Correlating fibronectin adsorption with endothelial cell adhesion and signaling on polymer substrates.
Fibronectin (Fn) adsorption was studied on different commercial polymer surface chemistries, including tissue culture polystyrene (TCPS), bacteriologic polystyrene (BPS), fluoropolymer Teflon AF, and poly-L-lactide (PLLA). Antibody probes detected the availability of Fn's cell binding domain on adsorbed Fn in the competitive presence and absence of bovine serum albumin (BSA). Domain availability was highest for Fn adsorbed on TCPS, especially in the presence of either serum albumin or dilute serum. Attachment and growth efficiencies for human umbilical venous endothelial cells (HUVECs) cultured on surfaces preadsorbed with Fn in serum and serum-free media correlated with antibody cell-binding domain availability: TCPS > BPS, Teflon AF > PLLA. Intracellular signaling from the GTPase, RhoA, was highest (RhoA:RhoGDI inhibitor ratio) in cells cultured on the Teflon AF surfaces, indicating that despite lower attached cell numbers on Teflon AF compared to TCPS, cell signaling remained activated after 24 h of growth. Up-regulated cellular Fn mRNA messages, assessed using RT-PCR techniques, supported HUVECs' producing the endogenous extracellular matrix (ECM) protein Fn in order to attach and survive on the suboptimal Teflon AF culture surfaces.
Koenig
Andrea L
AL
Department of Chemistry, Colorado State University, Fort Collins, Colorado 80523-1872, USA.
Gambillara
Veronica
V
Grainger
David W
DW
D000327:Adsorption; D001483:Base Sequence; D015153:Blotting, Western; D002448:Cell Adhesion; D002478:Cells, Cultured; D017931:DNA Primers; D004577:Electron Probe Microanalysis; D004730:Endothelium, Vascular / Q000737:chemistry / Q000166:cytology*; D005353:Fibronectins / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D007118:Immunoassay; D011108:Polymers / Q000737:chemistry*; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015398:Signal Transduction; D013499:Surface Properties
D017931:DNA Primers; D005353:Fibronectins; D011108:Polymers; D012333:RNA, Messenger