Influence of retinoic acid on the expression of cytokeratins, vimentin and ICAM-1 in human gingival epithelia in vitro

Phenotypic differences exist in vivo between junctional (JE) and oral gingival (OGE) epithelia and an in vitro system has been developed that maintains phenotypic differences. This system, which permits in vitro studies of factors that may influence the epithelial phenotype, was used to investigate the effects of retinoic acid (RA) on epithelial expression of various markers known to distinguish JE from OGE. Primary cultures of JE and OGE were initiated from defined gingival regions and were subcultured and grown for 48 h in 96-well plates or on multiple-well slides. Control cultures were grown in medium supplemented with delipidized serum and all-trans RA was added to experimental groups. Other cultures were grown in a defined RA-free medium. Cultures were examined using monoclonal antibodies against cytokeratins, vimentin, and ICAM-1 and binding displayed by indirect immunocytochemical staining. Staining reactions were assessed by direct microscopic observation and assayed by spectrophotometric quantitation. The results showed that RA had minor effects on the marker expression of JE but markedly enhanced expression of cytokeratins 8, 18, 19, vimentin and ICAM-1 in OGE. These markers, which normally distinguish JE from OGE, were expressed at levels approaching or exceeding those of control JE cultures. These observations indicate that RA responsive mechanisms affect the phenotypes expressed by epithelia in vitro and suggest that such mechanisms may be related to the different phenotypic patterns expressed by gingival epithelia in vivo.