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

Overview

Journal J Orofac Orthop

Specialty Dentistry

Date 2009 Dec 10

PMID 19997994

Citations 5

Authors

Eva Mussig

Pascal Tomakidi

Thorsten Steinberg

Affiliations

Soon will be listed here.

Abstract

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.

Citing Articles

Vacuum Ultraviolet (VUV) Light Photofunctionalization to Induce Human Oral Fibroblast Transmigration on Zirconia.

Suzumura T, Matsuura T, Komatsu K, Sugita Y, Maeda H, Ogawa T Cells. 2023; 12(21).

PMID: 37947620 PMC: 10647316. DOI: 10.3390/cells12212542.

A Temperature-Based Easy-Separable (TempEasy) 3D Hydrogel Coculture System.

Zhang Z, Chen W, Tiemessen D, Oosterwijk E, Kouwer P Adv Healthc Mater. 2022; 11(10):e2102389.

PMID: 35029325 PMC: 11469334. DOI: 10.1002/adhm.202102389.

FAK Shutdown: Consequences on Epithelial Morphogenesis and Biomarker Expression Involving an Innovative Biomaterial for Tissue Regeneration.

Wang X, Steinberg T, Dieterle M, Ramminger I, Husari A, Tomakidi P Int J Mol Sci. 2021; 22(18).

PMID: 34575938 PMC: 8470904. DOI: 10.3390/ijms22189774.

Synthetic Calcium Phosphate Ceramics as a Potential Treatment for Bisphosphonate-Related Osteonecrosis of the Jaw.

Paulo S, Laranjo M, Abrantes A, Casalta-Lopes J, Santos K, Goncalves A Materials (Basel). 2019; 12(11).

PMID: 31174333 PMC: 6601279. DOI: 10.3390/ma12111840.

Establishment of an oral infection model resembling the periodontal pocket in a perfusion bioreactor system.

Bao K, Papadimitropoulos A, Akgul B, Belibasakis G, Bostanci N Virulence. 2015; 6(3):265-73.

PMID: 25587671 PMC: 4601317. DOI: 10.4161/21505594.2014.978721.

References

Spirito F, Chavanas S, Prost-Squarcioni C, Pulkkinen L, Fraitag S, Bodemer C . Reduced expression of the epithelial adhesion ligand laminin 5 in the skin causes intradermal tissue separation. J Biol Chem. 2001; 276(22):18828-35. DOI: 10.1074/jbc.M100381200. View

Mohrdieck C, Wanner A, Roos W, Roth A, Sackmann E, Spatz J . A theoretical description of elastic pillar substrates in biophysical experiments. Chemphyschem. 2005; 6(8):1492-8. DOI: 10.1002/cphc.200500109. View

Mussig E, Tomakidi P, Steinberg T . Molecules contributing to the maintenance of periodontal tissues. Their possible association with orthodontic tooth movement. J Orofac Orthop. 2005; 66(6):422-33. DOI: 10.1007/s00056-005-0520-6. View

Tan J, Tien J, Pirone D, Gray D, Bhadriraju K, Chen C . Cells lying on a bed of microneedles: an approach to isolate mechanical force. Proc Natl Acad Sci U S A. 2003; 100(4):1484-9. PMC: 149857. DOI: 10.1073/pnas.0235407100. View

Scheideler L, Rupp F, Wendel H, Sathe S, Geis-Gerstorfer J . Photocoupling of fibronectin to titanium surfaces influences keratinocyte adhesion, pellicle formation and thrombogenicity. Dent Mater. 2006; 23(4):469-78. DOI: 10.1016/j.dental.2006.03.005. View

Ruhrberg C, Watt F . The plakin family: versatile organizers of cytoskeletal architecture. Curr Opin Genet Dev. 1997; 7(3):392-7. DOI: 10.1016/s0959-437x(97)80154-2. View

Nischt R, Schmidt C, Mirancea N, Baranowsky A, Mokkapati S, Smyth N . Lack of nidogen-1 and -2 prevents basement membrane assembly in skin-organotypic coculture. J Invest Dermatol. 2006; 127(3):545-54. DOI: 10.1038/sj.jid.5700562. View

du Roure O, Saez A, Buguin A, Austin R, Chavrier P, Silberzan P . Force mapping in epithelial cell migration. Proc Natl Acad Sci U S A. 2005; 102(7):2390-5. PMC: 548966. DOI: 10.1073/pnas.0408482102. View

Roesch-Ely M, Steinberg T, Bosch F, Mussig E, Whitaker N, Wiest T . Organotypic co-cultures allow for immortalized human gingival keratinocytes to reconstitute a gingival epithelial phenotype in vitro. Differentiation. 2006; 74(9-10):622-37. DOI: 10.1111/j.1432-0436.2006.00099.x. View

10.

Walsh P, Hakkinen L, Pernu H, Knuuttila M, Larjava H . Expression of fibronectin-binding integrins in gingival epithelium in drug-induced gingival overgrowth. J Periodontal Res. 2007; 42(2):144-51. DOI: 10.1111/j.1600-0765.2006.00927.x. View

11.

Werner S, Smola H . Paracrine regulation of keratinocyte proliferation and differentiation. Trends Cell Biol. 2001; 11(4):143-6. DOI: 10.1016/s0962-8924(01)01955-9. View

12.

Reuther T, Kohl A, Komposch G, Tomakidi P . Morphogenesis and proliferation in mono- and organotypic co-cultures of primary human periodontal ligament fibroblasts and alveolar bone cells. Cell Tissue Res. 2003; 312(2):189-96. DOI: 10.1007/s00441-003-0717-1. View

13.

Ye P, Chapple C, Kumar R, Hunter N . Expression patterns of E-cadherin, involucrin, and connexin gap junction proteins in the lining epithelia of inflamed gingiva. J Pathol. 2000; 192(1):58-66. DOI: 10.1002/1096-9896(2000)9999:9999<::AID-PATH673>3.0.CO;2-T. View

14.

Srisuwan T, Tilkorn D, Wilson J, Morrison W, Messer H, Thompson E . Molecular aspects of tissue engineering in the dental field. Periodontol 2000. 2006; 41:88-108. DOI: 10.1111/j.1600-0757.2006.00176.x. View

15.

Lemmon C, Sniadecki N, Alom Ruiz S, Tan J, Romer L, Chen C . Shear force at the cell-matrix interface: enhanced analysis for microfabricated post array detectors. Mech Chem Biosyst. 2006; 2(1):1-16. PMC: 1480360. View

16.

Vigneswaran N, Peters K, Hornstein O, Haneke E . Comparison of cytokeratin, filaggrin and involucrin profiles in oral leukoplakias and squamous carcinomas. J Oral Pathol Med. 1989; 18(7):377-90. DOI: 10.1111/j.1600-0714.1989.tb01569.x. View

17.

Steinberg T, Schulz S, Spatz J, Grabe N, Mussig E, Kohl A . Early keratinocyte differentiation on micropillar interfaces. Nano Lett. 2007; 7(2):287-94. DOI: 10.1021/nl062271z. View

18.

Bloor B, Tidman N, Leigh I, Odell E, Dogan B, Wollina U . Expression of keratin K2e in cutaneous and oral lesions: association with keratinocyte activation, proliferation, and keratinization. Am J Pathol. 2003; 162(3):963-75. PMC: 1868097. DOI: 10.1016/S0002-9440(10)63891-6. View

19.

Vasioukhin V, BOWERS E, Bauer C, Degenstein L, Fuchs E . Desmoplakin is essential in epidermal sheet formation. Nat Cell Biol. 2002; 3(12):1076-85. DOI: 10.1038/ncb1201-1076. View

20.

Haas K, Berndt A, Stiller K, Hyckel P, Kosmehl H . A comparative quantitative analysis of laminin-5 in the basement membrane of normal, hyperplastic, and malignant oral mucosa by confocal immunofluorescence imaging. J Histochem Cytochem. 2001; 49(10):1261-8. DOI: 10.1177/002215540104901008. View