Leptin-Induced Osteogenic Differentiation and Heterotopic Ossification of Achilles Tendon in TDSCsis inhibited by Rapamycin, an Inhibitor of mTORC1 Signaling Pathway

Background/Aims: Tendinopathy is a common sports injury that is manifested by the heterotopic ossification of tendon tissue. Tendon stem cells (TSCs) are prone to osteogenic differentiation under excessive tension. The underlying mechanisms remain poorly understood. Methods: Uniaxial mechanical tension (UMT) served to stretch rat tendon-derived stem cells (rTDSCs) at 8% elongation (frequency: 1 Hz; 48, 60, or 72 hours). Results: The osteogenic differentiation of rTDSCs appeared after UMT along with increased mRNA expression of the osteogenic genes Runx2, Dlx5, Alpl, and Col1a1 and increased Runx2 protein expression. Wnt5a, Wnt5b and P-JNK protein levels were also upregulated after UMT stimulation. The inhibition of JNK expression by SP600125 and JNK1-shRNA decreased UMT-induced Runx2 protein expression, and the activation of JNK expression by anisomycin and JNK1-cDNA increased UMT-induced Runx2 protein expression. When shRNA knocked down Wnt5a and Wnt5b expression in rTDSCs, the induction of Runx2 and P-JNK expression by UMT was reduced. The inhibition of Runx2 expression could be rescued by the activation of JNK expression by anisomycin. Conclusion: UMT induced the osteogenic differentiation of rTDSCs via the Wnt5a/Wnt5b/JNK signaling pathway. Accordingly, this pathway may influence the heterotopic ossification of tendon tissue subjected to excessive tension.

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Gene targeting of the transcription factor Mohawk in rats causes heterotopic ossification of Achilles tendon via failed tenogenesis

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1522054113 ◽

2016 ◽

Vol 113 (28) ◽

pp. 7840-7845 ◽

Cited By ~ 43

Author(s):

Hidetsugu Suzuki ◽

Yoshiaki Ito ◽

Masahiro Shinohara ◽

Satoshi Yamashita ◽

Shizuko Ichinose ◽

...

Keyword(s):

Achilles Tendon ◽

Osteogenic Differentiation ◽

Heterotopic Ossification ◽

Chondrogenic Differentiation ◽

Molecular Mechanisms ◽

Mechanical Load ◽

Adipogenic Differentiation ◽

Tendon Repair ◽

Mechanical Stretch ◽

Tendon Cells

Cell-based or pharmacological approaches for promoting tendon repair are currently not available because the molecular mechanisms of tendon development and healing are not well understood. Although analysis of knockout mice provides many critical insights, small animals such as mice have some limitations. In particular, precise physiological examination for mechanical load and the ability to obtain a sufficient number of primary tendon cells for molecular biology studies are challenging using mice. Here, we generated Mohawk (Mkx)−/− rats by using CRISPR/Cas9, which showed not only systemic hypoplasia of tendons similar to Mkx−/− mice, but also earlier heterotopic ossification of the Achilles tendon compared with Mkx−/− mice. Analysis of tendon-derived cells (TDCs) revealed that Mkx deficiency accelerated chondrogenic and osteogenic differentiation, whereas Mkx overexpression suppressed chondrogenic, osteogenic, and adipogenic differentiation. Furthermore, mechanical stretch stimulation of Mkx−/− TDCs led to chondrogenic differentiation, whereas the same stimulation in Mkx+/+ TDCs led to formation of tenocytes. ChIP-seq of Mkx overexpressing TDCs revealed significant peaks in tenogenic-related genes, such as collagen type (Col)1a1 and Col3a1, and chondrogenic differentiation-related genes, such as SRY-box (Sox)5, Sox6, and Sox9. Our results demonstrate that Mkx has a dual role, including accelerating tendon differentiation and preventing chondrogenic/osteogenic differentiation. This molecular network of Mkx provides a basis for tendon physiology and tissue engineering.

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Baicalin Promotes Osteogenic Differentiation of Human Cementoblast Lineage Cells Via the Wnt/β Catenin Signaling Pathway

Current Pharmaceutical Design ◽

10.2174/1381612824666181116103514 ◽

2019 ◽

Vol 24 (33) ◽

pp. 3980-3987 ◽

Cited By ~ 2

Author(s):

Aya Kimura ◽

Ryo Kunimatsu ◽

Yuki Yoshimi ◽

Yuji Tsuka ◽

Tetsuya Awada ◽

...

Keyword(s):

Osteogenic Differentiation ◽

Signaling Pathway

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Salidroside promoted osteogenic differentiation of adipose-derived stromal cells through Wnt/β-catenin signaling pathway

Journal of Orthopaedic Surgery and Research ◽

10.1186/s13018-021-02598-w ◽

2021 ◽

Vol 16 (1) ◽

Author(s):

Xiao-hua Li ◽

Fu-ling Chen ◽

Hong-lin Shen

Keyword(s):

Western Blot ◽

Osteogenic Differentiation ◽

Signaling Pathway ◽

Differentially Expressed Genes ◽

Stromal Cells ◽

Differentially Expressed ◽

Calcium Deposition ◽

Western Blot Assay ◽

Adipose Derived Stromal Cells ◽

Interfering Rna

Abstract Background Bone disease causes short-term or long-term physical pain and disability. It is necessary to explore new drug for bone-related disease. This study aimed to explore the role and mechanism of Salidroside in promoting osteogenic differentiation of adipose-derived stromal cells (ADSCs). Methods ADSCs were isolated and treated with different dose of Salidroside. Cell count kit-8 (CCK-8) assay was performed to assess the cell viability of ADSCs. Then, ALP and ARS staining were conducted to assess the early and late osteogenic capacity of ADSCs, respectively. Then, differentially expressed genes were obtained by R software. Then, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis of the differentially expressed genes were further analyzed. The expression of OCN, COL1A1, RUNX2, WNT3A, and β-catenin were measured by real-time PCR and Western blot analysis. Last, β-catenin was silenced by small interfering RNA. Results Salidroside significantly increased the ADSCs viability at a dose-response manner. Moreover, Salidroside enhanced osteogenic capacity of ADSCs, which are identified by enhanced ALP activity and calcium deposition. A total of 543 differentially expressed genes were identified between normal and Salidroside-treated ADSCs. Among these differentially expressed genes, 345 genes were upregulated and 198 genes were downregulated. Differentially expressed genes enriched in the Wnt/β-catenin signaling pathway. Western blot assay indicated that Salidroside enhanced the WNT3A and β-catenin expression. Silencing β-catenin partially reversed the promotion effects of Salidroside. PCR and Western blot results further confirmed these results. Conclusion Salidroside promoted osteogenic differentiation of ADSCs through Wnt/β-catenin signaling pathway.

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Quercetin promotes osteogenic differentiation and antioxidant responses of mouse bone mesenchymal stem cells through activation of the AMPK / SIRT1 signaling pathway

Phytotherapy Research ◽

10.1002/ptr.7010 ◽

2021 ◽

Author(s):

Nan Wang ◽

Luyao Wang ◽

Jihao Yang ◽

Zhihong Wang ◽

Liangxing Cheng

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Osteogenic Differentiation ◽

Signaling Pathway ◽

Bone Mesenchymal Stem Cells ◽

Antioxidant Responses

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BRD4 induces osteogenic differentiation of BMSCs via the Wnt/β-catenin signaling pathway

Tissue and Cell ◽

10.1016/j.tice.2021.101555 ◽

2021 ◽

pp. 101555

Author(s):

Kai Wang ◽

Zhiping Zhao ◽

Xiangyu Wang ◽

Yongtao Zhang

Keyword(s):

Osteogenic Differentiation ◽

Signaling Pathway

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RTEF-1 Inhibits Vascular Smooth Muscle Cell Calcification through Regulating Wnt/β-Catenin Signaling Pathway

Calcified Tissue International ◽

10.1007/s00223-021-00833-4 ◽

2021 ◽

Author(s):

Jingjing Cong ◽

Bei Cheng ◽

Jinyu Liu ◽

Ping He

Keyword(s):

Smooth Muscle ◽

Vascular Smooth Muscle ◽

Osteogenic Differentiation ◽

Signaling Pathway ◽

Glycogen Synthase ◽

Critical Role ◽

Transcriptional Enhancer ◽

Alizarin Red ◽

Gsk 3Β ◽

Enhancer Factor

AbstractVascular calcification (VC) is highly prevailing in cardiovascular disease, diabetes mellitus, and chronic kidney disease and, when present, is associated with cardiovascular events and mortality. The osteogenic differentiation of vascular smooth muscle cells (VSMCs) is regarded as the foundation for mediating VC. Related transcriptional enhancer factor (RTEF-1), also named as transcriptional enhanced associate domain (TEAD) 4 or transcriptional enhancer factor-3 (TEF-3), is a nuclear transcriptional factor with a potent effect on cardiovascular diseases, apart from its oncogenic role in the canonical Hippo pathway. However, the role and mechanism of RTEF-1 in VC, particularly in calcification of VSMCs, are poorly understood. Our results showed that RTEF-1 was reduced in calcified VSMCs. RTEF-1 significantly ameliorated β-glycerophosphate (β-GP)-induced VSMCs calcification, as detected by alizarin red staining and calcium content assay. Also, RTEF-1 reduced alkaline phosphatase (ALP) activity and decreased expressions of osteoblast markers such as Osteocalcin and Runt-related transcription factor-2 (Runx2), but increased expression of contractile protein, including SM α-actin (α-SMA). Additionally, RTEF-1 inhibited β-GP-activated Wnt/β-catenin pathway which plays a critical role in calcification and osteogenic differentiation of VSMCs. Specifically, RTEF-1 reduced the levels of Wnt3a, p-β-catenin (Ser675), glycogen synthase kinase-3β (GSK-3β), and p-GSK-3β (Ser9), but increased the levels of p-β-catenin (Ser33/37). Also, RTEF-1 increased the ratio of p-β-catenin (Ser33/37) to β-catenin proteins and decreased the ratio of p-GSK-3β (Ser9) to GSK-3β protein. LiCl, a Wnt/β-catenin signaling activator, was observed to reverse the protective effect of RTEF-1 overexpression on VSMCs calcification induced by β-GP. Accordingly, Dickkopf-1 (Dkk1), a Wnt antagonist, attenuated the role of RTEF-1 deficiency in β-GP-induced VSMCs calcification. Taken together, we concluded that RTEF-1 ameliorated β-GP-induced calcification and osteoblastic differentiation of VSMCs by inhibiting Wnt/β-catenin signaling pathway.

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