Lina Gölz1, Ruth Andrea Simonis2, Joana Reichelt2, Helmut Stark2, Matthias Frentzen3, Jean-Pierre Allam4, Rainer Probstmeier5, Jochen Winter3, Dominik Kraus6. 1. Department of Orthodontics, Center of Dento-Maxillo-Facial Medicine, University of Bonn, Bonn, Germany. 2. Department of Prosthodontics, Preclinical Education and Material Sciences, Center of Dento-Maxillo-Facial Medicine, University of Bonn, Bonn, Germany. 3. Department of Periodontology, Operative and Preventive Dentistry, University of Bonn, Bonn, Germany. 4. Department of Dermatology and Allergy, University of Bonn, Bonn, Germany. 5. Neuro- and Tumor Cell Biology Group, Department of Nuclear Medicine, University Hospital Bonn, Bonn, Germany. 6. Department of Prosthodontics, Preclinical Education and Material Sciences, Center of Dento-Maxillo-Facial Medicine, University of Bonn, Bonn, Germany. Electronic address: Dominik.Kraus@ukb.uni-bonn.de.
Abstract
OBJECTIVES: Resin infiltrants have been successfully used in dental medicine preventing the progression of tooth decay in an early phase of caries development. ICON(®) is an infiltrant of low-viscosity which penetrates via dentinal tubules into the lesion in dependence of the demineralization depth. Hence, we performed an in vitro study to determine the effect of ICON(®) on human dental pulp stem cells (hDPSCs). METHODS: Using explant technique, primary hDPSCs were collected from extracted teeth. Characterization and isolation were performed with typical mesenchymal stem cell markers (Stro-1, CD73, CD90, CD105) and hDPSCs differentiation was validated by immunofluorescence and flow cytometry. HDPSCs were stimulated with light-cured ICON(®) (lc) and non-light-cured ICON(®) (nc) conditioned media as well as different TEGDMA concentrations followed by the analysis of cytotoxicity, pro- and anti-inflammatory responses and differentiation using XTT assay, RT-PCR and ELISAs, respectively. RESULTS: Initial analysis demonstrated that hDPSCs express characteristic mesenchymal stem cell markers and differentiate into adipocytes, chondrocytes and osteoblasts. Notably, ICON(®) nc dramatically reduced cell viability (up to 98.9% after 48h), whereas ICON(®) lc showed only a modest cytotoxicity (10%). Data were in line with cytokine expression demonstrating increased levels of IL-6 and IL-8 as well as decreased IL-10 after ICON(®) nc exposure compared to ICON(®) lc. ICON(®) lc caused almost no alterations of DSPP, whereas ICON(®) nc markedly elevated DSPP mRNA levels (130.3-times). A concentration-dependent effect was observed in TEGDMA challenged hDPSCs. SIGNIFICANCE: ICON(®) is a successful minimal invasive technique. However, clinicians should strictly follow manufacturer's instructions to prevent adverse effects.
OBJECTIVES: Resin infiltrants have been successfully used in dental medicine preventing the progression of tooth decay in an early phase of caries development. ICON(®) is an infiltrant of low-viscosity which penetrates via dentinal tubules into the lesion in dependence of the demineralization depth. Hence, we performed an in vitro study to determine the effect of ICON(®) on human dental pulp stem cells (hDPSCs). METHODS: Using explant technique, primary hDPSCs were collected from extracted teeth. Characterization and isolation were performed with typical mesenchymal stem cell markers (Stro-1, CD73, CD90, CD105) and hDPSCs differentiation was validated by immunofluorescence and flow cytometry. HDPSCs were stimulated with light-cured ICON(®) (lc) and non-light-cured ICON(®) (nc) conditioned media as well as different TEGDMA concentrations followed by the analysis of cytotoxicity, pro- and anti-inflammatory responses and differentiation using XTT assay, RT-PCR and ELISAs, respectively. RESULTS: Initial analysis demonstrated that hDPSCs express characteristic mesenchymal stem cell markers and differentiate into adipocytes, chondrocytes and osteoblasts. Notably, ICON(®) nc dramatically reduced cell viability (up to 98.9% after 48h), whereas ICON(®) lc showed only a modest cytotoxicity (10%). Data were in line with cytokine expression demonstrating increased levels of IL-6 and IL-8 as well as decreased IL-10 after ICON(®) nc exposure compared to ICON(®) lc. ICON(®) lc caused almost no alterations of DSPP, whereas ICON(®) nc markedly elevated DSPP mRNA levels (130.3-times). A concentration-dependent effect was observed in TEGDMA challenged hDPSCs. SIGNIFICANCE: ICON(®) is a successful minimal invasive technique. However, clinicians should strictly follow manufacturer's instructions to prevent adverse effects.