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Biomedical subjects

Elisabeth Payan

Publications and source records attributed to Elisabeth Payan.

4 recordsLinked to original sources

New physically and chemically crosslinked hyaluronate (HA)-based hydrogels for cartilage repair.

When dissolved in aqueous solutions, sodium hyaluronate substituted with low amounts of alkyl chains [amphiphilic hyaluronate (HA)] can give rise to hydrogels thanks to intermolecular reversible hydrophobic interactions, leading to a three-dimensional (3D) network. Such hydrogels possess shear-thinning properties and can thus be injected in cartilage defect to promote chondrocyte proliferation and cartilage repair. However, these hydrogels are only physically crosslinked and can progressively loose their 3D structure when they are in contact with aqueous fluids. To overcome this drawback, HA derivatives substituted with dodecyl chains were chemically crosslinked by a difunctional reagent, tetraethylene glycol ditosylate (TEG-diOTs). To preserve the shear-thinning properties of amphiphilic HA, small amounts of TEG-diOTs were used so as to obtain a low chemical crosslinking ratio. After optimization of the synthesis parameters, aqueous solutions of the HA derivatives, crosslinked both physically and chemically, were obtained, with rheological properties improved compared to the amphiphilic polymers. As the hydrogels are aimed to cartilage repair, they were sterilized by wet heating; the effect of this treatment on the polymer characteristics was analyzed by different techniques. A similar study was carried out on HA derivatives stored under conditions mimicking physiological ones.

Animals↗

Imaging cell interactions with native and crosslinked polyelectrolyte multilayers.

The adhesion of primary chondrocytes to polyelectrolyte multilayer films, made of poly(l-lysine) (PLL) and hyaluronan (HA), was investigated for native and crosslinked films, either ending by PLL or HA. Crosslinking the film was achieved by means of a water-soluble carbodiimide in combination with N-hydroxysulfosuccinimide. The adhesion of macrophages and primary chondrocytes was investigated by microscopical techniques (optical, confocal, and atomic), providing useful information on the cell/film interface. Native films were found to be nonadhesive for the primary chondrocytes, but could be degraded by macrophages, as could be visualized by confocal laser scanning microscopy after film labeling. Confocal microscopy images show that these films can be deformed by the chondrocytes and that PLL diffuses at the chondrocyte membrane. In contrast, the cells adhered and proliferated well on the crosslinked films, which were not degraded by the macrophages. These results were confirmed by a MTT test over a 6-d period and by atomic force microscopy observations. We thus prove that chemical crosslinking can dramatically change cell adhesion properties, the cells being more stably anchored on the crosslinked films.

Animals↗

Layer by layer buildup of polysaccharide films: physical chemistry and cellular adhesion aspects.

The formation ofpolysaccharide films based on the alternate deposition of chitosan (CHI) and hyaluronan (HA) was investigated by several techniques. The multilayer buildup takes place in two stages: during the first stage, the surface is covered by isolated islets that grow and coalesce as the construction goes on. After several deposition steps, a continuous film is formed and the second stage of the buildup process takes place. The whole process is characterized by an exponential increase of the mass and thickness of the film with the number of deposition steps. This exponential growth mechanism is related to the ability of the polycation to diffuse "in" and "out" of the whole film at each deposition step. Using confocal laser microscopy and fluorescently labeled CHI, we show that such a diffusion behavior, already observed with poly(L-lysine) as a polycation, is also found with CHI, a polycation presenting a large persistence length. We also analyze the effect of the molecular weight (MW) of the diffusing polyelectrolyte (CHI) on the buildup process and observe a faster growth for low MW chitosan. The influence of the salt concentration during buildup is also investigated. Whereas the CHI/HA films grow rapidly at high salt concentration (0.15 M NaCl) with the formation of a uniform film after only a few deposition steps, it is very difficult to build the film at 10(-4) M NaCl. In this latter case, the deposited mass increases linearly with the number of deposition steps and the first deposition stage, where the surface is covered by islets, lasts at least up to 50 bilayer deposition steps. However, even at these low salt concentrations and in the islet configuration, CHI chains seem to diffuse in and out of the CHI/HA complexes. The linear mass increase of the film with the number of deposition steps despite the CHI diffusion is explained by a partial redissolution of the CHI/HA complexes forming the film during different steps of the buildup process. Finally, the uniform films built at high salt concentrations were also found to be chondrocyte resistant and, more interestingly, bacterial resistant. Therefore, the (CHI/HA) films may be used as an antimicrobial coating.

Animals↗

Induction of heat shock protein 70 (Hsp70) by proteasome inhibitor MG 132 protects articular chondrocytes from cellular death in vitro and in vivo.

The aim of this work was to determine whether Hsp70 overexpression via proteasome inhibitor MG132 was able to protect chondrocytes towards mono-iodoacetate (MIA) cytotoxicity both in vitro and in vivo. In vitro, overexpression of Hsp70 via MG132 was significantly able to protect chondrocytes from MIA toxicity (MTT/LDH analyses). Hsp70 essentially mediated this chondroprotective effect as demonstrated by antisense strategy. In vivo, chondrocytic overexpression of Hsp70, after a preventive intra-articular injection of MG132 in rat knee, was sufficient to decrease the severity of OA-induced MIA lesions, as demonstrated histologically and biochemically. In conclusion, intracellular overexpression of Hsp70, through proteasome inhibition, could be an interesting tool in protecting chondrocytes from cellular injuries, either necrotic or apoptotic in nature, and thus might be a novel chondroprotective modality in rat experimental OA.

Animals↗