Proteomic analysis of human skin fibroblasts grown on titanium: novel approach to study molecular biocompatibility.
Despite the growing use of titanium as an implant material, there is a lack of consensus on what constitutes the molecular basis of its biocompatibility. In the present study we compared the response of skin fibroblasts to two different growth supporting surfaces: commercially pure titanium (cpTi) and tissue culture polystyrene (TCPS). Proteins from extracts of whole cells and adsorbed serum were separated and identified by 2-dimensional gel electrophoresis. In all, 40 proteins (46 spots) were identified by matrix-assisted laser desorption/ionization and mass spectrometry, database searching, immunoblotting, running a standard, or a combination of these techniques. Many of the proteins collected from the two surfaces were found to derive from the serum used in the culture medium. The surface properties of titanium appeared to promote the formation of a more concentrated carpet of serum proteins. Several proteins from bovine or human serum, such as albumin, alpha2-HS-glycoprotein, alpha-fetoprotein, plasminogen, thrombospondin 1, and serotransferrin, along with a few unidentified serum components, were found to adsorb onto cpTi in comparatively high concentrations. The adsorption of serum proteins did not appear to be selective on either substrate. We found that among the major cellular proteins, fibronectin and a cytoskeletal protein (non-muscle myosin heavy chain type A) were expressed at lower levels by fibroblasts grown on cpTi compared to TCPS. By analyzing the changes in the entire proteome of cells in response to different growth substrates, we may gain a better understanding of the molecular basis of biocompatibility.
Derhami
K
K
Department of Dentistry, University of Alberta, Edmonton, Alberta T6G 2H7, Canada.
Zheng
J
J
Li
L
L
Wolfaardt
J F
JF
Scott
P G
PG
D000818:Animals; D001672:Biocompatible Materials; D015153:Blotting, Western; D002417:Cattle; D002448:Cell Adhesion / Q000187:drug effects; D002478:Cells, Cultured; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D005347:Fibroblasts / Q000166:cytology; D015894:Genome, Human; D006801:Humans; D007525:Isoelectric Focusing; D011137:Polystyrenes / Q000737:chemistry*; D020543:Proteome; D012709:Serum Albumin / Q000032:analysis; D012867:Skin / Q000166:cytology*; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014025:Titanium / Q000737:chemistry*
D001672:Biocompatible Materials; D011137:Polystyrenes; D020543:Proteome; D012709:Serum Albumin; D014025:Titanium