RT Journal Article
T1 Cell survival and differentiation with nanocrystalline glass-like carbon using substantia nigra dopaminergic cells derived from transgenic mouse embryos
A1 Rodriguez-Losada, Noela
A1 Romero, Pablo
A1 Estivill-Torrús, Guillermo
A1 Guzmán de Villoria, Roberto
A1 Aguirre, Jose A
K1 Animales
K1 Apoptosis
K1 Western blotting
K1 Carbono
K1 Línea celular
K1 Replicación del ADN
K1 Conductividad eléctrica
K1 Células madre embrionarias
K1 Citometría de flujo
K1 Técnica del anticuerpo fluorescente
K1 Grafito
K1 Histonas
K1 Ratones transgénicos
K1 Nanoestructuras
K1 Canales de potasio
K1 Antígeno nuclear de célula en proliferación
K1 Medicina regenerativa
K1 Serina
K1 Sustancia negra
K1 Sinaptofisina
K1 Ingeniería de tejidos
K1 Tirosina 3-Monooxigenasa
AB Regenerative medicine requires, in many cases, physical supports to facilitate appropriate cellular architecture, cell polarization and the improvement of the correct differentiation processes of embryonic stem cells, induced pluripotent cells or adult cells. Because the interest in carbon nanomaterials has grown within the last decade in light of a wide variety of applications, the aim of this study was to test and evaluate the suitability and cytocompatibility of a particular nanometer-thin nanocrystalline glass-like carbon film (NGLC) composed of curved graphene flakes joined by an amorphous carbon matrix. This material is a disordered structure with high transparency and electrical conductivity. For this purpose, we used a cell line (SN4741) from substantia nigra dopaminergic cells derived from transgenic mouse embryos. Cells were cultured either in a powder of increasing concentrations of NGLC microflakes (82±37μm) in the medium or on top of nanometer-thin films bathed in the same culture medium. The metabolism activity of SN4741 cells in presence of NGLC was assessed using methylthiazolyldiphenyl-tetrazolium (MTT) and apoptosis/necrosis flow cytometry assay respectively. Growth and proliferation as well as senescence were demonstrated by western blot (WB) of proliferating cell nuclear antigen (PCNA), monoclonal phosphorylate Histone 3 (serine 10) (PH3) and SMP30 marker. Specific dopaminergic differentiation was confirmed by the WB analysis of tyrosine hydroxylase (TH). Cell maturation and neural capability were characterized using specific markers (SYP: synaptophysin and GIRK2: G-protein-regulated inward-rectifier potassium channel 2 protein) via immunofluorescence and coexistence measurements. The results demonstrated cell positive biocompatibility with different concentrations of NGLC. The cells underwent a process of adaptation of SN4741 cells to NGLC where their metabolism decreases. This process is related to a decrease of PH3 expression and significant increase SMP30 related to senescence processes. After 7 days, the cells increased the expression of TH and PCNA that is related to processes of DNA replication. On the other hand, cells cultured on top of the film showed axonal-like alignment, edge orientation, and network-like images after 7 days. Neuronal capability was demonstrated to a certain extent through the analysis of significant coexistence between SYP and GIRK2. Furthermore, we found a direct relationship between the thickness of the films and cell maturation. Although these findings share certain similarities to our previous findings with graphene oxide and its derivatives, this particular nanomaterial possesses the advantages of high conductivity and transparency. In conclusion, NGLC could represent a new platform for biomedical applications, such as for use in neural tissue engineering and biocompatible devices.
PB Public Libray of Science
YR 2017
FD 2017-03-23
LK http://hdl.handle.net/10668/2664
UL http://hdl.handle.net/10668/2664
LA en
NO Rodriguez-Losada N, Romero P, Estivill-Torrús G, Guzmán de Villoria R, Aguirre JA. Cell survival and differentiation with nanocrystalline glass-like carbon using substantia nigra dopaminergic cells derived from transgenic mouse embryos. Plos One. 2017; 12(3):e0173978.
DS RISalud
RD Apr 11, 2025