title: MC3T3-E1 cell adhesion to hydroxyapatite with adsorbed bone sialoprotein, bone osteopontin, and bovine serum albumin.
abstract: Native bone tissue is composed of a complex matrix of collagen, non-collagenous proteins, and hydroxyapatite (HAP). Bone sialoprotein (BSP) and bone osteopontin (OPN) are members of the non-collagenous protein family termed the SIBLING (small integrin-binding ligand, N-linked glycoproteins) proteins, which are primarily found in mineralized tissues. Previously, OPN was shown to exhibit a preferential orientation for MC3T3-E1 cell adhesion when it was specifically bound to collagen, while the MC3T3-E1 cell adhesion was shown to be dependant on the conformational flexibility of BSP specifically bound to collagen. Additionally, OPN was shown to play a greater role than BSP for cell binding to collagen. In this work, the orientations and conformations of BSP and OPN specifically bound to HAP are probed under similar conditions. Radiolabeled adsorption isotherms were obtained for BSP and OPN on HAP formed from a simulated body fluid, and the results show that HAP has the capacity to bind significantly more BSP than OPN. An in vitro MC3T3-E1 cell adhesion assay was then performed to compare the cell binding ability of adsorbed BSP and OPN specifically bound to HAP. It was found that there is a preference for cell binding to HAP with adsorbed BSP as compared to OPN, but not to a statistically significant level. However, the maximum cell binding was observed on HAP substrates with adsorbed heat denatured bovine serum albumin (BSA). The influence of BSA on cell binding was shown to be concentration dependant and it is believed that the adsorbed BSA modulates the proliferation state of the bound cells.
authors.affiliation: Department of Chemical Engineering, University of Washington, Seattle, WA 98195, United States.
authors.forename: Matthew T
authors.initials: MT
authors.lastname: Bernards
authors.forename: Chunlin
authors.initials: C
authors.lastname: Qin
authors.forename: Shaoyi
authors.initials: S
authors.lastname: Jiang
mesh_terms: D000818:Animals; D001665:Binding Sites; D002417:Cattle; D002448:Cell Adhesion; D018929:Cell Culture Techniques; D002478:Cells, Cultured; D017886:Durapatite / Q000378:metabolism*; D008855:Microscopy, Electron, Scanning; D008954:Models, Biological; D010006:Osteoblasts / Q000378:metabolism*; D053495:Osteopontin / Q000737:chemistry / Q000378:metabolism*; D011485:Protein Binding; D011487:Protein Conformation; D051381:Rats; D012710:Serum Albumin, Bovine / Q000378:metabolism*; D012795:Sialoglycoproteins / Q000737:chemistry / Q000378:metabolism*; D013977:Tibia / Q000166:cytology; D013997:Time Factors
chemical_list: D012795:Sialoglycoproteins; D053495:Osteopontin; D012710:Serum Albumin, Bovine; D017886:Durapatite