scholarly journals BMSC-Derived Exosomes Ameliorate Osteoarthritis by Inhibiting Pyroptosis of Cartilage via Delivering miR-326 Targeting HDAC3 and STAT1//NF-κB p65 to Chondrocytes

2021 ◽  
Vol 2021 ◽  
pp. 1-26
Author(s):  
Honggang Xu ◽  
Bin Xu
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Animal Experiments ◽  
Luciferase Reporter ◽  
Specific Gene ◽  
Migration Ability ◽  
Marrow Mesenchymal Stem Cells ◽  
Oa Chondrocytes ◽  
Related Proteins ◽  
And Cartilage

Background. In the past decade, mesenchymal stem cells (MSCs) have been widely used for the treatment of osteoarthritis (OA), and noncoding RNAs in exosomes may play a major role. Aim. The present study is aimed at exploring the effect and mechanism of miR-326 in exosomes secreted by bone marrow mesenchymal stem cells (BMSCs) on pyroptosis of cartilage and OA improvement. Methods. Exosomes from BMSCs (BMSC-Exos) were isolated and identified to incubate with OA chondrocytes. Proliferation, migration, specific gene and miR-326 expression, and pyroptosis of chondrocytes were detected. BMSCs or chondrocytes were transfected with miR-326 mimics or inhibitors to investigate the effect of miR-326 in BMSC-Exos on pyroptosis of chondrocytes and the potential mechanism. Finally, a rat OA model was established to verify the effect and mechanism of miR-326 in BMSC-Exos on cartilage of pyroptosis. Results. Incubation with BMSC-Exos could significantly improve the survival rate, migration ability, and chondrocyte-specific genes (COL2A1, SOX9, Agg, and Prg4) and miR-326 expression of OA chondrocytes and significantly inhibit pyroptosis of chondrocytes by downregulation of the levels of inflammatory cytokines, Caspase-1 activity, and pyroptosis-related proteins such as GSDMD, NLRP3, ASC, IL-1β, and IL-18 ( P < 0.01 ). PKH26 labeling confirmed the uptake of BMSC-Exos by chondrocytes. Incubation with exosomes extracted from BMSCs overexpressing miR-326 can significantly repress the pyroptosis of chondrocytes, while knockdown of miR-326 had the opposite effect ( P < 0.01 ). The same result was also demonstrated by direct interference with the expression level of miR-326 in chondrocytes ( P < 0.01 ). In addition, we found that the overexpression of miR-326 significantly inhibited the expression of HDAC3 and NF-κB p65 and significantly promoted the expression of STAT1, acetylated STAT1, and acetylated NF-κB p65 in chondrocytes ( P < 0.01 ). The targeted relationship between miR-326 and HDAC3 was verified by dual-luciferase reporter assay. Animal experiments confirmed the mechanism by which miR-326 delivered by BMSC-Exos inhibits pyroptosis of cartilage by targeting HDAC3 and STAT1/NF-κB p65 signaling pathway. Conclusion. BMSC-Exos can deliver miR-326 to chondrocytes and cartilage and improve OA by targeting HDAC3 and STAT1//NF-κB p65 to inhibit pyroptosis of chondrocytes and cartilage. Our findings provide a new mechanism for BMSC-Exos to treat OA.

scholarly journals Protein Expression of Mesenchymal Stem Cells after Transfection of pcDNA3.1−-hVEGF165by Ultrasound-Targeted Microbubble Destruction

2011 ◽  
Vol 2011 ◽  
pp. 1-4 ◽  
Author(s):  
Zhaoxia Pu ◽  
Xiangdong You ◽  
Qiyuan Xu ◽  
Feng Gao ◽  
Xiaojie Xie ◽  
...  
Keyword(s):  
Drug Delivery ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Protein Expression ◽  
Specific Gene ◽  
Elisa Kit ◽  
The Third ◽  
Marrow Mesenchymal Stem Cells ◽  
Organ Specific ◽  
A New Technique

Ultrasound-targeted microbubble destruction (UTMD) has been proposed as a new technique for organ-specific gene transfer and drug delivery. This study was performed to investigate the effect of UTMD on marrow mesenchymal stem cells (MSCs) transfected with pcDNA3.1−-hVEGF165.pcDNA3.1−-hVEGF165were transfected into the third passage of MSCs, with or without UTMD under different ultrasound conditions. Protein expression was quantified by hVEGF165-ELISA kit after transfection for 24, 48, and 72 hours. UTMD-mediated transfection of MSCs yielded a significant protein expression. UTMD of mechanic index (MI) 0.6 for 90 seconds led to the highest level of protein expression.

scholarly journals MicroRNA-16, via FGF2 Regulation of the ERK/MAPK Pathway, Is Involved in the Magnesium-Promoted Osteogenic Differentiation of Mesenchymal Stem Cells

2020 ◽  
Vol 2020 ◽  
pp. 1-14 ◽  
Author(s):  
Hong Qi ◽  
Yang Liu ◽  
Lu Wu ◽  
Su Ni ◽  
Jing Sun ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Mapk Pathway ◽  
Luciferase Reporter ◽  
Erk Mapk ◽  
Marrow Mesenchymal Stem Cells ◽  
Elevated Expression ◽  
Reporter Assays ◽  
Induced Changes

microRNAs (miRNAs) participate in the osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs). However, few reports have discussed the effect of miRNAs on the magnesium chloride (MgCl2)-induced promotion of osteogenic differentiation of BMSCs, a process involved in the healing of bone tissue. As determined in the present investigation, MgCl2 decreased miR-16 levels; increased levels of fibroblast growth factor 2 (FGF2), p-p38, and p-ERK; and promoted the osteogenic differentiation of BMSCs. Enhancement of miR-16 levels by an miR-16 mimic blocked these MgCl2-induced changes. Moreover, luciferase reporter assays confirmed that miR-16 binds to the 3′UTR region of FGF2 mRNA. Down-regulation of FGF2 blocked the MgCl2-induced increases of p-p38 and p-ERK and the promotion of the osteogenic differentiation of BMSCs. Furthermore, over-expression of miR-16 attenuated the MgCl2-induced overproduction of p-p38 and p-ERK1/2 and the high levels of osteogenic differentiation, effects that were reversed by elevated expression of FGF2. In summary, the present findings provide a mechanism by which miR-16 regulates MgCl2-induced promotion of osteogenic differentiation by targeting FGF2-mediated activation of the ERK/MAPK pathway.

scholarly journals miR-142-5p in Bone Marrow-Derived Mesenchymal Stem Cells Promotes Osteoporosis Involving Targeting Adhesion Molecule VCAM-1 and Inhibiting Cell Migration

2018 ◽  
Vol 2018 ◽  
pp. 1-7 ◽  
Author(s):  
Zhaowei Teng ◽  
Xueguan Xie ◽  
Yun Zhu ◽  
Jianping Liu ◽  
Xingbo Hu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Adhesion Molecule ◽  
The Elderly ◽  
Luciferase Reporter ◽  
Sequencing Analysis ◽  
Migration Ability ◽  
Aberrant Expression ◽  
Migration Assay

Osteoporosis is a systemic bone metabolic disease that is highly prevalent in the elderly population, particularly in postmenopausal women, which results in enhanced bone fragility and an increased susceptibility to fractures. However, the underlying molecular pathogenesis mechanisms still remain to be further elucidated. In this study, in a rat ovariectomy- (OVX-) induced postmenopausal osteoporosis model, aberrant expression of a microRNA miR-142-5p and vascular cell adhesion molecule 1 (VCAM-1) was found by RNA sequencing analysis and qRT-PCR. Using a dual-luciferase reporter assay, we found that miR-142-5p can bind to and decrease expression of VCAM-1 mRNA. Such reduction was prohibited when the miR-142-5p binding site in VCAM-1 3′UTR was deleted, and Western blotting analyses validated the fact that miR-142-5p inhibited the expression of VCAM-1 protein. Bone marrow-derived mesenchymal stem cells (BMMSCs) transfected with miR-142-5p showed a significantly decreased migration ability in a Transwell migration assay. Collectively, these data indicated the important role of miR-142-5p in osteoporosis development involving targeting VCAM-1 and inhibiting BMMSC migration.

scholarly journals Human amnion-derived mesenchymal stem cells promote osteogenic differentiation of lipopolysaccharide-induced human bone marrow mesenchymal stem cells via ANRIL/miR-125a/APC axis

2020 ◽  
Author(s):  
Yuli Wang ◽  
Fengyi Lv ◽  
Lintong Huang ◽  
Hengwei Zhang ◽  
Bing Li ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Noncoding Rna ◽  
Human Bone ◽  
Human Bone Marrow ◽  
Luciferase Reporter ◽  
Human Amnion ◽  
Marrow Mesenchymal Stem Cells

Abstract Background and aim: Periodontitis is a chronic inflammatory disease inducing the absorption of alveolar bone and leading to tooth loss. Human amnion–derived mesenchymal stem cells (HAMSCs) have been used for studying inflammatory processes. This study aimed to explore the role of long noncoding RNA (lncRNA) antisense noncoding RNA in the INK4 locus (ANRIL) in HAMSC-driven osteogenesis in lipopolysaccharide (LPS)-induced human bone marrow mesenchymal stem cells (HBMSCs).Methods: The cells were incubated with a co-culture system. Reactive oxygen species (ROS) level and superoxide dismutase (SOD) activity were used to detect the oxidative stress level. Flow cytometry was performed to determine cell proliferation. The alkaline phosphatase (ALP) activity, Alizarin red assay, cell transfection, and rat mandibular defect model were used to evaluate the osteogenic differentiation. Quantitative real-time reverse transcription–polymerase chain reaction (RT-PCR), Western blot analysis, dual-luciferase reporter assay, and immunofluorescence staining were used to evaluate the molecular mechanisms.Results: This study showed that HAMSCs promoted the osteogenesis of LPS-induced HBMSCs, while the ANRIL level in HBMSCs decreased during co-culture. ANRIL had no significant influence on the proliferation of LPS-induced HBMSCs. However, its overexpression inhibited the HAMSC-driven osteogenesis in vivo and in vitro, whereas its knockdown reversed these effects. Mechanistically, this study found that downregulating ANRIL led to the overexpression of microRNA-125a (miR-125a), and further contributed to the competitive binding of miR-125a and adenomatous polyposis coli (APC), thus significantly activating the Wnt/β-catenin pathway.Conclusion: The study indicated that HAMSCs promoted the osteogenic differentiation of LPS-induced HBMSCs via the ANRIL/miR-125a/APC axis, and offered a novel approach for periodontitis therapy.

MicroRNA-21 Derived from Exosomes of Bone Marrow Mesenchymal Stem Cells (BMSCs) Alleviates Pancreatic Cancer by Downregulation of a Disintegrin and Metalloprotease 9 (ADAM9)

2021 ◽  
Vol 11 (12) ◽  
pp. 2346-2356
Author(s):  
Jie Zhong ◽  
Juncheng Tang ◽  
Kun Huang
Keyword(s):  
Pancreatic Cancer ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Specific Inhibitor ◽  
Underlying Mechanism ◽  
Luciferase Reporter ◽  
Invasion And Migration ◽  
Marrow Mesenchymal Stem Cells ◽  
Novel Strategy

We aimed to explore underlying mechanism by which microRNA-21 (miR-21) derived from bone marrow mesenchymal stem cells (BMSCs) extracted exosomes (exo) in pancreatic cancer (PC). Bioinformatics analysis identified candidate miRNAs and target mRNAs in PC those were verified by luciferase reporter assay. BMSCs and exo were isolated and co-cultivated with PC cells. PC cells were then treated with plasmids loaded with miR-21 or a disintegrin and metalloprotease 9 (ADAM9), followed by detection of invasion, metastasis and apoptosis through Transwell assay and flow cytometry. MiR-21 was downregulated in PC tissues and cells, while ADAM9 was upregulated and positively correlated with poor prognosis. Overexpression of miR-21 restrained the capacities of proliferation, invasion and migration of PC cells by inhibiting ADAM9 expression. Specific inhibitor GW4869 reduced release of exo and declined miR-21 expression. Treatment with BMSC-exo containing miR-21 suppressed the malignant characteristics of cancer cells. MiR-21 derived from exo of BMSCs inhibited PC progression by ADAM9 down-regulation, providing insight into novel strategy against PC.

Bone Marrow Mesenchymal Stem Cells (BMSC)-Originated miR-133 Impedes Migration and Invasiveness of Glioma Cells While Enhancing Apoptosis via Targeting the Receptor for Activated C Kinase 1 (RACK1)

2021 ◽  
Vol 11 (9) ◽  
pp. 1818-1824
Author(s):  
Jiangbo Xiong ◽  
Sheng Liu ◽  
Bin Xiang ◽  
Weibo Zhang ◽  
Jun Du ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Glioma Cells ◽  
Luciferase Reporter ◽  
Mechanistic Study ◽  
Migration And Invasion ◽  
Luciferase Reporter Gene ◽  
Gene Assay ◽  
Marrow Mesenchymal Stem Cells

This study aims to dissect the effects of bone marrow mesenchymal stem cells (BMSC) on the in vitro activity of glioma cells and the underlying mechanisms. The glioma cells were transfected with miR-133 mimics, RACK1-Vector, negative control (NC) and miR-133 mimic+RACK1-Vector, respectively, and then co-cultured with BMSC followed by analysis of miR-133 expression via PCR, apoptosis via flow cytometry, proliferation via CCK-8, invasion and migration via Transwell assay, the expression of proteins involved in apoptosis, anti-apoptosis, invasiveness and RACK1 by western blot, and the targeting relationship between miR-133 and RACK1 by dual-luciferase reporter gene assay. In comparison with normal glial cells, glioma cells exhibited a significantly diminished miR-133 level. miR-133 was upregulated in glioma cells after co-culture with BMSC, along with significantly restrained proliferation rate, migration and invasion activities as well as reduced protein levels (MMP-2, Vimentin, N-cadherin and MMP-9). Mechanistic study showed that miR-133 can retard the expression of RACK1, thereby impeding the invasion, migration and proliferation activities of cells while triggering cell apoptosis. In conclusion, BMSC-originated miR-133 can impede the migration and invasion while enhancing the apoptosis of glioma cells via targeting RACK1.

Mechanism of miR-506 Targeting Autophagy and Apoptosis-Associated Gene Beclin1 to Promote Bone Marrow Mesenchymal Stem Cells (BMSCs) Differentiation and Metastasis to Breast Cancer

2021 ◽  
Vol 11 (12) ◽  
pp. 2502-2506
Author(s):  
Qiumei Liu ◽  
Yanyan Wu ◽  
Jian Ye
Keyword(s):  
Breast Cancer ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Cancer Cells ◽  
Breast Cancer Cells ◽  
Luciferase Reporter ◽  
Differentiation Capacity ◽  
Marrow Mesenchymal Stem Cells ◽  
Metastasis To Breast

This study investigates miR-506 targeting the autophagy and apoptosis-related gene Beclin1 and analyzes the mechanism of its effect on bone marrow mesenchymal stem cells (BMSCs) differentiation and metastasis to breast cancer. Detection of miRNA-506 expression in BMSCs and breast cancer cells was done by Real-time PCR. A luciferase reporter system analyzed the targeting relationship between Beclin1 and miR-506. miR-NC group, BMSCs induction group, siRNA-NC group, and siRNA-Beclin1 group was set to measure Beclin1 expression, cell differentiation and migration by transwell assay, cell viability by MTT assay, proliferation by EdU staining and apoptosis and cycle by flow cell assay. miRNA-506 showed a high expression in breast cancer cells and low expression in BMSCs. miRNA-506 mimics significantly promote breast cancer cell proliferation which was inhibited by miRNA-506 inhibitors. The expression of Beclin1mRNA was significantly higher and miR-506 was lower in breast cancer cells. BMSCs induction significantly downregulated Beclin1 expression, increased miR-506 expression, and promoted cell invasive differentiation and metastatic capacity. In conclusion, elevated miR-506 expression was associated with decreased Beclin1 expression and increased metastatic differentiation capacity of breast cancer cells, which could effectively increase differentiation capacity and metastatic differentiation after induction by BMSCs.

scholarly journals Letter to the Editor: Is miR-199b-3p really involved in the migration ability of bone marrow mesenchymal stem cells?

2017 ◽  
Vol 101 (5) ◽  
pp. 1067-1067
Author(s):  
Blanca Torrejón ◽  
Ion Cristóbal ◽  
Juan Madoz-Gúrpide ◽  
Federico Rojo ◽  
Jesús García-Foncillas
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Migration Ability ◽  
Letter To The Editor ◽  
Marrow Mesenchymal Stem Cells

scholarly journals Preparing Sheep Bladder Scaffold and Examining the Differentiation of Mesenchymal Stem Cell into Myocytes on Scaffolds

2021 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Reza Najafi ◽  
Asadollah Asadi ◽  
Saber Zahri ◽  
Arash Abdolmaleki
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Maximum Level ◽  
Specific Gene ◽  
Biological Scaffolds ◽  
Marrow Mesenchymal Stem Cells ◽  
Physical And Chemical

Background: Tissue engineering may be used to repair, preserve, or improve tissues and organs. In this regard, acellular biological scaffolds are mainly used to reconstruct damaged tissues in regenerative medicine. Objectives: The present study examined the in vitro process of myocytes differentiated from bone marrow mesenchymal stem cells (BM‐MSCs) on the sheep bladder scaffold induced by 5-azacytidine. Methods: Decellularization was performed using a mixed method (physical and chemical) to prepare scaffolds kept at -20°C. The 5-azacytidine was used to induce BM‐MSCs to myocytes. Moreover, the muscle-specific gene expression (Desmin, α-Actinin, Myo D) was evaluated using the RT-PCR method. Results: It was revealed that BM‐MSCs on the scaffold had high proliferation and differentiation potentials. Desmin and α-Actinin gene expression marked the differentiation at the end of the fourth week. Moreover, the results of Masson’s trichrome staining at the end of the second, third and, fourth weeks also indicated that the first differentiation signs emerged at the end of the second week. Furthermore, differentiation reached its maximum level during the fourth week. Conclusions: According to the findings, combining physical and chemical methods was the best technique to prepare the bladder scaffold so that the bone marrow mesenchymal stem cells can be differentiated into myocytes on the bladder scaffold affected by 5-azacytidine (5 µmol), and As the induction time increases to day 28, myocyte cells become more developed.

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