Clinical oral investigations · 2025
The synergistic effect of low-intensity focused ultrasound and tideglusib on odontogenic differentiation of human dental pulp stem cells.
СтатьяПоддержка исследования: вне правительства СШАНазвание и аннотация: Перевод в очереди
Обработка статьи · этапы, ошибки и управление
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Maria Rajan Sheetal, Ouyang Wanlu, Idrees Majdy, Kujan Omar, Fawzy Amr
Аннотация
OBJECTIVES: This study investigates the synergistic effects of Tideglusib, a GSK-3 inhibitor, and low-intensity focused ultrasound (LIFU) on the proliferation and odontogenic differentiation of human dental pulp stem cells (DPSCs) and lipopolysaccharide (LPS)-induced inflammatory DPSCs (LPS-DPSCs).
MATERIALS AND METHODS: DPSCs were exposed to Tideglusib (100, 250, and 500 nM) and LIFU (250 kHz, 2.5 W and 5 W for 60s and 120s). Cell proliferation was evaluated using acid phosphatase (APH) assay, and optimal parameters were determined as 100 nM Tideglusib and LIFU (5 W) at 60s, 120s, 60s/100 nM, and 120s/100 nM. Inflammatory conditions were induced using LPS. Cell proliferation was assessed using APH assay and confocal laser scanning microscope (CLSM), while odontogenic differentiation was evaluated via Alizarin red staining, wound healing assay, and qRT-PCR targeting odontogenic markers (RUNX2, ALPL, BMP2, DMP1), Wnt pathway activation (AXIN2), pro-apoptotic (CASP3), and anti-apoptotic (BCL2L1) genes.
RESULTS: The combination of LIFU and tideglusib (5 W 120s/100 nM) significantly enhanced (p < 0.05) proliferation, mineralization, and wound closure in both DPSC and LPS-DPSC groups. CLSM confirmed increased cell density, corroborating the APH assay, and qRT-PCR revealed upregulation of odontogenic and Wnt signalling markers. In LPS-DPSCs, CASP3 was downregulated and BCL2L1 upregulated, indicating improved cell survival in an inflammatory microenvironment.
CONCLUSIONS: The synergistic use of LIFU and Tideglusib promotes DPSC proliferation, migration, wound closure, and odontogenic differentiation, even under inflammatory stress.
CLINICAL RELEVANCE: This pharmacomechanical approach offers a minimally invasive, biologically driven strategy for regenerative endodontic therapy, with strong translational potential in managing reversible pulpitis and promoting dentine repair.
MATERIALS AND METHODS: DPSCs were exposed to Tideglusib (100, 250, and 500 nM) and LIFU (250 kHz, 2.5 W and 5 W for 60s and 120s). Cell proliferation was evaluated using acid phosphatase (APH) assay, and optimal parameters were determined as 100 nM Tideglusib and LIFU (5 W) at 60s, 120s, 60s/100 nM, and 120s/100 nM. Inflammatory conditions were induced using LPS. Cell proliferation was assessed using APH assay and confocal laser scanning microscope (CLSM), while odontogenic differentiation was evaluated via Alizarin red staining, wound healing assay, and qRT-PCR targeting odontogenic markers (RUNX2, ALPL, BMP2, DMP1), Wnt pathway activation (AXIN2), pro-apoptotic (CASP3), and anti-apoptotic (BCL2L1) genes.
RESULTS: The combination of LIFU and tideglusib (5 W 120s/100 nM) significantly enhanced (p < 0.05) proliferation, mineralization, and wound closure in both DPSC and LPS-DPSC groups. CLSM confirmed increased cell density, corroborating the APH assay, and qRT-PCR revealed upregulation of odontogenic and Wnt signalling markers. In LPS-DPSCs, CASP3 was downregulated and BCL2L1 upregulated, indicating improved cell survival in an inflammatory microenvironment.
CONCLUSIONS: The synergistic use of LIFU and Tideglusib promotes DPSC proliferation, migration, wound closure, and odontogenic differentiation, even under inflammatory stress.
CLINICAL RELEVANCE: This pharmacomechanical approach offers a minimally invasive, biologically driven strategy for regenerative endodontic therapy, with strong translational potential in managing reversible pulpitis and promoting dentine repair.