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. 2022 Mar 30;19(7):4112.
doi: 10.3390/ijerph19074112.

Tolerance of Human Fibroblasts to Benfo-Oxythiamine In Vitro

Ming Yan  1   2 , Ralf Smeets  1   3 , Martin Gosau  1 , Tobias Vollkommer  1 , Sandra Fuest  3 , Eva Stetzer  4 , Lan Kluwe  1 , Johannes F Coy  4 , Simon Burg  1
Affiliations

Tolerance of Human Fibroblasts to Benfo-Oxythiamine In Vitro

Ming Yan et al. Int J Environ Res Public Health. .

Abstract

Objectives: To explore the potential application of B-OT in the aspiration tract.

Materials and methods: We conceived and optimized an in vitro model simulating the mouth-washing process to assess tolerance to B-OT on primary human gingival fibroblasts. Cells derived from 4 unrelated donors were flushed with medium containing drugs of various concentration for one minute twice daily for 3 days.

Results: No effect was seen on the cells up to 1000 µM B-OT. In addition, we treated the cells with B-OT permanently in medium, corresponding to a systemic treatment. No effect was seen by 10 µM B-OT and only a slight reduction (approximately 10%) was seen by 100 µM B-OT.

Conclusions: Our results suggest good tolerance of oral cells for B-OT, favoring the further development of this antiviral reagent as a mouth-washing solution and nasal spray.

Keywords: SARS-CoV-2; benfo-oxythiamine; fibroblast; virus replication.

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Conflict of interest statement

This study was supported by Benfovir AG (Darmstadt, Germany). Johannes F. Coy declares a conflicts of interest as he is the founder of TAVARTIS GmbH, which founded Benfovir AG. Johannes F. Coy owns TAVARTIS GmbH and is a major shareholder of Benfovir AG. Eva Stetzer declares a conflicts of interest as she is an employee of Benfovir AG.

Figures

Figure 1
Figure 1
An illustration of the procedure of the in vitro model for mouth washing. Cells were seeded into wells of a 96-plate plate. For each washing, the medium of each well containing cells was saved in an empty well; medium containing various drugs of various concentrations was added to the corresponding wells. After 1 min, the drug-containing medium was discarded and the saved medium was transferred back to the original wells. The treatment was carried out twice daily.
Figure 2
Figure 2
Relative viability of cells in 3 cultures of primary human gingival fibroblasts derived from 3 unrelated donor teeth. Fresh B-OT: fresh benfooxythamine; serum B-OT: benfooxythamine preincubated in serum-containing medium overnight at room temperature. OT: oxythiamine. Keto: Ketoconazole.
Figure 3
Figure 3
Conventional treatment with B-OT in the medium over the treatment period of 3 days. The gingival cells were derived from a 4th donor tooth. The flushing treatment was also carried out in parallel for comparison. In addition, we measured the cell viabilities also for treatment of 1 and 2 days in both the flushing and conventional mode.
See this image and copyright information in PMC

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