Gingival fibroblasts established on microstructured model surfaces: their influence on epithelial morphogenesis and other tissue-specific cell functions in a co-cultured epithelium: an in-vitro model.
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.


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.
Müssig
Eva
E
Department of Orthodontics, Albert Ludwig University, Freiburg, Germany. eva.muessig@uniklinik-freiburg.de
Tomakidi
Pascal
P
Steinberg
Thorsten
T
D001672:Biocompatible Materials / Q000737:chemistry*; D002448:Cell Adhesion / Q000502:physiology; D002454:Cell Differentiation / Q000502:physiology; D002478:Cells, Cultured; D004848:Epithelium / Q000033:anatomy & histology* / Q000254:growth & development*; D005347:Fibroblasts / Q000166:cytology* / Q000502:physiology*; D005881:Gingiva / Q000166:cytology* / Q000502:physiology*; D006801:Humans; D008422:Materials Testing; D023822:Tissue Engineering / Q000379:methods*
D001672:Biocompatible Materials