STAT3 phosphorylation inhibition for treating inflammation and new bone formation in ankylosing spondylitis

Rheumatology ◽  
2020 ◽  
Author(s):  
Sungsin Jo ◽  
Eun Jeong Won ◽  
Moon-Ju Kim ◽  
Yu Jeong Lee ◽  
So-Hee Jin ◽  
...  
Keyword(s):  
Bone Formation ◽  
Chronic Inflammation ◽  
Mononuclear Cells ◽  
Kinase Inhibitor ◽  
Janus Kinase ◽  
Micro Ct ◽  
New Bone Formation ◽  
Osteoprogenitor Cells ◽  
Suppression Effect ◽  
Stat3 Phosphorylation

Abstract Objective AS is a rheumatic disease characterized by chronic inflammation and bony ankylosis. This study was to evaluate whether a signal transducer and activator of transcription 3 phosphorylation inhibitor (stat3-p Inh) could treat both chronic inflammation and bone formation in AS. Methods Primary AS osteoprogenitor cells and spinal entheseal cells were examined for osteogenic differentiation. SF mononuclear cells (SFMCs) and lamina propria mononuclear cells (LPMCs) were obtained from AS patients. Inflammatory cytokine-producing cells were analysed using flow cytometry and ELISA. Female SKG mice were treated with stat3-p Inh, IL-17A blocker or vehicle. Inflammation and new bone formation were evaluated using immunohistochemistry, PET and micro-CT. Results In the SKG mouse model, stat3-p Inh significantly suppressed arthritis, enthesitis, spondylitis and ileitis. In experiments culturing SFMCs and LPMCs, the frequencies of IFN-γ-, IL-17A- and TNF-α-producing cells were significantly decreased after stat3-p Inh treatment. When comparing current treatments for AS, stat3-p Inh showed a comparable suppression effect on osteogenesis to Janus kinase inhibitor or IL-17A blocker in AS-osteoprogenitor cells. Stat3-p Inh suppressed differentiation and mineralization of AS-osteoprogenitor cells and entheseal cells toward osteoblasts. Micro-CT analysis of hind paws revealed less new bone formation in stat3-p Inh-treated mice than vehicle-treated mice (P = 0.005). Hind paw and spinal new bone formation were similar between stat3-p Inh- and anti-IL-17A-treated SKG mice (P = 0.874 and P = 0.117, respectively). Conclusion Stat-3p inhibition is a promising treatment for both inflammation and new bone formation in AS.

scholarly journals Micro-CT Assessment of LED Irradiation on New Bone Formation in an Experimental Rat Calvaria Periosteal Elevation Model

2014 ◽  
Vol 35 (2) ◽  
pp. 151-157 ◽  
Author(s):  
Noboru Kuboyama ◽  
K. Bhawal Ujjal ◽  
Ryoichiro Uchida ◽  
Akira Kaneda ◽  
Yoshimitsu Abiko
Keyword(s):  
Bone Formation ◽  
Micro Ct ◽  
New Bone Formation ◽  
Rat Calvaria ◽  
Elevation Model ◽  
Led Irradiation

Micro-CT Analysis of New Bone Formation Around Implant with Electrochemically Deposited Apatite

2003 ◽  
Vol 240-242 ◽  
pp. 611-614 ◽  
Author(s):  
Seiji Ban ◽  
Norihiro Arimoto ◽  
Shozo Tsuruta ◽  
Hiroyuki Arikawa ◽  
Takahito Kanie ◽  
...  
Keyword(s):  
Bone Formation ◽  
Micro Ct ◽  
New Bone Formation ◽  
Electrochemically Deposited ◽  
Ct Analysis

scholarly journals Dual Blockade of TNF and IL-17A Inhibits Inflammation and Structural Damage in a Rat Model of Spondyloarthritis

2022 ◽  
Vol 23 (2) ◽  
pp. 859
Author(s):  
Ihsan Hammoura ◽  
Renee H. Fiechter ◽  
Shaughn H. Bryant ◽  
Susan Westmoreland ◽  
Gillian Kingsbury ◽  
...  
Keyword(s):  
Bone Formation ◽  
Rat Model ◽  
Structural Damage ◽  
Structural Changes ◽  
Ct Imaging ◽  
Control Treatment ◽  
Micro Ct ◽  
New Bone Formation ◽  
Dual Blockade ◽  
Dual Inhibition

The tumor necrosis factor (TNF) and IL-23/IL-17 axes are the main therapeutic targets in spondyloarthritis. Despite the clinical efficacy of blocking either pathway, monotherapy does not induce remission in all patients and its effect on new bone formation remains unclear. We aimed to study the effect of TNF and IL-17A dual inhibition on clinical disease and structural damage using the HLA-B27/human β2-microglobulin transgenic rat model of SpA. Immunized rats were randomized according to arthritis severity, 1 week after arthritis incidence reached 50%, to be treated twice weekly for a period of 5 weeks with either a dual blockade therapy of an anti-TNF antibody and an anti-IL-17A antibody, a single therapy of either antibody, or PBS as vehicle control. Treatment-blinded observers assessed inflammation and structural damage clinically, histologically and by micro-CT imaging. Both single therapies as well as TNF and IL-17A dual blockade therapy reduced clinical spondylitis and peripheral arthritis effectively and similarly. Clinical improvement was confirmed for all treatments by a reduction of histological inflammation and pannus formation (p < 0.05) at the caudal spine. All treatments showed an improvement of structural changes at the axial and peripheral joints on micro-CT imaging, with a significant decrease for roughness (p < 0.05), which reflects both erosion and new bone formation, at the level of the caudal spine. The effect of dual blockade therapy on new bone formation was more prominent at the axial than the peripheral level. Collectively, our study showed that dual blockade therapy significantly reduces inflammation and structural changes, including new bone formation. However, we could not confirm a more pronounced effect of dual inhibition compared to single inhibition.

scholarly journals Demineralized Bone Matrix Coating Si-Ca-P Ceramic Does Not Improve the Osseointegration of the Scaffold

Materials ◽  
2018 ◽  
Vol 11 (9) ◽  
pp. 1580 ◽  
Author(s):  
Andrés Parrilla-Almansa ◽  
Nuria García-Carrillo ◽  
Patricia Ros-Tárraga ◽  
Carlos Martínez ◽  
Francisco Martínez-Martínez ◽  
...  
Keyword(s):  
Bone Formation ◽  
Bone Tissue ◽  
Demineralized Bone Matrix ◽  
Bone Matrix ◽  
Micro Ct ◽  
New Bone Formation ◽  
Implantation Time ◽  
Post Surgery ◽  
Tissue Responses ◽  
Demineralized Bone

The aim of this study was to manufacture and evaluate the effect of a biphasic calcium silicophosphate (CSP) scaffold ceramic, coated with a natural demineralized bone matrix (DBM), to evaluate the efficiency of this novel ceramic material in bone regeneration. The DBM-coated CSP ceramic was made by coating a CSP scaffold with gel DBM, produced by the partial sintering of different-sized porous granules. These scaffolds were used to reconstruct defects in rabbit tibiae, where CSP scaffolds acted as the control material. Micro-CT and histological analyses were performed to evaluate new bone formation at 1, 3, and 5 months post-surgery. The present research results showed a correlation among the data obtained by micro-CT and the histomorphological results, the gradual disintegration of the biomaterial, and the presence of free scaffold fragments dispersed inside the medullary cavity occupied by hematopoietic bone marrow over the 5-month study period. No difference was found between the DBM-coated and uncoated implants. The new bone tissue inside the implants increased with implantation time. Slightly less new bone formation was observed in the DBM-coated samples, but it was not statistically significant. Both the DBM-coated and the CSP scaffolds gave excellent bone tissue responses and good osteoconductivity.

New bone formation by murine osteoprogenitor cells cultured on corticocancellous allograft bone

2008 ◽  
Vol 26 (12) ◽  
pp. 1660-1664 ◽  
Author(s):  
Ehren R. Nelson ◽  
Zhinong Huang ◽  
Ting Ma ◽  
Derek Lindsey ◽  
Christopher Jacobs ◽  
...  
Keyword(s):  
Bone Formation ◽  
Allograft Bone ◽  
New Bone Formation ◽  
Osteoprogenitor Cells ◽  
Cells Cultured

scholarly journals In Vivo Bone Regeneration Induced by a Scaffold of Chitosan/Dicarboxylic Acid Seeded with Human Periodontal Ligament Cells

2019 ◽  
Vol 20 (19) ◽  
pp. 4883 ◽  
Author(s):  
Teerawat Sukpaita ◽  
Suwabun Chirachanchai ◽  
Pornchanok Suwattanachai ◽  
Vincent Everts ◽  
Atiphan Pimkhaokham ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bone Formation ◽  
Dicarboxylic Acid ◽  
Bone Regeneration ◽  
Periodontal Ligament ◽  
Periodontal Ligament Cells ◽  
Micro Ct ◽  
New Bone Formation ◽  
Calvarial Defects

Chitosan/dicarboxylic acid (CS/DA) scaffold has been developed as a bone tissue engineering material. This study evaluated a CS/DA scaffold with and without seeded primary human periodontal ligament cells (hPDLCs) in its capacity to regenerate bone in calvarial defects of mice. The osteogenic differentiation of hPDLCs was analyzed by bone nodule formation and gene expression. In vivo bone regeneration was analyzed in mice calvarial defects. Eighteen mice were divided into 3 groups: one group with empty defects, one group with defects with CS/DA scaffold, and a group with defects with CS/DA scaffold and with hPDLCs. After 6 and 12 weeks, new bone formation was assessed using microcomputed tomography (Micro-CT) and histology. CS/DA scaffold significantly promoted in vitro osteoblast-related gene expression (RUNX2, OSX, COL1, ALP, and OPN) by hPDLCs. Micro-CT revealed that CS/DA scaffolds significantly promoted in vivo bone regeneration both after 6 and 12 weeks (p < 0.05). Histological examination confirmed these findings. New bone formation was observed in defects with CS/DA scaffold; being similar with and without hPDLCs. CS/DA scaffolds can be used as a bone regenerative material with good osteoinductive/osteoconductive properties.

scholarly journals IL-23 induces the expression of pro-osteogenic factors in osteoclasts

2020 ◽  
Vol 45 (05) ◽  
pp. 467-474
Author(s):  
Dan-Dan Pang ◽  
Li Cai ◽  
Jing-Ru Zhang ◽  
Sheng-Ming Dai
Keyword(s):  
Ankylosing Spondylitis ◽  
Bone Formation ◽  
Western Blot ◽  
Mrna Expression ◽  
Mononuclear Cells ◽  
New Bone Formation ◽  
Expression Levels ◽  
Semaphorin 4D ◽  
Osteogenic Factors ◽  
Mrna Expression Levels

Abstract Background The mechanism for the new bone formation in ankylosing spondylitis (AS) is still unclear. Although it has been demonstrated that IL-23 plays a pivotal role in the pathophysiology of AS, IL-23 has no direct effects on osteoblasts but modulates the function of osteoclasts. Aims To explore whether IL-23 indirectly facilitates new bone formation through osteoclasts in AS, here we analyzed whether IL-23 enhances the expression levels of pro-osteogenic factors by osteoclasts. Methods Mononuclear cells were harvested from mouse bone marrow and cultured in the presence of M-CSF (50 ng/ml) and RANKL (30 ng/ml) to trigger the production of osteoclasts. Protein and mRNA expression levels of Semaphorin 4D, Ephrin B2, BMP2, BMP6, SPHK1, HtrA1 and Wnt10b were measured using Western blot and qRT-PCR. Results Primary mononuclear cells were transformed into osteoclasts with RANKL and M-CSF. The increased expression of NFATc1 and TRAP together with TRAP staining of>3 nuclei were used to identify mature osteoclasts. The mRNA expression levels of BMP2, Ephrin B2 and SPHK1 were enhanced by 1.46, 2.1 and 2.46 folds after exposure to IL-23. Confirmation of increased levels of Ephrin B2 and SPHK1 in IL-23-stimulated osteoclasts was provided by Western blot analysis. IL-23 had no effects on the expression of BMP6 or Wnt10b, or on the anti-osteogenic factors Semaphorin 4D or HtrA1. Conclusions IL-23 induces osteoclasts to express pro-osteogenic factors rather than anti-osteogenic factors, suggesting IL-23 might indirectly promote the differentiation of osteoblasts through activated osteoclasts in ankylosing spondylitis.

scholarly journals Fabrication and Evaluation of Porous Beta-Tricalcium Phosphate/Hydroxyapatite (60/40) Composite as a Bone Graft Extender Using Rat Calvarial Bone Defect Model

2013 ◽  
Vol 2013 ◽  
pp. 1-9 ◽  
Author(s):  
Jae Hyup Lee ◽  
Mi Young Ryu ◽  
Hae-Ri Baek ◽  
Kyung Mee Lee ◽  
Jun-Hyuk Seo ◽  
...  
Keyword(s):  
Bone Formation ◽  
Bone Graft ◽  
Tricalcium Phosphate ◽  
Calvarial Defect ◽  
Micro Ct ◽  
Porous Substrate ◽  
New Bone Formation ◽  
Defect Model ◽  
Beta Tricalcium Phosphate ◽  
Bone Graft Extender

Beta-tricalcium phosphate (β-TCP) and hydroxyapatite (HA) are widely used as bone graft extenders due to their osteoconductivity and high bioactivity. This study aims to evaluate the possibility of using porous substrate with composite ceramics (β-TCP: HA = 60% : 40%, 60TCP40HA) as a bone graft extender and comparing it with Bio-Oss. Interconnectivity and macroporosity ofβ-TCP porous substrate were 99.9% and 83%, respectively, and the macro-porosity of packed granule after crushing was 69%. Calvarial defect model with 8 mm diameter was generated with male Sprague-Dawley rats and 60TCP40HA was implanted. Bio-Oss was implanted for a control group and micro-CT and histology were performed at 4 and 8 weeks after implantation. The 60TCP40HA group showed better new bone formation than the Bio-Oss group and the bone formation at central area of bone defect was increased at 8 weeks in micro-CT and histology. The percent bone volume and trabecular number of the 60TCP40HA group were significantly higher than those of Bio-Oss group. This study confirms the usefulness of the porous 60TCP40HA composite as a bone graft extender by showing increased new bone formation in the calvarial defect model and improved bone formation both quantitatively and qualitatively when compared to Bio-Oss.

scholarly journals SAT0353 STAT3 PHOSPHORYLATION IS INVOLVED IN THE DEVELOPMENTS OF INFLAMMATORY ARTHRITIS, ENTHESITIS, AND NEW BONE FORMATION IN ANKYLOSING SPONDYLITIS

2020 ◽  
Vol 79 (Suppl 1) ◽  
pp. 1122.2-1123
Author(s):  
S. H. Jin ◽  
P. R. Park ◽  
M. J. Kim ◽  
Y. J. Lee ◽  
S. Jo ◽  
...  
Keyword(s):  
Bone Formation ◽  
Type I Collagen ◽  
Type I ◽  
New Bone Formation ◽  
Peripheral Arthritis ◽  
Alizarin Red ◽  
Vitro Experiment ◽  
In Vitro Experiment ◽  
Stat3 Phosphorylation

Background:Ankylosing Spondylitis (AS) is a chronic inflammatory rheumatic disease, which is characterized by the enthesitis, peripheral arthritis, and chronic inflammation of the spine, leading to bony ankylosis. Signal transducer and activator of transcription (STAT) family proteins are latent cytoplasmic transcription factors that convey signals to the nucleus. It is activated by IL-6, IL-23, and IL-22 through JAK-mediated phosphorylation. Moreover, genetic studies implicate interleukin-23 (IL-23) receptor signal, including STAT3 in the development of AS. IL-17A has recently emerged as a potential target that regulates the extensive inflammation and abnormal bone formation observed in AS. It was reported that STAT3 is a regulatory factor that induces Th17 cell development from naive CD4 T cells.Objectives:The aim of this study is to investigate whether the STAT3 phosphorylation (stat3-p) inhibitor has a therapeutic effect on inflammation and new bone formation in AS.Methods:Eight weeks after curdlan injection, SKG mice were treated with stat3-p inhibitor or mock as a control. Clinical and histologic scores for arthritis and enthesitis were evaluated. Synovial fluid mononuclear cells (SFMC) samples were obtained from AS patients. Inflammatory cytokine producing cells were analyzed using flow cytometry. Bone tissue samples were obtained from the facet joints of patients with AS at surgery. Primary bone-derived cells (BdCs) were isolated and cultured. The osteogenic differentiation was assessed in vitro for 3 weeks using ALP activity, Alizarin red S (ARS), Type I collagen, von kossa,and hydroxyapatitestains. Statistical analysis was performed using Prism 5.0 Software. A p < 0.05 was considered statistically significant.Results:The stat3-p inhibitor significantly suppressed peripheral arthritis and enthesitis in SKG mice (figure 1). Inflammatory infiltration around the tendon–bone insertion site and along the tendon, as well as bony involvement were all reduced in stat3-p inhibitor-treated mice compared to control mice. We found that the levels of IFN-±, IL-17, TNF-± were higher in AS Synovial fluid. A significantly decreased frequencies of IFN-±, IL-17, TNF-± producing cells in AS SFMC were shown after stat3-p inhibitor treatment (P < 0.01).In vitro experiment of bone formation, the stat3-p inhibitor suppressed ALP activity. In addition, there were significant decrease in Alizarin red S (ARS), Type I collagen, von kossa staining scores due to stat3-p inhibitor at a concentration of 5 μM.Light intensity of hydroxyapatitestaining was also decreased by stat3-p inhibitor in a dose dependent manner (figure 2). Intriguingly, the stat3-p inhibitor suppressed osteogenesis in both early phase and late phase in AS-BdCs, down-regulating osteoblast-involved genes.Conclusion:The stat3-p inhibitor had beneficial effects on reducing inflammation and new bone formation in AS animal model. In addition, stat3-p inhibitor suppressed bone formation in vitro experiment. These findings suggest that the stat3-p inhibitor could be a potential therapeutic agent for AS.References:[1]Arthritis Res Ther 2018;20:115.[2]Nat Med 2012;18:1069-76.[3]Rheumatology (Oxford) 2017;56:488-493.[4]Nat Rev Immunol. 2011;11:239–50.[5]J Exp Med 2005;201:949–60.Acknowledgments:NoneDisclosure of Interests:None declared

scholarly journals Clonorchis Sinensis-Derived Protein Attenuates Inflammation and New Bone Formation in Ankylosing Spondylitis

2020 ◽  
Author(s):  
Yu Jeong Lee ◽  
Moon-Ju Kim ◽  
Sungsin Jo ◽  
So-Hee Jin ◽  
Pu-Reum Park ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Ankylosing Spondylitis ◽  
Bone Formation ◽  
Mononuclear Cells ◽  
Clonorchis Sinensis ◽  
Clinical Signs ◽  
Protective Effects ◽  
New Bone Formation ◽  
Micro Computed Tomography ◽  
Peripheral Blood Mononuclear

Abstract Background: Helminth infections and their components have been shown to have potential to modulate immunity and attenuate immune response. The objective of this study was to evaluate potential protective effects of Clonorchis sinensis–derived protein (CSp) on ankylosing spondylitis (AS).Methods: Cytotoxicity of CSp at different doses was assessed by MTS and flow cytometry before performing experiments. Peripheral blood mononuclear cells (PBMCs) and Synovial fluid mononuclear cells (SFMCs) were obtained from AS patients. Inflammatory cytokine-producing cells were analyzed using flow cytometry. SKG mice were treated with CSp or vehicle. Inflammation and new bone formation were evaluated using immunohistochemistry, positron emission tomography (PET) and micro–computed tomography (CT).Results: Treatment with CSp resulted in no reduced cell viability of PBMCs or SFMCs. In experiments culturing PBMCs and SFMCs, the frequencies of IFN-g and IL-17A producing cells were significantly reduced after CSp treatment. In the SKG mouse model, CSp treatment significantly suppressed arthritis and enthesitis. Micro-CT analysis of hind paw revealed less new bone formation in CSp-treated mice than in vehicle-treated mice. Conclusions: We provide the first evidence demonstrating that CSp can ameliorate clinical signs and cytokine derrangements in AS. In addition, such CSp treatment could reduce new bone formation of AS.

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