title: Significant role of adhesion properties of primary osteoblast-like cells in early adhesion events for chondroitin sulfate and dermatan sulfate surface molecules.
abstract: The purpose of this study was to characterize the role of cell surface adhesive macromolecules through enzyme modulation and metabolic recovery prior to and during a kinetic cell adhesion assay. Primary rat calvarial osteoblast-like cells were derived from Sprague-Dawley calvarial plates. Cell adhesion kinetics was evaluated with the definition of first-order adhesion kinetics. Osteoblasts were incubated in an adhesion buffer for 1 h prior to a cell attachment assay using various enzymes to remove cell surface glycosaminoglycans (GAGs). A subtractive adhesion analysis was performed by plating cells at 5 x 10(4)/well for variable periods through 2 h. The medium was collected, the well surface washed and pooled, and the number of cells enumerated with a Coulter Counter. Cell adhesion demonstrated first-order logarithmic adhesion kinetics in the first 60 min. Scatchard analysis demonstrated a linear relationship. Preexposure of cells to various enzyme combinations demonstrated that 50% of the equilibrium adhesion was dependent on chondroitin sulfate or dermatan sulfate surface macromolecules. These results were confirmed with pretreatment with a metabolic inhibitor of GAG synthesis (beta-D-xyloside). These results suggest an important role for cell associated chondroitin sulfate and dermatan sulfate in cell adhesion in addition to Arg-Gly-Asp or integrin mediated adhesion events.
authors.affiliation: N447 Dows Institute for Dental Research and Department of Biological Sciences, The University of Iowa, Iowa City, Iowa 52242, USA.
authors.forename: C M
authors.initials: CM
authors.lastname: Stanford
authors.forename: M
authors.initials: M
authors.lastname: Solursh
authors.forename: J C
authors.initials: JC
authors.lastname: Keller
mesh_terms: D000818:Animals; D000831:Animals, Newborn; D002448:Cell Adhesion / Q000502:physiology*; D002462:Cell Membrane / Q000502:physiology; D002478:Cells, Cultured; D002809:Chondroitin Sulfates / Q000502:physiology*; D003871:Dermatan Sulfate / Q000502:physiology*; D007700:Kinetics; D010006:Osteoblasts / Q000737:chemistry* / Q000502:physiology*; D051381:Rats; D017207:Rats, Sprague-Dawley; D012886:Skull / Q000166:cytology / Q000502:physiology*; D013499:Surface Properties
chemical_list: D003871:Dermatan Sulfate; D002809:Chondroitin Sulfates