Primary osteoblasts adhesion onto RGD-functionalized and cross-linked polyelectrolyte multilayer films
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Primary osteoblasts adhesion onto RGD-functionalized and cross-linked polyelectrolyte multilayer films Catherine Picart1, Ludovic Richert1, René Elkaim2, Pierre Schaaf3, Jean-Claude Voegel1, Benoît Frisch4 1 Institut National de la Santé et de la Recherche Médicale, Unité 595, Faculté de Chirurgie Dentaire, Université Louis Pasteur, 11 rue Humann, 67085 Strasbourg Cedex, France 2 Parogène, 11 rue Humann, 67085 Strasbourg Cedex, France 3 Institut Charles Sadron, Centre National de la Recherche Scientifique, Université Louis Pasteur, 6 rue Boussingault, 67083 Strasbourg Cedex, France 4 Laboratoire de Chimie Bioorganique, CNRS/ULP, 74 route du Rhin, 67400 Illkirch, France ABSTRACT The adhesion of primary osteoblastic cells on top of biocompatible polyelectrolyte multilayer (PEM) films was investigated for native films and after changing the films properties either with a chemical stimulus (film functionalization), with a mechanical stimulus (film cross-linking), or with both stimuli combined. For the functionalization, a 15 amino acid peptide containing a –RGD- (-Arg-Gly-Asp) sequence was grafted to poly(Lglutamic) acid and deposited on top of poly(L-lysine)/poly(L-glutamic) (PLL/PGA), PLL/Poly(alginic), and PLL/Poly(galacturonic) films. The film buildup and the adsorption of the PGA-RGD was followed by Optical Waveguide Lightmode Spectroscopy and by Atomic Force Microscopy. The mechanical stimulus was achieved by crosslinking the films with a water soluble carbodiimide (EDC) in combination with N-hydroxysulfo-succinimide (sulfoNHS) to induce amide formation. Fourier Transform Infrared Spectroscopy evidenced the conversion of amine and carboxylic groups into amide groups. The alkaline phosphatase (ALP) activity test was used to assess osteoblast adhesion and proliferation on top of the different films over a period of eight days in culture. Whereas the native films are poorly adherent, the RGD-functionalized films exhibit an increased short time adhesion. The native films could also be successfully cross-linked thereby dramatically enhancing cell proliferation. The cells did not react similarly on the different types of films investigated : the cross-linked (PLL/Palg) and (PLL/Pgal) films were much more efficient than the native or functionalized films in terms of proliferation. On the other hand, for the (PLL/PGA) films, functionalization and film cross-linking had a similar long term effect. Very interestingly, for these latter films, both stimuli could be combined. INTRODUCTION Surface modifications of materials are of primary importance for biomedical applications. Among the different techniques used to modify surfaces, the deposition of polyelectrolyte multilayers (PEM) based on the alternate adsorption of polycations and polyanions has emerged as a very easy to handle and versatile tool [1] . Polyelectrolyte multilayers made of biopolymers or polyaminoacids are hydrogels and must be considered as “soft” and fragile materials. Most often, such film architectures are poorly adherent for both cell lineages
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