scholarly journals TLR7 Expression Is Associated with M2 Macrophage Subset in Calcific Aortic Valve Stenosis

Cells ◽  
2020 ◽  
Vol 9 (7) ◽  
pp. 1710 ◽  
Author(s):  
Glykeria Karadimou ◽  
Oscar Plunde ◽  
Sven-Christian Pawelzik ◽  
Miguel Carracedo ◽  
Per Eriksson ◽  
...  
Keyword(s):  
Aortic Valve ◽  
Aortic Valve Stenosis ◽  
Ex Vivo ◽  
M2 Macrophage ◽  
Mrna Levels ◽  
Aortic Valves ◽  
Age Related ◽  
Ex Vivo Tissue ◽  
Tlr7 Ligand

Calcific aortic valve stenosis (CAVS) is a common age-related disease characterized by active calcification of the leaflets of the aortic valve. How innate immune cells are involved in disease pathogenesis is not clear. In this study we investigate the role of the pattern recognition receptor Toll-like receptor 7 (TLR7) in CAVS, especially in relation to macrophage subtype. Human aortic valves were used for mRNA expression analysis, immunofluorescence staining, or ex vivo tissue assays. Response to TLR7 agonist in primary macrophages and valvular interstitial cells (VICs) were investigated in vitro. In the aortic valve, TLR7 correlated with M2 macrophage markers on mRNA levels. Expression was higher in the calcified part compared with the intermediate and healthy parts. TLR7+ cells were co-stained with M2-type macrophage receptors CD163 and CD206. Ex vivo stimulation of valve tissue with the TLR7 ligand imiquimod significantly increased secretion of IL-10, TNF-α, and GM-CSF. Primary macrophages responded to imiquimod with increased secretion of IL-10 while isolated VICs did not respond. In summary, in human aortic valves TLR7 expression is associated with M2 macrophages markers. Ex vivo tissue challenge with TLR7 ligand led to secretion of immunomodulatory cytokine IL-10. These results connect TLR7 activation in CAVS to reduced inflammation and improved clearance.

scholarly journals Toll-like-receptor-3 function is critical for aortic valve stenosis development in mice

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
S.T Niepmann ◽  
A.S Boucher ◽  
M Bulic ◽  
P.R Goody ◽  
F Jansen ◽  
...  
Keyword(s):  
Aortic Valve ◽  
Aortic Valve Stenosis ◽  
Histological Analysis ◽  
Peak Velocity ◽  
Macrophage Infiltration ◽  
Funding Source ◽  
Aortic Valve Area ◽  
Mrna Levels ◽  
Toll Like Receptor ◽  
Age Related

Abstract Background Aortic valve stenosis (AS) is the most common valve diseases in the western world. After having been considered a passive degenerative process, which develops as an inevitable consequence of age-related valvular degeneration, basic research of the last two decades has led to a paradigm shift. It is now believed that AS pathophysiology is driven by distinct molecular and cellular mechanisms which include inflammatory pathways. In recent years, Toll-like-receptor-3 (TLR3) has emerged as a major regulator of vascular inflammation. TLR3 is a lysosomal pattern recognition receptor that recognizes single and double stranded RNA. Its activation leads to expression of pro-inflammatory cytokines via NFkb activation. The role of TLR3 in the development of AS has never been investigated. Methods Severe AS was induced in Wildtype-, ApoE- and TLR3/ApoE−/− mice. For this, a coronary springwire was used to induce an endothelial injury under echocardiographic guidance. Stenosis development was confirmed via ultrasound examinations. To inhibit TLR-3 activation, TLR3/RNA- Complex inhibitor C4a was injected every 48h after wire injury in WT mice. Valves were explanted and stained with hematoxylin/eosin (valve thickening) or anti-68 (macrophage infiltration). Valves from patients who received aortic valve replacement due to AS or aortic regurgitation (AR) were collected and mRNA levels of TLR3 and MyD88 were measured with use of quantitative-PCR. Results To evaluate weather TLR3 effects AS development in mice, we subjected TLR3/ApoE double- and ApoE knockout mice to our model of wire-induced AS. Surprisingly, TLR3 deficient mice failed to develop AS after wire injury. Peak velocity measurements showed no increase and histological analysis showed lower aortic valve area and macrophage infiltration compared to control mice. In order to pharmacological inhibit TLR3, WT mice were treated with C4a after wire injury. Compared to PBS control, C4a mice also did not develop AS upon wire injury. Trans-aortic valve peak velocity levels were significantly lower in C4a mice. Histological analysis underlined these results and showed thinner aortic valves and decreased macrophage infiltration in C4a mice comparted to control animals. To confirm our hypothesis, the expression of TLR3 and its downstream effector MyD88 were measured in human aortic valve specimens. qPCR analysis revealed decreased TLR3 and MyD88 expression in patients with AS compared to patients with AR. Conclusion In the presented study, we present first data that theTLR3 has a crucial role in the development of AS in mice. The exact downstream effects after TLR3 activation in AS need to be further investigated. Funding Acknowledgement Type of funding source: Public grant(s) – National budget only. Main funding source(s): Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)

scholarly journals New ex vivo calcification model for intact murine aortic valves

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
B Kruithof ◽  
V Van De Pol ◽  
T Los ◽  
K Lodder ◽  
M.C De Ruiter ◽  
...  
Keyword(s):  
Aortic Valve ◽  
Culture Media ◽  
Ex Vivo ◽  
Funding Source ◽  
Aortic Valves ◽  
Aortic Sinus ◽  
Intact Mouse ◽  
Jude Medical ◽  
Endochondral Calcification

Abstract Introduction Calcific aortic valve disease (CAVD) is a common, progressive disease of the aortic valves for which no medical treatment exists and surgery is the only therapeutic solution. The discovery of novel pharmacological treatment for CAVD has been hampered due to the lack of suitable test-systems. Purpose We aimed to establish an ex vivo calcification model for intact mouse aortic valves. Methods We used the ex vivo flow model for mouse hearts (Miniature Tissue Culture System) to induce calcification with culture media supplemented with 1) β-glycerophosphate, ascorbic acid, and dexamethasone (OSM) or 2) inorganic phosphates (PI) and compared this with in vitro calcification of mouse valvular interstitial cells (mVICs). Results Ex vivo cultured aortic valve leaflets calcified in the presence of PI, but not OSM. RUNX was upregulated in both OSM and PI conditions, whereas alkaline phosphatase (ALP) and cyclooxygenase-2 (COX2) were differentially expressed. Apoptosis was not observed, together indicating that osteogenic calcification took place. Interestingly, both OSM and PI were able to induce calcification in vitro, revealing in vitro-ex vivo differences. In addition to the calcification in the aortic valves, endochondral calcification was present in the sinus near the hinge of the aortic valve in both PI and OSM conditions as evidenced by the expression of collagen II, aggrecan and ALP. Conclusions Together these data show that we have established an ex vivo calcification model for mouse aortic valves. Osteogenic calcification is induced in the aortic leaflets, whereas endochondral calcification is induced in the aortic sinus, reflecting the calcification found in human CAVD. Using this model, we can now study the initiation and progression of aortic valve calcification. Funding Acknowledgement Type of funding source: Other. Main funding source(s): Dutch Heart Foundation, grant number 2013T093. Grants received from GE Healthcare, Lantheus medical imaging, St Jude Medical, Medtronic, Boston Scientific, Biotronik, and Edwards Lifesciences

scholarly journals Semicarbazide-Sensitive Amine Oxidase Increases in Calcific Aortic Valve Stenosis and Contributes to Valvular Interstitial Cell Calcification

2020 ◽  
Vol 2020 ◽  
pp. 1-9 ◽  
Author(s):  
Nathalie Mercier ◽  
Sven-Christian Pawelzik ◽  
John Pirault ◽  
Miguel Carracedo ◽  
Oscar Persson ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Aortic Valve ◽  
Aortic Valve Stenosis ◽  
Interstitial Cell ◽  
Amine Oxidase ◽  
Primary Amines ◽  
Common Disease ◽  
Mrna Levels ◽  
Aortic Valves ◽  
Valve Calcification

Introduction. Calcific aortic valve stenosis (CAVS) is a common disease associated with aging. Oxidative stress participates in the valve calcification process in CAVS. Semicarbazide-sensitive amine oxidase (SSAO), also referred to as vascular adhesion protein 1 (VAP-1), transforms primary amines into aldehydes, generating hydrogen peroxide and ammonia. SSAO is expressed in calcified aortic valves, but its role in valve calcification has remained largely unexplored. The aims of this study were to characterize the expression and the activity of SSAO during aortic valve calcification and to establish the effects of SSAO inhibition on human valvular interstitial cell (VIC) calcification. Methods. Human aortic valves from n=80 patients were used for mRNA extraction and expression analysis, Western blot, SSAO activity determination, immunohistochemistry, and the isolation of primary VIC cultures. Results. SSAO mRNA, protein, and activity were increased with increasing calcification within human aortic valves and localized in the vicinity of the calcified zones. The valvular SSAO upregulation was consistent after stratification of the subjects according to cardiovascular and CAVS risk factors associated with increased oxidative stress: body mass index, diabetes, and smoking. SSAO mRNA levels were significantly associated with poly(ADP-ribose) polymerase 1 (PARP1) in calcified tissue. Calcification of VIC was inhibited in the presence of the specific SSAO inhibitor LJP1586. Conclusion. The association of SSAO expression and activity with valvular calcification and oxidative stress as well as the decreased VIC calcification by SSAO inhibition points to SSAO as a possible marker and therapeutic target to be further explored in CAVS.

Hemodynamic and Structural Implications of Asymmetric Deployment of Transcatheter Aortic Valves: An In Vitro Study

2013 ◽  
Author(s):  
Paul S. Gunning ◽  
Neelakantan Saikrishnan ◽  
Ajit P. Yoganathan ◽  
Laoise M. McNamara
Keyword(s):  
Aortic Valve ◽  
Heart Surgery ◽  
Open Heart Surgery ◽  
Postoperative Mortality ◽  
In Vitro Study ◽  
Conventional Surgery ◽  
Aortic Valves ◽  
Transcatheter Aortic Valve ◽  
Age Related

Aortic stenosis is an age related degenerative disease of the aortic valve that causes narrowing of the valve and aortic regurgitation [1]. Transcatheter Aortic Valve (TAV) implantation is a percutaneous alternative to open heart surgery, which allows for the treatment of a cohort of patients for whom conventional surgery is deemed excessively risky due to high postoperative mortality rates or pre-existing illness [2].

Platelet membrane-coated nanoparticles target sclerotic aortic valves in ApoE−/− mice by multiple binding mechanisms under pathological shear stress

2020 ◽  
Vol 41 (Supplement_2) ◽  
Author(s):  
H Yang ◽  
Y Song ◽  
Z Huang ◽  
J Qian ◽  
Z Pang ◽  
...  
Keyword(s):  
Shear Stress ◽  
Aortic Valve ◽  
Aortic Valve Disease ◽  
Binding Capacity ◽  
Platelet Membrane ◽  
Valve Disease ◽  
Funding Source ◽  
Aortic Valves ◽  
Coated Nanoparticles

Abstract Background Aortic valve disease is the most common valvular heart disease leading to valve replacement. The efficacy of pharmacological therapy for aortic valve disease is limited by the high mechanical stress at the aortic valves impairing the binding rate. We aimed to identify nanoparticle coating with entire platelet membranes to fully mimic their inherent multiple adhesion mechanisms and target the sclerotic aortic valve of apolipoprotein E-deficient (ApoE−/−) mice based on their multiple sites binding capacity under high shear stress. Methods Considering the potent interaction of platelet membrane glycoproteins with components present in sclerotic aortic valves, platelet membrane-coated nanoparticles (PNPs) were synthetized and the binding capacity under high shear stress was evaluated in vitro and in vivo. Results Compared with PNPs bound intensity in the static station, 161%, 59%, and 39% of attached PNPs remained adherent on VWF-, collagen-, and fibrin-coated surfaces under shear stress of 25dyn/cm2 respectively. PNPs demonstrated effectively adhering to von Willebrand factor, collagen and fibrin under shear stresses in vitro. In an aortic valve disease model established in ApoE−/− mice, PNPs group exhibited significant increase of accumulation in the aortic valves compared with PBS and control NP group. PNPs displayed high degrees of proximity or co-localization with vWF, collagen and fibrin, which exhibited good targeting to sclerotic aortic valves by mimicking platelet multiple adhesive mechanisms. Conclusion PNPs could provide a promising platform for the molecular diagnosis and targeting treatment of aortic valve disease. Targeting combination Funding Acknowledgement Type of funding source: Foundation. Main funding source(s): National Natural Science Foundation of China

scholarly journals Age-related decline in the expression of GDF9 and BMP15 genes in follicle fluid and granulosa cells derived from poor ovarian responders

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Yan Gong ◽  
Jesse Li-Ling ◽  
Dongsheng Xiong ◽  
Jiajing Wei ◽  
Taiqing Zhong ◽  
...  
Keyword(s):  
Granulosa Cells ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mrna Levels ◽  
Clinical Trial Registry ◽  
Altered Expression ◽  
Vitro Fertilization ◽  
Age Related ◽  
Tertiary Teaching ◽  
Poor Ovarian Responders

Abstract Background Growth differentiation factor 9 (GDF9) and bone morphogenetic protein 15 (BMP15) genes play important roles in folliculogenesis. Altered expression of the two have been found among patients with poor ovarian response (POR). In this prospective cohort study, we have determined the expression of the GDF9 and BMP15 genes in follicle fluid (FF) and granulosa cells (GCs) derived from poor ovarian responders grouped by age, and explored its correlation with the outcome of in vitro fertilization and embryo transfer (IVF-ET) treatment. Methods A total of 196 patients with POR were enrolled from a tertiary teaching hospital. The patients were diagnosed by the Bologna criteria and sub-divided into group A (< 35 year old), group B (35–40 year old), and group C (> 40 year old). A GnRH antagonist protocol was conducted for all patients, and FF and GCs were collected after oocyte retrieval. Expression of the GDF9 and BMP15 genes in the FF and GCs was determined with enzyme-linked immunosorbent assay (ELISA), quantitative real-time polymerase chain reaction (qRT-PCR) and Western blotting. Results Compared with group C, groups A and B had significantly more two pronuclei (2PN) oocytes and transplantable embryos, in addition with higher rates of implantation and clinical pregnancy (P <  0.05). The expression level of GDF9 and BMP15 genes in the FF and GCs differed significantly among the three groups (P <  0.05), showing a trend of decline along with age. The ratio of GDF9/BMP15 mRNA levels were similar among the three groups (P > 0.05). The relative levels of GDF9 and BMP15 proteins in GCs have correlated with the relative mRNA levels in GCs and protein concentrations in FF (P <  0.05). Conclusions For poor ovarian responders, in particular those over 40, the expression of GDF9 and BMP15 is declined along with increased age and in accompany with poorer oocyte quality and IVF outcome, whilst the ratio of GDF9/BMP15 mRNA levels remained relatively constant. Trial registration Chinese Clinical Trial Registry Center (ChiCTR1800016107). Registered on 11 May 2018.

scholarly journals Granulosal and thecal expression of bone morphogenetic protein- and activin-binding protein mRNA transcripts during bovine follicle development and factors modulating their expression in vitro

Reproduction ◽  
2011 ◽  
Vol 142 (4) ◽  
pp. 581-591 ◽  
Author(s):  
Claire Glister ◽  
Leanne Satchell ◽  
Phil G Knight
Keyword(s):  
Bone Morphogenetic Proteins ◽  
Transforming Growth Factor ◽  
Ex Vivo ◽  
Mrna Level ◽  
Transcript Abundance ◽  
Transforming Growth Factor Β ◽  
Follicle Development ◽  
Mrna Levels ◽  
Theca Interna

Evidence supports local roles for transforming growth factor β superfamily members including activins and bone morphogenetic proteins (BMP) in follicle development. Access of these ligands to signalling receptors is likely modulated by extracellular binding proteins (BP). In this study, we comparedex vivoexpression of four BPs (chordin, gremlin, noggin and follistatin) in granulosal (GC) and theca interna (TC) compartments of developing bovine antral follicles (1–18 mm). Effects of FSH and IGF on BMP and BP expression by cultured GC, and effects of LH and BMPs on BP expression by cultured TC were also examined. Follicular expression of all four BP transcripts was higher in GC than TC compartments (P<0.001) a finding confirmed by immunohistochemistry. Follicle category affected (P<0.01) gremlin and follistatin mRNA abundance, with a significant cell-type×follicle category interaction for chordin, follistatin and noggin. Noggin transcript abundance was lower (P<0.05) in GC of large ‘E-active’ than ‘E-inactive’ follicles while follistatin mRNA level was higher (P<0.01). FSH enhanced CYP19, FSHR, INHBA and follistatin by GC without affecting BMP or BMP–BP expression. IGF increased CYP19 and follistatin, reduced BMP4, noggin and gremlin but did not affect chordin orFSHRmRNA levels. LH increased TC androgen secretion but had no effect on BMP or BP expression. BMPs uniformly suppressed TC androgen production whilst increasing chordin, noggin and gremlin mRNA levels up to 20-fold (P<0.01). These findings support the hypothesis that extracellular BP, mostly from GC, contribute to the regulation of intrafollicular BMP/activin signalling. Enhancement of thecal BP expression by BMP implies an autoregulatory feedback role to prevent excessive signalling.

scholarly journals Skin and hair on-a-chip: in vitro skin models versus ex vivo tissue maintenance with dynamic perfusion

Lab on a Chip ◽  
2013 ◽  
Vol 13 (18) ◽  
pp. 3555 ◽  
Author(s):  
Beren Ataç ◽  
Ilka Wagner ◽  
Reyk Horland ◽  
Roland Lauster ◽  
Uwe Marx ◽  
...  
Keyword(s):  
Ex Vivo ◽  
Ex Vivo Tissue

scholarly journals Effects of CREG1 on Age-Associated Metabolic Phenotypes and Renal Senescence in Mice

2021 ◽  
Vol 22 (3) ◽  
pp. 1276
Author(s):  
Michihiro Hashimoto ◽  
Ayumi Goto ◽  
Yuki Endo ◽  
Masataka Sugimoto ◽  
Jun Ueda ◽  
...  
Keyword(s):  
Renal Dysfunction ◽  
Body Weight Gain ◽  
Mrna Levels ◽  
Wild Type ◽  
Metabolic Phenotypes ◽  
Age Related ◽  
Secretory Phenotype ◽  
Filtering Function ◽  
Renal Senescence

Cellular repressor of E1A-stimulated genes 1 (CREG1) is a secreted glycoprotein that accelerates p16-dependent cellular senescence in vitro. We recently reported the ability of CREG1 to stimulate brown adipogenesis using adipocyte P2-CREG1-transgenic (Tg) mice; however, little is known about the effect of CREG1 on aging-associated phenotypes. In this study, we investigated the effects of CREG1 on age-related obesity and renal dysfunction in Tg mice. Increased brown fat formation was detected in aged Tg mice, in which age-associated metabolic phenotypes such as body weight gain and increases in blood glucose were improved compared with those in wild-type (WT) mice. Blood CREG1 levels increased significantly in WT mice with age, whereas the age-related increase was suppressed, and its levels were reduced, in the livers and kidneys of Tg mice relative to those in WT mice at 25 months. Intriguingly, the mRNA levels of Ink4a, Arf, and senescence-associated secretory phenotype (SASP)-related genes and p38MAPK activity were significantly lowered in the aged kidneys of Tg mice, in which the morphological abnormalities of glomeruli as well as filtering function seen in WT kidneys were alleviated. These results suggest the involvement of CREG1 in kidney aging and its potential as a target for improving age-related renal dysfunction.

Ozaki procedure: Ex vivo simulation

2021 ◽  
Keyword(s):  
Aortic Valve ◽  
Ex Vivo ◽  
Aortic Valves ◽  
Autologous Pericardium ◽  
Video Tutorial ◽  
Novel Technique ◽  
Valve Reconstruction

Replacements for diseased aortic valves are limited. Repair of the aortic valve is performed by only a few surgeons. A novel technique of aortic valve reconstruction using autologous pericardium shows promising results. In this video tutorial, we demonstrate the Ozaki procedure using an ex vivo low fidelity simulation.

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