The role of 3D-microscopy in the study of chondrocyte-matrix interaction (alginate bead or sponge, rat femoral head cap, human osteoarthritic cartilage) and pharmacological application

The potentialities of a new non-invasive optical scanning microscopy techni que were evaluated through 3D analysis of chondrocyte-matrix interactions. Five different 2D or 3D culture systems were used: (1) MonoLayer (ML) of hu man chondrosarcoma cell line; (2) rat or human chondrocytes encapsulated in Alginate Bead (AB); (3) human chondrocytes encapsulated in Alginate Sponge (AS); (4) Rat Femoral Head Cap (RFHC); (5) slices of knee human Osteoarthr itic Cartilage (HOAC). Chondrocytes ML, AB, RFHC were incubated For 24 h in vitro in the presence of recombinant human interleukinl-beta (rhILI-beta) and the effects on cyroskeleton organisation (F-actin filament), Focal Adhe sion Kinase (FAK) expression (tyrosine kinase), collagenase B expression (m etalloprotease) were studied. Furthermore, the production of intracellular IL1-beta by LPS-or rhIL1-beta-stimulated chondrocytes was shown to be partl y suppressed by rhein (active metabolite of diacerhein) in all culture syst ems. This high resolution light microscopy gave complementary information t hat could be important for a better understanding of the interaction of cho ndrocytes with the extracellular matrix in a variety of-culture devices.