The effect of functionalized self-assembling peptide scaffolds on human aortic endothelial cell function.
A class of designed self-assembling peptide nanofiber scaffolds with more than 99% water content has been shown to be a good biological material for cell culture. Here, we report the functionalization of one of these peptide scaffolds, RAD16-I (AcN-RADARADARADARADA-CONH2), by direct solid phase synthesis extension at the amino terminal with three short-sequence motifs. These motifs are present in two major protein components of the basement membrane, laminin 1 (YIGSR, RYVVLPR) and collagen IV (TAGSCLRKFSTM). These motifs have been previously shown to promote specific biological activities including endothelial cell adhesion, spreading, and tubular formation. Therefore, the generic functionalized peptide developed was AcN-X-GG-RADARADARADARADA-CONH2 with each motif represented by "X". We show in this work that these tailor-made peptide scaffolds enhance the formation of confluent cell monolayers of human aortic endothelial cells (HAEC) in culture. Moreover, additional assays designed to evaluate endothelial cell function showed that HAEC monolayers obtained on these scaffolds not only maintained LDL uptake activity but also enhanced nitric oxide release and elevated laminin 1 and collagen IV deposition. These results suggest that this new scaffold provide a better physiological substrate for endothelial cell culture and suggest its further application for biomedical research, cancer biology and regenerative biology.
Genové
Elsa
E
Center for Biomedical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139, USA.
Shen
Colette
C
Zhang
Shuguang
S
Semino
Carlos E
CE
D020816:Amino Acid Motifs; D001011:Aorta / Q000166:cytology* / Q000187:drug effects; D001485:Basement Membrane / Q000378:metabolism; D001667:Binding, Competitive; D001672:Biocompatible Materials / Q000737:chemistry; D032701:Biomimetics; D015153:Blotting, Western; D002448:Cell Adhesion; D049109:Cell Proliferation; D002942:Circular Dichroism; D024141:Collagen Type IV / Q000378:metabolism; D004591:Electrophoresis, Polyacrylamide Gel; D004730:Endothelium, Vascular / Q000166:cytology* / Q000187:drug effects; D005109:Extracellular Matrix / Q000378:metabolism; D006801:Humans; D007797:Laminin / Q000378:metabolism; D008077:Lipoproteins, LDL / Q000378:metabolism; D018625:Microscopy, Atomic Force; D009569:Nitric Oxide / Q000378:metabolism; D010455:Peptides / Q000737:chemistry*
D001672:Biocompatible Materials; D024141:Collagen Type IV; D007797:Laminin; D008077:Lipoproteins, LDL; D010455:Peptides; C121346:laminin 1; D009569:Nitric Oxide