Significant role of adhesion properties of primary osteoblast-like cells in early adhesion events for chondroitin sulfate and dermatan sulfate surface molecules
✍ Scribed by Stanford, Clark M. ;Solursh, Michael ;Keller, John C.
- Publisher
- John Wiley and Sons
- Year
- 1999
- Tongue
- English
- Weight
- 162 KB
- Volume
- 47
- Category
- Article
- ISSN
- 0021-9304
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✦ Synopsis
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 × 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 (-Dxyloside). 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.