Liposomal Nanoparticle-Mediated miR-27b Influences Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells via Peroxisome Proliferator-Activated Receptor Gamma in a Microgravity Environment

2020 ◽  
Vol 12 (11) ◽  
pp. 1301-1308
Author(s):  
Zhiwei He ◽  
Yan Zhu ◽  
Gentao Fan ◽  
Hongbo Qian
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Simulated Microgravity ◽  
Mrna Level ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells ◽  
Group A ◽  
Group B ◽  
Group D

This study was aimed at analyzing the effects of liposomal nanoparticle-based miR-27b on PPARγ and osteogenic differentiation of bone marrow mesenchymal stem cells under microgravity. The rat bone marrow mesenchymal stem cells were set as the research object, and the gyroscope was employed for simulation of microgravity. The cells were randomized into four groups, including the experimental group A (simulated microgravity+liposomal nanoparticle-mediated miR-27b transfection group), as well as the control groups: group B (simulated microgravity+negative control group), group C (simulated microgravity+transfection reagent group) and group D (normal gravity+liposomal nanoparticle-mediatedmiR-27b transfection group). After a two-week osteogenic induction in vitro, staining was performed to assess the lipogenesis rate of the samples. In addition, ALP activity and PPARγ mRNA level was detected. The number of alizarin staining-positive osteogenic nodules and ALP activity (0.21±0.44 King unit) in group A was significantly diminished compared to those in group B, C, and D. Moreover, its lipogenesis rate (9.31±1.02%) and PPARγ mRNA level (1.86±0.39) were significantly higher than those in group B, C, and D (P < 0.05). The number of alizarin staining-positive osteogenic nodules and ALP activity (0.96±0.18 King unit) in group D were significantly reduced in comparison with those in groups B and C, while the lipogenesis rate (4.86±0.77%) and PPARγ mRNA level (0.93±0.34) were significantly higher than those in group B and C (P < 0.05) without difference between group B and group C (P > 0.05). Under a microgravity condition, liposomal nanoparticle-mediated miR-27b can impede the differentiation of BMSCs into osteoblasts via regulating PPARγ signal transduction.

miRNA-126 Inhibits Osteogenic Differentiation of Rat Bone Marrow Mesenchymal Stem Cells (BMSCs)

2022 ◽  
Vol 12 (4) ◽  
pp. 794-799
Author(s):  
Le Chang ◽  
Wei Duan ◽  
Chuang Wang ◽  
Jian Zhang
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Mrna Level ◽  
Alizarin Red ◽  
Runx2 Expression ◽  
Alp Activity ◽  
Smad4 Protein ◽  
Marrow Mesenchymal Stem Cells

This study was to determine whether microRNA (miRNA)-126 regulates osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). Rat BMSCs were extracted and stimulated for osteogenic differentiation. Functional experiments were conducted to assess miR-126’s impact on BMSCs differentiation. Western blot and RT-qPCR determined miR-126 expression. ALP activity detection and alizarin red staining detection were also performed. After osteogenic differentiation of BMSCs, miR-126 expression was gradually decreased over time. Overexpression of miR-26 decreased ALP activity, Notch signaling activity as well as declined Runx2 expression and calcium Salt nodules after treatment. Importantly, we found that Smad4 serves as a target of miR-126 while upregulation of the miRNA was accompanied with the decreased Smad4 protein expression without affecting the Smad4 mRNA level. In conclusion, miR-126 restrains osteogenic differentiation through inhibition of SMAD4 signaling, providing a novel insight into the mechanism.

Progranulin Promotes Osteogenic Differentiation and Osteosynthesis of Bone Marrow Mesenchymal Stem Cells and Alleviates Osteoporosis Through Phosphatidylinositol 3-Kinase/Protein Kinase B/Peroxisome Proliferator-Activated Receptor γ Pathway

2020 ◽  
Vol 10 (12) ◽  
pp. 1865-1870
Author(s):  
Yang Ying ◽  
Binghao Zhao ◽  
Wei Qian ◽  
Li Xu
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Mrna Level ◽  
Control Group ◽  
Peroxisome Proliferator ◽  
Mineral Density ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

Bone marrow mesenchymal stem cells (BMSCs) have self-renewal potential with multi-directional differentiation. Progranulin prevents bone degradation, inhibits inflammation and protects bone tissue. However, the role of Progranulin in osteoporotic BMSCs is unclear. Osteoporosis (OP) rat models were prepared by ovarian removal and treated with different doses (5 and 10 μM) of Progranulin followed by analysis of BMP-2 level by ELISA, bone mineral density and ALP activity. OP rat BMSCs were isolated and assigned into control group and Progranulin group followed by analysis of Progranulin level by ELISA, cell proliferation by MTT assay, RUNX2 and COL1A1 mRNA level by Real time PCR, and PI3K/Akt/PPARγ signaling protein level by Western blot. Progranulin treatment of OP rats dose-dependently increased BMP-2 expression, bone density and ALP activity. Compared with OP group, there were significant differences (P <0.05). Progranulin expression and BMSCs proliferation was increased, and RUNX2 and COL1A1 mRNA expression was elevated in Progranulin-treated OP group along with increased PI3K/Akt expression and decreased PPARγ protein expression. Compared with OP group, the difference was statistically significant, and the change was more significant with increasing concentration (P <0.05). Progranulin promotes BMSCs osteogenic differentiation and proliferation by regulating PI3K/Akt/PPARγ signaling pathway, which is beneficial for OP rats’ bone synthesis.

scholarly journals The Effect of Bone Marrow Mesenchymal Stem Cells Application on Distracted Bone Quality during Rapid Rate of Distraction Osteogenesis

2018 ◽  
Vol 11 (3) ◽  
pp. 192-198
Author(s):  
Marwa El Kassaby ◽  
Khaled Abd El Kader ◽  
Nahed Khamis ◽  
Alaa Al Hammoud ◽  
Alaa Ben Talb ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Bone Quality ◽  
Distraction Osteogenesis ◽  
Rapid Rate ◽  
Marrow Mesenchymal Stem Cells ◽  
Group A ◽  
Group B ◽  
Post Hoc ◽  
Group D

Distraction osteogenesis (DO) bone regenerate usually suffers from an inferior quality especially with rapid rate. This study was conducted to investigate the effect of mesenchymal stem cells (MSCs) application on different rates of distraction bone quality. Twenty-four goats were divided into group A with standard DO and group B with rapid distraction osteogenesis (RDO) both aided by MSCs. Group C with standard DO and group (D) with RDO were controls. Kruskal–Wallis test and Conover's post hoc analysis was used to evaluate significance ( p = 0.05). Histomorphometry showed a strongly significant (SS) increase ( p = 0.00036) in trabecular bone (TB) in group A (TB = 174.7 μm, SD = 33.5) and group B (TB = 166.8 μm, SD = 14) compared with group C (TB = 115.4 μm, SD = 19.6) and group D (TB = 86.1 μm, SD = 9.3). There was SS decrease ( p = 0.00093) in osteoid percentage (OP) in group A (OP = 13.4%, SD = 2) and group B (OP = 11.5%, SD = 6.5) compared with group C (OP = 27.3, SD = 3.5) and group D (OP = 26.2%, SD = 2.6). Energy dispersive X-ray showed a nonsignificant increase ( p = 0.11) in calcification (Ca2+%) in group A (Ca2+% = 17.6%, SD = 4.9) and group B (Ca2+% = 17.6%, SD = 4.3) compared with group C (Ca2+ % = 14.2%, SD = 6.7) and group D (Ca2+% = 11.5%, SD = 2.4). MSCs application improved microscopic bone quality during standard DO and RDO. However, macroscopic bone quality improvement still needs further investigation.

Interleukin-6 Promotes Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells by Activating Wnt Signaling Pathway

2019 ◽  
Vol 9 (9) ◽  
pp. 1261-1265
Author(s):  
Hai Nan ◽  
Yun Zhang
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Wnt Signaling ◽  
Osteogenic Differentiation ◽  
Mrna Level ◽  
Control Group ◽  
Blocking Group ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

Bone marrow mesenchymal stem cells (BMSCs) can differentiate into adipocytes, osteoblasts. Osteoporosis is closely related to BMSCs osteogenic differentiation, and IL-6 is closely related to osteoporosis. This study assessed the effect of IL-6 on BMSCs osteogenic differentiation. Rat BMSCs were cultured and osteogenic induction of BMSCs was performed in the presence of different concentrations (0, 10, 100 ng/ml) of IL-6 followed by analysis of IL-6 level by ELISA, ALP activity by the instructions of the alkaline phosphatase (ALP) detection kit, IL-6, Runx2 and OCN mRNA level, and level of β-catenin by Western blot. Compared with 0 d, IL-6 protein content and IL-6 mRNA expression in cell culture medium was increased significantly on day 7 d, 14 d and 21 d. Compared with 0 ng/ml group, 10, 100 ng/ml IL-6 group showed significantly increased ALP activity and Runx2 and OCN mRNA level in a dose-response relationship. β-catenin was increased significantly in 100 ng/ml IL-6 group. No difference of ALP activity and the expression of osteogenic differentiationmarkers was found between blocking group and control group, which was significantly lower than those in experimental group. IL-6 can promote BMSCs osteogenic differentiation through Wnt signaling.

Effect of Yes Associated Protein on Osteogenic/Adipogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Under Oxidative Stress

2021 ◽  
Vol 11 (8) ◽  
pp. 1636-1642
Author(s):  
Yonghuan Zhou ◽  
Guotang Lan ◽  
Yan Zhou ◽  
Tianhao Qu ◽  
Qing Xiong
Keyword(s):  
Oxidative Stress ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Mrna Expression ◽  
Adipogenic Differentiation ◽  
Mrna Level ◽  
Control Group ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

Oxidative stress affects bone marrow mesenchymal stem cells (BMSCs). YAP is an effector in Hippo signaling, but its’ role in BMSCs osteogenesis/adipogenesis under oxidative stress has not been reported. Mice BMSCs were isolated and assigned into 3 groups, normal control group; oxidative stress group; and YAP group (transfected with YAP plasmid) followed by analysis of YAP expression by Real time PCR. After 14 days of osteogenesis or adipogenic induction, RUNX2, OPN, FABP4 and PPARγ2 mRNA level was measured along with ROS and SOD activities, ALP activity and Wnt5 expression by western blot. Under oxidative stress, YAP expression significantly decreased, RUNX2 and OPN mRNA expression decreased, ROS expression increased, SOD activity decreased, FABP4 and PPARγ2 protein expression increased, ALP activity and Wnt5 expression decreased (P <0.05). YAP plasmid transfection could significantly up-regulate YAP, RUNX2 and OPN mRNA level, decrease ROS, increase SOD and ALP activity, reduce FABP4 and PPARγ2 mRNA expression and increase Wnt5 expression (P <0.05). YAP level in BMSCs is decreased under oxidative stress. Up-regulating YAP can improve the redox balance, promote BMSCs osteogenic differentiation under oxidative stress and inhibit their differentiation to adipocytes.

Effect of αCGRP on Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Through Smad/Runx2 Signaling Pathway

2020 ◽  
Vol 10 (6) ◽  
pp. 868-873
Author(s):  
Shengxiang Huang ◽  
Haibo Mei ◽  
Rongguo He ◽  
Kun Liu ◽  
Jin Tang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Proliferation ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Real Time ◽  
Real Time Pcr ◽  
Caspase 3 ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

The α-calcitonin gene-related peptide (α-CGRP) regulates bone metabolism and has potential applications in enhancing bone remodeling in vivo. However, α-CGRP's role in bone marrow mesenchymal stem cells (BMSCs) osteogenic differentiation remain unclear. Rat BMSCs were separated into control group, α-CGRP group and α-CGRP siRNA group, in which BMSCs were transfected with α-CGRP plasmid and α-CGRP siRNA respectively followed by analysis of α-CGRP level by real time PCR and ELISA, cell proliferation by MTT assay, Caspase 3 activity, ALP activity, formation of calcified nodules by alizarin red staining, Smad1 and Smad7 level by Western blot and Runx2 by real time PCR. αCGRP transfection into BMSCs significantly up-regulated CGRP, which could promote cell proliferation, inhibit Caspase 3 activity, promote ALP activity, increase calcified nodules formation and upregulate Smad1, Smad7 and Runx2 compared to control (P < 0.05); transfection of αCGRP siRNA significantly down-regulated CGRP in BMSCs, inhibited cell proliferation, promoted Caspase 3 activity, inhibited ALP activity, inhibited calcified nodules formation and downregulate Smad1, Smad7 and Runx2 (P < 0.05). αCGRP overexpression promotes the Smad/Runx2 signaling, which in turn promotes BMSCs proliferation and osteogenesis. Decreased αCGRP level inhibits Smad/Runx2 signaling, promotes BMSCs apoptosis, inhibits proliferation and osteogenic differentiation.

Effect of SOX9 on Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Through WNTβ/Catenin Pathway

2019 ◽  
Vol 9 (10) ◽  
pp. 1429-1434
Author(s):  
Qing Yang ◽  
Cheng Li ◽  
Manli Yan ◽  
Chunhua Fang
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Proliferation ◽  
Mesenchymal Stem Cells ◽  
Protein Expression ◽  
Osteogenic Differentiation ◽  
Real Time ◽  
Real Time Pcr ◽  
Alp Activity ◽  
Marrow Mesenchymal Stem Cells

Bone marrow mesenchymal stem cells (BMSCs) can be differentiated into different types of cells. SOX9 involves in the development and progression of various diseases. Our study aims to assess SOX9's effect on osteogenic differentiation of BMSCs and its related regulatory mechanisms. Rat BMSCs were isolated and randomly divided into control group, SOX9 group and SOX9 siRNA group, which was transfected with pcDNA-SOX9 plasmid or SOX9 siRNA respectively followed by analysis of SOX9 expression by Real time PCR, cell proliferation by MTT assay, Caspase3 and ALP activity, GSK-3β expression and Wntβ/Catenin Signaling pathway protein expression by Western blot, and expression of osteogenic genes Runx2 and BMP-2 by Real time PCR. Transfection of pcDNA-SOX9 plasmid into BMSCs significantly inhibited cell proliferation, promoted Caspase3 activity, decreased ALP activity and downregulated Runx2 and BMP-2, increased GSK-3β expression and decreased Wntβ/Catenin expression protein expression (P< 0.05). SOX9 siRNA transfection significantly promoted cell proliferation, inhibited Caspase3 activity, increased ALP activity and upregulated Runx2 and BMP-2, downregulated GSK-3β and increased Wntβ/Catenin expression. SOX9 regulates BMSCs proliferation and osteogenic differentiation through Wntβ/Catenin signaling pathway.

Effect of Substance P on Bone Marrow Mesenchymal Stem Cells Differentiation in High Glucose Environment

2020 ◽  
Vol 10 (2) ◽  
pp. 252-258
Author(s):  
HeTong Yu ◽  
Yanjun Li ◽  
Xiaowei Ren ◽  
Huanhuan Zhao ◽  
Chong Nan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Proliferation ◽  
Mesenchymal Stem Cells ◽  
Substance P ◽  
High Glucose ◽  
Mtor Signaling ◽  
Alp Activity ◽  
Ampk Phosphorylation ◽  
Marrow Mesenchymal Stem Cells

Bone marrow mesenchymal stem cells (BMSCs) can be used to treat bone defects. The neuropeptide substance P (SP) plays an important role in a variety of life activities. However, the effect of SP on BMSCs differentiation in high glucose environment remains unclear. Rat BMSCs were isolated and divided into control group; high glucose group; and SP group. The secretion of SP was detected by ELISA; cell proliferation was detected by MTT assay; apoptosis activity was detected by Cas-pase3 activity kit. Real time PCR was performed to measure Bax and Bcl-2 expression. Alizarin red staining was to detect calcified nodule formation. Western blot was done to measure AMPK/mTOR signaling protein expression. In high glucose environment, SP secretion was significantly decreased, along with increased cell proliferation, Caspase3 activity and Bax expression. Meanwhile, Bcl-2 expression, ALP activity and calcified nodules formation was significantly decreased with reduced AMPK phosphorylation and increased mTOR expression (P < 0.05). SP addition in high glucose environment significantly promoted SP secretion and cell proliferation, decreased Caspase3 activity and Bax expression, increased Bcl-2 expression, ALP activity and calcification nodules formation with increased AMPK phosphorylation and decreased mTOR expression (P < 0.05). In high glucose environment, SP secretion is decreased in BMSCs. Up-regulation of SP in BMSCs cells in high glucose environment inhibit the apoptosis of BMSCs and promote cell proliferation and osteogenesis by regulating AMPK/mTOR signaling pathway.

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