In vitro Cytokeratin Expression Profiling of Human Oral Mucosa Substitutes Developed by Tissue Engineering

Abstract

In this work we performed a study of cytokeratin (CK) expression profiling on human artificial oral mucosa developed in vitro by tissue engineering at different stages of maturation (from immature to well-developed stages) at the protein and mRNA levels. Human artificial oral mucosa was generated in the laboratory using fibrin-agarose biomaterials. As controls, we used human native normal oral mucosa and embryonic oral tissues. Our results demonstrated that human embryonic oral tissues tended to express CK8 and CK19. In contrast, monolayered bioengineered oral mucosa did not show any CK expression by immunohistochemistry whereas bilayered and multilayered artificial oral mucosa showed several markers of stratified epithelia, but did not express CK10. These results suggest that the CK expression pattern is strongly dependent on the maturation state of the artificial tissues and that the CK expression profile of our model of artificial oral mucosa was partially similar to that of the non-keratinized human adult oral mucosa. However, the expression of CK8 by the artificial oral mucosa suggests that these samples correspond to an early stage of development while kept in vitro.

Sequential keratinocytic differentiation and maturation in a three‐dimensional model of human artificial oral mucosa

“…Most of the works focused on the study of AOM have used partial and non‐autologous oral mucosa models to identify epithelial patterns such as DG3, laminin and others . On the one hand, our results confirmed the epithelial phenotype of the bioengineered epithelial layer as determined by the positive expression of pancytokeratin in all AOM samples as we previously demonstrated . On the other hand, laminin expression required 3 wk of in vitro development and the use of air–liquid culture conditions.…”

Tissue Engineering of Lips and Muco-Cutaneous Junctions: In Vitro Development of Tissue Engineered Constructs of Oral Mucosa and Skin for Lip Reconstruction

“…Additional confirmation that the 3D construct has the features of a lip was found when keratin 10 was not found in the oral section of the construct and keratin 19 was only expressed in the basal layer of the oral section of the construct as seen in Figure 6. These temporary and/or reversible changes in the keratinization profiles of natural and bioengineered oral mucosa constructs have also been previously reported 33 and also have been shown in mice. 34 While our focus was in the fabrication of a construct incorporating labial mucosa cells, a preliminary experiment using cells taken from the alveolar ridge area of the mouth, which is keratinized and similar to skin, indicated that it is also possible to make tissue engineered M/C junctions with skin and keratinized mucosa (Fig.…”

Ex vivo construction of a novel model of bioengineered bladder mucosa: A preliminary study

“…When the stromal cells were cultured within the fibrin‐agarose biomaterial, adequate cell integration in the biomaterial was observed as shown by histological and cell viability analyses. These results confirm our previous results suggesting that fibrin‐agarose biomaterials are highly biocompatible ex vivo . Furthermore, the presence of abundant intracellular synthesis organelles as determined by TEM suggests that the cells immersed in the scaffold were not only viable, but also metabolically active, and supports the validness of this biomaterial as an ex vivo cell culturing scaffold.…”