Neuronal cell attachment to fluorinated ethylene propylene films with covalently immobilized laminin oligopeptides YIGSR and IKVAV. II.
Material surfaces that can mediate cellular interactions by the coupling of specific cell membrane receptors may allow for the design of a biomaterial that can control cell attachment, differentiation, and tissue organization. Cell adhesion proteins have been shown to contain minimum oligopeptide sequences that are recognized by cell surface receptors and can be covalently immobilized on material surfaces. In this study, cell attachment to fluorinated ethylene propylene (FEP) films functionalized with the laminin-derived oligopeptides, YIGSR and a 19-mer IKVAV-containing sequence, was assessed using NG108-15 neuroblastoma and PC12 cells. A radiofrequency glow discharge (RFGD) process that replaces the FEP surface fluorine atoms with reactive hydroxyl functionalities was used to activate the film surfaces. The oligopeptides were then covalently coupled to the surface by their C-terminus using a standard nucleophilic substitution reaction. The covalent attachment of the oligopeptides to the FEP surface was verified using electron spectroscopy for chemical analysis (ESCA). Receptor-mediated NG108-15 cell attachment on the YIGSR-modified films was determined using competitive binding assays. Average cell attachment on the oligopeptide immobilized films in medium containing soluble CDPGYIGSR was reduced by approximately a factor of 2, compared to cell attachment in serum-free medium alone. No significant decrease in cell attachment was noted in medium containing the mock oligopeptide sequence CDPGYIGSK. FEP films immobilized with the 19-mer IKVAV sequence demonstrated a higher percentage of receptor mediated cell attachment on the film surfaces.(ABSTRACT TRUNCATED AT 250 WORDS)
Ranieri
J P
JP
Division of Surgical Research, Centre Hospitalier Universitaire Vaudois, Lausanne University Medical School, Switzerland.
Bellamkonda
R
R
Bekos
E J
EJ
Vargo
T G
TG
Gardella
J A
JA
Aebischer
P
P
D000595:Amino Acid Sequence; D000818:Animals; D002448:Cell Adhesion; D002460:Cell Line; D016895:Culture Media, Serum-Free; D006801:Humans; D007797:Laminin / Q000737:chemistry*; D008565:Membrane Proteins / Q000378:metabolism; D008969:Molecular Sequence Data; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D009842:Oligopeptides / Q000737:chemistry*; D016716:PC12 Cells; D011138:Polytetrafluoroethylene / Q000031:analogs & derivatives*; D051381:Rats; D005465:Fluorocarbon Polymers
D016895:Culture Media, Serum-Free; D007797:Laminin; D008565:Membrane Proteins; D009842:Oligopeptides; D011138:Polytetrafluoroethylene; C096305:fluorinated ethylene propylene; D005465:Fluorocarbon Polymers