Wnt-GSK3β/β-Catenin Regulates the Differentiation of Dental Pulp Stem Cells into Bladder Smooth Muscle Cells

Abstract

Smooth muscle cell- (SMC-) based tissue engineering provides a promising therapeutic strategy for SMC-related disorders. It has been demonstrated that human dental pulp stem cells (DPSCs) possess the potential to differentiate into mature bladder SMCs by induction with condition medium (CM) from bladder SMC culture, in combination with the transforming growth factor-β1 (TGF-β1). However, the molecular mechanism of SMC differentiation from DPSCs has not been fully uncovered. The canonical Wnt signaling (also known as Wnt/β-catenin) pathway plays an essential role in stem cell fate decision. The aim of this study is to explore the regulation via GSK3βand associated downstream effectors for SMC differentiation from DPSCs. We characterized one of our DPSC clones with the best proliferation and differentiation abilities. This stem cell clone has shown the capacity to generate a smooth muscle layer-like phenotype after an extended differentiation duration using the SMC induction protocol we established before. We further found that Wnt-GSK3β/β-catenin signaling is involved in the process of SMC differentiation from DPSCs, as well as a serial of growth factors, including TGF-β1, basic fibroblast growth factor (bFGF), epidermal growth factor (EGF), hepatocyte growth factor (HGF), platelet-derived growth factor-homodimer polypeptide of B chain (BB) (PDGF-BB), and vascular endothelial growth factor (VEGF). Pharmacological inhibition on the canonical Wnt-GSK3β/β-catenin pathway significantly downregulated GSK3βphosphorylation andβ-catenin activation, which in consequence reduced the augmented expression of the growth factors (including TGF-β1, HGF, PDGF-BB, and VEGF) as well as SMC markers (especially myosin) at a late stage of SMC differentiation. These results suggest that the canonical Wnt-GSK3β/β-catenin pathway contributes to DPSC differentiation into mature SMCs through the coordination of different growth factors.

Between Fate Choice and Self-Renewal—Heterogeneity of Adult Neural Crest-Derived Stem Cells

“…Next to periodontal ligament, dental tissues particularly including the dental pulp are developmentally derived from the neural crest ( Chai et al, 2000 ). Human DPSCs were reported to be positive for the NCSC-marker Nestin ( Arthur et al, 2008 ) and to form dentin and dental pulp tissue in vivo , but are also able to give rise to osteoblasts, chondrocytes, adipocytes and even to NC-related melanocytes and functionally active neurons ( Gronthos et al, 2000 ; Arthur et al, 2008 ; Stevens et al, 2008 ; Jiang et al, 2019 ). In accordance with our own observations in human ITSCs, single cell-derived clonal cultures of murine DPSCs revealed highly heterogeneous gene expression levels of Nestin between individual stem cell clones.…”

Application of Adult and Pluripotent Stem Cells in Interstitial Cystitis/Bladder Pain Syndrome Therapy: Methods and Perspectives

“…Thereafter, the authors confirmed the upregulation of SMC-related genes and proteins, namely alpha smooth muscle actin (α-SMA), desmin, and calponin. In 2019, Jiang et al proved the crucial role of the canonical Wnt-GSK3β/β-catenin pathway in the differentiation of DPSCs into bladder SMCs [161]. However, these findings need to be verified in IC/BPS animal models.…”

“…n.a. [160,161] -Cell injection into the bladder (2.0 × 10 6 cells in PBS). -DPSC-derived CM contains high level of FGF-2, VEGF, C-C and C-X-C chemokine families.…”

Integration of Regenerative Dentistry Into the Dental Undergraduate Curriculum

“…While these are some examples of many aiming to regenerate specific dental tissues, other groups have focused on exploiting the regenerative potential of DPSCs, a highly pluripotent and accessible cell type. These studies have so far primarily focused on neural repair, smooth muscles regeneration, bladder tissue repair, retinal tissue repair, and bone regeneration, to name a few (25)(26)(27)(28).…”