Sanchez-Quevedo, M.C.Alaminos, M.Capitan, L.M.Moreu, G.Garzon, I.Crespo, P.V.Campos, A2012-12-032012-12-032007-06Sanchez-Quevedo MC, Alaminos M, Capitan LM, Moreu G, Garzon I et al.Histological and histochemical evaluation of human oral mucosa constructs developed by tissue engineering. Histol Histopathol. 2007 Jun;22(6):631-400213-3911http://hdl.handle.net/10668/683Journal Article; Research Support, Non-U.S. Gov't;Reconstruction of large oral mucosa defects is often challenging, since the shortage of healthy oral mucosa to replace the excised tissues is very common. In this context, tissue engineering techniques may provide a source of autologous tissues available for transplant in these patients. In this work, we developed a new model of artificial oral mucosa generated by tissue engineering using a fibrin-agarose scaffold. For that purpose, we generated primary cultures of human oral mucosa fibroblasts and keratinocytes from small biopsies of normal oral mucosa using enzymatic treatments. Then we determined the viability of the cultured cells by electron probe quantitative X-ray microanalysis, and we demonstrated that most of the cells in the primary cultures were alive and had high K/Na ratios. Once cell viability was determined, we used the cultured fibroblasts and keratinocytes to develop an artificial oral mucosa construct by using a fibrin-agarose extracellular matrix and a sequential culture technique using porous culture inserts. Histological analysis of the artificial tissues showed high similarities with normal oral mucosa controls. The epithelium of the oral substitutes had several layers, with desmosomes and apical microvilli and microplicae. Both the controls and the oral mucosa substitutes showed high suprabasal expression of cytokeratin 13 and low expression of cytokeratin 10. All these results suggest that our model of oral mucosa using fibrin-agarose scaffolds show several similarities with native human oral mucosa.enTécnicas de Cultivo de CélulaSupervivencia CelularCélulas CultivadasMicroanálisis por Sonda ElectrónicaFibrinaFibroblastosExpresión GénicaPerfilación de la Expresión GénicaHumanosHidrogelesInmunohistoquímicaQueratinocitosMicroscopía ElectrónicaMucosa BucalAnálisis de Secuencia en Orden de OligonucleótidoSefarosaIngeniería de TejidosFibrin-agaroseConstructsTissue engineeringMedical Subject Headings::Phenomena and Processes::Cell Physiological Phenomena::Cell Physiological Processes::Cell SurvivalMedical Subject Headings::Anatomy::Cells::Cells, CulturedMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Diagnosis::Diagnostic Techniques and Procedures::Diagnostic Imaging::Microscopy::Microscopy, Electron::Electron Probe MicroanalysisMedical Subject Headings::Chemicals and Drugs::Amino Acids, Peptides, and Proteins::Proteins::Blood Proteins::FibrinMedical Subject Headings::Anatomy::Cells::Connective Tissue Cells::FibroblastsMedical Subject Headings::Phenomena and Processes::Genetic Phenomena::Genetic Processes::Gene ExpressionMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Genetic Techniques::Gene Expression ProfilingMedical Subject Headings::Organisms::Eukaryota::Animals::Chordata::Vertebrates::Mammals::Primates::Haplorhini::Catarrhini::Hominidae::HumansMedical Subject Headings::Chemicals and Drugs::Complex Mixtures::Colloids::Gels::HydrogelsMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Diagnosis::Diagnostic Techniques and Procedures::Clinical Laboratory Techniques::Cytological Techniques::Histocytochemistry::ImmunohistochemistryMedical Subject Headings::Anatomy::Cells::Epithelial Cells::KeratinocytesMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Diagnosis::Diagnostic Techniques and Procedures::Diagnostic Imaging::Microscopy::Microscopy, ElectronMedical Subject Headings::Anatomy::Tissues::Membranes::Mucous Membrane::Mouth MucosaMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Chemistry Techniques, Analytical::Microchip Analytical Procedures::Microarray Analysis::Oligonucleotide Array Sequence AnalysisMedical Subject Headings::Chemicals and Drugs::Carbohydrates::Polysaccharides::SepharoseMedical Subject Headings::Analytical, Diagnostic and Therapeutic Techniques and Equipment::Investigative Techniques::Culture Techniques::Cell Engineering::Tissue Engineeringresearch article17357093open access1699-5848