Evaluation of L929 fibroblast attachment and proliferation on Arg-Gly-Asp-Ser (RGDS)-immobilized chitosan in serum-containing/serum-free cultures

J Biosci Bioeng. 2007 Jul;104(1):69-77. doi: 10.1263/jbb.104.69.

Abstract

In this study, chitosan membranes prepared by the solvent casting method were modified with the Arg-Gly-Asp-Ser (RGDS) sequence of fibronectin using the photochemical immobilization technique. The results obtained from attenuated total reflection-Fourier transform infrared spectra and X-ray photoelectron spectroscopy studies confirmed the successful immobilization of RGDS on chitosan membranes. The immobilized peptide concentration was determined by ninhydrin analysis on the order of 10(-7) mol/cm(2). In vitro cell culture studies were performed with L929 mouse fibroblasts to investigate the effect of biomodification on fibroblast cell behaviour in serum-free and 10% serum-containing media. The results obtained from cell culture studies pointed out the specific interactions between biosignal RGDS molecules and fibroblast cells. A triggered cell attachment and proliferation were observed on RGDS-modified chitosan membranes that were more distinguishable in serum-free medium. In addition, the photochemical immobilization technique was realized in the presence of a photomask that was used to immobilize the RGDS molecules in a defined micropattern. L929 mouse fibroblasts attached on the RGDS-micropatterned areas indicating integrin-mediated interactions.

Publication types

  • Evaluation Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Adhesion
  • Cell Culture Techniques
  • Cell Proliferation
  • Cells, Immobilized / chemistry
  • Cells, Immobilized / cytology
  • Cells, Immobilized / drug effects
  • Chitosan / chemistry*
  • Culture Media
  • Culture Media, Serum-Free / metabolism
  • Fibroblasts / chemistry
  • Fibroblasts / cytology
  • Fibroblasts / drug effects*
  • Membranes, Artificial*
  • Mice
  • Microscopy, Atomic Force
  • Oligopeptides / chemistry
  • Oligopeptides / pharmacology*
  • Spectroscopy, Fourier Transform Infrared
  • Tissue Engineering / methods*

Substances

  • Culture Media
  • Culture Media, Serum-Free
  • Membranes, Artificial
  • Oligopeptides
  • Chitosan
  • arginyl-glycyl-aspartyl-serine