OBJECTIVE: The objective of this study was to investigate how gingival fibroblasts cultured on microstructured model surfaces affect epithelial morphogenesis and other cell functions in a cocultured epithelium while conducting a molecular analysis of interactions between biomaterials employing periodontal tissue cells. MATERIALS AND METHODS: We are the first to have successfully co-cultured gingival fibroblasts together with gingival keratinocytes on biofunctionalized, microstructured model surfaces and, in the resulting co-cultured epithelium, examined the molecules of tissue homeostasis, the differentiation marker keratin (K) 1/10, and involucrin after 1- and 2-week periods of cultivation. Desmoplakin was perceived as evidence of cell-to-cell contact and thus as proof of epithelial integrity. We also analyzed the basement membrane component laminin-5. The aforementioned co-culture model without gingival fibroblasts was used as a control set-up. RESULTS: On the protein level, indirect immunofluorescence demonstrated the presence of K1/10, involucrin and the basement membrane component laminin-5 in the co-cultured epithelium in both culture periods. Furthermore, we observed that these epithelial markers had become re-oriented toward the suprabasal cell layers which, in turn, reflects the native in-vivo gingival epithelium. We identified cell-to-cell adhesion as a function of desmoplakin after just 1 week. In the mRNA analysis using quantitative RT-PCR after 2 weeks of cultivation, we noted a considerable rise in relative gene expression that was time-dependent for the early keratinocyte differentiation marker K1 and late marker involucrin. CONCLUSIONS: Our findings demonstrate that gingival fibroblasts on microstructured model surfaces are vitally important for tissue- specific cell functions such as epithelial morphogenesis and other biological cell functions such as differentiation and epithelial integrity. These study findings can thus contribute to the optimization and/or new development of biomaterials currently used in dental medicine.
OBJECTIVE: The objective of this study was to investigate how gingival fibroblasts cultured on microstructured model surfaces affect epithelial morphogenesis and other cell functions in a cocultured epithelium while conducting a molecular analysis of interactions between biomaterials employing periodontal tissue cells. MATERIALS AND METHODS: We are the first to have successfully co-cultured gingival fibroblasts together with gingival keratinocytes on biofunctionalized, microstructured model surfaces and, in the resulting co-cultured epithelium, examined the molecules of tissue homeostasis, the differentiation marker keratin (K) 1/10, and involucrin after 1- and 2-week periods of cultivation. Desmoplakin was perceived as evidence of cell-to-cell contact and thus as proof of epithelial integrity. We also analyzed the basement membrane component laminin-5. The aforementioned co-culture model without gingival fibroblasts was used as a control set-up. RESULTS: On the protein level, indirect immunofluorescence demonstrated the presence of K1/10, involucrin and the basement membrane component laminin-5 in the co-cultured epithelium in both culture periods. Furthermore, we observed that these epithelial markers had become re-oriented toward the suprabasal cell layers which, in turn, reflects the native in-vivo gingival epithelium. We identified cell-to-cell adhesion as a function of desmoplakin after just 1 week. In the mRNA analysis using quantitative RT-PCR after 2 weeks of cultivation, we noted a considerable rise in relative gene expression that was time-dependent for the early keratinocyte differentiation marker K1 and late marker involucrin. CONCLUSIONS: Our findings demonstrate that gingival fibroblasts on microstructured model surfaces are vitally important for tissue- specific cell functions such as epithelial morphogenesis and other biological cell functions such as differentiation and epithelial integrity. These study findings can thus contribute to the optimization and/or new development of biomaterials currently used in dental medicine.
Authors: F Spirito; S Chavanas; C Prost-Squarcioni; L Pulkkinen; S Fraitag; C Bodemer; J P Ortonne; G Meneguzzi Journal: J Biol Chem Date: 2001-03-14 Impact factor: 5.157
Authors: Camilla Mohrdieck; Alexander Wanner; Wouter Roos; Alexander Roth; Erich Sackmann; Joachim P Spatz; Eduard Arzt Journal: Chemphyschem Date: 2005-08-12 Impact factor: 3.102
Authors: Tanida Srisuwan; Daniel J Tilkorn; Jeremy L Wilson; Wayne A Morrison; Harold M Messer; Erik W Thompson; Keren M Abberton Journal: Periodontol 2000 Date: 2006 Impact factor: 7.589
Authors: Siri Paulo; Mafalda Laranjo; Ana M Abrantes; João Casalta-Lopes; Kathleen Santos; Ana C Gonçalves; Anabela Baptista Paula; Carlos Miguel Marto; Ana Bela Sarmento-Ribeiro; Eunice Carrilho; Arménio Serra; Maria F Botelho; Manuel M Ferreira Journal: Materials (Basel) Date: 2019-06-06 Impact factor: 3.623