Tissue-engineered oral epithelium (ΤΕΟΕ) was developed after comparing various culture conditions, including submerged (SUB) and air–liquid interface (ALI) human cell expansion options. Barrier formation was evaluated via transepithelial electrical resistance (TEER) and calcein permeation via spectrofluorometry. TEOE was further assessed for long-term viability via live/dead staining and development of intercellular connections via transmission electron microscopy. Tissue architecture was evaluated via histochemistry and the expression of pancytokeratin (pCK) via immunohistochemistry. The effect of two commonly used dental resinous monomers on TEOE was evaluated for alterations in cell viability and barrier permeability. ALI/keratinocyte growth factor-supplemented (ALI-KGS) culture conditions led to the formation of an 8–20-layer thick, intercellularly connected epithelial barrier. TEER values of ALI-KGS-developed TEOE decreased compared with all other tested conditions, and the established epithelium intensively expressed pCK. Exposure to dental monomers affected the integrity and architecture of TEOE and induced cellular vacuolation, implicating hydropic degeneration. Despite structural modifications, the permeability of TEOE was not substantially affected after exposure to the monomers. In conclusion, the biological properties of the TEOE mimicking the physiological functional conditions and its value as biocompatibility assessment tool for dental materials were characterized.
Impact Statement
In this study, a tissue-engineered oral epithelium (TEOE) was developed to create a reliable and consistent in vitro-to-in vivo extrapolation model and serves as a high-throughput biocompatibility assessment tool for dental materials. TEOE can accurately replicate the physiology (architecture, intercellular connectivity, and viability), as well as the functionality (barrier formation and permeability) of the human oral epithelium. The optimal microenvironmental culture conditions investigated resulted in a TEOE that consistently recapitulates responses to two commonly used resinous dental materials enabling large-scale evaluation of new formulations before moving to more complex dynamic culture systems.
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