Effect of blasting treatment and Fn coating on MG63 adhesion and differentiation on titanium: a gene expression study using real-time RT-PCR.
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.
Biomaterials, Biomechanics, and Tissue Engineering Group, Department of Material Science and Metallurgy, Universitat Politècnica de Catalunya, Av. Diagonal 647, 08028, Barcelona, Spain. marta.pegueroles@upc.edu
M
M
Pegueroles
A
A
Aguirre
E
E
Engel
G
G
Pavon
F J
FJ
Gil
J A
JA
Planell
V
V
Migonney
C
C
Aparicio
D000469:Alkaline Phosphatase / Q000378:metabolism; D001665:Binding Sites; D001672:Biocompatible Materials / Q000737:chemistry*; D002448:Cell Adhesion; D002454:Cell Differentiation; D002460:Cell Line; D005353:Fibronectins / Q000737:chemistry / Q000378:metabolism*; D006801:Humans; D039201:Integrin alpha3beta1 / Q000378:metabolism; D039302:Integrin alphaVbeta3 / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism; D015675:Osteocalcin / Q000378:metabolism; D010641:Phenotype; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D014025:Titanium / Q000737:chemistry*
D001672:Biocompatible Materials; D005353:Fibronectins; D039201:Integrin alpha3beta1; D039302:Integrin alphaVbeta3; D015675:Osteocalcin; D014025:Titanium; D000469:Alkaline Phosphatase
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