Stromal Cell-Derived Factor-1β Potentiates Bone Morphogenetic Protein-2-Stimulated Osteoinduction of Genetically Engineered Bone Marrow-Derived Mesenchymal Stem Cells<i>In Vitro</i>

Herberg, Samuel; Fulzele, Sadanand; Yang, Ningning; Shi, Xingming; Hess, Matthew; Periyasamy‐Thandavan, Sudharsan; Hamrick, Mark W.; Isales, Carlos M.; Hill, William

doi:10.1089/ten.tea.2012.0085

Cited by 44 publications

(113 citation statements)

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“…An imbalance of SDF-1α in MDS-MSC might be involved in the proliferation defects of MSC and the increased senescence of these cells: decreased SDF-1α gene expression in MSC or disturbances in its intracellular pathways leads to impaired cell survival, cytoskeletal disorganization and defective differentiation. 36,37 SCF gene expression was down-regulated in MDS-MSC. It has been reported that SCF levels are elevated in bone marrow plasma of MDS patients, but with no statistical significance and without identification of the specific source of the cytokine.…”

Section: Discussionmentioning

confidence: 92%

Mesenchymal stromal cells from patients with myelodyplastic syndrome display distinct functional alterations that are modulated by lenalidomide

et al. 2013

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ABSTRACTtypes including MDS associated with single del(5q), and characterized their clonogenicity and differentiation potential. Methods Human samplesTwenty MDS patients were studied; their characteristics are detailed in Online Supplementary Table S1. Our control cohort consisted of one male and five female healthy individuals undergoing orthopedic surgery with a median age of 60 years (range, 56-65). MSC were obtained as previously described. 18 Patients were classified for the study as "lower risk" with and without del(5q) (LR, LR-5q ) (<5% bone marrow blasts and IPSS-low/int-1), and as "higher-risk" (HR) (>5% bone marrow blasts and IPSS-int-2/high). Magnetically sorted HSPC were obtained from peripheral blood of healthy individuals after mobilization. Samples were collected after approval by our Institutional Review Board, and written informed consent was obtained. A detailed description is available in the Online Supplementary Material. Lenalidomide treatmentLEN was kindly provided by Celgene (Munich, Germany) and prepared using 10% dimethyl-sulfoxide as a vehicle. The experiments were performed with 0.1 μM and 1 μM of the drug. Characterization of the mesenchymal stromal cellsClonality, self-renewal, generation of single cell-derived colonies (SCD), expression of cell surface and adhesion molecules with flow cytometry, differentiation potential, proliferation, viability, apoptosis, senescence, and cell cycle were studied. Detailed information about the experiments is provided in the Online Supplementary Material. Co-culture of myelodysplastic syndrome-mesenchymal stromal cells with hematopoietic stem and progenitor cells, clonogenicity and apoptosis assayThe capacity of MSC to support HSPC was tested using a 4-week cobblestone area-forming (CAF-C) assay and a 14-day colony-forming unit granulocyte-erythrocytemonocyte/macrophage (CFU-GEMM) assay as described in the Online Supplementary Material. HSPC or the cell line KG1-α were labeled with carboxyfluorescein-diacetate-succinimidyl-ester (CFSE) prior to co-culture with MSC for 72 h, or left unlabeled and co-cultured for 24 h, with or without the addition of 100 nM recombinant tumor necrosis factor alpha (TNF-α, Peprotech). Apoptosis and proliferation were studied by annexin-V staining and CFSE dilution using flow cytometry. Cytokine measurement and transwell migration assaySupernatant from MSC was obtained after culture with or without LEN and used for enzyme-linked immunoabsorbent assay (ELISA) determinations of SDF-1α, angiopoietin-1 (ANG1), and stem cell factor (SCF) levels. Supernatant was also used for a 4-h transwell migration assay with 1x10 5 HSPC, as described previously. 18 The CXCR4 antagonist AMD3100 (Sigma) was used at a concentration of 10 μM.RNA extraction, complementary DNA synthesis and real time polymerase chain reaction mRNA extracted with Trizol reagent (Invitrogen) was transcribed into complementary-DNA and used for real time polymerase chain reaction (RT-PCR) analysis for fatty acid binding protein 4 (FABP4), SDF-1α , ANG1, and SCF. Ab...

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Section: Discussionmentioning

confidence: 92%

Mesenchymal stromal cells from patients with myelodyplastic syndrome display distinct functional alterations that are modulated by lenalidomide

et al. 2013

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“…[26][27][28][29] Previous studies have suggested that the CXCR4/SDF-1 axis functions in postnatal bone formation by regulating osteoblast development in cooperation with BMP signaling, and that CXCR4 acts as an endogenous signaling component necessary for bone formation. 29 We 30 and others 31 have shown that bone marrow endothelial and stromal cells, and osteoblasts express both major SDF-1 splice variants, and that unexpectedly the beta isoform may be present at a high level in bone tissues. 31 We have also shown that, independent of SDF-1a, SDF-1b enhances BMP-2-stimulated mineralization, mRNA and protein expression of key osteogenic markers, regulates BMP-2 signal transduction via Erk1/2 phosphorylation, and promotes the migratory response of CXCR4-expressing BMSCs in vitro, 30 suggesting an SDF-1b autocrine and paracrine activity.…”

mentioning

confidence: 93%

“…29 We 30 and others 31 have shown that bone marrow endothelial and stromal cells, and osteoblasts express both major SDF-1 splice variants, and that unexpectedly the beta isoform may be present at a high level in bone tissues. 31 We have also shown that, independent of SDF-1a, SDF-1b enhances BMP-2-stimulated mineralization, mRNA and protein expression of key osteogenic markers, regulates BMP-2 signal transduction via Erk1/2 phosphorylation, and promotes the migratory response of CXCR4-expressing BMSCs in vitro, 30 suggesting an SDF-1b autocrine and paracrine activity. An important characteristic of SDF-1b is its protection from proteolytic cleavage at the C-terminus, distinguishing SDF-1b from SDF-1a.…”

mentioning

confidence: 93%

Low-Dose Bone Morphogenetic Protein-2/Stromal Cell-Derived Factor-1β Cotherapy Induces Bone Regeneration in Critical-Size Rat Calvarial Defects

Herberg

Susin

Peláez

et al. 2014

Tissue Engineering Part A

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Increasing evidence suggests that stromal cell-derived factor-1 (SDF-1/CXCL12) is involved in bone formation, though underlying molecular mechanisms remain to be fully elucidated. Also, contributions of SDF-1b, the second most abundant splice variant, as an osteogenic mediator remain obscure. We have shown that SDF-1b enhances osteogenesis by regulating bone morphogenetic protein-2 (BMP-2) signaling in vitro. Here we investigate the dose-dependent contribution of SDF-1b to suboptimal BMP-2-induced local bone formation; that is, a dose that alone would be too low to significantly induce bone formation. We utilized a critical-size rat calvarial defect model and tested the hypotheses that SDF-1b potentiates BMP-2 osteoinduction and that blocking SDF-1 signaling reduces the osteogenic potential of BMP-2 in vivo. In preliminary studies, radiographic analysis at 4 weeks postsurgery revealed a dose-dependent relationship in BMP-2-induced new bone formation. We then found that codelivery of SDF-1b potentiates suboptimal BMP-2 (0.5 mg) osteoinduction in a dose-dependent order, reaching comparable levels to the optimal BMP-2 dose (5.0 mg) without apparent adverse effects. Blocking the CXC chemokine receptor 4 (CXCR4)/SDF-1 signaling axis using AMD3100 attenuated the osteoinductive potential of the optimal BMP-2 dose, confirmed by qualitative histologic analysis. In conclusion, SDF-1b provides potent synergistic effects that support BMP-induced local bone formation and thus appears a suitable candidate for optimization of bone augmentation using significantly lower amounts of BMP-2 in spine, orthopedic, and craniofacial settings.

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“…These growth factors include: BMP-2, Vascular Endothelial Growth Factor (VEGF), Nellike Protein-1 (NELL-1), Platelet Rich Plasma (PRP), Erythropoietin (EPO) and stromal cell-derived factor 1 alpha (SDF-1a), to name a few [5,6,7] . Among all of these substances, BMP-2 has by far been the most extensively studied.…”

Section: Introductionmentioning

confidence: 99%

A mouse femoral defect model demonstrates the clinically relevant side effects of BMP-2

Shen

Velasco

Khadarian

et al. 2015

International Journal of Orthopaedics

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AIM: Bone morphogenetic protein-2 (BMP-2) is an FDA approved, osteoinductive growth factor for bone regeneration. However, increasing awareness of the side effects of BMP-2, including inflammation, bone resorption and fat formation, require the development of animal surgical models that replicate these adverse events. MATERIAL AND METHODS:We devised a mouse femoral uni-cortical defect (UCD) model to study the clinical side effects of BMP-2. UCDs (3×1×1 mm) were drilled in one femur of mice and filled with poly(D,L-lactide-co-glycolide) (PLGA) scaffolding coated with PBS or rhBMP-2 (0.3mg/ml or 0.6mg/ml). Post mortem analyses were performed at four weeks post-surgery. RESULTS: 100% successful surgery was achieved in mouse femoral UCD without surgical complication. Micro-CT analyses of the high dose rhBMP-2 group revealed bone cyst formation with ~2.5 fold increase in bone volume and more than 50% decrease in bone mineral density and trabecular number compared to PBS control. Tartrate-resistant acid phosphatase (TRAP) staining revealed a significant increase in osteoclast number, while immunostaining for fatty acid binding protein 4 (FABP4) and peroxisome proliferatoractivated receptor gamma (PPAR-γ) confirmed significant increases in adipocyte formation with rhBMP-2 in a dose dependent manner. Lastly, rhBMP-2 treatment caused significant increases in inflammation, as determined by immunostaining for tumor necrosis factor-α (TNF-α) and interleukin 6 (IL6). CONCLUSION: Here, we established a mouse UCD model that faithfully replicates the clinically reported side effects of rhBMP-2. This model may facilitate future studies to improve upon current efforts in rhBMP-2 based bone repair.

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Stromal Cell-Derived Factor-1β Potentiates Bone Morphogenetic Protein-2-Stimulated Osteoinduction of Genetically Engineered Bone Marrow-Derived Mesenchymal Stem CellsIn Vitro

Cited by 44 publications

References 45 publications

Mesenchymal stromal cells from patients with myelodyplastic syndrome display distinct functional alterations that are modulated by lenalidomide

Mesenchymal stromal cells from patients with myelodyplastic syndrome display distinct functional alterations that are modulated by lenalidomide

Low-Dose Bone Morphogenetic Protein-2/Stromal Cell-Derived Factor-1β Cotherapy Induces Bone Regeneration in Critical-Size Rat Calvarial Defects

A mouse femoral defect model demonstrates the clinically relevant side effects of BMP-2

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