Pulsed‐Electromagnetic‐Field‐Assisted Reduced Graphene Oxide Substrates for Multidifferentiation of Human Mesenchymal Stem Cells

Electromagnetic fields (EMFs) can modulate cell proliferation, DNA replication, wound healing, cytokine expression, and the differentiation of mesenchymal stem cells (MSCs). Graphene, a 2D crystal of sp(2) -hybridized carbon atoms, has entered the spotlight in cell and tissue engineering research. However, a combination of graphene and EMFs has never been applied in tissue engineering. This study combines reduced graphene oxide (RGO) and pulsed EMFs (PEMFs) on the osteogenesis and neurogenesis of MSCs. First, the chemical properties of RGO are measured. After evaluation, the RGO is adsorbed onto glass, and its morphological and electrical properties are investigated. Next, an in vitro study is conducted using human alveolar bone marrow stem cells (hABMSCs). Their cell viability, cell adhesion, and extracellular matrix (ECM) formation are increased by RGO and PEMFs. The combination of RGO and PEMFs enhances osteogenic differentiation. Together, RGO and PEMFs enhance the neurogenic and adipogenic differentiation of hABMSCs. Moreover, in a DNA microarray analysis, the combination of RGO and PEMFs synergically increases ECM formation, membrane proteins, and metabolism. The combination of RGO and PEMFs is expected to be an efficient platform for stem cell and tissue engineering.

[1]  N. Peres,et al.  Fine Structure Constant Defines Visual Transparency of Graphene , 2008, Science.

[2]  Seunghun Hong,et al.  Carbon nanotube monolayer cues for osteogenesis of mesenchymal stem cells. , 2011, Small.

[3]  H. Emamy,et al.  Genotoxicity of graphene nanoribbons in human mesenchymal stem cells , 2013 .

[4]  Yu-Sheng Hsiao,et al.  Multifunctional Graphene–PEDOT Microelectrodes for On‐Chip Manipulation of Human Mesenchymal Stem Cells , 2013 .

[5]  Nigel J. Cassidy,et al.  Electrical stimulation: a novel tool for tissue engineering. , 2013, Tissue engineering. Part B, Reviews.

[6]  N. Portwood,et al.  Effects of a low-intensity electromagnetic field on fibroblast migration and proliferation , 2011, Electromagnetic biology and medicine.

[7]  S. Mikhalovsky,et al.  Therapeutic potential of electromagnetic fields for tissue engineering and wound healing , 2014, Cell proliferation.

[8]  R. Car,et al.  Oxygen-driven unzipping of graphitic materials. , 2006, Physical review letters.

[9]  N. Gadegaard,et al.  Nanoscale surfaces for the long-term maintenance of mesenchymal stem cell phenotype and multipotency. , 2011, Nature materials.

[10]  N. Wertheimer,et al.  Electrical wiring configurations and childhood cancer. , 1979, American journal of epidemiology.

[11]  Dong Jun Lee,et al.  Transparent and Stretchable Interactive Human Machine Interface Based on Patterned Graphene Heterostructures , 2015 .

[12]  O. Akhavan,et al.  Differentiation of human neural stem cells into neural networks on graphene nanogrids. , 2013, Journal of materials chemistry. B.

[13]  X. Qu,et al.  New Horizons for Diagnostics and Therapeutic Applications of Graphene and Graphene Oxide , 2013, Advanced materials.

[14]  A. Pilla.  Nonthermal electromagnetic fields: From first messenger to therapeutic applications , 2013, Electromagnetic biology and medicine.

[15]  Elena García-Gareta,et al.  Osteoinduction of bone grafting materials for bone repair and regeneration. , 2015, Bone.

[16]  Abdul Rahman Mohamed,et al.  Surface charge modification via protonation of graphitic carbon nitride (g-C3N4) for electrostatic self-assembly construction of 2D/2D reduced graphene oxide (rGO)/g-C3N4 nanostructures toward enhanced photocatalytic reduction of carbon dioxide to methane , 2015 .

[17]  Chwee Teck Lim,et al.  Origin of enhanced stem cell growth and differentiation on graphene and graphene oxide. , 2011, ACS nano.

[18]  Ki-Taek Lim,et al.  Graphene-incorporated chitosan substrata for adhesion and differentiation of human mesenchymal stem cells. , 2013, Journal of materials chemistry. B.

[19]  Kye-Yong Song,et al.  Neural stimulation on human bone marrow‐derived mesenchymal stem cells by extremely low frequency electromagnetic fields , 2012, Biotechnology progress.

[20]  B. Hong,et al.  Monolayer Graphene-Directed Growth and Neuronal Differentiation of Mesenchymal Stem Cells. , 2015, Journal of biomedical nanotechnology.

[21]  George J. Christ,et al.  The effect of low-frequency electromagnetic field on human bone marrow stem/progenitor cell differentiation , 2015, Stem cell research.

[22]  Jun Zhang,et al.  Hydrogen Adsorption of Mg-Doped Graphene Oxide: A First-Principles Study , 2013 .

[23]  Kyoung Soon Choi,et al.  Bioactive effects of graphene oxide cell culture substratum on structure and function of human adipose-derived stem cells. , 2013, Journal of biomedical materials research. Part A.

[24]  A. Patruno,et al.  Extremely low frequency electromagnetic field and wound healing: implication of cytokines as biological mediators. , 2013, European cytokine network.

[25]  J. Park,et al.  Electromagnetic fields mediate efficient cell reprogramming into a pluripotent state. , 2014, ACS nano.

[26]  FoleyBriana,et al.  Development of a Human Adipose-Derived Stem Cell Model for Characterization of Chemical Modulation of Adipogenesis , 2015 .

[27]  R. Leighton,et al.  The Feynman Lectures on Physics; Vol. I , 1965 .

[28]  G. Costin,et al.  Trends in wound repair: cellular and molecular basis of regenerative therapy using electromagnetic fields. , 2012, Current molecular medicine.

[29]  K. Yeung,et al.  Silver nanoparticles promote osteogenesis of mesenchymal stem cells and improve bone fracture healing in osteogenesis mechanism mouse model. , 2015, Nanomedicine : nanotechnology, biology, and medicine.

[30]  Yu-Sheng Hsiao,et al.  Electrodes: Multifunctional Graphene–PEDOT Microelectrodes for On‐Chip Manipulation of Human Mesenchymal Stem Cells (Adv. Funct. Mater. 37/2013) , 2013 .

[31]  M I Katsnelson,et al.  Magnetic correlations at graphene edges: basis for novel spintronics devices. , 2007, Physical review letters.

[32]  W. Jang,et al.  Extremely low frequency electromagnetic fields enhance neuronal differentiation of human mesenchymal stem cells on graphene-based substrates , 2015 .

[33]  W T Lawrence,et al.  Physiology of the acute wound. , 1998, Clinics in plastic surgery.

[34]  N. Zagris.  Extracellular matrix in development of the early embryo. , 2001, Micron.

[35]  Vinayak Sant,et al.  Graphene-based nanomaterials for drug delivery and tissue engineering. , 2014, Journal of controlled release : official journal of the Controlled Release Society.

[36]  Byung-Soo Kim,et al.  Graphene‒Regulated Cardiomyogenic Differentiation Process of Mesenchymal Stem Cells by Enhancing the Expression of Extracellular Matrix Proteins and Cell Signaling Molecules , 2014, Advanced healthcare materials.

[37]  Wei Gao,et al.  New insights into the structure and reduction of graphite oxide. , 2009, Nature chemistry.

[38]  Lu Wang,et al.  Graphene oxide as an ideal substrate for hydrogen storage. , 2009, ACS nano.

[39]  D. Narmoneva,et al.  Regulation of endothelial MAPK/ERK signalling and capillary morphogenesis by low-amplitude electric field , 2013, Journal of the Royal Society Interface.

[40]  Jingyu Sun,et al.  Direct low-temperature synthesis of graphene on various glasses by plasma-enhanced chemical vapor deposition for versatile, cost-effective electrodes , 2015, Nano Research.

[41]  K. Lim,et al.  Regeneration of chronic tympanic membrane perforation using an EGF-releasing chitosan patch. , 2013, Tissue engineering. Part A.

[42]  Jijun Zhao,et al.  Graphene oxide: A promising nanomaterial for energy and environmental applications , 2015 .

[43]  C. Yi,et al.  Gold nanoparticles promote osteogenic differentiation of mesenchymal stem cells through p38 MAPK pathway. , 2010, ACS nano.

[44]  D. O’Carroll,et al.  Neurogenic potential of dental pulp stem cells isolated from murine incisors , 2014, Stem Cell Research & Therapy.

[45]  W. Que,et al.  Lithium-assisted exfoliation of pristine graphite for few-layer graphene nanosheets , 2015, Nano Research.

[46]  Moon Gyu Sung,et al.  Enhanced Differentiation of Human Neural Stem Cells into Neurons on Graphene , 2011, Advanced materials.

[47]  Xiaogang Qu,et al.  3D Graphene Oxide–Polymer Hydrogel: Near‐Infrared Light‐Triggered Active Scaffold for Reversible Cell Capture and On‐Demand Release , 2013, Advanced materials.

[48]  Kwang S. Kim,et al.  Large-scale pattern growth of graphene films for stretchable transparent electrodes , 2009, Nature.

[49]  M. Mahmoudi,et al.  Graphene: promises, facts, opportunities, and challenges in nanomedicine. , 2013, Chemical reviews.