Camphorquinone alters the expression of extracellular proteases in a 3D co-culture model of the oral mucosa.

Renke Perduns, Joachim Volk, Melanie Plum, André Jochums, Frank Gutzki, Volkhard Kaever, Werner Geurtsen

Dental materials : official publication of the Academy of Dental Materials 2021 Feb

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Objective of our investigation was to determine the influence of CQ on the expression of antioxidant proteins and extracellular proteases in a 3D co-culture model (3DCCM) of the oral mucosa and to analyze the distribution and stability of CQ within 3D-CCMs. 3D-CCMs consist of confluent keratinocytes (OKF6/TERT2) on cell culture inserts on top of human gingival fibroblasts (HGFs) in collagen. The treatment was carried out by adding CQ to the cell culture inserts at two time points with declining concentrations. Mass spectrometry was used to analyze the CQ concentration above and underneath the OKF6/TERT2-layer. The expression of antioxidant genes was analyzed by qRT-PCR and western blot. The regulation of extracellular proteases from different families was analyzed by qRT-PCR and Proteome Profiler arrays. GC/MS analysis showed that CQ was evenly distributed within the model. Heme oxygenase-1, NAD(P)H quinone dehydrogenase 1 (NQO1), and superoxide dismutase 1 were induced on the mRNA and protein level in OKF6/TERT2 cells. In HGFs, only the transcription of NQO1 was induced. The transcription of extracellular proteases was increased mainly in OKF6/TERT2 cells 72 h after the initial treatment. The quantity of ten out of 25 analyzed extracellular proteases in the cell culture supernatant above and six underneath the keratinocyte-layer were modulated by CQ. Despite its high reactivity, CQ is able to penetrate a dense keratinocyte-layer, presumably across plasma membranes. CQ initially induced the cellular defense machinery against oxidative stress and altered the expression of extracellular proteases. We assume a relationship between both processes. Copyright © 2020 The Academy of Dental Materials. Published by Elsevier Inc. All rights reserved.

Citation

Renke Perduns, Joachim Volk, Melanie Plum, André Jochums, Frank Gutzki, Volkhard Kaever, Werner Geurtsen. Camphorquinone alters the expression of extracellular proteases in a 3D co-culture model of the oral mucosa. Dental materials : official publication of the Academy of Dental Materials. 2021 Feb;37(2):236-248