Evaluation of Resveratrol and BioDendrimer-Resveratrol in Models of Dental Pulp Inflammation – In-vitro

Abstract

Vital pulp therapy (VPT) aims to preserve dental pulp tissues and maintain tooth function, while alleviating dental pain. However, persistent pain may necessitate more invasive treatments like root canal therapy (RCT). Current clinical VPT protocols lack options for treatments that might enhance tissue healing and improve post-operative pain outcomes. Resveratrol (RESV), a naturally occurring bioactive compounds, that exists in 72 plant species such as grapevines, blueberries, and others. RESV is known for its anti-inflammatory, antimicrobial, and neuroprotective effects, and it holds promise for VPT. Our aim is to determine the effects of two formulations of resveratrol (resveratrol (RESV) and biodendrimer- resveratrol (BD- RESV) on cytokine expression, mineralization and migration of dental pulp stem cells (DPSCs) challenged with LPS as an in-vitro model of pulpitis. Commercially available (Lansa) DPSCs from healthy adult teeth were grown using a-Minimum Essential Medium (Sigma-Aldrich, St Louis, MO) supplemented with other agents. Cells were maintained at 37 °C / 5% CO2 and plated for experiments at passage 2 or 3. Cells were treated for 2 to 7 days with varying doses of RESV (0.001-100μM), BD-RESV (0.001-100μM) and challenged with LPS or vehicle control (0.001-100μg/ml). Cell viability was assessed using the MTT assay on day 7, cell migration and cytokine expression (qRT-PCR) were evaluated on day 2 and mineralization was evaluated with alizarin red staining on day 28. We found that BD-RESV decreased LPS -induced DPSC cytotoxicity with statistically significant effect on LPS dose of 1μg/ml (p< 0.05), while RESV had no effect. However, both RESV and BD-RESV significantly prevented LPS induced upregulation of IL1-β, IL-6, IL-8 mRNA (p< 0.05). RESV and BD-RESV also induced the formation of calcium nodules (alizarin Staining) with the most significant effect observed at RESV 5μM and BD-10 μM (p< 0.05) and significantly increased DPSC migration (p< 0.05). Resv and Biodendrimer demonstrated a similar ability to mitigate LPS-induced cytokine upregulation and induce mineralization and DPSC migration suggesting its properties are particularly relevant to improve tissue healing in dental pulpas an inflammatory response modulator. However, further in-vivo research that incorporates RESV and BD-RESV as a direct pulp capping material in pulp exposure model of pulpitis is necessary to confirm that RESV's would be useful as a VPT supplementary component.