The Effect of Rosmarinic Acid in Neural Differentiation of Wartons Jelly-derived Mesenchymal Stem Cells in Two Dimensional and Three Dimensional Cultures using Chitosan-based Hydrogel

Numerous studies have shown the positive effects of rosmarinic acid on the nervous system. Rosmarinic acid as a herbal compound with anti-inflammatory effects can prevent the destructive effect of inflammation on the nervous system. Furthermore, various studies have emphasized the advantages of three dimensional (3D) culture over the two dimensional (2D) culture of cells. In this study, thermosensitive chitosan-based hydrogel as a 3D scaffold with the combination of chitosan (CH), beta-glycerol phosphate(βGP) and hydroxyl ethyl cellulose (HEC) CH-βGP-HEC loaded with rosmarinic acid was used to induce neuronal differentiation in human Wharton jelly stem cells. Also, cells were divided into eight groups in order to evaluate the effect of 3D cell culture and to compare gene expression in different induction conditions. The results of gene expression analysis showed the highest expression of neuronal markers in WJMSCs cultured in CH-βGP-HEC loaded with differentiation medium and rosmarinic acid. According to the results of gene expression, rosmarinic acid alone have positively effect on the induction of expression of neural markers. This positive effect is enhanced by cell culture in 3D conditions. This study shows that rosmarinic acid can be considered an inexpensive and available compound for use in neural tissue engineering. The results of this study indicate that rosmarinic acid can be considered a cheap and available compound for use in neural tissue engineering. The results of our study also emphasize the need to study stem cell differentiation under 3D culture conditions.