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An Inhibitory Role of Osthole in Rat MSCs Osteogenic Differentiation and Proliferation via Wnt/β-Catenin and
Background/Aims:
Bone marrow-derived mesenchymal stem cells (MSCs) are responsible for new bone formation during adulthood. Accumulating evidences showed that Osthole promotes the osteogenic differentiation in primary osteoblasts. The aim of this study was to investigate whether Osthole exhibits a potential to stimulate the osteogenic differentiation of MSCs and the underlying mechanism.
Methods:
MSCs were treated with a gradient concentration of Osthole (6.25 µM, 12.5 µM, and 25 µM). Cell proliferation was assessed by western blotting with the proliferating cell nuclear antigen (PCNA) and Cyclin D1 antibodies, fluorescence activated cell sorting (FACS), and cell counting kit 8 (CCK8). MSCs were cultured in osteogenesis-induced medium for one or two weeks. The osteogenic differentiation of MSCs was estimated by Alkaline Phosphatase (ALP) staining, Alizarin red staining, Calcium influx, and quantitative PCR (qPCR). The underlying mechanism of Osthole-induced osteogenesis was further evaluated by western blotting with antibodies in Wnt/β-catenin, PI3K/Akt, BMPs/smad1/5/8, and MAPK signaling pathways.
Results:
Osthole inhibited proliferation of rat MSCs in a dose-dependent manner. Osthole suppressed osteogenic differentiation of rat MSCs by down-regulating the activities of Wnt/β-catenin and Erk1/2-MAPK signaling.
Conclusions:
Osthole inhibits the proliferation and osteogenic differentiation of rat MSCs, which might be mediated through blocking the Wnt/β-catenin and Erk1/2-MAPK signaling pathways.
Insights
Osthole inhibits mesenchymal stem cell (MSC) proliferation and osteogenic differentiation. This effect may involve blocking the Wnt/β-catenin and Erk1/2-MAPK signaling pathways, impacting bone formation.
Area of Science:
- Stem cell biology
- Bone biology
- Pharmacology
Background:
- Mesenchymal stem cells (MSCs) are crucial for adult bone formation.
- Osthole has been shown to promote osteogenic differentiation in primary osteoblasts.
- Investigating Osthole's effect on MSCs is important for understanding bone regeneration.
Purpose of the Study:
- To determine if Osthole stimulates osteogenic differentiation of MSCs.
- To elucidate the underlying molecular mechanisms of Osthole's action on MSCs.
- To assess Osthole's impact on MSC proliferation and differentiation pathways.
Main Methods:
- Rat MSCs were treated with varying Osthole concentrations (6.25–25 µM).
- Proliferation was measured using PCNA, Cyclin D1, FACS, and CCK8 assays.
- Osteogenic differentiation was assessed via ALP staining, Alizarin red staining, calcium influx, and qPCR.
- Signaling pathway involvement (Wnt/β-catenin, PI3K/Akt, BMPs/smad1/5/8, MAPK) was analyzed by western blotting.
Main Results:
- Osthole inhibited rat MSC proliferation in a dose-dependent manner.
- Osthole suppressed osteogenic differentiation of rat MSCs.
- This suppression was linked to downregulated Wnt/β-catenin and Erk1/2-MAPK signaling.
Conclusions:
- Osthole inhibits both proliferation and osteogenic differentiation of rat MSCs.
- The observed effects are potentially mediated by the blockade of Wnt/β-catenin and Erk1/2-MAPK pathways.
- These findings suggest Osthole may not be a suitable agent for promoting MSC-based bone formation.
