Evaluating the Effect of Eugenol on the Expression of Genes Involved in the Immunomodulatoty Potency of Mouse Mesenchymal Stem Cells In Vitro

The immunomodulatory ability of mesenchymal stem cells (MSCs) has attracted interest as a unique property thatmakes them interesting tools for the treatment of inflammatory and autoimmune diseases. Eugenol is a volatilecompound from the phenylpropanoids class of chemical compounds. Despite extensive investigations on thebiological and pharmacological properties of Eugenol, its effect on stem cells, especially, on MSCs remains to beclarified. Therefore, this study was designed to evaluate the effect of Eugenol on the expression of genes (Tlr3,Tlr4, Ccl2, and Ccl3) involved in immunomodulatory potency of mouse bone-marrow derived MSCs byquantitative real-time PCR (qRT-PCR). To do so, MSCs were isolated from 4-8 weeks old mouse bone marrow(BM). The effect of Eugenol on the viability of BM-MSCs was evaluated by MTT assay at 24, 48, and 72h aftertreatment. The results showed that Eugenol reduced the number of BM-MSCs in a dose- and time-dependentmanner. In addition, the half maximum inhibitory concentration of Eugenol on MSCs was 400μg/ml at 24 and 48hand 200μg/ml at 72h after treatment. Moreover, about 90% of MSCs were alive at the concentration of 12.5μg/ml24h after treatment. The qRT-PCR results indicated that Tlr3, Tlr4, and Ccl3 genes were up-regulated 1.6-, 1.8-,and 2.2-fold, respectively, in Eugenol-treated BM-MSCs compared to untreated controls (Fold change > 1.5; P≤0.05). In conclusion, we suggest that Eugenol may somewhat regulate the immunomodulatory potency of MSCsand thus this study provides a background for further studies on the effect of Eugenol on MSCs characteristics andfunctions, which may finally improve their potency for cell-based therapy applications.

Keywords:

Eugenol , Mesenchymal stem cells , Immunomodulatory potency , Mouse