scholarly journals Apoptotic mesenchymal stromal cells support osteoclastogenesis while inhibiting multinucleated giant cells formation in vitro

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Paul Humbert ◽  
Meadhbh Á. Brennan ◽  
Julien De Lima ◽  
Régis Brion ◽  
Annie Adrait ◽  
...  
Keyword(s):  
Bone Regeneration ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Quantitative Proteomics ◽  
Giant Cells ◽  
Multinucleated Giant Cells ◽  
Short Term ◽  
Before And After ◽  
Induced Apoptosis

AbstractIn bone regeneration induced by the combination of mesenchymal stromal cells (MSCs) and calcium-phosphate (CaP) materials, osteoclasts emerge as a pivotal cell linking inflammation and bone formation. Favorable outcomes are observed despite short-term engraftments of implanted MSCs, highlighting their major paracrine function and the possible implication of cell death in modulating their secretions. In this work, we focused on the communication from MSCs towards osteoclasts-like cells in vitro. MSCs seeded on a CaP biomaterial or undergoing induced apoptosis produced a conditioned media favoring the development of osteoclasts from human CD14+ monocytes. On the contrary, MSCs’ apoptotic secretion inhibited the development of inflammatory multinucleated giant cells formed after IL-4 stimulation. Components of MSCs’ secretome before and after apoptotic stress were compared using mass spectrometry-based quantitative proteomics and a complementary immunoassay for major cytokines. CXCR-1 and CXCR-2 ligands, primarily IL-8/CXCL-8 but also the growth-regulated proteins CXCL-1, -2 or -3, were suggested as the major players of MSCs’ pro-osteoclastic effect. These findings support the hypothesis that osteoclasts are key players in bone regeneration and suggest that apoptosis plays an important role in MSCs’ effectiveness.

scholarly journals Mesenchymal stromal cells' secretome enhances osteoclastogenesis but reduces multinucleated giant cells formation in vitro

Bone Reports ◽  
2020 ◽  
Vol 13 ◽  
pp. 100371
Author(s):  
Paul Humbert ◽  
Julien De Lima ◽  
Meadhbh Á. Brennan ◽  
Frédéric Blanchard ◽  
Pierre Layrolle
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Giant Cells ◽  
Multinucleated Giant Cells

scholarly journals Molecular Imaging for Comparison of Different Growth Factors on Bone Marrow-Derived Mesenchymal Stromal Cells’ Survival and ProliferationIn Vivo

2016 ◽  
Vol 2016 ◽  
pp. 1-10 ◽  
Author(s):  
Hongyu Qiao ◽  
Ran Zhang ◽  
Lina Gao ◽  
Yanjie Guo ◽  
Jinda Wang ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Growth Factors ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Bioluminescence Imaging ◽  
Firefly Luciferase ◽  
Control Group ◽  
Induced Apoptosis

Introduction. Bone marrow-derived mesenchymal stromal cells (BMSCs) have emerged as promising cell candidates but with poor survival after transplantation. This study was designed to investigate the efficacy of VEGF, bFGF, and IGF-1 on BMSCs’ viability and proliferation bothin vivoandin vitrousing bioluminescence imaging (BLI).Methods. BMSCs were isolated fromβ-actin-Fluc+transgenic FVB mice, which constitutively express firefly luciferase. Apoptosis was induced by hypoxia preconditioning for up to 24 h followed by flow cytometry and TUNEL assay. 106BMSCs with/without growth factors were injected subcutaneously into wild type FVB mice’s backs. Survival of BMSCs was longitudinally monitored using bioluminescence imaging (BLI) for 5 weeks. Protein expression of Akt, p-Akt, PARP, and caspase-3 was detected by Western blot.Results. Hypoxia-induced apoptosis was significantly attenuated by bFGF and IGF-1 compared with VEGF and control groupin vitro(P<0.05). When combined with matrigel, IGF-1 showed the most beneficial effects in protecting BMSCs from apoptosisin vivo.The phosphorylation of Akt had a higher ratio in the cells from IGF-1 group.Conclusion. IGF-1 could protect BMSCs from hypoxia-induced apoptosis through activation of p-Akt/Akt pathway.

In vitro and in vivo assessment of CaP materials for bone regenerative therapy. The role of multinucleated giant cells/osteoclasts in bone regeneration

2019 ◽  
Vol 108 (1) ◽  
pp. 282-297 ◽  
Author(s):  
Ana Carolina Cestari Bighetti ◽  
Tania Mary Cestari ◽  
Paula Sanches Santos ◽  
Ricardo Vinicius Nunes Arantes ◽  
Suelen Paini ◽  
...  
Keyword(s):  
Bone Regeneration ◽  
Giant Cells ◽  
Regenerative Therapy ◽  
Multinucleated Giant Cells ◽  
In Vivo Assessment

scholarly journals Crosstalk of Endothelial and Mesenchymal Stromal Cells under Tissue-Related O2

2021 ◽  
Vol 1 (2) ◽  
pp. 116-136
Author(s):  
Olga Zhidkova ◽  
Elena Andreeva ◽  
Mariia Ezdakova ◽  
Ludmila Buravkova
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Reciprocal Effects ◽  
Hypoxic Stress ◽  
Protective Effects ◽  
Short Term ◽  
Tnf Α ◽  
Vitro Analysis ◽  
In Vitro Analysis

Mesenchymal stromal cells (MSCs) are considered a valuable tool for cell therapy. After systemic administration, the outcome of MSCs and endothelial cells (ECs) interactions strongly depend on the local microenvironment and tissue O2 levels in particular. In vitro analysis of EC effects on MSC regenerative potential in co-culture was performed after short-term interaction at “physiological” hypoxia (5% O2) and acute hypoxic stress (0.1% O2). At 5% O2, MSCs retained stromal phenotype and CFU-f numbers, osteogenic RUNX2 was upregulated. A shift in the expression of adhesion molecules, and an increase in transcription/synthesis of IL-6, IL-8 contributed to facilitation of directed migration of MSCs. In the presence of MSCs, manifestations of oxidative stress in ECs were attenuated, and a decrease in adhesion of PBMCs to TNF-α-activated ECs was observed. Under 0.1% O2, reciprocal effects of ECs and MSCs were similar to those at 5% O2. Meanwhile, upregulation of RUNX2 was canceled, IL-6 decreased, and IL-8 significantly increased. “Protective” effects of MSCs on TNF-α-ECs were less pronounced, manifested as NOS3 downregulation and intracellular NO elevation. Therefore, interaction with ECs at “physiological” hypoxia enhanced pro-regenerative capacities of MSCs including migration and anti-inflammatory modulation of ECs. Under acute hypoxic stress, the stimulating effects of ECs on MSCs and the “protective” potential of MSCs towards TNF-α-ECs were attenuated.

scholarly journals Bioactive and Topographically-Modified Electrospun Membranes for the Creation of New Bone Regeneration Models

Processes ◽  
2020 ◽  
Vol 8 (11) ◽  
pp. 1341
Author(s):  
Dina Abdelmoneim ◽  
Ghsaq M. Alhamdani ◽  
Thomas E. Paterson ◽  
Martin E. Santocildes Romero ◽  
Beatriz J. C. Monteiro ◽  
...  
Keyword(s):  
Stem Cell ◽  
Bone Regeneration ◽  
Cell Fate ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Bone Tissue Regeneration ◽  
Ageing Population ◽  
Native Bone ◽  
The Creation

Bone injuries that arise from trauma, cancer treatment, or infection are a major and growing global challenge. An increasingly ageing population plays a key role in this, since a growing number of fractures are due to diseases such as osteoporosis, which place a burden on healthcare systems. Current reparative strategies do not sufficiently consider cell-substrate interactions that are found in healthy tissues; therefore, the need for more complex models is clear. The creation of in vitro defined 3D microenvironments is an emerging topographically-orientated approach that provides opportunities to apply knowledge of cell migration and differentiation mechanisms to the creation of new cell substrates. Moreover, introducing biofunctional agents within in vitro models for bone regeneration has allowed, to a certain degree, the control of cell fate towards osteogenic pathways. In this research, we applied three methods for functionalizing spatially-confined electrospun artificial microenvironments that presented relevant components of the native bone stem cell niche. The biological and osteogenic behaviors of mesenchymal stromal cells (MSCs) were investigated on electrospun micro-fabricated scaffolds functionalized with extracellular matrix (ECM) proteins (collagen I), glycosaminoglycans (heparin), and ceramic-based materials (bioglass). Collagen, heparin, and bioglass (BG) were successfully included in the models without modifying the fibrous structures offered by the polycaprolactone (PCL) scaffolds. Mesenchymal stromal cells (MSCs) were successfully seeded in all the biofunctional scaffolds and they showed an increase in alkaline phosphatase production when exposed to PCL/BG composites. This research demonstrates the feasibility of manufacturing smart and hierarchical artificial microenvironments for studying stem cell behavior and ultimately the potential of incorporating these artificial microenvironments into multifunctional membranes for bone tissue regeneration

scholarly journals The Functions of Clusterin Expression in Renal Mesenchymal Stromal Cells: Regulation of Cell Proliferation and Macrophage Activation

2021 ◽  
Author(s):  
xiaodong weng ◽  
Jing Li ◽  
Qiunong Guan ◽  
Haimei Zhao ◽  
Zihuan Wang ◽  
...  
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Renal Tissue ◽  
Renal Protection ◽  
Phagocytic Capacity ◽  
Induced Apoptosis ◽  
Clusterin Expression

Abstract BackgroundThe expression of clusterin (CLU) in mice increases resistance to renal ischemia-reperfusion injury and promotes renal tissue repair. However, the mechanisms underlying of the renal protection of CLU remain largely unknown. Mesenchymal stromal cells (MSCs), found in different compartments of the kidney, may contribute to kidney cell turnover and injury repair. This study investigated the in vitro functions of CLU in kidney mesenchymal stromal cells (KMSCs). MethodsKMSCs were isolated by digestion of kidney tissues with collagenase type 1 and growth in plastic culture plates. Cell surface markers, apoptosis and phagocytosis were determined by flow cytometry, and CLU protein by Western blot. ResultsKMSCs isolated from both wild type (WT) and CLU knockout (KO) mice positively expressed CD133, Sca-1, CD44, and CD117, and negatively of CD34, CD45, CD163, CD41, CD276, CD138 and CD79a. There was no difference in trilineage differentiation to chondrocytes, adipocytes and osteocytes between WT and KO KMSCs. CLU protein was expressed in and secreted by WT KMSCs, and it was up-regulated in response to hypoxia, but the degrees of hypoxia-induced apoptosis in WT KMSCs were not significantly different from those in KO KMSCs. The WT KMSCs proliferated faster than KO KMSCs in cultures. Furthermore, the incubation of macrophages with CLU-containing culture medium from WT KMSCs increased the CD206 expression in the macrophages and their phagocytic capacity. ConclusionOur data for the first time demonstrate the functions of CLU in the promotion of KMSCs proliferation, and may be required for KMSCs-regulated macrophage M2 polarization and phagocytic activity.

scholarly journals Platelet-derived extracellular vesicles promote osteoinduction of mesenchymal stromal cells

2020 ◽  
Vol 9 (10) ◽  
pp. 667-674
Author(s):  
Miquel Antich-Rosselló ◽  
Maria Antònia Forteza-Genestra ◽  
Javier Calvo ◽  
Antoni Gayà ◽  
Marta Monjo ◽  
...  
Keyword(s):  
Bone Regeneration ◽  
Extracellular Vesicles ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Messenger Rna ◽  
Platelet Rich Plasma ◽  
Size Exclusion ◽  
Human Umbilical Cord ◽  
Mrna Expression Analysis

Aims Platelet concentrates, like platelet-rich plasma (PRP) and platelet lysate (PL), are widely used in regenerative medicine, especially in bone regeneration. However, the lack of standard procedures and controls leads to high variability in the obtained results, limiting their regular clinical use. Here, we propose the use of platelet-derived extracellular vesicles (EVs) as an off-the-shelf alternative for PRP and PL for bone regeneration. In this article, we evaluate the effect of PL-derived EVs on the biocompatibility and differentiation of mesenchymal stromal cells (MSCs). Methods EVs were obtained first by ultracentrifugation (UC) and then by size exclusion chromatography (SEC) from non-activated PL. EVs were characterized by transmission electron microscopy, nanoparticle tracking analysis, and the expression of CD9 and CD63 markers by western blot. The effect of the obtained EVs on osteoinduction was evaluated in vitro on human umbilical cord MSCs by messenger RNA (mRNA) expression analysis of bone markers, alkaline phosphatase activity (ALP), and calcium (Ca2+) content. Results Osteogenic differentiation of MSCs was confirmed when treated with UC-isolated EVs. In order to disprove that the effect was due to co-isolated proteins, EVs were isolated by SEC. Purer EVs were obtained and proved to maintain the differentiation effect on MSCs and showed a dose-dependent response. Conclusion PL-derived EVs present an osteogenic capability comparable to PL treatments, emerging as an alternative able to overcome PL and PRP limitations. Cite this article: Bone Joint Res 2020;9(10):667–674.

scholarly journals Bone regeneration in rat calvarial defects using dissociated or spheroid mesenchymal stromal cells in scaffold-hydrogel constructs

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Siddharth Shanbhag ◽  
Salwa Suliman ◽  
Samih Mohamed-Ahmed ◽  
Carina Kampleitner ◽  
Mohamed Nageeb Hassan ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bone Regeneration ◽  
Osteogenic Differentiation ◽  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Micro Ct ◽  
In Vitro Mineralization ◽  
Calvarial Defects

Abstract Background Three-dimensional (3D) spheroid culture can promote the osteogenic differentiation of bone marrow mesenchymal stromal cells (BMSC). 3D printing offers the possibility to produce customized scaffolds for complex bone defects. The aim of this study was to compare the potential of human BMSC cultured as 2D monolayers or 3D spheroids encapsulated in constructs of 3D-printed poly-L-lactide-co-trimethylene carbonate scaffolds and modified human platelet lysate hydrogels (PLATMC-HPLG) for bone regeneration. Methods PLATMC-HPLG constructs with 2D or 3D BMSC were assessed for osteogenic differentiation based on gene expression and in vitro mineralization. Subsequently, PLATMC-HPLG constructs with 2D or 3D BMSC were implanted in rat calvarial defects for 12 weeks; cell-free constructs served as controls. Bone regeneration was assessed via in vivo computed tomography (CT), ex vivo micro-CT and histology. Results Osteogenic gene expression was significantly enhanced in 3D versus 2D BMSC prior to, but not after, encapsulation in PLATMC-HPLG constructs. A trend for greater in vitro mineralization was observed in constructs with 3D versus 2D BMSC (p > 0.05). In vivo CT revealed comparable bone formation after 4, 8 and 12 weeks in all groups. After 12 weeks, micro-CT revealed substantial regeneration in 2D BMSC (62.47 ± 19.46%), 3D BMSC (51.01 ± 24.43%) and cell-free PLATMC-HPLG constructs (43.20 ± 30.09%) (p > 0.05). A similar trend was observed in the histological analysis. Conclusion Despite a trend for superior in vitro mineralization, constructs with 3D and 2D BMSC performed similarly in vivo. Regardless of monolayer or spheroid cell culture, PLATMC-HPLG constructs represent promising scaffolds for bone tissue engineering applications.

scholarly journals The Functions of Clusterin Expression in Renal Mesenchymal Stromal Cells: Regulation of Cell Proliferation and Macrophage Activation

2021 ◽  
Author(s):  
xiaodong weng ◽  
Jing Li ◽  
Qiunong Guan ◽  
Haimei Zhao ◽  
Zihuan Wang ◽  
...  
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Stromal Cells ◽  
Ischemia Reperfusion Injury ◽  
Ischemia Reperfusion ◽  
Renal Tissue ◽  
Renal Protection ◽  
Phagocytic Capacity ◽  
Induced Apoptosis ◽  
Clusterin Expression

Abstract The expression of clusterin (CLU) in mice increases resistance to renal ischemia-reperfusion injury and promotes renal tissue repair. However, the mechanisms underlying of the renal protection of CLU remain largely unknown. Mesenchymal stromal cells (MSCs), found in different compartments of the kidney, may contribute to kidney cell turnover and injury repair. This study investigated the in vitro functions of CLU in kidney mesenchymal stromal cells (KMSCs). KMSCs were isolated by digestion of kidney tissues with collagenase type 1 and growth in plastic culture plates. Cell surface markers, apoptosis and phagocytosis were determined by flow cytometry, and CLU protein by Western blot. Here, we showed that KMSCs isolated from both wild type (WT) and CLU knockout (KO) mice positively expressed CD133, Sca-1, CD44, and CD117, and negatively of CD34, CD45, CD163, CD41, CD276, CD138 and CD79a. There was no difference in trilineage differentiation to chondrocytes, adipocytes and osteocytes between WT and KO KMSCs. CLU protein was expressed in and secreted by WT KMSCs, and it was up-regulated in response to hypoxia. However, lack of CLU expression did not negatively affect hypoxia-induced apoptosis in cultured KMSCs. The WT KMSCs proliferated faster than KO KMSCs in cultures. Furthermore, the incubation of macrophages with CLU-containing culture medium from WT KMSCs increased the CD206 expression in the macrophages and their phagocytic capacity. In conclusion, our data for the first time demonstrate the functions of CLU in the promotion of KMSCs proliferation, and may be required for KMSCs-regulated macrophage M2 polarization and phagocytic activity.

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