Human urine-derived stem cells can be induced into osteogenic lineage by silicate bioceramics via activation of the Wnt/β-catenin signaling pathway
Introduction
Recent studies have demonstrated that urine-derived stem cells (USCs) possess self-renewal and multilineage differentiation potential [1], [2], [3]. Several investigations used USCs as cell source to form engineered urethral tissue [4], [5]. For example, USCs seeded in bacterial cellulose scaffolds can form multilayered urothelium and cell matrix [4]. Cells differentiated from USCs were seeded on a modified 3D porous small intestinal submucosa in order to form engineered urethral tissue. Results showed that the cells formed multiple uniform layers on the scaffolds, which was similar to that of native urinary tract tissue [5]. As USCs can be obtained using non-invasive and simple methods, they represent a promising alternative stem cell population for tissue regeneration. However, whether USCs could serve as cell source for bone tissue regeneration is still unknown. In our previous study, we have demonstrated that USCs share similar biological characteristics with adipose stem cells (ASCs) and could further differentiate into an osteogenic lineage in vitro [2]. These evidences suggested that USCs might be a good cell source for bone tissue engineering. In addition, to better apply USCs in bone tissue engineering, it is critical to develop bioactive material that could facilitate the proliferation and osteogenic differentiation of USCs.
Silicates, including silicate bioglass and silicate bioceramics, have been reported to be able to stimulate osteoblastic differentiation and bone regeneration in many studies [6], [7], [8]. Previous studies have shown that Si plays an important role during skeletal development and regeneration [9], [10]. Si has been widely incorporated into bioceramics to enhance their bioactivity [7], [11], [12], [13], [14]. Calcium silicate (CS), one of the silicate bioceramics with simple components, has shown a great osteostimulatory effect without the need of additional exogenous growth factors [15], [16]. It has been demonstrated that CS can stimulate osteogenic differentiation of several kind of adult stem cells including human bone marrow mesenchymal stem cells (BMSCs), orofacial mesenchymal stem cells as well as tooth germ stem cells [17], [18], [19]. In addition, several studies have shown that CS ion extracts or composite scaffolds containing CS possess pro-angiogenic properties [13], [20], which may further stimulate bone regeneration through enhancing vascularization and angiogenesis to deliver oxygen and nutrients to seeded cells in bone tissue engineering. Therefore, scaffolds containing CS show great potential for bone tissue regeneration.
To develop an ideal silicate-based biomaterial, it is important to clarify the molecular mechanism through which the biomaterial regulates stem cell differentiation. The canonical Wnt/β-catenin signaling cascade plays a critical role in cell proliferation, differentiation and apoptosis [21]. It has been demonstrated that the Wnt/β-catenin signaling pathway is involved in bone formation [22]. Recently, Zhou et al. reported that the Wnt/β-catenin signaling pathway was involved in the regulation of bredigite bioceramic-induced osteogenic differentiation of periodontal ligament cells [23]. Han et al. demonstrated that Si ions stimulated the osteogenic differentiation of BMSCs and that the Wnt/β-catenin signaling pathway played an important role in this process [24]. Therefore, we hypothesize that Wnt/β-catenin signaling pathway may also play an important role on proliferation and differentiation of USCs with the presence of CS. However, little is known about the involvement of Wnt/β-catenin signaling in the USCs osteogenic differentiation upon CS interaction.
This study was conducted to investigate the application potential of USCs as cell source for bone tissue engineering by studying the in vitro and in vivo osteogenic differentiation of USCs when they were cultured with CS ion extracts or on scaffolds containing CS and to explore the role of the Wnt/β-catenin signaling pathway during this process.
Section snippets
Synthesis of CS powders
Chemical co-precipitation method was used to obtain CS powders [15], [16]. Briefly, chemical co-precipitation happened during continuous mixing of an aqueous solution of Na2SiO3 (1 mol/L) and aqueous solution of Ca(NO3)2 (1 mol/L) at ambient temperature overnight (molar ratio: Na2SiO3:Ca(NO3)2 = 1:1). Then, CS suspension was obtained after the reacted mixture were filtered and washed with deionized water and ethanol. After that, CS suspension was dried at 80 °C overnight and calcined at 800 °C
Analysis of Ca, P and Si concentrations in the CS extracts
To investigate the effects of ionic products of CS on osteogenic differentiation of USCs, the ion concentrations of Ca, P and Si in CS ion extracts diluted at different ratios with USC growth medium were examined by ICP-AES. The result was listed in Table 1. As shown, Ca and P ion concentration in the CS extracts diluted with control medium at 1/64, 1/128 and 1/256 were similar to those in the control medium. However, the concentrations of Si ions in the CS extracts diluted with control medium
Discussion
The present study demonstrated that USCs could be induced to osteogenic lineage by CS ion extracts or scaffolds containing CS. Particularly, Wnt/β-catenin signaling pathway plays an important role in this process. PLGA/CS scaffolds in combination with USCs significantly promoted the mineralization of the scaffolds and blood vessels invasion to the scaffolds in nude mice as compared to the pure PLGA scaffolds and the scaffolds without USCs. The results revealed that CS has stimulatory effects on
Conclusion
In the present study, we not only explored the application potential of USCs as cell source for bone tissue engineering but also investigated the effects of CS on proliferation and osteogenic differentiation of USCs in vitro and in vivo and further proposed the mechanisms through which CS stimulated osteogenic differentiation of USCs. In vitro study suggested that the ions extracted from CS were capable of inducing the osteogenic differentiation of USCs through the Wnt/β-catenin signaling
Acknowledgments
This study was financially supported by the National High Technology Research and Development Program of China (2012AA020506), National Natural Science Foundation of China (Grant No.: 31200714 and 31470918), Nature Science Foundation of Shanghai Municipal (Grant No.: 12ZR1413900), the Shanghai Pujiang Talent Program (Grant no. 13PJ1404100) and “Priority Among Priorities” Clinical Medical Center Construction Project of the Shanghai Municipal.
References (59)
- et al.
Urine derived cells are a potential source for urological tissue reconstruction
J Urol
(2008) - et al.
Tissue-engineered conduit using urine-derived stem cells seeded bacterial cellulose polymer in urinary reconstruction and diversion
Biomaterials
(2010) - et al.
Human urine-derived stem cells seeded in a modified 3D porous small intestinal submucosa scaffold for urethral tissue engineering
Biomaterials
(2011) - et al.
Osteogenesis and angiogenesis induced by porous β-CaSiO3/PDLGA composite scaffold via activation of AMPK/ERK1/2 and PI3K/Akt pathways
Biomaterials
(2013) - et al.
Influences of ionic dissolution products of dicalcium silicate coating on osteoblastic proliferation, differentiation and gene expression
Acta Biomater
(2009) - et al.
Thorough analysis of silicon substitution in biphasic calcium phosphate bioceramics: a multi-technique study
Acta Biomater
(2010) - et al.
Synthesis and characterization of single-phase silicon-substituted alpha-tricalcium phosphate
Biomaterials
(2006) - et al.
Silicate bioceramics enhanced vascularization and osteogenesis through stimulating interactions between endothelia cells and bone marrow stromal cells
Biomaterials
(2014) - et al.
Fabrication and characterization of bioactive wollastonite/PHBV composite scaffolds
Biomaterials
(2004) - et al.
Apatite formation on bioactive calcium-silicate cements for dentistry affects surface topography and human marrow stromal cells proliferation
Dent Mater
(2010)
Stimulation of proangiogenesis by calcium silicate bioactive ceramic
Acta Biomater
Canonical WNT signaling promotes osteogenesis by directly stimulating Runx2 gene expression
J Biol Chem
The stimulation of proliferation and differentiation of periodontal ligament cells by the ionic products from Ca7Si2P2O16 bioceramics
Acta Biomater
Stimulatory effects of the ionic products from Ca–Mg–Si bioceramics on both osteogenesis and angiogenesis in vitro
Acta Biomater
Gene array analysis of Wnt-regulated genes in C3H10T1/2 cells
Bone
Fabrication and characterization of β-dicalcium silicate/poly(d,l-lactic acid) composite scaffolds
Mater Lett
Effects of the controlled-released TGF-β1 from chitosan microspheres on chondrocytes cultured in a collagen/chitosan/glycosaminoglycan scaffold
Biomaterials
Lentivirus-mediated gene transfer induces long-term transgene expression of BMP-2 in vitro and new bone formation in vivo
Mol Ther
In vitro degradation of porous degradable and bioactive PHBV/wollastonite composite scaffolds
Polym Degrad Stab
Bone grafts engineered from human adipose-derived stem cells in dynamic 3D-environments
Biomaterials
Human embryonic stem cell encapsulation in alginate microbeads in macroporous calcium phosphate cement for bone tissue engineering
Acta Biomater
Future prospects for regenerated heart using induced pluripotent stem cells
J Pharmacol Sci
Bioactive glass in tissue engineering
Acta Biomater
Histone demethylase Jmjd3 regulates osteoblast differentiation via transcription factors Runx2 and osterix
J Biol Chem
A review of the biological response to ionic dissolution products from bioactive glasses and glass-ceramics
Biomaterials
Nodule formation and mineralisation of human primary osteoblasts cultured on a porous bioactive glass scaffold
Biomaterials
The cementogenic differentiation of periodontal ligament cells via the activation of Wnt/β-catenin signalling pathway by Li+ ions released from bioactive scaffolds
Biomaterials
The repair of large segmental bone defects in the rabbit with vascularized tissue engineered bone
Biomaterials
Coating of VEGF-releasing scaffolds with bioactive glass for angiogenesis and bone regeneration
Biomaterials
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These authors contributed equally to this work.