scholarly journals Adenovirus-Mediated Expression of BMP-2 and BFGF in Bone Marrow Mesenchymal Stem Cells Combined with Demineralized Bone Matrix For Repair of Femoral Head Osteonecrosis in Beagle Dogs

2017 ◽  
Vol 43 (4) ◽  
pp. 1648-1662 ◽  
Author(s):  
Wu-Xun Peng ◽  
Lei Wang
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Femoral Head ◽  
Bending Strength ◽  
Demineralized Bone Matrix ◽  
Bone Matrix ◽  
Beagle Dogs ◽  
X Ray ◽  
Marrow Mesenchymal Stem Cells

Background: This study investigated the effect of using adenovirus-mediated expression of bone morphogenetic protein 2 (Ad-BMP-2) and basic fibroblast growth factor (bFGF) in bone marrow mesenchymal stem cells (BMSCs) in combination with a demineralized bone matrix (DBM) to repair osteonecrosis of the femoral head (ONFH) in Beagle dogs. Methods: A total of 30 Beagle dogs were selected for the isolation of BMSCs, which were cultured and transfected with the recombinant adenovirus vector Ad-BMP2-bFGF-GFP (carrying BMP-2 and bFGF) or a control adenovirus plasmid (encoding green fluorescent protein (Ad-GFP)). The expression of the transfected BMP-2 and bFGF proteins was detected by Western blotting. After transfection, the BMSCs were induced to undergo osteoblastic differentiation. The DBM was prepared to construct a DBM/BMSC complex. Beagle models of canine femoral head defects and necrosis were established and divided into control, DBM, DBM/BMSC, vector Ad-BMP2-bFGF-GFP and Ad-GFP groups. The composite graft was then implanted, and new bone morphology was visualized via X-ray at 3, 6 and 12 weeks after the operation. Hematoxylin and eosin (HE) staining and Masson’s trichrome staining were used to identify new bone formation. Immunohistochemistry was performed to calculate the density of new blood vessels. The compressive and bending strength of the BMSCs was evaluated at 12 weeks after the operation. Results: BMSCs were successfully isolated. The protein expression of BMP-2 and bFGF was significantly higher in the Ad-BMP-2/bFGF group than the normal and Ad-GFP groups. Compared with the control group, at 12 weeks after the operation, the DBM, DBM/BMSC, vector Ad-BMP2-bFGF-GFP and Ad-GFP groups showed a larger area of new bone, higher X-ray scores, greater neovascularization density, and increased compressive and bending strength. The most significant modifications occurred in thevector Ad-BMP2-bFGF-GFP group. Conclusion: The results indicate that the use of Ad-BMP-2/bFGF-modified BMSCs in conjunction with DBM could successfully repair ONFH in a dog model by promoting bone formation and angiogenesis.

scholarly journals Role of FGF-2 Transfected Bone Marrow Mesenchymal Stem Cells in Engineered Bone Tissue for Repair of Avascular Necrosis of Femoral Head in Rabbits

2018 ◽  
Vol 48 (2) ◽  
pp. 773-784 ◽  
Author(s):  
Fei Zhang ◽  
Wu-xun  Peng ◽  
Lei Wang ◽  
Jian Zhang ◽  
Wen-tao Dong ◽  
...  
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Femoral Head ◽  
Avascular Necrosis ◽  
Therapeutic Effect ◽  
The Other ◽  
X Ray ◽  
Marrow Mesenchymal Stem Cells

Background/Aims: Avascular necrosis of the femoral head (ANFH) is the focus and difficulty of orthopedic diseases. Recently, tissue engineering bone for this disease has shown a good therapeutic effect. The aim of the present study was to investigate the therapeutic effect of basic fibroblast growth factor (FGF-2) as cytokines transfected bone marrow mesenchymal stem cells (BMSCs) in constructing tissue-engineered bone for avascular necrosis of the femoral head. Methods: The FGF-2 gene overexpressed lentivirus-transfected rBMSCs with xenogeneic antigen-extracted cancellous bone (XACB) to construct tissue engineered bone, and the model of early avascular necrosis of the femoral head was established by lipopolysaccharide (LPS) combined with hormone. The models were randomly divided into five groups: A (control), B (XACB), C (XACB+rBMSCs), D (XACB+rBMSCs+Lv-GFP), and E (XACB+rBMSCs+Lv-FGF-2/GFP) groups. The therapeutic effect of the tissue engineered bone for the avascular necrosis of the femoral head was evaluated by gross anatomy, X-ray examination, immunohistochemistry and H&E staining. Results: The FGF-2 gene was transfected into rBMSCs (Multiplicity of infection [MOI] = 100) by lentivirus, and its efficiency was above 95%. The rBMSCs were successfully transfected overexpressing FGF-2 by qPCR and western blot. After tissue engineering bone implantation, X-ray examination and gross specimen observation revealed that the repair area in the E group was > 80% at six weeks, the defect was completely repaired at 12 weeks, and osteogenesis was stronger, when compared with the other groups. For the X-ray score, vascular area density and new bone formation area were higher, when compared with the other groups, and the difference was statistically significant (P< 0.05). Conclusion: FGF-2 gene overexpression lentivirus transfection BMSCs combined with XACB to construct tissue engineered bone can effectively promote vascular regeneration, and improve the repair effect of avascular necrosis of the femoral head.

Plasma Cell Proliferation Is Reduced in Myeloma-Induced Hypercalcemia and in Co-Culture with Normal Healthy BM-MSCs

2020 ◽  
Author(s):  
Nader Vazifeh Shiran ◽  
Saeid Abroun
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Demineralized Bone Matrix ◽  
Bone Matrix ◽  
Inhibitory Effects ◽  
Myeloma Cells ◽  
Flow Cytometric ◽  
Marrow Mesenchymal Stem Cells ◽  
Induced Apoptosis

Abstract Objective In multiple myeloma (MM), stimulation of osteoclasts and bone marrow (BM) lesions lead to hypercalcemia, renal failure, and anemia. Co-culture of the myeloma cells in both hypocalcemia and hypercalcemia concentrations with bone marrow-mesenchymal stem cells were evaluated. Materials and Methods Viability and survival of myeloma cells were assessed by microculture tetrazolium test and flow cytometric assays. Mesenchymal stem cells (MSCs) were extracted from normal and myeloma patients and were co-cultured with myeloma cells. Results Myeloma cells showed less survival in both hypocalcaemia and hypercalcemia conditions (P &lt;.01). The paracrine and juxtacrine conditions of demineralized bone matrix-induced hypercalcemia increased the proliferation and survival of the cells (P &lt;.05). Unlike myeloma MSCs, normal MSCs reduced the survival of and induced apoptosis in myeloma cells (P &lt;.1). Conclusion Normal healthy-MSCs do not protect myeloma cells, but inhibit them. However, increasing the ratio of myeloma cells to MSCs reduces their inhibitory effects of MSCs and leads to their myelomatous transformation.

scholarly journals Residual γH2AX foci induced by low dose x-ray radiation in bone marrow mesenchymal stem cells do not cause accelerated senescence in the progeny of irradiated cells

Aging ◽  
2017 ◽  
Vol 9 (11) ◽  
pp. 2397-2410 ◽  
Author(s):  
Margarita Pustovalova ◽  
Тatiana A. Astrelina ◽  
Anna Grekhova ◽  
Natalia Vorobyeva ◽  
Anastasia Tsvetkova ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Low Dose ◽  
X Ray ◽  
Γh2ax Foci ◽  
Marrow Mesenchymal Stem Cells ◽  
Accelerated Senescence
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