Camphorquinone alters the expression of extracellular proteases in a 3D co-culture model of the oral mucosa. Issue 2 (February 2021)

• Record Type: Journal Article
• Authors: Perduns, Renke; Volk, Joachim; Plum, Melanie; Jochums, André; Gutzki, Frank; Kaever, Volkhard; Geurtsen, Werner
• Is Part Of: Dental materials. Volume 37:Issue 2(2021)
• Page Start: 236
• Page End: 248
• Publication Date: 2021-02
• Subjects: Camphorquinone -- Oral keratinocytes -- Gingival fibroblasts -- 3D co-culture model -- Redox-regulated gene expression -- Extracellular proteases; Dentistry -- Periodicals; Dental materials -- Periodicals; 617.695
• DOI: 10.1016/j.dental.2020.11.005
• Languages: English
• ISSNs: 0109-5641

Abstract

Highlights: Camphorquinone (CQ) induces antioxidant enzymes in a 3D model of the oral mucosa. CQ is able to pass a dense layer of oral keratinocytes and is metabolized by cells. CQ alters the transcription and abundance of extracellular proteases. Abstract: Objective: 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. Methods: 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. Results: 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. Significance: 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.