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 GSK3β and 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 GSK3β phosphorylation and β-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.
Highlights
A range of injuries or diseases including cancer, benign bladder contracture, and congenital anomalies can result in the damage or loss of the bladder [1,2,3]
The Wnt-Glycogen synthase kinase 3β (GSK3β)/β-catenin signaling pathway has been previously reported to play an important role in transforming growth factor-β1 (TGF-β1)-induced Mesenchymal stem cells (MSCs) differentiation. To investigate whether this canonical Wnt signaling is involved in the bladder SMC differentiation from dental pulp stem cells (DPSCs), we evaluated the protein level of phosphorylation GSK3β (p-GSK3β), total GSK3β (t-GSK3β), and active β-catenin in A32 in response to the induction of differentiation medium by western blotting
We have demonstrated that Wnt-GSK3β/β-catenin signaling as well as several key growth factors was required in the process of human DPSCs differentiating into human bladder SMCs
Summary
A range of injuries or diseases including cancer, benign bladder contracture, and congenital anomalies (such as bladder exstrophy and myelomeningocele) can result in the damage or loss of the bladder [1,2,3]. Patients with those illnesses require bladder augmentation cystoplasty or replacement. Stem Cells International smooth muscle cell-based tissue engineering becomes one of the most promising remedies for restoring bladder organ function of the urinary system [6, 7]. Search for an alternative cell source is essential for bladder tissue engineering, which could provide a new way to overcome the shortcomings of the aforementioned methods in the future
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