers and location. In particular, single-cell arraying and positioning has come into focus as a basis of such applications. An ideal substrate would combine biocompatibility with favorable attributes such as pattern stability and easy processing. Here, we present a simple yet effective approach to single-cell arraying based on a graphene oxide (GO) surface carrying protein (fibronectin) microarrays to define cell adhesion points. These capture NIH-3T3 cells, resulting in cell arrays, which are benchmarked against analogous arrays on silanized glass samples. We reveal significant improvement in cell-capture performance by the GO coating with regards to overall cell adhesion and single-cell feature occupancy. This overall improvement of cell-arraying combined with retained transparency of substrate for microscopy and good biocompatibility makes this graphene-based approach attractive for single-cell experiments.

SUBMITTER: Kumar R

PROVIDER: S-EPMC8913813 | biostudies-literature | 2022 Mar

REPOSITORIES: biostudies-literature

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Protein spot arrays on graphene oxide coatings for efficient single-cell capture.

Kumar R R Llewellyn S S Vasantham S K SK Nie Kaiwen K Sekula-Neuner S S Vijayaraghavan A A Hirtz M M

Scientific reports 20220310 1

Biomedical applications such as cell screening or cell-cell interaction studies require placement and adhesion of cells on surfaces with controlled numbers and location. In particular, single-cell arraying and positioning has come into focus as a basis of such applications. An ideal substrate would combine biocompatibility with favorable attributes such as pattern stability and easy processing. Here, we present a simple yet effective approach to single-cell arraying based on a graphene oxide (GO ...[more]

PMID: 35273174

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Project description:Graphene oxide (GO) is a promising material for bone tissue engineering, but the validation of its molecular biological effects, especially in the context of clinically applied materials, is still limited. In this study, we compare the effects of graphene oxide framework structures (F-GO) and reduced graphene oxide-based framework structures (F-rGO) as scaffold material with a special focus on vascularization associated processes and mechanisms in the bone. Highly porous networks of zinc oxide tetrapods serving as sacrificial templates were used to create F-GO and F-rGO with porosities >99% consisting of hollow interconnected microtubes. Framework materials were seeded with human mesenchymal stem cells (MSC), and the cell response was evaluated by confocal laser scanning microscopy (CLSM), deoxyribonucleic acid (DNA) quantification, real-time polymerase chain reaction (RT-PCR), enzyme-linked immunosorbent assay (ELISA), and alkaline phosphatase activity (ALP) to define their impact on cellular adhesion, osteogenic differentiation, and secretion of vascular growth factors. F-GO based scaffolds improved adhesion and growth of MSC as indicated by CLSM and DNA quantification. Further, F-GO showed a better vascular endothelial growth factor (VEGF) binding capacity and improved cell growth as well as the formation of microvascular capillary-like structures in co-cultures with outgrowth endothelial cells (OEC). These results clearly favored non-reduced graphene oxide in the form of F-GO for bone regeneration applications. To study GO in the context of a clinically used implant material, we coated a commercially available xenograft (Bio-Oss® block) with GO and compared the growth of MSC in monoculture and in coculture with OEC to the native scaffold. We observed a significantly improved growth of MSC and formation of prevascular structures on coated Bio-Oss®, again associated with a higher VEGF binding capacity. We conclude that graphene oxide coating of this clinically used, but highly debiologized bone graft improves MSC cell adhesion and vascularization.

Project description:Magnesium (Mg) is a promising biodegradable implant material because of its appropriate mechanical properties and safe degradation products. However, in vivo corrosion speed and hydrogen gas production need to be controlled for uses in biomedical applications. Here we report the development of a conducting polymer 3,4-ethylenedioxythiphene (PEDOT) and graphene oxide (GO) composite coating as a corrosion control layer. PEDOT/GO was electropolymerized on Mg samples in ethanol media. The coated Mg samples were subjected to various corrosion tests. The PEDOT/GO coating significantly reduced the rate of corrosion as evidenced by lower Mg ion concentration and pH of the corrosion media. In addition, the coating decreased the evolved hydrogen. Electrochemical analysis of the corroding samples showed more positive corrosion potential, a decreased corrosion current, and an increase in the polarization resistance. PEDOT/GO corrosion protection is attributed to three factors; an initial passive layer preventing solution ingress, buildup of negative charges in the film, and formation of corrosion protective Mg phosphate layer through redox coupling with Mg corrosion. To explore the biocompatibility of the coated implants in vitro, corrosion media from PEDOT/GO coated or uncoated Mg samples were exposed to cultured neurons where PEDOT/GO coated samples showed decreased toxicity. These results suggest that PEDOT/GO coating will be an effective treatment for controlling corrosion of Mg based medical implants.Statement of significanceCoating Mg substrates with a PEDOT/GO composite coating showed a significant decrease in corrosion rate. While conducting polymer coatings have been used to prevent corrosion on various metals, there has been little work on the use of these coatings for Mg. Additionally, to our knowledge, there has not been a report of the combined used of conducting polymer and GO as a corrosion control layer. Corrosion control is attributed to an initial barrier layer followed by electrochemical coupling of the PEDOT/GO coating with the substrate to facilitate the formation of a protective phosphate layer. This coupling also resulted in a decrease in hydrogen produced during corrosion, which could further improve the host tissue integration of Mg implants. This work elaborates on the potential for electroactive polymers to serve as corrosion control methods.

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Protein spot arrays on graphene oxide coatings for efficient single-cell capture.

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Protein spot arrays on graphene oxide coatings for efficient single-cell capture.

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