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Changes in actin and tubulin expression in osteogenic cells cultured on bioactive glass‐based surfaces
Author(s) -
Martins Carolina Scanavez,
Ferraz Emanuela Prado,
CastroRaucci Larissa Moreira Spinola De,
Teixeira Lucas Novaes,
Maximiano William Marcatti Amarú,
Rosa Adalberto Luiz,
De Oliveira Paulo Tambasco
Publication year - 2015
Publication title -
microscopy research and technique
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.536
H-Index - 118
eISSN - 1097-0029
pISSN - 1059-910X
DOI - 10.1002/jemt.22583
Subject(s) - borosilicate glass , tubulin , cytoskeleton , fluorescence microscope , actin , microtubule , microbiology and biotechnology , messenger rna , cell , bioactive glass , fluorescence , chemistry , biophysics , biology , biochemistry , materials science , gene , physics , organic chemistry , quantum mechanics , composite material
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. Microsc. Res. Tech. 78:1046–1053, 2015 . © 2015 Wiley Periodicals, Inc.