Effects of ethanol on human periodontal ligament fibroblasts subjected to static compressive force.

Alcohol (Fayetteville, N.Y.) (2018-10-20)
Agnes Schröder, Erika Calvano Küchler, Marjorie Omori, Gerrit Spanier, Peter Proff, Christian Kirschneck

Consumption of toxic substances such as alcohol is widespread in the general population and thus also in patients receiving orthodontic treatment. Since human periodontal ligament (hPDL) fibroblasts play a key role in orthodontic tooth movement (OTM) by expressing cytokines and chemokines, we wanted to clarify whether ethanol modulates the physiological activity and expression pattern of hPDL fibroblasts during static compressive force application. We pre-incubated hPDL fibroblasts for 24 h with different ethanol concentrations, corresponding to casual (0.041% blood alcohol concentration [BAC], % by volume) and excessive (0.179%) alcohol consumption. At each ethanol concentration, we incubated the cells for another 48 h with and without an additional physiological compressive force of 2 g/cm2 occurring during orthodontic tooth movement in compression areas of the periodontal ligament. Thereafter, we analyzed expression and secretion of genes and proteins involved in OTM regulation by RT-qPCR and ELISA. We also performed co-culture experiments to observe hPDL-fibroblast-mediated osteoclastogenesis. We observed no effects of ethanol on cytotoxicity or cell viability of hPDL fibroblasts in the applied concentrations. Ethanol showed an enhancing effect on angiogenesis and activity of alkaline phosphatase. Simultaneously, ethanol reduced the induction of IL-6 and increased prostaglandin E2 synthesis as well as hPDL-fibroblast-mediated osteoclastogenesis without affecting the RANK-L/OPG-system. hPDL fibroblasts thus seem to be a cell type quite resistant to ethanol, as no cytotoxic effects or influence on cell viability were detected. High ethanol concentrations, however, seem to promote bone formation and angiogenesis. Ethanol at 0.179% also enhanced hPDL-induced osteoclastogenesis, indicating increased bone resorption and thus tooth movement velocity to be expected during orthodontic therapy.

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Triton X-100, laboratory grade
Antibiotic Antimycotic Solution (100×), Stabilized, with 10,000 units penicillin, 10 mg streptomycin and 25 μg amphotericin B per mL, 0.1 μm filtered, BioReagent, suitable for cell culture
Triton X-100, BioXtra
L-Ascorbic acid 2-phosphate sesquimagnesium salt hydrate, ≥95%
CelLytic M, Cell Lysis Reagent, Suitable for Mammalian cell lysis and protein solubilization.
Human VEGF-A ELISA Kit, for serum, plasma, cell culture supernatants and urine