IADR Abstract Archives
Hippo and Wnt Pathways Modulated Mineralization of Decellularized ECM-derived hDPSCs
Objectives: Extracellular matrix (ECM) is an intricate structure providing the microenvironment niche that influences stem cell differentiation. This study aimed to compare the efficacy of decellularized ECM derived from human dental pulp stem cells (dECM_DPSCs) and gingival-derived mesenchymal stem cells (dECM_GSCs) as an osteogenic inductive scaffold for reseeded GSCs.
Methods: ECM was constructed from DPSCs and GSCs that were cultured in a growth medium supplemented with L-ascorbic acid (N-ECM) or osteogenic induction medium (OM-ECM). Matrisome profile was analyzed using proteomic analysis. GSCs were reseeded on dECM_DPSCs and dECM_GSCs to evaluate the osteogenic differentiation potential of each type of dECM. Global differential gene expression and potential regulatory pathways were examined using bioinformatic analysis. The Mann Whitney U test was used to verify significant differences for two independent group comparisons. For three or more group comparisons, statistical differences were assessed using the Kruskal Wallis test followed by a pairwise comparison. The significance level was set at 5% (p < 0.05).
Results: GSCs reseeded on N- and OM-dECM_DPSCs exhibited a significant increase in cell proliferation and mineral deposits compared to those derived from GSCs, even though. The proteomic analysis demonstrated that common and signature matrisome proteins from dECM_DPSCs and dECM_GSCs were related to osteogenic differentiation. In the GSC reseeding experiment, RNA sequencing data revealed that dECM_DPSCs upregulated genes related to the Hippo and Wnt signaling pathways. Hippo and Wnt inhibitors significantly decreased the osteogenic differentiation potential of reseeded GSCs that were cultured in growth medium. Further, GSCs reseeded on dECM_DPSC and cultured in growth medium exhibited significant upregulation of genes related to Hippo and Wnt signaling.
Conclusions: dECM_DPSCs contained mineralization-associated factors that superiorly promoted GSCs osteogenic differentiation through Hippo and Wnt signaling pathways. These results emphasized the promising translational application of dECM_DPSCs as a bio-scaffold rich in favorable regenerative microenvironment for tissue engineering.
Methods: ECM was constructed from DPSCs and GSCs that were cultured in a growth medium supplemented with L-ascorbic acid (N-ECM) or osteogenic induction medium (OM-ECM). Matrisome profile was analyzed using proteomic analysis. GSCs were reseeded on dECM_DPSCs and dECM_GSCs to evaluate the osteogenic differentiation potential of each type of dECM. Global differential gene expression and potential regulatory pathways were examined using bioinformatic analysis. The Mann Whitney U test was used to verify significant differences for two independent group comparisons. For three or more group comparisons, statistical differences were assessed using the Kruskal Wallis test followed by a pairwise comparison. The significance level was set at 5% (p < 0.05).
Results: GSCs reseeded on N- and OM-dECM_DPSCs exhibited a significant increase in cell proliferation and mineral deposits compared to those derived from GSCs, even though. The proteomic analysis demonstrated that common and signature matrisome proteins from dECM_DPSCs and dECM_GSCs were related to osteogenic differentiation. In the GSC reseeding experiment, RNA sequencing data revealed that dECM_DPSCs upregulated genes related to the Hippo and Wnt signaling pathways. Hippo and Wnt inhibitors significantly decreased the osteogenic differentiation potential of reseeded GSCs that were cultured in growth medium. Further, GSCs reseeded on dECM_DPSC and cultured in growth medium exhibited significant upregulation of genes related to Hippo and Wnt signaling.
Conclusions: dECM_DPSCs contained mineralization-associated factors that superiorly promoted GSCs osteogenic differentiation through Hippo and Wnt signaling pathways. These results emphasized the promising translational application of dECM_DPSCs as a bio-scaffold rich in favorable regenerative microenvironment for tissue engineering.