scholarly journals A NOTCH1/LSD1/BMP2 co-regulatory network mediated by miR-137 negatively regulates osteogenesis of human adipose-derived stem cells

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Cong Fan ◽  
Xiaohan Ma ◽  
Yuejun Wang ◽  
Longwei Lv ◽  
Yuan Zhu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Osteoblastic Differentiation ◽  
Negative Control ◽  
Bone Morphogenetic Protein 2 ◽  
Luciferase Reporter ◽  
Adipose Derived Stem Cells ◽  
Differentiation Potential ◽  
Lineage Differentiation ◽  
Morphogenetic Protein ◽  
Genes Expression

Abstract Background MicroRNAs have been recognized as critical regulators for the osteoblastic lineage differentiation of human adipose-derived stem cells (hASCs). Previously, we have displayed that silencing of miR-137 enhances the osteoblastic differentiation potential of hASCs partly through the coordination of lysine-specific histone demethylase 1 (LSD1), bone morphogenetic protein 2 (BMP2), and mothers against decapentaplegic homolog 4 (SMAD4). However, still numerous molecules involved in the osteogenic regulation of miR-137 remain unknown. This study aimed to further elucidate the epigenetic mechanisms of miR-137 on the osteogenic differentiation of hASCs. Methods Dual-luciferase reporter assay was performed to validate the binding to the 3′ untranslated region (3′ UTR) of NOTCH1 by miR-137. To further identify the role of NOTCH1 in miR-137-modulated osteogenesis, tangeretin (an inhibitor of NOTCH1) was applied to treat hASCs which were transfected with miR-137 knockdown lentiviruses, then together with negative control (NC), miR-137 overexpression and miR-137 knockdown groups, the osteogenic capacity and possible downstream signals were examined. Interrelationships between signaling pathways of NOTCH1-hairy and enhancer of split 1 (HES1), LSD1 and BMP2-SMADs were thoroughly investigated with separate knockdown of NOTCH1, LSD1, BMP2, and HES1. Results We confirmed that miR-137 directly targeted the 3′ UTR of NOTCH1 while positively regulated HES1. Tangeretin reversed the effects of miR-137 knockdown on osteogenic promotion and downstream genes expression. After knocking down NOTCH1 or BMP2 individually, we found that these two signals formed a positive feedback loop as well as activated LSD1 and HES1. In addition, LSD1 knockdown induced NOTCH1 expression while suppressed HES1. Conclusions Collectively, we proposed a NOTCH1/LSD1/BMP2 co-regulatory signaling network to elucidate the modulation of miR-137 on the osteoblastic differentiation of hASCs, thus providing mechanism-based rationale for miRNA-targeted therapy of bone defect.

scholarly journals A NOTCH1/LSD1/BMP2 Co-Regulatory Network Mediated by miR-137 Negatively Regulates Osteogenesis of Human Adipose-Derived Stem Cells

2020 ◽  
Author(s):  
Cong Fan ◽  
Xiaohan Ma ◽  
Yuejun Wang ◽  
Longwei Lv ◽  
Yuan Zhu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Osteoblastic Differentiation ◽  
Bone Morphogenetic Protein 2 ◽  
Luciferase Reporter ◽  
Adipose Derived Stem Cells ◽  
Degree Relative ◽  
Matrix Mineralization ◽  
Differentiation Potential ◽  
Lineage Differentiation ◽  
Relative Expression Level

Abstract Background: MicroRNAs have been recognized as critical regulators for the osteoblastic lineage differentiation of human adipose-derived stem cells (hASCs). Previously, we have displayed that silencing of miR-137 enhances the osteoblastic differentiation potential of hASCs partly through the coordination of lysine-specific histone demethylase 1 (LSD1), bone morphogenetic protein 2 (BMP2), and mothers against decapentaplegic homolog 4 (SMAD4). However, still numerous molecules involved in the osteogenic regulation of miR-137 remain unknown. This study aimed to further elucidate the epigenetic mechanisms of miR-137 on the osteogenic differentiation of hASCs.Methods: hASCs transfected with miR-137 overexpression or knockdown lentiviruses were used to assess the osteogenic capacity by testing the alkaline phosphatase activity, matrix mineralization degree, relative expression level of osteogenesis-associated genes and ectopic osteogenesis in nude mice. Dual-luciferase reporter assay was performed to examine the targeting of the 3' untranslated region (3' UTR) of NOTCH1 by miR-137. Interrelationships of signaling pathways of NOTCH1-hairy and enhancer of split 1 (HES1), LSD1, and BMP2-SMAD4 were thoroughly investigated by separate knockdown of NOTCH1, LSD1, and BMP2.Results: We confirmed that miR-137 directly targeted the 3' UTR of NOTCH1 while positively regulated HES1. After knocking down NOTCH1 or BMP2 individually, we found that these two signals formed a positive feedback loop and activated LSD1. In addition, LSD1 knockdown induced the expression of NOTCH1 while suppressed HES1.Conclusions: Collectively, we proposed a NOTCH1/LSD1/BMP2 co-regulatory signaling network to elucidate the modulation of miR-137 on the osteoblastic differentiation of hASCs, thus providing mechanism-based rationale for miRNA-targeted therapy of bone defect.

scholarly journals Synergistic Effects of Orbital Shear Stress onIn VitroGrowth and Osteogenic Differentiation of Human Alveolar Bone-Derived Mesenchymal Stem Cells

2014 ◽  
Vol 2014 ◽  
pp. 1-18 ◽  
Author(s):  
Ki Taek Lim ◽  
Jin Hexiu ◽  
Jangho Kim ◽  
Hoon Seonwoo ◽  
Pill-Hoon Choung ◽  
...  
Keyword(s):  
Stem Cells ◽  
Shear Stress ◽  
Mesenchymal Stem Cells ◽  
Alveolar Bone ◽  
Synergistic Effects ◽  
Osteoblastic Differentiation ◽  
Bone Morphogenetic Protein 2 ◽  
Control Group ◽  
Protein Levels ◽  
Morphogenetic Protein

Cellular behavior is dependent on a variety of physical cues required for normal tissue function. In order to mimic native tissue environments, human alveolar bone-derived mesenchymal stem cells (hABMSCs) were exposed to orbital shear stress (OSS) in a low-speed orbital shaker. The synergistic effects of OSS on proliferation and differentiation of hABMSCs were investigated. In particular, we induced the osteoblastic differentiation of hABMSCs cultured in the absence of OM by exposing hABMSCs to OSS (0.86–1.51 dyne/cm2). Activation of Cx43 was associated with exposure of hABMSCs to OSS. The viability of cells stimulated for 10, 30, 60, 120, and 180 min/day increased by approximately 10% compared with that of control. The OSS groups with stimulation of 10, 30, and 60 min/day had more intense mineralized nodules compared with the control group. In quantification of vascular endothelial growth factor (VEGF) and bone morphogenetic protein-2 (BMP-2) protein, VEGF protein levels under stimulation for 10, 60, and 180 min/day and BMP-2 levels under stimulation for 60, 120, and 180 min/day were significantly different compared with those of the control. In conclusion, the results indicated that exposing hABMSCs to OSS enhanced their differentiation and maturation.

Human adipose-derived stem cells isolated from young and elderly women: their differentiation potential and scaffold interaction during in vitro osteoblastic differentiation

Cytotherapy ◽  
2009 ◽  
Vol 11 (6) ◽  
pp. 793-803 ◽  
Author(s):  
Laura de Girolamo ◽  
Silvia Lopa ◽  
Elena Arrigoni ◽  
Matteo F. Sartori ◽  
Franz W. Baruffaldi Preis ◽  
...  
Keyword(s):  
Stem Cells ◽  
Elderly Women ◽  
Osteoblastic Differentiation ◽  
Adipose Derived Stem Cells ◽  
Differentiation Potential

184 Effect of selenium on the differentiation of porcine adipose-derived stem cells into osteoblasts

2019 ◽  
Vol 31 (1) ◽  
pp. 216
Author(s):  
L. Siegel ◽  
T. Bane ◽  
J. Bertels ◽  
K. Ratz ◽  
M. Rubessa ◽  
...  
Keyword(s):  
Stem Cells ◽  
Statistical Difference ◽  
Bone Growth ◽  
Negative Control ◽  
Nodule Formation ◽  
Adipose Derived Stem Cells ◽  
Osteogenic Medium ◽  
Alizarin Red ◽  
Differentiation Potential ◽  
Cell Therapies

Bone damage is a common occurrence and although traditional bone healing methods work well, they are not always able to fix all issues pertaining to the injury. However, recent research on stem cell therapies using adipose-derived stem cells (ASC) has shown that ASC that differentiate into osteoblasts can be used as a potentially better solution for bone injuries. Selenium is a nutritional trace element that has been found to be essential in the production of selenoproteins and bone growth. Selenium plays an integral role in osteoblast cell differentiation and proliferation. The aim of this experiment was to test whether adding selenium to osteogenic influences the differentiation potential of ASC into osteoblasts. Porcine ASC were isolated as described (Monaco et al. 2009, Open Tissue Eng. Regen. Med. J. 2, 20-33). Seven different treatments were given to the cells: a negative control of DMEM, a positive control of osteogenic medium, and 5 concentrations of selenium in the osteogenic medium (10, 5, 1, 0.5, and 0.1 µM). Medium was changed twice a week for 4 weeks, at the end of which the cells were stained with Alizarin Red S stain. The number of osteoblastic nodules in each well were counted and divided into the categories of “forming” and “formed.” A generalized linear model (GLM) procedure (SPSS Inc./IBM Corp., Chicago, IL, USA) was used to analyse the data. The least statistical difference (l.s.d.) post hoc test was used to perform statistical multiple comparison with an α-level of 0.05. The results showed (Table 1) that medium containing selenium concentrations of 0.1, 0.5, and 1.0 µM were able to develop more nodules than the osteogenic medium. There was no statistical difference in the total amount of forming and formed nodules in those 4 groups; however, there was a statistical tendency for a difference (P=0.06) between the medium with 1 µM selenium and the osteogenic medium. Medium containing 10 or 5 µM selenium had lower nodule counts (both forming and formed) than osteogenic medium. There was no statistical difference between 10 µM and DMEM. These results suggest that low concentrations of selenium have a positive effect on nodule formation, whereas higher concentrations are detrimental. Table 1.The average number (standard deviations in parentheses) of formed, forming, and total osteoblastic nodules in each of the different treatments

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