Neuronal cell attachment to fluorinated ethylene propylene films with covalently immobilized laminin oligopeptides YIGSR and IKVAV. II

Abstract

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)

Organic–Inorganic Surface Modifications for Titanium Implant Surfaces

“…Additionally, various other peptide sequences have been immobilized onto implant materials (Table III) (99,112,(116)(117)(118)(119)(120). To provide a stable link, peptide sequences are usually covalently attached to the titanium surface, e.g.…”

Peptide-based biopolymers in biomedicine and biotechnology

“…In some cases, the RGD sequences can also promote other cellular functions such as matrix mineralization of osteogenic cells [359,360] and neurite outgrowth [358]. Other peptide sequences representing important domains of laminin and fibronectin such as IKVAV, YIGSR, and KNEED also promote the attachment of neuronal and endothelial cells [8][9][10][11]. At the tissue level, the peptide sequence GHK was found to promote dermal wound healing and tissue repair [12].…”

“…Myc/c-myc (EQKLISEED) 10 aa Antibody • Ligand density characterization of peptide-modified biomaterials [371] • Biomimetic peptides [359,372] • The effect of peptide surface density on mineralization of a matrix deposited by osteogenic cells [359,360] • Enhancement of the growth of human endothelial cells by surface roughness at nanometer scale [361] • Adhesion of α 5 β 1 receptors to biomimetic substrates constructed from peptide amphiphiles [349] • Biomimetic peptide surfaces that regulate adhesion, spreading, cytoskeletal organization, and mineralization of the matrix deposited by osteoblast-like cells [359,360] RGD-linked polymers • RGD-peptide-modified poly(lactic acid-co-lysine) [354] • Acrylic terpolymers with RGD peptides [356] • Diblock copolymers modified with cyclic RGD peptides [355] • Polyurethanes [357] • Synthetic hydrogels [350,363,370] • Immobilized RGD peptides on surface-grafted dextran [351] • Collagen by covalent grafting with RGD peptides [352] • Hyaluronan hydrogel [350] • RGD-grafted poly-l-lysine-graft-(polyethylene glycol) copolymers block [353] • Neurite outgrowth on well-characterized surfaces [ • Neuronal cell attachment [10] • Immobilization of laminin peptide in molecularly aligned chitosan by covalent bonding [11] …”

“…Despite this important limitation, peptide based materials offer many potential advantages over synthetic materials. First, short peptide motifs such as RGD, KNEED, and IKVAV that are ubiquitous ligands for cell receptors and mediate various cell behaviors such as attachment and spreading [8][9][10][11][12] can be appended to or embedded within repetitive polypeptide materials with greater ease than in synthetic materials. Second, selfassembly and directed-assembly of peptides have recently gained research interest as viable ways to generate functional biomaterials.…”

Biology on a Chip: Microfabrication for Studying the Behavior of Cultured Cells

“…Since then many studies in neuronal patterning have been performed in a variety of labs, using various methods, including surface modification by silane chemistry 113 137 Ti, 140 polystyrene, 130,131 Teflon (fluorinated ethylene propylene), [132][133][134][141][142][143][144] and biodegradable polymers. 145 These studies mainly yielded insights into the adhesion and guidance of neuronal growth by insoluble factors and could prove valuable for selective neuronal immobilization on future microsensors.…”