Behaviors of NIH-3T3 Fibroblasts on Graphene/Carbon Nanotubes: Proliferation, Focal Adhesion, and Gene Transfection Studies

Abstract

Carbon-based materials, including graphene and carbon nanotubes, have been considered attractive candidates for biomedical applications such as scaffolds in tissue engineering, substrates for stem cell differentiation, and components of implant devices. Despite the potential biomedical applications of these materials, only limited information is available regarding the cellular events, including cell viability, adhesion, and spreading, that occur when mammalian cells interface with carbon-based nanomaterials. Here, we report behaviors of mammalian cells, specifically NIH-3T3 fibroblast cells, grown on supported thin films of graphene and carbon nanotubes to investigate biocompatibility of the artificial surface. Proliferation assay, cell shape analysis, focal adhesion study, and quantitative measurements of cell adhesion-related gene expression levels by RT-PCR reveal that the fibroblast cells grow well, with different numbers and sizes of focal adhesions, on graphene- and carbon nanotube-coated substrates. Interestingly, the gene transfection efficiency of cells grown on the substrates was improved up to 250% that of cells grown on a cover glass. The present study suggests that these nanomaterials hold high potential for bioapplications showing high biocompatibility, especially as surface coating materials for implants, without inducing notable deleterious effects while enhancing some cellular functions (i.e., gene transfection and expression).

Graphene Enhances Cellular Proliferation through Activating the Epidermal Growth Factor Receptor

“…11−14 For instance, Ryoo et al showed that graphene substrates improved gene transfection efficacy in NIH-3T3 fibroblasts without inducing notable deleterious effects. 14 Chang et al found that graphene oxide (GO) did not enter A549 cells and showed no obvious toxicity to A549 cells. 15 On the contrary, Wang et al demonstrated that water-soluble GO had obvious toxicity to human fibroblast cells, such as decreasing cell adhesion, inducing cell apoptosis, and entering into lysosomes, mitochondria, endoplasm, and cell nucleus when the dose was higher than 50 μg/mL.…”

Graphene and graphene oxide as nanomaterials for medicine and biology application

“…NIH-3T3 fibroblasts behaviour was studied in different substrates covered with carbon nanomaterial, for example, GO, carbon nanotubes and RGO. The substrates coated with carbon nanomaterial had enhanced gene transfection efficiency and high biocompatibility [99]. Graphene/chitosan films demonstrated promising application to enhance and repair tissue functions in tissue engineering [100].…”

In Vitro Toxicology Testing: It’s Time to Report the Sex of Cells

“…In a 2015 report by Wan and colleagues, 19 researchers used quantitative Polymerase Chain Reaction (qPCR) -a very common molecular method -to newly identify NIH/3T3 (ATCC No. : CRL-1658) murine embryonic fibroblast cells as female; 3T3 cells have long been used in toxicological experiments, most recently to assess developmental toxicity to therapeutic drugs 20 and phototoxic potential, 21 but also to document cell behavior on nanotubes, 22 response to nanoparticles, 23 and antioxidant response. 24 Importantly, sex determination using qPCR is now added to the list of several other available tools that can be used to determine the sex type of cultured cells, including immunodetection of H-Y antigen, 25 nested PCR, 26 Southern blot, 27 and enzymatic assays.…”