The behavior of osteoblast-like cells on various substrates with functional blocking of integrin-beta1 and integrin-beta3.
This study was designed to examine the influence of integrin subunit-beta1 and subunit-beta3 on the behavior of primary osteoblast-like cells, cultured on calcium phosphate (CaP)-coated and non coated titanium (Ti). Osteoblast-like cells were incubated with specific monoclonal antibodies against integrin-beta1 and integrin-beta3 to block the integrin function. Subsequently, cells were seeded on Ti discs, either non coated or provided with a 2 microm carbonated hydroxyapatite coating using Electrostatic Spray Deposition. Results showed that on CaP coatings, cellular attachment was decreased after a pre-treatment with either anti-integrin-beta1 or anti-integrin-beta3 antibodies. On Ti, cell adhesion was only slightly affected after a pre-treatment with anti-integrin-beta3 antibodies. Scanning electron microscopy showed that on both types of substrate, cellular morphology was not changed after a pre-treatment with either antibody. With quantitative PCR, it was shown for both substrates that mRNA expression of integrin-beta1 was increased after a pre-treatment with either anti-integrin-beta1 or anti-integrin-beta3 antibodies. Furthermore, after a pre-treatment with either antibody, mRNA expression of integrin-beta3 and ALP was decreased, on both types of substrate. In conclusion, osteoblast-like cells have the ability to compensate to great extent for the blocking strategy as applied here. Still, integrin-beta1 and beta3 seem to play different roles in attachment, proliferation, and differentiation of osteoblast-like cells, and responses on CaP-coated substrates differ to non coated Ti. Furthermore, the influence on ALP expression suggests involvement of both integrin subunits in signal transduction for cellular differentiation.
Siebers
M C
MC
Department of Periodontology and Biomaterials, College of Dental Science 309, Radboud University Nijmegen Medical Centre, Nijmegen, The Netherlands.
Walboomers
X F
XF
van den Dolder
J
J
Leeuwenburgh
S C G
SC
Wolke
J G C
JG
Jansen
J A
JA
D000818:Animals; D000906:Antibodies; D001672:Biocompatible Materials / Q000737:chemistry*; D002130:Calcium Phosphates / Q000737:chemistry; D002448:Cell Adhesion; D002478:Cells, Cultured; D005786:Gene Expression Regulation; D019012:Integrin beta1 / Q000235:genetics / Q000378:metabolism*; D039661:Integrin beta3 / Q000235:genetics / Q000378:metabolism*; D008855:Microscopy, Electron, Scanning; D010006:Osteoblasts / Q000166:cytology* / Q000502:physiology*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D014025:Titanium / Q000737:chemistry
D000906:Antibodies; D001672:Biocompatible Materials; D002130:Calcium Phosphates; D019012:Integrin beta1; D039661:Integrin beta3; D012333:RNA, Messenger; C020243:calcium phosphate; D014025:Titanium