21 results on '"Menzel, Ursula"'
Search Results
2. Identification of circulating miRNAs as fracture-related biomarkers.
- Author
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Della Bella, Elena, Menzel, Ursula, Naros, Andreas, Kubosch, Eva Johanna, Alini, Mauro, and Stoddart, Martin J.
- Subjects
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FRACTURE healing , *MESENCHYMAL stem cells , *MICRORNA , *BIOMARKERS , *RNA sequencing , *PROGENITOR cells - Abstract
Fracture non-unions affect many patients worldwide, however, known risk factors alone do not predict individual risk. The identification of novel biomarkers is crucial for early diagnosis and timely patient treatment. This study focused on the identification of microRNA (miRNA) related to the process of fracture healing. Serum of fracture patients and healthy volunteers was screened by RNA sequencing to identify differentially expressed miRNA at various times after injury. The results were correlated to miRNA in the conditioned medium of human bone marrow mesenchymal stromal cells (BMSCs) during in vitro osteogenic differentiation. hsa-miR-1246, hsa-miR-335-5p, and miR-193a-5p were identified both in vitro and in fracture patients and their functional role in direct BMSC osteogenic differentiation was assessed. The results showed no influence of the downregulation of the three miRNAs during in vitro osteogenesis. However, miR-1246 may be involved in cell proliferation and recruitment of progenitor cells. Further studies should be performed to assess the role of these miRNA in other processes relevant to fracture healing. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
3. Three-dimensional culture and characterization of mononuclear cells from human bone marrow
- Author
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Bara, Jennifer J., Herrmann, Marietta, Menzel, Ursula, Benneker, Lorin, Alini, Mauro, and Stoddart, Martin J.
- Published
- 2015
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4. CD34/CD133 enriched bone marrow progenitor cells promote neovascularization of tissue engineered constructs in vivo
- Author
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Herrmann, Marietta, Binder, Andreas, Menzel, Ursula, Zeiter, Stephan, Alini, Mauro, and Verrier, Sophie
- Published
- 2014
- Full Text
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5. Replication allows inactivation of a knocked-in locus control region in inappropriate cell lineages
- Author
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Menzel, Ursula, Ktistaki, Eleni, Tolaini, Mauro, Veiga-Fernandes, Henrique, and Kioussis, Dimitris
- Published
- 2010
6. HP1-β Is Required for Development of the Cerebral Neocortex and Neuromuscular Junctions
- Author
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Aucott, Rebecca, Bullwinkel, Jörn, Yu, Yang, Shi, Wei, Billur, Mustafa, Brown, Jeremy P., Menzel, Ursula, Kioussis, Dimitris, Wang, Guozheng, Reisert, Ingrid, Weimer, Jörg, Pandita, Raj K., Sharma, Girdhar G., Pandita, Tej K., Fundele, Reinald, and Singh, Prim B.
- Published
- 2008
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7. HP1-[beta] is required for development of the cerebral neocortex and neuromuscular junctions
- Author
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Aucott, Rebecca, Bullwinkel, Jorn, Yu, Yang, Shi, Wei, Billur, Mustafa, Brown, Jeremy P., Menzel, Ursula, Kioussis, Dimitris, Wang, Guozheng, Reisert, Ingrid, Weimer, Jorg, Pandita, Raj K., Sharma, Girdhar G., Pandita, Tej K., Fundele, Reinald, and Singh, Prim B.
- Subjects
Cerebral cortex -- Growth ,Cerebral cortex -- Abnormalities ,Neuromuscular junction -- Growth ,Proteins -- Identification and classification ,Proteins -- Health aspects ,Company growth ,Biological sciences - Abstract
HP1 proteins are thought to be modulators of chromatin organization in all mammals, yet their exact physiological function remains unknown. In a first attempt to elucidate the function of these proteins in vivo, we disrupted the murine Cbx1 gene, which encodes the HP1-[beta] isotype, and show that the [Cbx1.sup.-/-] null mutation leads to perinatal lethality. The newborn mice succumbed to acute respiratory failure, whose likely cause is the defective development of neuromuscular junctions within the endplate of the diaphragm. We also observe aberrant cerebral cortex development in [Cbx1.sup.-/-] mutant brains, which have reduced proliferation of neuronal precursors, widespread cell death, and edema. In vitro cultures of neurospheres from [Cbx1.sup.-/-] mutant brains reveal a dramatic genomic instability. Our results demonstrate that HP1 proteins are not functionally redundant and that they are likely to regulate lineage-specific changes in hetero-chromatin organization.
- Published
- 2008
8. Mesenchymal Stem Cell Homing Into Intervertebral Discs Enhances the Tie2-positive Progenitor Cell Population, Prevents Cell Death, and Induces a Proliferative Response.
- Author
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Wangler, Sebastian, Peroglio, Marianna, Menzel, Ursula, Benneker, Lorin M., Haglund, Lisbet, Daisuke Sakai, Mauro Alini, Grad, Sibylle, Sakai, Daisuke, and Alini, Mauro
- Published
- 2019
- Full Text
- View/download PDF
9. Shear and Dynamic Compression Modulates the Inflammatory Phenotype of Human Monocytes in vitro.
- Author
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Fahy, Niamh, Menzel, Ursula, Alini, Mauro, and Stoddart, Martin J.
- Subjects
MONOCYTES ,FRACTURE healing ,MACROPHAGES ,INFLAMMATION ,IMMUNOREGULATION ,TUMOR necrosis factors ,SHEAR (Mechanics) ,COMPRESSION loads - Abstract
Monocytes and their derived macrophages are found at the site of remodeling tissue, such as fracture hematoma, that is exposed to mechanical forces and have been previously implicated in the reparative response. However, the mechanoresponsive of monocytes and macrophages to skeletal tissue-associated mechanical forces and their subsequent contribution to skeletal repair remains unclear. The aim of this study was to investigate the potential of skeletal tissue-associated loading conditions to modulate human monocyte activation and phenotype. Primary human monocytes or the human monocyte reporter cell line, THP1-Blue, were encapsulated in agarose and exposed to a combination of shear and compressive loading for 1 h a day for 3 consecutive days. Exposure of monocytes to mechanical loading conditions increased their pro-inflammatory gene and protein expression. Exposure of undifferentiated monocytes to mechanical loading conditions significantly upregulated gene expression levels of interleukin(IL)-6 and IL-8 compared to free swelling controls. Additionally, multiaxial loading of unstimulated monocytes resulted in increased protein secretion of TNF-α (17.1 ± 8.9 vs. 8 ± 7.4 pg/ml) and MIP-1α (636.8 ± 471.1 vs. 124.1 ± 40.1 pg/ml), as well as IL-13 (42.1 ± 19.8 vs. 21.7 ± 13.6) compared monocytes cultured under free-swelling conditions. This modulatory effect was observed irrespective of previous activation with the M1/pro-inflammatory differentiation stimuli lipopolysaccharide and interferon-γ or the M2/anti-inflammatory differentiation factor interleukin-4. Furthermore, mechanical shear and compression were found to differentially regulate nitric oxide synthase 2 (NOS2) and IL-12B gene expression as well as inflammatory protein production by THP1-Blue monocytes. The findings of this study indicate that human monocytes are responsive to mechanical stimuli, with a modulatory effect of shear and compressive loading observed toward pro-inflammatory mediator production. This may play a role in healing pathways that are mechanically regulated. An in depth understanding of the impact of skeletal tissue-associated mechanical loading on monocyte behavior may identify novel targets to maximize inflammation-mediated repair mechanisms. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
10. Isolation of high-quality RNA from intervertebral disc tissue via pronase predigestion and tissue pulverization.
- Author
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Caprez, Stephanie, Menzel, Ursula, Li, Zhen, Grad, Sibylle, Alini, Mauro, and Peroglio, Marianna
- Abstract
The isolation of high-quality RNA from the intervertebral disc and especially from the nucleus pulposus is challenging due to the low cellularity and high proteoglycan content of this tissue. In this study, we report a simple modification of the standard guanidinium thiocyanate-phenol-chloroform extraction method, which involves enzymatic predigestion of the tissue prior to standard RNA isolation. Yield, purity and integrity of RNA isolated from bovine nucleus pulposus, inner annulus fibrosus and outer annulus fibrosus were compared among complete matrix digestion, predigestion and pulverization, pulverization alone, and pulverization followed by on-column purification. With predigestion, the average yield of RNA obtained from bovine nucleus pulposus was 8.82 ± 2.05 ng/mg of wet tissue with 260/280 and 260/230 optical density ratios of 1.91 ± 0.15 and 1.84 ± 0.30, respectively. RIN analysis indicated that RNA quality was best preserved with the predigestion method (RNA integrity number > 7), and the extracted RNA was suitable for real-time polymerase chain reaction. This method is of importance for gene expression studies on intervertebral disc development, degeneration and repair, and we anticipate that it may be further applied to other tissues rich in proteoglycans. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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- View/download PDF
11. Modulation of the Murine CD8 Gene Complex Following the Targeted Integration of Human CD2-Locus Control Region Sequences.
- Author
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Menzel, Ursula, Kosteas, Theodoros, MaurovTolaini, Killeen, Nigel, Roderick, Kathleen, and Kioussis, Dimitris
- Subjects
- *
T cells , *LYMPHOCYTES , *LABORATORY mice , *GENE expression , *LOCUS (Genetics) - Abstract
The human CD2 (hCD2) locus control region (LCR) inserted in the mouse CD8 gene complex activates expression of the CD8 genes in T cell subsets in which the CD8 locus is normally silenced (e.g., CD4+ single-positive T cells). In this article, we show that, in conditional mCD8/hCD2-LCR (CD8/LCR) knock-in mice, the continuous presence of the hCD2-LCR is required for this effect. Deletion of the inserted hCD2-LCR in a developmental stage and cell lineage-specific manner revealed that the temporary presence of the LCR during early development does not permanently alter the expression pattern of the CD8 genes. As a result, cells that have been affected by the insertion of the LCR can convert to their destined phenotype once the LCR is removed. DNaseI hypersensitive sites 1 and 2 of the hCD2-LCR influence the expression of the CD8 genes in a similar manner as does the full LCR, whereas insertion of hypersensitive site 3 alone of the LCR does not result in a changed expression pattern. This analysis revealed a dynamic interaction between the hCD2-LCR and the endogenous regulatory elements of the CD8 genes. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
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12. Fate mapping of IL-17-producing T cells in inflammatory responses.
- Author
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Hirota, Keiji, Duarte, João H, Veldhoen, Marc, Hornsby, Eve, Li, Ying, Cua, Daniel J, Ahlfors, Helena, Wilhelm, Christoph, Tolaini, Mauro, Menzel, Ursula, Garefalaki, Anna, Potocnik, Alexandre J, and Stockinger, Brigitta
- Subjects
GENE mapping ,T cells ,INFLAMMATION ,INTERLEUKINS ,PHYSIOLOGICAL adaptation ,ENCEPHALOMYELITIS ,INTERFERONS ,SPINAL cord ,CYTOKINES - Abstract
Here we describe a reporter mouse strain designed to map the fate of cells that have activated interleukin 17A (IL-17A). We found that IL-17-producing helper T cells (T
H 17 cells) had distinct plasticity in different inflammatory settings. Chronic inflammatory conditions in experimental autoimmune encephalomyelitis (EAE) caused a switch to alternative cytokines in TH 17 cells, whereas acute cutaneous infection with Candida albicans did not result in the deviation of TH 17 cells to the production of alternative cytokines, although IL-17A production was shut off in the course of the infection. During the development of EAE, interferon-γ (IFN-γ) and other proinflammatory cytokines in the spinal cord were produced almost exclusively by cells that had produced IL-17 before their conversion by IL-23 ('ex-TH 17 cells'). Thus, this model allows the actual functional fate of effector T cells to be related to TH 17 developmental origin regardless of IL-17 expression. [ABSTRACT FROM AUTHOR]- Published
- 2011
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13. MII Has a Critical Role in Fetal and Adult Hematopoletic Stem Cell Self-Renewal.
- Author
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McMahon, Kathryn A., Hiew, Samantha Y.-L., Hadjur, Suzana, Veiga-Fernandes, Henrique, Menzel, Ursula, Price, Amanda J., Kioussis, Dimitris, Williams, Owen, and Brady, Hugh J. M.
- Subjects
CANCER genes ,CANCER genetics ,LEUKEMIA in children ,CELL proliferation ,HEMATOPOIETIC stem cells ,HEMATOPOIESIS - Abstract
The Mixed Lineage Leukemia (MIl) gene is a homolog of Drosophila Trithorax commonly re-arranged in infant leukemia. Comprehensive analysis of the role of MlI in hematopoiesis in fetal and adult knockout mice has been prevented by the lethality of MIl
-/- mice. We have established a conditional deletion model that allows us to study adult hematopoiesis in the absence of MIl. In this study, M-/- embryos survive to E16.5 and have reduced numbers of HSCs. The quiescent fraction of these HSCs is greatly reduced, and they are unable to compete with wild-type cells in transplantation assays. Mice with MIl expression conditionally deleted in the hematopoietic system have grossly normal hematopolesis in bone marrow, thymus, and spleen. However, transplanted MII-deficient bone marrow cells are highly compromised in their ability to competitively reconstitute irradiated recipients. These results suggest a critical role for MIl in regulating stem cell self-renewal. [ABSTRACT FROM AUTHOR]- Published
- 2007
- Full Text
- View/download PDF
14. In Vitro Evaluation of a Nanoparticle-Based mRNA Delivery System for Cells in the Joint.
- Author
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Sturm, Lisa, Schwemberger, Bettina, Menzel, Ursula, Häckel, Sonja, Albers, Christoph E., Plank, Christian, Rip, Jaap, Alini, Mauro, Traweger, Andreas, Grad, Sibylle, and Basoli, Valentina
- Subjects
MESENCHYMAL stem cells ,MESSENGER RNA ,PROGENITOR cells ,EXTRACELLULAR matrix - Abstract
Biodegradable and bioresponsive polymer-based nanoparticles (NPs) can be used for oligonucleotide delivery, making them a promising candidate for mRNA-based therapeutics. In this study, we evaluated and optimized the efficiency of a cationic, hyperbranched poly(amidoamine)s-based nanoparticle system to deliver tdTomato mRNA to primary human bone marrow stromal cells (hBMSC), human synovial derived stem cells (hSDSC), bovine chondrocytes (bCH), and rat tendon derived stem/progenitor cells (rTDSPC). Transfection efficiencies varied among the cell types tested (bCH 28.4% ± 22.87, rTDSPC 18.13% ± 12.07, hBMSC 18.23% ± 14.80, hSDSC 26.63% ± 8.81) and while an increase of NPs with a constant amount of mRNA generally improved the transfection efficiency, an increase of the mRNA loading ratio (2:50, 4:50, or 6:50 w/w mRNA:NPs) had no impact. However, metabolic activity of bCHs and rTDSPCs was significantly reduced when using higher amounts of NPs, indicating a dose-dependent cytotoxic response. Finally, we demonstrate the feasibility of transfecting extracellular matrix-rich 3D cell culture constructs using the nanoparticle system, making it a promising transfection strategy for musculoskeletal tissues that exhibit a complex, dense extracellular matrix. [ABSTRACT FROM AUTHOR]
- Published
- 2021
- Full Text
- View/download PDF
15. Differential Regulation of circRNA, miRNA, and piRNA during Early Osteogenic and Chondrogenic Differentiation of Human Mesenchymal Stromal Cells.
- Author
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Della Bella, Elena, Menzel, Ursula, Basoli, Valentina, Tourbier, Céline, Alini, Mauro, and Stoddart, Martin J.
- Subjects
- *
LINCRNA , *CIRCULAR RNA , *MESENCHYMAL stem cells , *NUCLEOTIDE sequence , *MICRORNA , *STROMAL cells , *NON-coding RNA - Abstract
The goal of the present study is to identify the differential expression of circular RNA (circRNA), miRNA, and piwi-interacting RNA (piRNA) after lineage commitment towards osteo- and chondrogenesis of human bone marrow mesenchymal stromal cells (hMSCs). The cells were maintained for 7 days in either osteogenic or chondrogenic medium. RNA sequencing was performed to assess the expression of miRNA and piRNA, while RNA hybridization arrays were used to identify which circRNA were differentially expressed. qPCR validation of a selection of targets for both osteogenic and chondrogenic differentiation was carried out. The differential expression of several circRNA, miRNA, and piRNA was identified and validated. The expression of total and circular isoforms of FKBP5 was upregulated both in osteo- and chondrogenesis and it was influenced by the presence of dexamethasone. ZEB1, FADS2, and SMYD3 were also identified as regulated in differentiation and/or by dexamethasone. In conclusion, we have identified a set of different non-coding RNAs that are differentially regulated in early osteogenic and chondrogenic differentiation, paving the way for further investigation to understand how dexamethasone controls the expression of those genes and what their function is in MSC differentiation. [ABSTRACT FROM AUTHOR]
- Published
- 2020
- Full Text
- View/download PDF
16. Phenotypic Characterization of Bone Marrow Mononuclear Cells and Derived Stromal Cell Populations from Human Iliac Crest, Vertebral Body and Femoral Head.
- Author
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Herrmann, Marietta, Hildebrand, Maria, Menzel, Ursula, Fahy, Niamh, Alini, Mauro, Lang, Siegmund, Benneker, Lorin, Verrier, Sophie, Stoddart, Martin J., and Bara, Jennifer J.
- Subjects
B cells ,STROMAL cells ,CELL populations ,POPULATION ,IDIOPATHIC femoral necrosis ,BONE marrow cells ,ILIUM - Abstract
(1) In vitro, bone marrow-derived stromal cells (BMSCs) demonstrate inter-donor phenotypic variability, which presents challenges for the development of regenerative therapies. Here, we investigated whether the frequency of putative BMSC sub-populations within the freshly isolated mononuclear cell fraction of bone marrow is phenotypically predictive for the in vitro derived stromal cell culture. (2) Vertebral body, iliac crest, and femoral head bone marrow were acquired from 33 patients (10 female and 23 male, age range 14–91). BMSC sub-populations were identified within freshly isolated mononuclear cell fractions based on cell-surface marker profiles. Stromal cells were expanded in monolayer on tissue culture plastic. Phenotypic assessment of in vitro derived cell cultures was performed by examining growth kinetics, chondrogenic, osteogenic, and adipogenic differentiation. (3) Gender, donor age, and anatomical site were neither predictive for the total yield nor the population doubling time of in vitro derived BMSC cultures. The abundance of freshly isolated progenitor sub-populations (CD45−CD34−CD73+, CD45−CD34−CD146+, NG2+CD146+) was not phenotypically predictive of derived stromal cell cultures in terms of growth kinetics nor plasticity. BMSCs derived from iliac crest and vertebral body bone marrow were more responsive to chondrogenic induction, forming superior cartilaginous tissue in vitro, compared to those isolated from femoral head. (4) The identification of discrete progenitor populations in bone marrow by current cell-surface marker profiling is not predictive for subsequently derived in vitro BMSC cultures. Overall, the iliac crest and the vertebral body offer a more reliable tissue source of stromal progenitor cells for cartilage repair strategies compared to femoral head. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
17. Monitoring live human mesenchymal stromal cell differentiation and subsequent selection using fluorescent RNA-based probes.
- Author
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Li, Bojun, Menzel, Ursula, Loebel, Claudia, Schmal, Hagen, Alini, Mauro, and Stoddart, Martin J.
- Published
- 2016
- Full Text
- View/download PDF
18. Pre-TCR Signaling and CD8 Gene Bivalent Chromatin Resolution during Thymocyte Development.
- Author
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Harker, Nicola, Garefalaki, Anna, Menzel, Ursula, Ktistaki, Eleni, Naito, Taku, Georgopoulos, Katia, and Kioussis, Dimitris
- Subjects
- *
IMMUNOGENETICS , *CHROMATIN , *HETEROCHROMATIC genes , *HETEROCHROMATIN , *IMMUNOREGULATION - Abstract
The CD8 gene is silent in CD4-CD8- double-negative thymocytes, expressed in CD4+CD8+ double-positive cells, and silenced in cells committing to the CD4+ single-positive (SP) lineage, remaining active in the CD8+ SP lineage. In this study, we show that the chromatin of the CD8 locus is remodeled in C57BL/6 and B6/J Rag1-/- MOM double-negative thymocytes as indicated by DNaseI hypersensitivity and widespread bivalent chromatin marks. Pre-TCR signaling coincides with chromatin bivalency resolution into monovalent activating modifications in double-positive and CD8 SP cells. Shortly after commitment to CD4 SP cell lineage, monovalent repressive characteristics and chromatin inaccessibility are established. Differential binding of Ikaros, NuRD, and heterochromatin protein 1a on the locus during these processes may participate in the complex regulation of CD8. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
19. Five Days Granulocyte Colony-Stimulating Factor Treatment Increases Bone Formation and Reduces Gap Size of a Rat Segmental Bone Defect: A Pilot Study.
- Author
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Herrmann M, Zeiter S, Eberli U, Hildebrand M, Camenisch K, Menzel U, Alini M, Verrier S, and Stadelmann VA
- Abstract
Bone is an organ with high natural regenerative capacity and most fractures heal spontaneously when appropriate fracture fixation is provided. However, additional treatment is required for patients with large segmental defects exceeding the endogenous healing potential and for patients suffering from fracture non-unions. These cases are often associated with insufficient vascularization. Transplantation of CD34+ endothelial progenitor cells (EPCs) has been successfully applied to promote neovascularization of bone defects, however including extensive ex vivo manipulation of cells. Here, we hypothesized, that treatment with granulocyte colony-stimulating factor (G-CSF) may improve bone healing by mobilization of CD34+ progenitor cells into the circulation, which in turn may facilitate vascularization at the defect site. In this pilot study, we aimed to characterize the different cell populations mobilized by G-CSF and investigate the influence of cell mobilization on the healing of a critical size femoral defect in rats. Cell mobilization was investigated by flow cytometry at different time points after five consecutive daily G-CSF injections. In a pilot study, bone healing of a 4.5-mm critical femoral defect in F344 rats was compared between a saline-treated control group and a G-CSF treatment group. In vivo microcomputed tomography and histology were applied to compare bone formation in both treatment groups. Our data revealed that leukocyte counts show a peak increase at the first day after the last G-CSF injection. In addition, we found that CD34+ progenitor cells, including EPCs, were significantly enriched at day 1, and further increased at day 5 and day 11. Upregulation of monocytes, granulocytes and macrophages peaked at day 1. G-CSF treatment significantly increased bone volume and bone density in the defect, which was confirmed by histology. Our data show that different cell populations are mobilized by G-CSF treatment in cell specific patterns. Although in this pilot study no bridging of the critical defect was observed, significantly improved bone formation by G-CSF treatment was clearly shown.
- Published
- 2018
- Full Text
- View/download PDF
20. Corrigendum: Monitoring live human mesenchymal stromal cell differentiation and subsequent selection using fluorescent RNA-based probes.
- Author
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Li B, Menzel U, Loebel C, Schmal H, Alini M, and Stoddart MJ
- Published
- 2016
- Full Text
- View/download PDF
21. An In Vitro Investigation of Platelet-Rich Plasma-Gel as a Cell and Growth Factor Delivery Vehicle for Tissue Engineering.
- Author
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Jalowiec JM, D'Este M, Bara JJ, Denom J, Menzel U, Alini M, Verrier S, and Herrmann M
- Subjects
- Biocompatible Materials chemical synthesis, Cell Survival, Cells, Cultured, Diffusion, Drug Implants administration & dosage, Elastic Modulus, Gels chemistry, Humans, Intercellular Signaling Peptides and Proteins chemistry, Materials Testing, Mesenchymal Stem Cells cytology, Mesenchymal Stem Cells physiology, Viscosity, Drug Implants chemical synthesis, Intercellular Signaling Peptides and Proteins administration & dosage, Mesenchymal Stem Cell Transplantation instrumentation, Platelet-Rich Plasma chemistry, Tissue Engineering instrumentation, Tissue Scaffolds
- Abstract
Platelet-rich plasma (PRP) has been used for different applications in human and veterinary medicine. Many studies have shown promising therapeutic effects of PRP; however, there are still many controversies regarding its composition, properties, and clinical efficacy. The aim of this study was to evaluate the influence of different platelet concentrations on the rheological properties and growth factor (GF) release profile of PRP-gels. In addition, the viability of incorporated bone marrow-derived human mesenchymal stem cells (MSCs) was investigated. PRP (containing 1000 × 10(3), 2000 × 10(3), and 10,000 × 10(3) platelets/μL) was prepared from human platelet concentrates. Platelet activation and gelification were achieved by addition of human thrombin. Viscoelastic properties of PRP-gels were evaluated by rheological studies. The release of GFs and inflammatory proteins was measured using a membrane-based protein array and enzyme-linked immunosorbent assay. MSC viability and proliferation in PRP-gels were assessed over 7 days by cell viability staining. Cell proliferation was examined using DNA quantification. Regardless of the platelet content, all tested PRP-gels showed effective cross-linking. A positive correlation between protein release and the platelet concentration was observed at all time points. Among the detected proteins, the chemokine CCL5 was the most abundant. The greatest release appeared within the first 4 h after gelification. MSCs could be successfully cultured in PRP-gels over 7 days, with the highest cell viability and DNA content found in PRP-gels with 1000 × 10(3) platelets/μL. The results of this study suggest that PRP-gels represent a suitable carrier for both cell and GF delivery for tissue engineering. Notably, a platelet concentration of 1000 × 10(3) platelets/μL appeared to provide the most favorable environment for MSCs. Thus, the platelet concentration is an important consideration for the clinical application of PRP-gels.
- Published
- 2016
- Full Text
- View/download PDF
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