Gingival Fibroblasts Established on Microstructured Model Surfaces: Their Influence on Epithelial Morphogenesis and other Tissue-specific Cell Functions in a Co-cultured Epithelium

Department of Orthodontics, Albert Ludwig University, Freiburg, Germany.
Fortschritte der Kieferorthopädie (Impact Factor: 0.83). 09/2009; 70(5):351-62. DOI: 10.1007/s00056-009-0905-z
Source: PubMed


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.
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.
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.
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.

Download full-text


Available from: Thorsten Steinberg
  • [Show abstract] [Hide abstract]
    ABSTRACT: Most natural tissues consist of multi-cellular systems made up of two or more cell types. However, some of these tissues may not regenerate themselves following tissue injury or disease without some form of intervention, such as from the use of tissue engineered constructs. Recent studies have increasingly used co-cultures in tissue engineering applications as these systems better model the natural tissues, both physically and biologically. This review aims to identify the challenges of using co-culture systems and to highlight different approaches with respect to the use of biomaterials in the use of such systems. The application of co-culture systems to stimulate a desired biological response and examples of studies within particular tissue engineering disciplines are summarized. A description of different analytical co-culture systems is also discussed and the role of biomaterials in the future of co-culture research are elaborated on. Understanding the complex cell-cell and cell-biomaterial interactions involved in co-culture systems will ultimately lead the field towards biomaterial concepts and designs with specific biochemical, electrical, and mechanical characteristics that are tailored towards the needs of distinct co-culture systems.
    No preview · Article · Mar 2014 · Biomaterials
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Abstract Periodontal infection involves complex interplay between oral biofilm, gingival tissues and cells of the immune system in a dynamic microenvironment. A humanized in vitro model that reduces the need for experimental animal models, while recapitulating key biological events in a periodontal pocket, would constitute a technical advancement in the study of periodontal disease. The aim of this study was to use a dynamic perfusion bioreactor to develop a gingival epithelial-fibroblast-monocyte organotypic co-culture on collagen sponges. An 11 species subgingival biofilm was used to challenge the generated tissue in the bioreactor for a period of 24h. The histological and scanning electron microscopy analysis displayed an epithelial-like layer on the surface of the collagen sponge, supported by the underlying growth of gingival fibroblasts, while monocytic cells were also found within the sponge mass. Bacterial quantification of the biofilm showed that in the presence of the organotypic tissue, the growth of selected biofilm species, especially for Campylobacter rectus, Actinomyces oris, Streptococcus anginosus, Veillonella dispar, and Porphyromonas gingivalis, was suppressed, indicating a potential antimicrobial effect by the tissue. Multiplex analysis of cytokine secretion showed that interleukin (IL)-1 beta, IL-2, IL-4, and tumor necrosis factor (TNF)-alpha levels in cell culture supernatants were significantly up-regulated in presence of the biofilm, indicating a positive inflammatory response of the organotypic tissue to the biofilm challenge. In conclusion, this novel host-biofilm interaction organotypic model might have an important impact on the study of periodontal infections, by minimizing the need for the use of experimental animal models.
    Full-text · Article · Jan 2015 · Virulence