Matrix expression and proliferation of primary gingival fibroblasts in a three-dimensional cell culture model

The growth of cultured primary human gingival fibroblasts and the three-dim ensional arrangement of the extracellular matrix in a polyester carrier sys tem was investigated using various histological techniques, The results wer e compared with monolayer cultures. Collagen types I, III, V, and VI were i nvestigated by conventional and fluorescence microscopy, scanning and trans mission electron microscopy, and confocal laser scanning microscopy. Human gingival fibroblasts were obtained from tissue biopsies of five donors and were cultivated up to 5 weeks under three-dimensional culture conditions. T he cells displayed an elongated, spindle-like or stellate morphology resemb ling the in vivo situation. Collagen type I revealed thick fiber bundles, a nd collagens type Zn and V were distributed as fine fibrils or small bundle s throughout the culture system. Frequently, the fibers were oriented paral lel to the long axis of the cells, Type VI collagen formed thin fibers and revealed a reticular pattern. In histological sections the cultured cells e xhibited a morphology clearly different from that of cells cultured in mono layers, Their shape and spatial distribution resembled that of cells in tis sue biopsies more closely The culture system presented here promotes a dyna mic model for performing studies for instance on the interactions of cultur ed cells with extracellular matrix molecules, on the pathogenesis of inflam matory processes or on the interactions with biomaterials, thus providing q ualitative and quantitative information.