Inhibition of transforming growth factor‐β signaling pathway enhances the osteogenic differentiation of unrestricted somatic stem cells

Hedayati, S M; Tafreshi, Azita Parvaneh; Moradi, N.; Zeynali, Bahman

doi:10.1002/jcb.27209

Cited by 9 publications

(6 citation statements)

References 27 publications

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“…hMSCs were cultured for 21 days under conditions that promote osteogenic differentiation. SB431542, a specific inhibitor of TGF-β receptor kinase, was added to ODM after 7 days of osteogenic differentiation because it enhances osteogenic differentiation of some somatic stem cells [40,41]. Although osteogenic differentiation of hMSCs was observed in the absence of SB431542, osteogenic differentiation of hMSCs was obvious in the presence of SB-431542 at 21 days of osteogenic differentiation by increased calcium deposition ( Fig.…”

Section: Identification Of Target Antigens Of Mr14-e5 Er7-a7 Er7-a8mentioning

confidence: 99%

Expression dynamics of integrin α2, α3, and αV upon osteogenic differentiation of human mesenchymal stem cells

et al. 2020

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Background: The differentiation of human mesenchymal stem cells (hMSCs) into osteoblasts (OBs) is a prerequisite for bone formation. However, little is known about the definitive surface markers for OBs during osteogenesis. Methods: To study the surface markers on OBs, we generated and used monoclonal antibodies (MAbs) against surface molecules on transforming growth factor-β1 (TGF-β1)-treated cancer cells. The generated MAbs were further selected toward expression changes on hMSCs cultured with TGF-β1/bone morphogenetic protein-2 (BMP-2) or osteogenic differentiation medium (ODM) by flow cytometry. Immunoprecipitation and mass spectrometry were performed to identify target antigens of selected MAbs. Expression changes of the target antigens were evaluated in hMSCs, human periodontal ligament cells (hPDLCs), and human dental pulp cells (hDPCs) during osteogenic and adipogenic differentiation by quantitative polymerase chain reaction (qPCR) and flow cytometry. hMSCs were also sorted by the MAbs using magnetic-activated cell sorting system, and osteogenic potential of sorted cells was evaluated via Alizarin Red S (ARS) staining and qPCR. Results: The binding reactivity of MR14-E5, one of the MAbs, was downregulated in hMSCs with ODM while the binding reactivity of ER7-A7, ER7-A8, and MR1-B1 MAbs was upregulated. Mass spectrometry and overexpression identified that MR14-E5, ER7-A7/ER7-A8, and MR1-B1 recognized integrin α2, α3, and αV, respectively. Upon osteogenic differentiation of hMSCs, the expression of integrin α2 was drastically downregulated, but the expression of integrin α3 and αV was upregulated in accordance with upregulation of osteogenic markers. Expression of integrin α3 and αV was also upregulated in hPDLCs and hDPCs during osteogenic differentiation. Cell sorting showed that integrin αV-high hMSCs have a greater osteogenic potential than integrin αV-low hMSCs upon the osteogenic differentiation of hMSCs. Cell sorting further revealed that the surface expression of integrin αV is more dramatically induced even in integrin αV-low hMSCs.

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Section: Identification Of Target Antigens Of Mr14-e5 Er7-a7 Er7-a8mentioning

confidence: 99%

Expression dynamics of integrin α2, α3, and αV upon osteogenic differentiation of human mesenchymal stem cells

et al. 2020

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“…The activated SMAD2/3 proteins form a complex with the SMAD4 protein, migrate to the nucleus, and activate the Runx2, Dlx5, Osterix, Col2, and aggrecan proteins’ coding genes. These genes are required for osteoblastogenesis [ 33 ]. Activation of TGF receptors can also activate Ras protein which activates the protein kinase activity of Raf.…”

Section: Resultsmentioning

confidence: 99%

“…Activation of TGF receptors can also activate Ras protein which activates the protein kinase activity of Raf. Raf kinase phosphorylates and activates mitogen-activated protein kinase (MEK1/2), which phosphorylates and activates extracellular signal-regulated kinases (ERK1/2) [ 33 , 34 ]. Phosphorylated ERK1/2 then migrate to the nucleus, reducing SMAD2/3/4 transcription as well as cell growth and proliferation.…”

Section: Resultsmentioning

confidence: 99%

“…TGF-β genes, in particular TGF-β1, due to their importance in the regulation of bone metabolism, are considered leading candidates for increasing individual susceptibility to OPLL [ 25 ]. TGF-β is present in the ossified matrix and chondrocytes of the adjacent cartilaginous regions of OPLL, but not in the mesenchymal stem cells and unossified ligament, indicating that it may stimulate bone formation at a later stage of ectopic ossification [ 25 , 33 ]. The gene encoding TGF-β1 location is 19q13.2 and consists of 52,325 base pairs (bp).…”

Section: Resultsmentioning

confidence: 99%

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Molecular and Genetic Mechanisms of Spinal Stenosis Formation: Systematic Review

Byvaltsev

Калинин

Hernandez

et al. 2022

IJMS

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Spinal stenosis (SS) is a multifactorial polyetiological condition characterized by the narrowing of the spinal canal. This condition is a common source of pain among people over 50 years old. We perform a systematic review of molecular and genetic mechanisms that cause SS. The five main mechanisms of SS were found to be ossification of the posterior longitudinal ligament (OPLL), hypertrophy and ossification of the ligamentum flavum (HLF/OLF), facet joint (FJ) osteoarthritis, herniation of the intervertebral disc (IVD), and achondroplasia. FJ osteoarthritis, OPLL, and HLF/OLFLF/OLF have all been associated with an over-abundance of transforming growth factor beta and genes related to this phenomenon. OPLL has also been associated with increased bone morphogenetic protein 2. FJ osteoarthritis is additionally associated with Wnt/β-catenin signaling and genes. IVD herniation is associated with collagen type I alpha 1 and 2 gene mutations and subsequent protein dysregulation. Finally, achondroplasia is associated with fibroblast growth factor receptor 3 gene mutations and fibroblast growth factor signaling. Although most publications lack data on a direct relationship between the mutation and SS formation, it is clear that genetics has a direct impact on the formation of any pathology, including SS. Further studies are necessary to understand the genetic and molecular changes associated with SS.

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“…Small molecule SB431542, as a potent and specific TGF-β type I receptor inhibitor, had been reported to potentiate endogenous bone regeneration [2]. In addition, SB431542 treatment shows a potential to enhance osteogenic differentiation of cord blood-derived unrestricted somatic stem cells and periodontal ligament cells depending on different context [24, 25]. However, whether SB431542 could be used to promote GMSCs osteogenesis is unknown.…”

Section: Introductionmentioning

confidence: 99%

Small molecule inhibitor of TGF-β signaling enables robust osteogenesis of autologous GMSCs to successfully repair minipig severe maxillofacial bone defects

et al. 2019

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Background Clinically, for stem cell-based therapy (SCBT), autologous stem cells are considered better than allogenic stem cells because of little immune rejection and no risk of communicable disease infection. However, severe maxillofacial bone defects restoration needs sufficient autologous stem cells, and this remains a challenge worldwide. Human gingival mesenchymal stem cells (hGMSCs) derived from clinically discarded, easily obtainable, and self-healing autologous gingival tissues, have higher proliferation rate compared with autologous bone marrow mesenchymal stem cells (hBMSCs). But for clinical bone regeneration purpose, GMSCs have inferior osteogenic differentiation capability. In this study, a TGF-β signaling inhibitor SB431542 was used to enhance GMSCs osteogenesis in vitro and to repair minipig severe maxillofacial bone defects. Methods hGMSCs were isolated and cultured from clinically discarded gingival tissues. The effects of SB431542 on proliferation, apoptosis, and osteogenic differentiation of hGMSCs were analyzed in vitro, and then, SB431542-treated hGMSCs composited with Bio-Oss® were transplanted into immunocompromised mice subcutaneously to explore osteogenic differentiation in vivo. After that, SB431542-treated autologous pig GMSCs (pGMSCs) composited with Bio-Oss® were transplanted into circular confined defects (5 mm × 12 mm) in minipigs maxillary to investigate severe bone defect regeneration. Minipigs were sacrificed at 2 months and nude mice at 8 weeks to retrieve specimens for histological or micro-CT or CBCT analysis. Effects of SB431542 on TGF-β and BMP signaling in hGMSCs were investigated by Western Blot or qRT-PCR. Results One micromolar of SB431542 treatment induced a robust osteogenesis of hGMSCs in vitro, without adverse effect on apoptosis and growth. In vivo, 1 μM SB431542 treatment also enabled striking osteogenesis of hGMSCs subcutaneously in nude mice and advanced new bone formation of pGMSCs in minipig maxillary bone defect model. In addition, SB431542-treated hGMSCs markedly increased bone-related proteins expression, and BMP2 and BMP4 gene expression. Conversely, SMAD3 protein-dependent TGF-β signal pathway phosphorylation was decreased. Conclusions Our study show that osteogenic differentiation of GMSCs treated with TGF-β signaling inhibitor SB431542 was increased, and SB431542-treated autologous pig GMSCs could successfully repair minipig severe maxillofacial bone defects. This preclinical study brings about a promising large bone regeneration therapeutic potential of autologous GMSCs induced by SB431542 in clinic settings. Electronic supplementary material The online version of this article (10.1186/s13287-019-1281-2) contains supplementary material, which is available to authorized users.

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Inhibition of transforming growth factor‐β signaling pathway enhances the osteogenic differentiation of unrestricted somatic stem cells

Cited by 9 publications

References 27 publications

Expression dynamics of integrin α2, α3, and αV upon osteogenic differentiation of human mesenchymal stem cells

Expression dynamics of integrin α2, α3, and αV upon osteogenic differentiation of human mesenchymal stem cells

Molecular and Genetic Mechanisms of Spinal Stenosis Formation: Systematic Review

Small molecule inhibitor of TGF-β signaling enables robust osteogenesis of autologous GMSCs to successfully repair minipig severe maxillofacial bone defects

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