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.

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.

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 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.

scholarly journals Extracellular IL-37 promotes osteogenic differentiation of human bone marrow mesenchymal stem cells via activation of the PI3K/AKT signaling pathway

2019 ◽  
Vol 10 (10) ◽  
Author(s):  
Chenyi Ye ◽  
Wei Zhang ◽  
Kai Hang ◽  
Mo Chen ◽  
Weiduo Hou ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Signaling Pathway ◽  
Akt Signaling ◽  
Specific Gene ◽  
Akt Signaling Pathway ◽  
Calvarial Bone ◽  
Marrow Mesenchymal Stem Cells ◽  
Orthopedic Diseases

Abstract Interleukin (IL)-37, a pivotal anti-inflammatory cytokine and a fundamental inhibitor of innate immunity, has recently been shown to be abnormally expressed in several autoimmune-related orthopedic diseases, including rheumatoid arthritis, ankylosing spondylitis, and osteoporosis. However, the role of IL-37 during osteogenic differentiation of mesenchymal stem cells (MSCs) remains largely unknown. In this study, extracellular IL-37 significantly increased osteoblast-specific gene expression, the number of mineral deposits, and alkaline phosphatase activity of MSCs. Moreover, a signaling pathway was activated in the presence of IL-37. The enhanced osteogenic differentiation of MSCs due to supplementation of IL-37 was partially rescued by the presence of a PI3K/AKT signaling inhibitor. Using a rat calvarial bone defect model, IL-37 significantly improved bone healing. Collectively, these findings indicate that extracellular IL-37 enhanced osteogenesis of MSCs, at least in part by activation of the PI3K/AKT signaling pathway.

scholarly journals Effect of 5-azacytidine on the Protein Expression of Porcine Bone Marrow Mesenchymal Stem Cells in vitro

2006 ◽  
Vol 4 (1) ◽  
pp. 18-25 ◽  
Author(s):  
Neng-Sheng Ye ◽  
Rong-Li Zhang ◽  
Yan-Feng Zhao ◽  
Xue Feng ◽  
Yi-Ming Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Protein Expression ◽  
Marrow Mesenchymal Stem Cells

scholarly journals Exosomes from Bone Marrow Mesenchymal Stem Cells Inhibit Neuronal Apoptosis and Promote Motor Function Recovery via the Wnt/β-catenin Signaling Pathway

2019 ◽  
Vol 28 (11) ◽  
pp. 1373-1383 ◽  
Author(s):  
Ci Li ◽  
Guangjun Jiao ◽  
Wenliang Wu ◽  
Hongliang Wang ◽  
Shanwu Ren ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Protein Expression ◽  
Signaling Pathway ◽  
Neuronal Apoptosis ◽  
Marrow Mesenchymal Stem Cells

Severe spinal cord injury (SCI) is caused by external mechanical injury, resulting in unrecoverable neurological injury. Recent studies have shown that exosomes derived from bone marrow mesenchymal stem cells (BMSCs-Exos) might be valuable paracrine molecules in the treatment of SCI. In this study, we designed SCI models in vivo and in vitro and then investigated the possible mechanism of successful repair by BMSCs-Exos. In vivo, we established one Sham group and two SCI model groups. The Basso, Beattie, Bresnahan (BBB) scores showed that BMSCs-Exos could effectively promote the recovery of spinal cord function. The results of the Nissl staining, immunohistochemistry, and TUNEL/NeuN/DAPI double staining showed that BMSCs-Exos inhibited neuronal apoptosis. Western blot analysis showed that the protein expression level of Bcl-2 was significantly increased in the BMSCs-Exos group compared with the PBS group, while the protein expression levels of Bax, cleaved caspase-3, and cleaved caspase-9 were significantly decreased. The results of western bolt and qRT-PCR demonstrated that BMSCs-Exos could activate the Wnt/β-catenin signaling pathway effectively. In vitro, we found that inhibition of the Wnt/β-catenin signaling pathway could promote neuronal apoptosis following lipopolysaccharide (LPS) induction. These results demonstrated that BMSCs-Exos may be a promising therapeutic for SCI by activating the Wnt/β-catenin signaling pathway.

scholarly journals Bone marrow mesenchymal stem cells stimulated by bFGF up-regulated protein expression in comparison with periodontal fibroblasts in vitro

2014 ◽  
Vol 59 (3) ◽  
pp. 268-276 ◽  
Author(s):  
Renato Colenci ◽  
Luciana Reichert da Silva Assunção ◽  
Suely Regina Mogami Bomfim ◽  
Marjorie de Assis Golim ◽  
Elenice Deffune ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Protein Expression ◽  
Marrow Mesenchymal Stem Cells
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