Growth of human endothelial cells on different concentrations of Gly-Arg-Gly-Asp grafted chitosan surface.
To improve the adhesion and growth of endothelial cells on chitosan, different concentrations of cell adhesive peptide, Gly-Arg-Gly-Asp (GRGD), were photochemically grafted to its surface. After different concentrations of GRGD-SANPAH (N-Succinimidyl-6-4'-azido-2'-nitrophenylamino]-hexanoate) solutions were formed, they were grafted to chitosan surfaces by performing surface adsorption of the peptide solutions and subsequent ultraviolet (UV) irradiation to induce photoreaction. The grafted efficiency for GRGD peptide to chitosan surface was higher than 80%, as semiquantitatively analyzed by a high performance liquid chromatography (HPLC). FTIR spectra and electron spectroscopy for chemical analysis (ESCA) also confirmed that GRGD was successfully grafted to form chitosan-GRGD surfaces. For example, the composition fraction of nitrogen (N1s) for chitosan-GRGD surface grafted at 0.05 M GRGD was 6.8% instead of 3.2% for chitosan only. Human umbilical vein endothelial cells (HUVECs) adhered well and grew on different GRGD grafted concentrations of chitosan-GRGD surfaces after 36 hrs of incubation, but not on the chitosan surface. Furthermore, viability for the growth of HUVECs was increased with an increase in initial concentration of GRGD grafted chitosan-GRGD surfaces analyzed by MTT assay. In conclusion, an increased concentration of GRGD grafted on the chitosan-GRGD surface leads to more HUVECs adhered and grown on the surface.
Chung
Tze-Wen
TW
Department of Biomedical Engineering, Chung-Yuan Christian University, Chung-Li, Taiwan, ROC. twchung@yuntech.edu.tw
Lu
Ya-Fen
YF
Wang
Hsin-Ya
HY
Chen
Weng-Pin
WP
Wang
Shoei-Shen
SS
Lin
Yeong-Shang
YS
Chu
Shu-Hsun
SH
D001704:Biopolymers; D002448:Cell Adhesion; D002455:Cell Division; D002470:Cell Survival; D002478:Cells, Cultured; D002686:Chitin / Q000031:analogs & derivatives*; D048271:Chitosan; D002851:Chromatography, High Pressure Liquid; D004577:Electron Probe Microanalysis; D004730:Endothelium, Vascular / Q000166:cytology*; D006801:Humans; D009842:Oligopeptides; D017550:Spectroscopy, Fourier Transform Infrared; D013499:Surface Properties; D023822:Tissue Engineering
D001704:Biopolymers; D009842:Oligopeptides; D002686:Chitin; D048271:Chitosan; C064597:glycyl-arginyl-glycyl-aspartic acid