Adhesion and growth of vascular smooth muscle cells in cultures on bioactive RGD peptide-carrying polylactides
The surface of poly(l-lactide) (PLLA) films deposited on glass coverslips was modified with poly(dl-lactide) (PDLLA), or 1:4 mixtures of PDLLA and PDLLA-b-PEO block copolymers, in which either none, 5% or 20% of the copolymer molecules carried a synthetic extracellular matrix-derived ligand for integrin adhesion receptors, the GRGDSG oligopeptide, attached to the end of the PEO chain. The materials, perspective for vascular tissue engineering, were seeded with rat aortic smooth muscle cells (11,000 cells/cm2) and the adhesion, spreading, DNA synthesis and proliferation of these cells was followed on inert and bioactive surfaces. In 24-h-old cultures in serum-supplemented media, the number of cells adhering to the PDLLA-b-PEO copolymer was almost eight times lower than that on the control PDLLA surface. On the surfaces containing 5% and 20% GRGDSG-PEO-b-PDLLA copolymer, the number of cells increased 6- and 3-fold respectively, compared to the PDLLA-b-PEO copolymer alone. On PDLLA-b-PEO copolymer alone, the cells were typically round and non-spread, whereas on GRGDSG-modified surfaces the cell spreading areas approached those found on PDLLA, reaching values of 991 μm2 and 611 μm2 for 5% and 20% GRGDSG respectively, compared to 958 μm2 for PDLLA. The cells on GRGDSG-grafted copolymers were able to form vinculin-containing focal adhesion plaques, to synthesize DNA and even proliferate in a serum-free medium, which indicates specific binding to the GRGDSG sequences through their adhesion receptors.
This study was supported by the Academy of Sciences of the Czech Republic (grant No. IAA4050202). We also thank Mrs. Ivana Zajanová for her excellent technical assistance. Mr. Benjamin J. Watson-Jones (Preklad Centrum, Prague) and Mr. Robin Healey (Czech Technical University, Prague) gratefully are acknowledged for the language revision of the manuscript.
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