Small molecule drug TD-198946 enhances osteogenic differentiation of rat bone marrow mesenchymal stem cells

Yang Chao; Luo Zongping

doi:10.12307/2025.060

Abstract：BACKGROUND:TD-198946 is a small molecule drug known to induce stem cells to form cartilage;however,its effect on osteogenic differentiation remains unclear. OBJECTIVE:To investigate the effects of the small molecule drug TD-198946 on promoting osteoblastic differentiation of rat bone marrow mesenchymal stem cells and its mechanism of action. METHODS:Bone marrow mesenchymal stem cells (BMSCs) were extracted from SD rats.CCK-8 assay was used to evaluate the effect of different concentrations of TD-198946 on the proliferation of bone marrow mesenchymal stem cells to determine the optimal concentration of TD-198946.Then,the optimal concentration of TD-198946 and osteogenic medium were added to induce osteogenic differentiation of bone marrow mesenchymal stem cells.On day 3,qRT-PCR was used to detect the expression of alkaline phosphatase,Runt-related transcription factor 2,osteopontin,osteocalcin,and type Ⅰ collagen genes.On day 7 of osteogenic induction,alkaline phosphatase staining and western blot assay were performed to detect the expression of Runt-related transcription factor 2,type Ⅰ collagen,AKT,p-AKT,PI3K,and p-PI3K proteins.On day 21 of osteogenic induction,Alizarin red staining was conducted. RESULTS AND CONCLUSION:(1) CCK-8 assay results revealed that 100 nmol/L TD-198946 could promote the proliferation of bone marrow mesenchymal stem cells.(2) The results of alkaline phosphatase staining and alizarin red staining showed that TD-198946 could promote osteogenic differentiation of rat bone marrow mesenchymal stem cells.(3) qRT-PCR results showed that TD-198946 could promote the expression of osteogenic genes alkaline phosphatase,Runt-related transcription factor 2,osteopontin,osteocalcin,and type Ⅰ collagen.(4) Western blot assay results showed that TD-198946 could enhance the expression of Runt-related transcription factor 2,type Ⅰ collagen,p-PI3K,and p-AKT.The results indicate that the small molecule drug TD-198946 may induce osteogenic differentiation of rat bone marrow mesenchymal stem cells by activating the PI3K/AKT signaling pathway.