Evaluation the Effects of both Aqueous and Alcoholic Extract Derived from the Root of Rubia Tinctorum on the Osteogenic Differentiation of Rat Mesenchymal Stem Cells

Background and purpose: The osteogenic differentiation of MSCs plays a key role in bone tissue engineering and regenerative medicine, making it essential to explore natural compounds that may enhance this process. This aim this study was to investigate the osteogenic potential of both aqueous and alcoholic extracts derived from the root of Rubia tinctorum on rat bone marrow-derived mesenchymal stem cells (BM-MSCs). Materials and methods: Aqueous and alcoholic extracts were prepared from the root of Rubia tinctorum. Rat BM-MSCs were isolated and cultured and their osteogenic differentiation was assessed in the presence of varying concentrations of the extracts using alizarin red staining and alkaline phosphatase (ALP) activity. Cytotoxic effect of root extract of Rubia tinctorum was evaluated using MTT assay. Results: The MTT assay demonstrated that the root extract of Rubia tinctorum was cytotoxic at 1000 µg/ml. Additionally, the alizarin red assay showed that the most significant color intensity at 1 and 10 µg/ml concentrations on the 14th day. In terms of bone differentiation induction which measured via the alkaline phosphatase test, both the aqueous and alcoholic extracts at 1 and 10 µg/ml concentrations led to substantial increases (14.9- and 14.57-fold for the aqueous extract and approximately 7.37- and 7-fold for the alcoholic extract, respectively) in alkaline phosphatase activity compared to the negative control. Conclusion: This study sheds light on the osteogenic effects of Rubia tinctorum root extracts, emphasizing their potential as natural agents in promoting bone regeneration and tissue engineering applications. Further investigations into the underlying molecular mechanisms and in vivo studies are warranted to comprehensively evaluate the feasibility of utilizing these extracts for therapeutic bone regeneration purposes