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.
Richert
Ludovic
L
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.
Schneider
Aurore
A
Vautier
Dominique
D
Vodouhe
Constant
C
Jessel
Nadia
N
Payan
Elisabeth
E
Schaaf
Pierre
P
Voegel
Jean-Claude
JC
Picart
Catherine
C
D000818:Animals; D002448:Cell Adhesion; D002470:Cell Survival; D002478:Cells, Cultured; D019902:Chondrocytes / Q000166:cytology*; D003432:Cross-Linking Reagents; D006820:Hyaluronic Acid; D008264:Macrophages / Q000166:cytology*; D018625:Microscopy, Atomic Force; D018613:Microscopy, Confocal; D011107:Polylysine; D051381:Rats; D017208:Rats, Wistar
D003432:Cross-Linking Reagents; D011107:Polylysine; D006820:Hyaluronic Acid