Adhesion of MC3T3-E1 cells to bone sialoprotein and bone osteopontin specifically bound to collagen I.
Bone sialoprotein (BSP) and bone osteopontin (OPN) are members of the SIBLING (small integrin-binding ligand, N-linked glycoproteins) family of proteins commonly 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. In this work, the orientation of BSP under similar circumstances is examined and compared with OPN. Radiolabeled adsorption isotherms were obtained for BSP bound to both tissue culture polystyrene (TCPS) and collagen-coated TCPS. The results show that collagen has the capacity to bind almost twice as much OPN under identical conditions. An in vitro MC3T3-E1 cell adhesion assay was then performed to compare the cell binding ability of BSP on either TCPS or collagen-coated TCPS with identical amounts of adsorbed protein. It was found that there is no significant difference in the cell binding ability of BSP on either of the substrates. For cell binding studies on collagen-coated TCPS, it was shown that there are a greater number of cells bound to substrates with adsorbed OPN as compared with BSP. The preferable orientation of OPN for cell binding coupled with the higher binding capability of collagen for OPN indicates that OPN is more important than BSP for osteoblast adhesion to the collagen matrix. In addition, a cell inhibition assay was performed to show that all of the cell binding that occurred throughout these studies was dependent upon integrin interactions with the RGD cell binding moiety.
Department of Chemical Engineering, University of Washington, Seattle, Washington 98195, USA.
Matthew T
MT
Bernards
Chunlin
C
Qin
Buddy D
BD
Ratner
Shaoyi
S
Jiang
D000327:Adsorption; D000818:Animals; D001842:Bone and Bones / Q000378:metabolism*; D002448:Cell Adhesion; D002452:Cell Count; D002460:Cell Line; D024042:Collagen Type I / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D058574:Integrin-Binding Sialoprotein; D007457:Iodine Radioisotopes; D051379:Mice; D053495:Osteopontin / Q000378:metabolism*; D011137:Polystyrenes / Q000378:metabolism; D011485:Protein Binding; D051381:Rats; D012795:Sialoglycoproteins / Q000378:metabolism*
D024042:Collagen Type I; C546872:Ibsp protein, mouse; D058574:Integrin-Binding Sialoprotein; D007457:Iodine Radioisotopes; D011137:Polystyrenes; D012795:Sialoglycoproteins; D053495:Osteopontin