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

Multinucleated rat alveolar macrophages express functional receptors for calcitonin

1991 ◽  
Vol 261 (6) ◽  
pp. F1026-F1032 ◽  
Author(s):  
A. Vignery ◽  
M. J. Raymond ◽  
H. Y. Qian ◽  
F. Wang ◽  
S. A. Rosenzweig
Keyword(s):  
Plasma Membrane ◽  
Alveolar Macrophages ◽  
Giant Cells ◽  
Mononuclear Phagocytes ◽  
Inflammatory Reactions ◽  
Calcitonin Gene ◽  
Novel Model

The fusion of mononuclear phagocytes occurs spontaneously in vivo and leads to the differentiation of either multinucleated giant cells or osteoclasts in chronic inflammatory sites or in bone, respectively. Although osteoclasts are responsible for resorbing bone, the functional role of giant cells in chronic inflammatory reactions and tumors remains poorly understood. We recently reported that the plasma membrane of multinucleated macrophages is, like that of osteoclasts, enriched in Na-K-adenosinetriphosphatases (ATPases). We also observed that the localization of their Na-K-ATPases is restricted to the nonadherent domain of the plasma membrane of cells both in vivo and in vitro, thus imposing a functional polarity on their organization. By following this observation, we wished to investigate whether these cells also expressed, like osteoclasts, functional receptors for calcitonin (CT). To this end, alveolar macrophages were fused in vitro, and both their structural and functional association with CT was analyzed and compared with those of mononucleated peritoneal and alveolar macrophages. Evidence is presented that multinucleated alveolar macrophages express a high copy number of functional receptors for CT. Our results also indicate that alveolar macrophages, much like peritoneal, express functional receptors for calcitonin gene-related peptide. It is suggested that multinucleated rat alveolar macrophages offer a novel model system to study CT receptors and that calcitonin may control local immune reactions where giant cells differentiate.

Osseoinduction Evaluation of Hydroxyapatite and Zinc Containing Hydroxyapatite Granules in Rabbits

2011 ◽  
Vol 493-494 ◽  
pp. 252-257 ◽  
Author(s):  
L. Nascimento ◽  
M. Medeiros ◽  
J. Calasans-Maia ◽  
A. Alves ◽  
Antonella M. Rossi ◽  
...  
Keyword(s):  
Histological Analysis ◽  
Bone Matrix ◽  
Fibrous Tissue ◽  
Particle Diameter ◽  
Inflammatory Cells ◽  
Giant Cells ◽  
Multinucleated Giant Cells ◽  
Ethics Commission

This study investigated the osteoinductive potential of granules of stoichiometric hydroxyapatite (HA) and 0.5% zinc containing hydroxyapatite (ZnHA) in intramuscular (IM) site of rabbit’s abdomen. The biomaterials were both used in granular form, with 75% porosity and particle diameter between 450 and 500μm, sintered at 1100°C. Both materials performed adequately on a multiparametric in vitro cytocompatibility assay, indicating their suitability for in vivo testing. After approval by the Ethics Commission on Teaching and Research in Animals, fifteen rabbits were submitted to general anesthesia, incision and tissue dilatation, and a small site was created for HA (right incision) and ZnHA (left incision) intramuscular implantation. The animals were killed after 2, 4 and 12 weeks for biomaterials and surrounding tissues removal. Histological analysis after 2 weeks revealed the presence of granulation tissue surrounding biomaterials with multinucleated giant cells and no newly formed bone for both materials. After 4 weeks there was fibrous tissue involving the material and few inflammatory cells. Following 12 weeks it was observed the presence of connective tissue surrounding the biomaterial, cellularized enough for the two experimental groups, but it was not observed the presence of bone matrix associated with the biomaterials. We conclude that both biomaterials are cytocompatible and did not present the property of osseoinduction after 12 weeks of implantation.

scholarly journals Mid-Gestation lethality of Atxn2l-Ablated Mice

2020 ◽  
Vol 21 (14) ◽  
pp. 5124 ◽  
Author(s):  
Jana Key ◽  
Patrick N. Harter ◽  
Nesli-Ece Sen ◽  
Elise Gradhand ◽  
Georg Auburger ◽  
...  
Keyword(s):  
Giant Cells ◽  
Neural Cells ◽  
Rna Surveillance ◽  
Extended Lifespan ◽  
Ataxin 2 ◽  
Locomotor Deficits ◽  
Lateral Sclerosis

Depletion of yeast/fly Ataxin-2 rescues TDP-43 overexpression toxicity. In mouse models of Amyotrophic Lateral Sclerosis via TDP-43 overexpression, depletion of its ortholog ATXN2 mitigated motor neuron degeneration and extended lifespan from 25 days to >300 days. There is another ortholog in mammals, named ATXN2L (Ataxin-2-like), which is almost uncharacterized but also functions in RNA surveillance at stress granules. We generated mice with Crispr/Cas9-mediated deletion of Atxn2l exons 5-8, studying homozygotes prenatally and heterozygotes during aging. Our novel findings indicate that ATXN2L absence triggers mid-gestational embryonic lethality, affecting female animals more strongly. Weight and development stages of homozygous mutants were reduced. Placenta phenotypes were not apparent, but brain histology showed lamination defects and apoptosis. Aged heterozygotes showed no locomotor deficits or weight loss over 12 months. Null mutants in vivo displayed compensatory efforts to maximize Atxn2l expression, which were prevented upon nutrient abundance in vitro. Mouse embryonal fibroblast cells revealed more multinucleated giant cells upon ATXN2L deficiency. In addition, in human neural cells, transcript levels of ATXN2L were induced upon starvation and glucose and amino acids exposure, but this induction was partially prevented by serum or low cholesterol administration. Neither ATXN2L depletion triggered dysregulation of ATXN2, nor a converse effect was observed. Overall, this essential role of ATXN2L for embryogenesis raises questions about its role in neurodegenerative diseases and neuroprotective therapies.

scholarly journals Role of Mesenchymal Stem Cell-Conditioned Medium (MSC-CM) in the Bone Regeneration: A Systematic Review from 2007-2018

2019 ◽  
pp. 1-16
Author(s):  
Ismail Hadisoebroto Dilogo ◽  
Jessica Fiolin
Keyword(s):  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Bone Regeneration ◽  
Conditioned Medium ◽  
Bone Healing ◽  
In Vitro Studies ◽  
In Vivo Studies

Background: The therapeutic value of mesenchymal stem cells (MSCs) in tissue engineering and regenerative medicine is attributable in part to paracrine pathways triggered by several secreted factors secreted into culture media. The secreted factor here is known as the conditioned medium (CM) or secretome. Objectives: This review is aimed to investigate and summarise the in-vitro, pre-clinical in-vivo studies regarding the role of CM-MSC in bone regeneration from 2007 until 2018 Data Sources: A systematic literature search on PubMed, MEDLINE, OVID, Scopus and Cochrane library was carried out by using search terms: Secretome, conditioned medium, mesenchymal stem cell, bone healing, osteogenic, osteogenesis. Methods: A total of 611 articles were reviewed. Ten articles were identified as relevant for this systematic literature review. Results: Three tables of studies were constructed for in vitro studies and in-vivo studies. Conclusion: All of the included in-vitro studies and in-vivo studies have shown a promoting effect of bone regeneration at various stages. Although there are no clinical studies regarding the use of CM-MSC in the human bone regeneration that have been conducted, transplantation of secretome has shown a promising result in the acceleration of bone healing process.

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 Transition of podosomes into zipper-like structures in macrophage-derived multinucleated giant cells

2020 ◽  
Vol 31 (18) ◽  
pp. 2002-2020
Author(s):  
Arnat Balabiyev ◽  
Nataly P. Podolnikova ◽  
Aibek Mursalimov ◽  
David Lowry ◽  
Jason M. Newbern ◽  
...  
Keyword(s):  
Plasma Membranes ◽  
Giant Cells ◽  
Multinucleated Giant Cells ◽  
E Cadherin

Our study reveals previously unrecognized actin-based zipper-like structures (ZLSs) formed between macrophage-derived multinucleated giant cells undergoing fusion in vivo and in vitro. It is shown that podosomes are precursors of these structures. The transition of podosomes into ZLSs is induced by bridging plasma membranes by E-cadherin and nectin-2.

Morphological assessment of the development of multinucleated giant cells in glioma by using mitosis-specific phosphorylated antibodies

2003 ◽  
Vol 98 (4) ◽  
pp. 854-859 ◽  
Author(s):  
Kenkou Maeda ◽  
Masaaki Mizuno ◽  
Toshihiko Wakabayashi ◽  
Syuntarou Takasu ◽  
Tetsurou Nagasaka ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Giant Cells ◽  
Laser Scanning Confocal Microscopy ◽  
Multinucleated Giant Cells ◽  
Content Type ◽  
Scanning Confocal Microscopy ◽  
Drug Treatments ◽  
Fine Print

Object. The nature and origin of multinucleated giant cells in glioma have not been made clear. To investigate the phosphorylation of intermediate filaments, the authors studied multinucleated giant cells in vitro and in vivo by using mitosis-specific phosphorylated antibodies. Methods. Cultured human glioma cells were immunostained with monoclonal antibodies (mAbs) 4A4, KT13, and TM71, which recognized the phosphorylation of vimentin at Ser55, glial fibrillary acidic protein at Ser13, and vimentin at Ser71, respectively. Subsequently, the nature of multinucleated giant cells was investigated using laser scanning confocal microscopy. In addition, paraffin-embedded tissue sections obtained in three patients with giant cell glioblastoma were also investigated. Multinucleated giant cells were immunoreacted with the mAb 4A4 and not with KT13 and TM71 in vitro and in vivo. In addition, the authors obtained these results in multinucleated giant cells under natural conditions, without drug treatments. Conclusions. Findings in this investigation indicated that multinucleated giant cells are those remaining in mitosis between metaphase and telophase, undergoing neither fusion nor degeneration.

197. In vitro and in vivo assessment of bone regeneration after local steroid administration using a rodent posterolateral fusion model

2020 ◽  
Vol 20 (9) ◽  
pp. S97-S98
Author(s):  
Mark A. Plantz ◽  
Abhishek S. Kannan ◽  
Silvia Minardi ◽  
David J. Ellenbogen ◽  
Joseph G. Lyons ◽  
...  
Keyword(s):  
Bone Regeneration ◽  
Posterolateral Fusion ◽  
Fusion Model ◽  
Steroid Administration ◽  
In Vivo Assessment

scholarly journals miR-129-5p Promotes Osteogenic Differentiation of BMSCs and Bone Regeneration via Repressing Dkk3

2021 ◽  
Vol 2021 ◽  
pp. 1-18
Author(s):  
Changming Zhao ◽  
Yulin Gu ◽  
Yan Wang ◽  
Qiaozhen Qin ◽  
Ting Wang ◽  
...  
Keyword(s):  
Western Blot ◽  
Bone Regeneration ◽  
Osteogenic Differentiation ◽  
Luciferase Reporter ◽  
Luciferase Reporter Gene ◽  
Alizarin Red ◽  
Gene Assay

Objective. Accumulating evidence indicates that microRNAs (miRNAs) play crucial roles in osteogenic differentiation. However, the associated mechanisms remain elusive. This paper is aimed at exploring the role of miR-129-5p in regulating bone marrow mesenchymal stem cell (BMSC) differentiation and bone regeneration in vivo and in vitro. Methods. BMSCs were transduced by miR-129-5p mimic, miR-129-5p inhibitor, and negative control lentivirus. The ability of BMSC differentiation to osteoblast was tested by alkaline phosphatase (ALP) and alizarin red staining (ARS). The expression of osteogenic genes (Runx2, Bmp2, and OCN) was examined via quantitative RT-PCR and western blot. A mouse model of calvaria defect was investigated by Micro-CT, immunohistochemistry, and histological examination. The luciferase reporter gene assay was performed to confirm the binding between Dkk3 and miR-129-5p. For the transfection experiments, lipofectamine 3000 was used to transfect pcDNA-Dkk3 into BMSCs to overexpress Dkk3. Coimmunoprecipitation and immunofluorescent localization assay were included for exploring the role of Dkk3 and β-catenin. Results. miR-129-5p was induced in BMSCs and MSC cell line C3H10T1/2 cells under osteogenic medium. Overexpression of miR-129-5p significantly promoted osteogenic differentiation of BMSCs in vitro. Moreover, BMSCs transduced with miR-129-5p mimic exhibited better bone regeneration compared with BMSCs transduced with control counterpart in vivo. Luciferase and western blot data showed that Dickkopf3 (Dkk3) is a target gene of miR-129-5p and the expression of Dkk3 was inhibited in BMSCs transduced with miR-129-5p mimic but enhanced in BMSCs transduced with miR-129-5p inhibitor. In addition, Dkk3 interacted with β-catenin directly. Conclusions. miR-129-5p promotes osteogenic differentiation of BMSCs and bone regeneration, and miR-129-5p/Dkk3 axis may be new potential targets for the treatment of bone defect and bone loss.

scholarly journals Bio-Absorption of Human Dentin-Derived Biomaterial in Sheep Critical-Size Iliac Defects

Materials ◽  
2021 ◽  
Vol 14 (1) ◽  
pp. 223
Author(s):  
Md Arafat Kabir ◽  
Masaru Murata ◽  
Mamata Shakya ◽  
Katsuhisa Yamada ◽  
Toshiyuki Akazawa
Keyword(s):  
Bone Regeneration ◽  
Critical Size ◽  
Morphological Changes ◽  
Giant Cells ◽  
Human Tooth ◽  
Micro Ct ◽  
Multinucleated Giant Cells ◽  
Custom Made ◽  
Histological Images

The aim of this study was to evaluate the bio-absorption and bone regeneration of human tooth-derived dentin scaffold, entitled as perforated root-demineralized dentin matrix (PR-DDM), after in vivo implantation into the critical-size iliac defects. The dentin scaffolds were prepared from human vital, non-functional teeth. Thirty artificial macro-pores (Ø 1 mm) were added after removing the enamel portion. The modified teeth were supersonically demineralized in 0.34 N HNO3 for 30 min. The microstructure was observed by scanning electron microscope (SEM). The 3D micro-CT and histological analysis were carried out to evaluate the bio-absorption of PR-DDM at 2 and 4 months. A smooth dentin collagen surface with symmetrical macro-pores and tube-type dentinal tubules (Ø 1–2 µm) with micro-cracks were observed on the perforated region. A significant number of custom-made macro-pores disappeared, and the size of the macro-pores became significantly wider at 4 months compared with the 2 months (p < 0.05) evaluated by 3D micro-CT. Histological images revealed the presence of multinucleated giant cells attached to the scalloped border of the PR-DDM. The morphological changes due to bio-absorption by the cellular phagocytes were comparable to the 3D micro-CT and histological images at 2 and 4 months. Altogether, the results demonstrated that the PR-DDM block was gradually absorbed by multinucleated giant cells and regenerated bone. Human PR-DDM might serve as a unique scaffold for extraoral bone regeneration.

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