Influence of resveratrol application with pulp-capping materials on the genetic expression levels of stem cells.

Abstract

To evaluate in a laboratory setting the response of human mesenchymal stem cells (MSCs) to pulp-capping materials with and without resveratrol (RSV). Five materials, Calcimol LC, Life, TheraCal LC, ProRoot MTA and Biodentine, were prepared according to the manufacturers' instructions. Human MSCs were then exposed to these materials, with and without RSV, for 24h (n=8). Cell viability was evaluated using the MTT assay, and total cell death was quantified by annexin V-FITC staining with flow cytometry. The expression levels of the IL-8, IL-10, HBD-2 and BCL-2 genes were investigated using real-time polymerase chain reaction (RT-PCR). Data obtained from MTT test were analysed using one-way anova, and Tukey's multiple-comparison test. The paired Student t test was employed to compare the effects of materials on gene expression (significance level of 5%). The group cell viabilities were Calcimol LC 53%, Life 43%, TheraCal LC 78%, ProRoot MTA 75% and Biodentine 78%. Calcimol LC and Life exhibited significant differences compared with the control groups (P<0.05). The percentages of necrotic/late apoptotic cells associated with Calcimol LC and TheraCal LC were greater than in the other materials. However, when RSV was added to wells containing materials, cell viability increased to Calcimol LC 63%, Life 52%, TheraCal LC 82%, ProRoot MTA 91% and Biodentine 96%, and the percentages of early apoptotic and late apoptotic/necrotic cells decreased. Calcimol LC+RSV and Life+RSV differed significantly from the control group (P<0.05). The expression of IL-8 gene was high for all materials, ProRoot MTA caused significant overexpression, and the addition of RSV reduced the expression of IL-8 in the Calcimol LC, TheraCal LC and ProRoot MTA groups and led to increased expression of IL-10 in the Calcimol LC, Life and Biodentine groups. HBD-2 and BCL-2 exhibited increased expression in ProRoot MTA with RSV (P<0.05). The addition of RSV exerted a protective effect on MSCs and regulated the inflammatory process by altering the expression levels of pro- and anti-inflammatory genes.