Effect of blasting treatment and Fn coating on MG63 adhesion and differentiation on titanium: a gene expression study using real-time RT-PCR

Abstract

Biomaterial surface properties, via alterations in the adsorbed protein layer, and the presence of specific functional groups can influence integrin binding specificity, thereby modulating cell adhesion and differentiation processes. The adsorption of fibronectin, a protein directly involved in osteoblast adhesion to the extracellular matrix, has been related to different physical and chemical properties of biomaterial surfaces. This study used blasting particles of different sizes and chemical compositions to evaluate the response of MG63 osteoblast-like cells on smooth and blasted titanium surfaces, with and without fibronectin coatings, by means of real-time reverse transcription-polymerase chain reaction (qRT-PCR) assays. This response included (a) expression of the α(5), α(v) and α(3) integrin subunits, which can bind to fibronectin through the RGD binding site, and (b) expression of alkaline phosphatase (ALP) and osteocalcin (OC) as cell-differentiation markers. ALP activity and synthesis of OC were also tested. Cells on SiC-blasted Ti surfaces expressed higher amounts of the α(5) mRNA gene than cells on Al(2)O(3)-blasted Ti surfaces. This may be related to the fact that SiC-blasted surfaces adsorbed higher amounts of fibronectin due to their higher surface free energy and therefore provided a higher number of specific cell-binding sites. Fn-coated Ti surfaces decreased α(5) mRNA gene expression, by favoring the formation of other integrins involved in adhesion over α(5)β(1). The changes in α(5) mRNA expression induced by the presence of fibronectin coatings may moreover influence the osteoblast differentiation pathway, as fibronectin coatings on Ti surfaces also decreased both ALP mRNA expression and ALP activity after 14 and 21 days of cell culture.

Differential Effects of Tissue Culture Coating Substrates on Prostate Cancer Cell Adherence, Morphology and Behavior

“…Vergani and collaborators showed that modifications of cell shape directly reflected on the nucleus and the nuclear architecture, followed by chromatin condensation, and finally affecting the transcriptional profile of genes. [49] In addition, the integrins presented by the cells are an effect of the surface substrate which can control the expression levels of their subunits [50].…”

In search of an osteoblast cell model for in vitro research

“…• arrested in pre-osteoblast state • inconsistent regarding cell mineralisation Heremans et al, 1978Clover et al, 1992Olivares-Navarrete et al, 2008Kumarasuriyar et al, 2009Saldana et al, 2011Pierschbacher et al, 1988…”

“…This is surprising, considering that these animals are frequently used as large animal in vivo models, and other skeletal cells from these animals serve as established in vitro models. For instance, chondrocytes or intervertebral disc cells from bovine specimens are widely used in tissue engineering research and for investigating basic biological mechanisms (Dowthwaite et al, 2004;Sakai et al, 2009;Collin et al, 2011;Hoshiba et al, 2011;Peroglio et al, 2011). However, studies focused on bovine osteoblast cells are limited.…”

Potential mechanism for osseointegration of dental implants in Zucker diabetic fatty rats

“…12 It has been reported 20 that both periodontal and gingival fibroblasts express ␣5␤1 integrin subunits and adhere to laminin, vitronectin, and bone sialoprotein as part of the cementum. Pegueroles et al, 21 reported that the changes in ␣5 and ␣ v mRNA expression induced by the presence of fibronectin coatings can influence the osteoblasts' differentiation pathway.…”