PDGF-AA guides cell crosstalk between human dental pulp stem cells in vitro via the PDGFR-/PI3K/Akt axis

AimTo explore the influence of PDGF-AA on cell communication between human dental pulp stem cells (DPSCs) by characterizing gap junction intercellular communication (GJIC) and its potential biomechanical mechanism.MethodologyQuantitative real-time PCR was used to measure connexin family member expression in DPSCs. Cell migration and CCK-8 assays were utilized to examine the influence of PDGF-AA on DPSC migration and proliferation. A scrape loading/dye transfer assay was applied to evaluate GJIC triggered by PDGF-AA, a PI3K/Akt signalling pathway blocker (LY294002) and a PDGFR-alpha blocker (AG1296). Western blotting and immunofluorescence were used to test the expression and distribution of the Cx43 and p-Akt proteins in DPSCs. Scanning electron microscopy (SEM) and immunofluorescence were used to observe the morphology of GJIC in DPSCs.ResultsPDGF-AA promoted gap junction formation and intercellular communication between human dental pulp stem cells. PDGF-AA upregulates the expression of Cx43 to enhance gap junction formation and intercellular communication. PDGF-AA binds to PDGFR-alpha and activates PI3K/Akt signalling to regulate cell communication.ConclusionsThis research demonstrated that PDGF-AA can enhance Cx43-mediated GJIC in DPSCs via the PDGFR-alpha/PI3K/Akt axis, which provides new cues for dental pulp regeneration from the perspective of intercellular communication.