The detachment strength and morphology of bone cells contacting materials modified with a peptide sequence found within bone sialoprotein.
Adhesion, spreading, and focal contact formation of primary bone-derived cells on quartz surfaces grafted with a 15 amino acid peptide that contained a -RGD-(-Arg-Gly-Asp-) sequence unique to bone sialoprotein was investigated. The peptide surfaces were fabricated by using a heterbifunctional crosslinker, sulfosuccinimidyal 4-(N-maleimidomethyl)cyclohexane-1-carboxylate, to link the peptide to amine functionalized quartz surfaces. Contact angle measurements, spectroscopic ellipsometry, and X-ray photoelectron spectroscopy were used to confirm the chemistry and thickness of the overlayers. A radial flow apparatus was used to characterize cell detachment from peptide-grafted surfaces. After 20 min of cell incubation, the strength of cell adhesion was significantly (p < 0.05) higher on the -RGD- compared to -RGE- (control) surfaces. Furthermore, the mean area of cells contacting the -RGD- was significantly (p < 0.05) higher than -RGE- surfaces. Vinculin staining showed formation of small focal contact patches on the periphery of bone cells incubated for 2 h on the -RGD- surfaces; however, few or no focal contacts were formed by cells seeded on the -RGE-grafted surfaces. The methods of peptide immobilization utilized in this study can be applied to implants, biosensors, and diagnostic devices that require specificity in cell adhesion.
Rezania
A
A
Division of Biological Materials, Northwestern University Dental School, Chicago, Illinois 60611-3008, USA.
Thomas
C H
CH
Branger
A B
AB
Waters
C M
CM
Healy
K E
KE
D000595:Amino Acid Sequence; D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry*; D001842:Bone and Bones / Q000166:cytology* / Q000502:physiology; D002448:Cell Adhesion; D002478:Cells, Cultured; D003432:Cross-Linking Reagents; D058574:Integrin-Binding Sialoprotein; D008024:Ligands; D008422:Materials Testing; D008856:Microscopy, Fluorescence; D008858:Microscopy, Phase-Contrast; D009842:Oligopeptides / Q000737:chemistry / Q000235:genetics; D051381:Rats; D012795:Sialoglycoproteins / Q000737:chemistry*; D013499:Surface Properties
D001672:Biocompatible Materials; D003432:Cross-Linking Reagents; D058574:Integrin-Binding Sialoprotein; D008024:Ligands; D009842:Oligopeptides; D012795:Sialoglycoproteins; C047981:arginyl-glycyl-aspartic acid