title: Changes in actin and tubulin expression in osteogenic cells cultured on bioactive glass-based surfaces.
abstract: The present study evaluated whether the changes in the labeling pattern of cytoskeletal proteins in osteogenic cells cultured on bioactive glass-based materials are due to altered mRNA and protein levels. Primary rat-derived osteogenic cells were plated on Bioglass® 45S5, Biosilicate®, and borosilicate (bioinert control). The following parameters were assayed: (i) qualitative epifluorescence analysis of actin and tubulin; (ii) quantitative mRNA and protein expression for actin and tubulin by real-time PCR and ELISA, respectively, and (iii) qualitative analysis of cell morphology by scanning electron microscopy (SEM). At days 3 and 7, the cells grown on borosilicate showed typical actin and tubulin labeling patterns, whereas those on the bioactive materials showed roundish areas devoid of fluorescence signals. The cultures grown on bioactive materials showed significant changes in actin and tubulin mRNA expression that were not reflected in the corresponding protein levels. A positive correlation between the mRNA and protein as well as an association between epifluorescence imaging and quantitative data were only detected for the borosilicate. SEM imaging of the cultures on the bioactive surfaces revealed cells partly or totally coated with material aggregates, whose characteristics resembled the substrate topography. The culturing of osteogenic cells on Bioglass® 45S5 and Biosilicate® affect actin and tubulin mRNA expression but not the corresponding protein levels. Changes in the labeling pattern of these proteins should then be attributed, at least in part, to the presence of a physical barrier on the cell surface as a result of the material surface reactions, thus limiting fluorescence signals.
authors.affiliation: Cell Culture Laboratory, Faculty of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: Carolina Scanavez
authors.initials: CS
authors.lastname: Martins
authors.affiliation: Cell Culture Laboratory, Faculty of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: Emanuela Prado
authors.initials: EP
authors.lastname: Ferraz
authors.affiliation: Cell Culture Laboratory, Faculty of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: Larissa Moreira Spinola
authors.initials: LM
authors.lastname: De Castro-Raucci
authors.affiliation: Cell Culture Laboratory, Faculty of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: Lucas Novaes
authors.initials: LN
authors.lastname: Teixeira
authors.affiliation: Cell Culture Laboratory, Faculty of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: William Marcatti Amarú
authors.initials: WM
authors.lastname: Maximiano
authors.affiliation: Cell Culture Laboratory, Faculty of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: Adalberto Luiz
authors.initials: AL
authors.lastname: Rosa
authors.affiliation: Cell Culture Laboratory, Faculty of Dentistry of Ribeirão Preto, University of São Paulo, Ribeirão Preto, SP, Brazil.
authors.forename: Paulo Tambasco
authors.initials: PT
authors.lastname: De Oliveira
mesh_terms: D000199:Actins / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics; D000818:Animals; D002478:Cells, Cultured; D002516:Ceramics; D004280:Dobutamine; D020869:Gene Expression Profiling; D005898:Glass; D008855:Microscopy, Electron, Scanning; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism*; D017208:Rats, Wistar; D060888:Real-Time Polymerase Chain Reaction; D013997:Time Factors; D014404:Tubulin / Q000032:analysis / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D000199:Actins; D014404:Tubulin; C425330:bioactive glass 45S5; D004280:Dobutamine
keywords: bioactive materials; cell culture; cytoskeleton; glass ceramics; osteoblastic cells