Layer-Number-Dependent Effects of Graphene Oxide on the Pluripotency of Mouse Embryonic Stem Cells Through the Regulation of the Interaction Between the Extracellular Matrix and Integrins

Abstract

Introduction Embryonic stem cells (ESCs) possess great application prospects in biological research and regenerative medicine, so it is important to obtain ESCs with excellent and stable cellular states during in vitro expansion. The feeder layer culture system with the addition of leukemia inhibitory factor (LIF) is currently applied in ESC cultures, but it has a series of disadvantages that could influence the culture efficiency and quality of the ESCs. With the development of nanotechnology, many studies have applied nanomaterials to optimize the stem cell culture system and regulate the fate of stem cells. In this study, we investigated the layer-number-dependent biofunction of graphene oxide (GO) on the pluripotency of ESCs from mice (mESCs). Methods Single-layer GO (SGO) and multi-layer GO (MGO) were characterized and their effects on the cytotoxicity and self-renewal of mESCs were detected in vitro. The differentiation potentials of mESCs were identified through the formation of embryoid bodies and teratomas. The regulatory mechanism of GO was verified by blocking the target receptors on the surface of mESCs using antibodies. Results Both SGO and MGO were biocompatible with mESCs, but only MGO effectively sustained their self-renewal and differentiation potential. In addition, GO influenced the cellular activities of mESCs by regulating the interactions between extracellular matrix proteins and integrins. Conclusion This work demonstrates the layer-number-dependent effects of GO on regulating the cell behavior of mESCs and reveals the extracellular regulatory mechanism of this process.

Ion elemental-optimized layered double hydroxide nanoparticles promote chondrogenic differentiation and intervertebral disc regeneration of mesenchymal stem cells through focal adhesion signaling pathway

“…Binding of integrin receptors to cell adhesion ligands of ECM allows integrin receptors to activate cascade signals to connect the cytoskeleton and ECM [ 14 , 67 ]. Cell behavior can be feasibly regulated by adjusting the density of adhesion ligands in nanomaterials [ 69 , 70 ]. In the earlier materials screening of four different LDH nanoparticles, MgAl–Cl and MgFe–NO 3 LDH showed superior effects than other groups, thus were selected for further cell function validation.…”

Is Graphene Shortening the Path toward Spinal Cord Regeneration?

“…This outcome was recently correlated to the number of layers of GO nanosheets as well as to their preferential distribution at the outside of the cell membrane, where the multilayered GO configuration (thickness of 3.2 nm) was able to better adsorb ECM components like fibronectin and, therefore, compromise its interaction with integrin receptors. 89 Contrarily, it was highlighted that the lower surface energy of monolayered GO (thickness of 0.7−1.2 nm) was insufficient to substantially restrain fibronectin−integrin coupling, thus promoting a decrease of the differentiation potential of mESCs. The preferential attachment of GO nanosheets onto the outer of the plasmatic membrane of human fetal NSCs (hfNSCs) was also pointed out by Kim et al 90 as a key event to enhance cell−cell and cell−matrix interactions responsible for regulating the differentiation potential of neurospheres in vitro.…”

“…This outcome was recently correlated to the number of layers of GO nanosheets as well as to their preferential distribution at the outside of the cell membrane, where the multilayered GO configuration (thickness of 3.2 nm) was able to better adsorb ECM components like fibronectin and, therefore, compromise its interaction with integrin receptors. 89 Contrarily, it was highlighted that the lower surface energy of monolayered GO (thickness of 0.7–1.2 nm) was insufficient to substantially restrain fibronectin–integrin coupling, thus promoting a decrease of the differentiation potential of mESCs.…”

Impact of Graphene Derivatives as Artificial Extracellular Matrices on Mesenchymal Stem Cells

“…Rat bone marrow MSCs proliferation is influenced by the concentration of GO used in several treatments [155]. MSCs appeared to be highly proliferative when the concentration of GO was up to 0.1 g/mL, compared to the control group.…”