Selective neuronal cell attachment to a covalently patterned monoamine on fluorinated ethylene propylene films

Abstract

The patterned covalent surface addition of a monoamine to fluorinated ethylene propylene films (FEP) controls both cellular attachment and differentiation in defined media conditions. A radio frequency glow discharge (RFGD) process was used to replace FEP surface fluorine atoms with hydroxyl groups. The primary amine was then covalently attached by polymerizing aminopropyl-triethoxysilane (APTES) via the hydroxyl functionalities. The selective attachment of cells to the APTES regions was determined to be dependent upon the initial adsorption of albumin to the patterned FEP membrane. Albumin was determined to enhance cellular attachment to the APTES regions and prevent attachment to the unmodified FEP areas for both an NB2a neuroblastoma cell line and primary rat endothelial cells. If albumin were not preadsorbed onto the membrane, selective attachment to the modified regions would not occur. Radiolabeling albumin with 125I demonstrated the preference of albumin for adsorption onto the monoamine surface where the cells preferentially attached. Both hydrophobic and ionic forces contributed to the adsorption process. Although selective cellular attachment to the patterned APTES regions could be achieved by albumin preadsorption to the surface, the neuroblastoma cells did not significantly differentiate unless additional serum components were supplemented to the media.

Relative importance of surface wettability and charged functional groups on NIH 3T3 fibroblast attachment, spreading, and cytoskeletal organization

“…Ranieri et al also have observed loss of cell attachment to hydrophobic fluorinated polymers preadsorbed with BSA. 30 One of the primary challenges in developing tissueengineered devices is culturing anchorage-dependent cell populations on synthetic materials while maintaining their differentiated properties and functions. Several recent studies have suggested that cell spreading, shape, and cytoskeletal organization have a significant influence on cell differentiation.…”

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.…”

Fibronectin-pluronic coadsorption on a polystyrene surface with increasing hydrophobicity: Relationship to cell adhesion

“…14 For example, proteins such as fibronectin 15 or laminin, 16,17 peptides, 18 or amine groups 19 promoted neuronal cell adhesion and neurite outgrowth, while albumin 15,17 or alkane Therefore, to orientate cellular growth in vitro, many authors seeded cells on heterogeneous patchwise substrates presenting adhesive and antiadhesive areas, and observed preferential or exclusive cell adhesion on adhesive areas. 18,19,21,22 Among the well-known antiadhesive molecules, pluronic F68 is a tribloc surfactant composed of polyethylene oxide (PEO)-polypropylene oxide (PPO)-polyethylene oxide, 23 which adsorbs preferentially on hydrophobic surfaces 24 and prevents protein adsorption 25,26 and cell adhesion. 27 When adsorbed in competition with proteins such as collagen, it strongly reduces protein adsorption.…”