Cell survival and differentiation with nanocrystalline glass-like carbon using substantia nigra dopaminergic cells derived from transgenic mouse embryos
| Autor: | Jose A. Aguirre, Guillermo Estivill-Torrús, Roberto Guzman de Villoria, Pablo Romero, Noela Rodriguez-Losada |
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| Přispěvatelé: | [Rodriguez-Losada, Noela] Univ Malaga, Fac Med, Dept Human Physiol, Malaga, Spain, [Aguirre, Jose A.] Univ Malaga, Fac Med, Dept Human Physiol, Malaga, Spain, [Rodriguez-Losada, Noela] Biomed Biomed Res Inst Malaga IBIMA, Campus Teatinos,Blvd Louis Pasteur, Malaga, Spain, [Aguirre, Jose A.] Biomed Biomed Res Inst Malaga IBIMA, Campus Teatinos,Blvd Louis Pasteur, Malaga, Spain, [Romero, Pablo] IMDEA Mat Inst, C Eric Kandel 2, Madrid, Spain, [Guzman de Villoria, Roberto] IMDEA Mat Inst, C Eric Kandel 2, Madrid, Spain, [Estivill-Torrus, Guillermo] Reg Univ Hosp Malaga, Biomed Res Inst Malaga IBIMA, Unidad Clin Neurociencia, Av Carlos Haya S-N, Malaga, Spain, [Guzman de Villoria, Roberto] FIDAMC Fdn Res Dev & Applicat Composite Mat, Avda Rita Levi Montalcini 29, Madrid, Spain, University of Malaga and Campus de Exceiencia Internacional Andalucia Tech (UMA), Junta de Andalucia Government, Spain, Spanish Ministry of Science and Innovation through the Ramon y Cajal Fellowship, [Rodríguez-Losada,N, Aguirre,JA] Department of Human Physiology, Faculty of Medicine, University of Malaga and Biomedicine Biomedical Research Institute of Malaga (IBIMA), Malaga, Spain. [Romero,P, Guzmán de Villoria,R] IMDEA Material Institute, Getafe, Madrid, Spain, [Estivill-Torrús,G] Unidad de Clínica de Neurociencia, Biomedical Research Institute of Malaga (IBIMA), Regional University Hospital Malaga, Málaga, Spain. [Guzmán de Villoria,R] Getafe, Madrid, Spain., This work was supported by 1) Fundsfrom the University of Malaga and Campus de Excelencia Internacional Andalucı´a Tech (UMA), 2)Junta de Andalucia Government, Spain (PAIDICTS156), 3) Spanish Ministry of Science andInnovation through the Ramon y Cajal Fellowship(RG). |
| Jazyk: | angličtina |
| Rok vydání: | 2017 |
| Předmět: |
Sinaptofisina
Analytical Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Clinical Laboratory Techniques::Cytological Techniques::Histocytochemistry::Immunohistochemistry::Fluorescent Antibody Technique [Medical Subject Headings] Technology and Food and Beverages::Technology Industry and Agriculture::Manufactured Materials::Nanostructures [Medical Subject Headings] Apoptosis Ratones transgénicos Mice Medicina regenerativa Anatomy::Cells::Cells Cultured::Cell Line [Medical Subject Headings] Phenomena and Processes::Physical Phenomena::Magnetic Phenomena::Electromagnetic Phenomena::Electricity::Electric Conductivity [Medical Subject Headings] Organisms::Eukaryota::Animals [Medical Subject Headings] Enzyme assays Nanotechnology Organisms::Eukaryota::Animals::Animal Population Groups::Animals Genetically Modified::Mice Transgenic [Medical Subject Headings] lcsh:Science Phenomena and Processes::Chemical Phenomena::Biochemical Phenomena::Biochemical Processes::DNA Replication [Medical Subject Headings] Analytical Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Culture Techniques::Cell Engineering::Tissue Engineering [Medical Subject Headings] Bioassays and physiological analysis Tissue Scaffolds Línea celular Chemicals and Drugs::Amino Acids Peptides and Proteins::Proteins::Carrier Proteins::Membrane Transport Proteins::Ion Channels::Potassium Channels [Medical Subject Headings] Cell biology Substantia Nigra Analytical Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Clinical Laboratory Techniques::Cytological Techniques::Cell Separation::Flow Cytometry [Medical Subject Headings] Antígeno nuclear de célula en proliferación Disciplines and Occupations::Health Occupations::Medicine::Regenerative Medicine [Medical Subject Headings] Anatomy::Nervous System::Central Nervous System::Brain::Mesencephalon::Tegmentum Mesencephali::Substantia Nigra [Medical Subject Headings] Physical Sciences Engineering and Technology Biological Cultures Cell Physiology Materials by Structure Histone h3 Materials Science Blotting Western Chemicals and Drugs::Amino Acids Peptides and Proteins::Proteins::Metalloproteins::Iron-Binding Proteins::Nonheme Iron Proteins::Tyrosine 3-Monooxygenase [Medical Subject Headings] 03 medical and health sciences Anatomy::Cells::Stem Cells::Embryonic Stem Cells [Medical Subject Headings] Botany Carbono Immunoassays Chemicals and Drugs::Amino Acids Peptides and Proteins::Proteins::Membrane Proteins::Vesicular Transport Proteins::Synaptophysin [Medical Subject Headings] Materials by Attribute Chemicals and Drugs::Amino Acids Peptides and Proteins::Proteins::Nuclear Proteins::Proliferating Cell Nuclear Antigen [Medical Subject Headings] Dopaminergic Neurons lcsh:R Biology and Life Sciences Chemicals and Drugs::Amino Acids Peptides and Proteins::Amino Acids::Amino Acids Neutral::Serine [Medical Subject Headings] Public repository Cell Metabolism Chemicals and Drugs::Inorganic Chemicals::Elements::Carbon::Graphite [Medical Subject Headings] 030104 developmental biology nervous system Biofilms Animales Nanoparticles lcsh:Q Conductividad eléctrica Citometría de flujo Developmental Biology 0301 basic medicine Immunofluorescence Chemicals and Drugs::Amino Acids Peptides and Proteins::Proteins::Nuclear Proteins::Histones [Medical Subject Headings] Canales de potasio Sustancia negra lcsh:Medicine Growth Western blotting Histonas Colorimetric assays Serina Thin Films Microscopy Multidisciplinary MTT assay Cell Death Embryo Cell Differentiation Chemicals and Drugs::Inorganic Chemicals::Elements::Carbon [Medical Subject Headings] Cell Processes Adhesion Research Article Genetically modified mouse Células madre embrionarias Cell Survival Substantia nigra Mice Transgenic Biology Research and Analysis Methods Técnica del anticuerpo fluorescente Tirosina 3-Monooxigenasa Cell Line Coatings Replicación del ADN Dopaminergic Cell Animals Cell survival Nanomaterials Nanoestructuras Cell Biology Cell Cultures Phenomena and Processes::Cell Physiological Phenomena::Cell Physiological Processes::Cell Death::Apoptosis [Medical Subject Headings] Grafito Analytical Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Chemistry Techniques Analytical::Electrophoresis::Blotting Western [Medical Subject Headings] Biochemical analysis Immunologic Techniques Ingeniería de tejidos Graphene |
| Zdroj: | PLoS ONE Repositorio Institucional de la Consejería de Sanidad de la Comunidad de Madrid Consejería de Sanidad de la Comunidad de Madrid PLoS ONE, Vol 12, Iss 3, p e0173978 (2017) BASE-Bielefeld Academic Search Engine |
| Popis: | 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 +/- 37 mu m) 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. |
| Databáze: | OpenAIRE |
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