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Six dentin adhesives were tested in vitro regarding their cytotoxicity on human fibroblasts. The adhesives Hybrid Bond, One-up Bond F Plus, AdheSE, Clearfil SE Bond, Optibond Solo Plus and Syntac were eluted with culture medium as single or sequentially applied adhesive part for 24 h. 75 Petri dishes were produced per group. They were evaluated triangulated, comprising the quantitative evaluation (105 ones) to determine “viable”, “dead” and “debris” cells with the use of a cell-counter and the reactivity index was also identified based on the qualitative assessment (420 ones). One-up Bond F Plus, AdheSE and Clearfil SE Bond showed a statistical difference of viable cells to the cell control. For One-up Bond F Plus, statistically, differences compared to hybrid bond and Syntac were also found. All the adhesives except One-up Bond F Plus showed significant differences between single and sequentially applied adhesive part regarding the quantitative evaluation. The test material showed a moderate grade of cytotoxicity. As a result, a statistically significant difference of the cytotoxicity between the self-etch and etch-and-rinse adhesives cannot be demonstrated regarding the qualitative evaluation and the reactivity index, but the differences between sequentially applied and single applied components can be proved.
Although vaccination campaigns are currently being rolled out to prevent coronavirus disease (COVID-19), antivirals will remain an important adjunct to vaccination. Antivirals against coronaviruses do not exist, hence global drug repurposing efforts have been carried out to identify agents that may provide clinical benefit to patients with COVID-19. Itraconazole, an antifungal agent, has been reported to have activity against animal coronaviruses. Using cell-based phenotypic assays, the in vitro antiviral activity of itraconazole and 17-OH itraconazole was assessed against clinical isolates from a German and Belgian patient infected with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Itraconazole demonstrated antiviral activity in human Caco-2 cells (EC50 = 2.3 µM; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay). Similarly, its primary metabolite, 17-OH itraconazole, showed inhibition of SARS-CoV-2 activity (EC50 = 3.6 µM). Remdesivir inhibited viral replication with an EC50 = 0.4 µM. Itraconazole and 17-OH itraconazole resulted in a viral yield reduction in vitro of approximately 2-log10 and approximately 1-log10, as measured in both Caco-2 cells and VeroE6-eGFP cells, respectively. The viral yield reduction brought about by remdesivir or GS-441524 (parent nucleoside of the antiviral prodrug remdesivir; positive control) was more pronounced, with an approximately 3-log10 drop and >4-log10 drop in Caco-2 cells and VeroE6-eGFP cells, respectively. Itraconazole and 17-OH itraconazole exert in vitro low micromolar activity against SARS-CoV-2. Despite the in vitro antiviral activity, itraconazole did not result in a beneficial effect in hospitalized COVID-19 patients in a clinical study (EudraCT Number: 2020-001243-15).