scholarly journals Histological and molecular characterization of human aortic stenosis: a matter of sex

2021 ◽  
Vol 42 (Supplement_1) ◽  
Author(s):  
L Matilla ◽  
E Jover ◽  
V Arrieta ◽  
A Garcia-Pena ◽  
A Fernandez-Celis ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Aortic Stenosis ◽  
Valve Replacement ◽  
Ex Vivo ◽  
Tissue Inhibitor Of Metalloproteinase ◽  
Heart Valve Disease ◽  
Type I ◽  
Differentiation Markers ◽  
Alizarin Red ◽  
Surgical Valve Replacement

Abstract Introduction Aortic stenosis (AS) is the most common heart valve disease and it is strongly prevalent with elderly. AS is a progressive, degenerative disease associated with fibrosis and calcification of the valve leaflets. Surgical valve replacement is the only treatment available. Molecular, cellular and interstitial events activate multifactorial and complex cues with a significant contribution by valve interstitial cells (VICs). Despite male sex is a risk factor for developing AS, there is scant information on sex-specific differences in aortic valve (AV) biology or pathology. Purpose The aim of our study was to analyse sex-specific differences in aortic valves from AS patients. Methods 185 patients with severe AS undergoing surgical valve replacement were recruited. 149 AVs (66 women; 83 men) were used for ex vivo analyses. Human VICs were isolated from 36 AVs (12 women; 24 men) for in vitro experiments. AVs structure were evaluated by haematoxylin-eosin, Movat, Alizarin Red, Congo red and Alcian blue/Sirius Red staining and immunohistochemistry. Western blot, ELISA and zymography were used for molecular biology studies. Results AVs from men presented increased inflammatory infiltrates (CD68 and CD45 positive cells) as compared to women. Complementarily, AVs from men exhibited higher levels of the inflammatory molecules interleukin (IL)-6 and IL-1b and RANTES. In line with these results, oxidative stress markers (eNOS, myeloperoxidase, malondialdehyde and nitrotyrosine) were upregulated in male AVs. Concerning, fibrosis, increased levels of collagen type I, fibronectin and syndecan-1 were found in AVs from men. Extracellular matrix (ECM) remodelling was characterized by reduced metalloproteinase-1 expression and increased tissue inhibitor of metalloproteinase-2 expression in male AVs. Importantly, calcification and osteogenic markers (bone morphogenetic protein-9, periostin, osteocalcin and Sox-9) was greatly enhanced in men AVs as compared to women. These findings were confirmed in isolated VICs. At baseline, male VICs presented higher myofibroblast-like phenotype than female VICs. In line with our ex vivo results, male VICs exhibited increased inflammatory, oxidative stress, fibrotic and osteogenic differentiation markers. Conclusions Our results suggest that the mechanisms driving the AV pathogenesis could be different in men and women patients with the same AS severity. Male AVs and isolated VICs presented more inflammation, oxidative stress, fibrosis and ECM remodelling including extensive calcification as compared to female. A better knowledge of the pathophysiological pathways in AVs and VICs will allow developing sex-specific options for AS treatment. FUNDunding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Instituto de Salud Carlos III

Economic assessment of traditional surgical valve replacement versus use of transfemoral intervention in degenerative aortic stenosis

2021 ◽  
Vol 112 (3) ◽  
Author(s):  
Christoph EDLINGER ◽  
Florian KRIZANIC ◽  
Christian BUTTER ◽  
Marwin BANNEHR ◽  
Michael NEUSS ◽  
...  
Keyword(s):  
Aortic Stenosis ◽  
Valve Replacement ◽  
Economic Assessment ◽  
Surgical Valve Replacement

scholarly journals Betulin Promotes Differentiation of Human Osteoblasts In Vitro and Exerts an Osteoinductive Effect on the hFOB 1.19 Cell Line Through Activation of JNK, ERK1/2, and mTOR Kinases

Molecules ◽  
2019 ◽  
Vol 24 (14) ◽  
pp. 2637 ◽  
Author(s):  
Magdalena Mizerska-Kowalska ◽  
Adrianna Sławińska-Brych ◽  
Katarzyna Kaławaj ◽  
Aleksandra Żurek ◽  
Beata Pawińska ◽  
...  
Keyword(s):  
Cell Lines ◽  
Osteoblast Differentiation ◽  
Mrna Level ◽  
Osteogenic Potential ◽  
Type I ◽  
Differentiation Markers ◽  
Alizarin Red ◽  
Human Osteoblasts ◽  
Osteogenic Supplement

Although betulin (BET), a naturally occurring pentacyclic triterpene, has a variety of biological activities, its osteogenic potential has not been investigated so far. The aim of this study was to assess the effect of BET on differentiation of human osteoblasts (hFOB 1.19 and Saos-2 cells) in vitro in osteogenic (with ascorbic acid as an osteogenic supplement) and osteoinductive (without an additional osteogenic supplement) conditions. Osteoblast differentiation was evaluated based on the mRNA expression (RT-qPCR) of Runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), type I collagen-α1 (COL1A1), and osteopontin (OPN). Additionally, ALP activity and production of COL1A1 (western blot analysis) and OPN (ELISA) were evaluated. The level of mineralization (calcium accumulation) was determined with Alizarin red S staining. BET upregulated the mRNA level of RUNX2 and the expression of other osteoblast differentiation markers in both cell lines (except the influence of BET on ALP expression/activity in the Saos-2 cells). Moreover, it increased mineralization in both cell lines in the osteogenic conditions. BET also increased the mRNA level of osteoblast differentiation markers in both cell lines (except for ALP in the Saos-2 cells) in the osteoinductive conditions, which was accompanied with increased matrix mineralization. The osteoinductive activity of BET in the hFOB 1.19 cells was probably mediated via activation of MAPKs (JNK and ERK1/2) and mTOR, as the specific inhibitors of these kinases abolished the BET-induced osteoblast differentiation. Our results suggest that BET has the potential to enhance osteogenesis.

Prosthesis-patient mismatch and left ventricle systolic strain in patients with severe degenerative aortic stenosis, who are undergoing surgical valve replacement

2015 ◽  
pp. 537-546
Author(s):  
Katarzyna Mizia-Stec ◽  
Magdalena Mizia ◽  
Agnieszka Sikora-Puz ◽  
Klaudia Gieszczyk-Strózik ◽  
Tomasz Bochenek ◽  
...  
Keyword(s):  
Left Ventricle ◽  
Aortic Stenosis ◽  
Valve Replacement ◽  
Systolic Strain ◽  
Surgical Valve Replacement

scholarly journals Dihydromyricetin ameliorates vascular calcification in chronic kidney disease by targeting AKT signaling

2021 ◽  
Author(s):  
Liyun Feng ◽  
Dongdong Que ◽  
Zehua Li ◽  
Xinglong Zhong ◽  
Jing Yan ◽  
...  
Keyword(s):  
Chronic Kidney Disease ◽  
Kidney Disease ◽  
Vascular Calcification ◽  
Type I Collagen ◽  
Ex Vivo ◽  
Therapeutic Option ◽  
Akt Signaling ◽  
Type I ◽  
Dependent Manner ◽  
Alizarin Red

Vascular calcification is highly prevalent in chronic kidney disease (CKD), and characterized by trans-differentiation from contractile vascular smooth muscle cells (VSMCs) into an osteogenic phenotype. However, no effective and therapeutic option to prevent vascular calcification is yet available. Dihydromyricetin (DMY), a bioactive flavonoid isolated from Ampelopsis grossedentata, has been found to inhibit VSMCs proliferation and the injury-induced neointimal formation. However, whether DMY has an effect on osteogenic differentiation of VSMCs and vascular calcification is still unclear. In this study, we sought to investigate the effect of DMY on vascular calcification in CKD and the underlying mechanism. DMY treatment significantly attenuated calcium/phosphate-induced calcification of rat and human VSMCs in a dose-dependent manner, as shown by alizarin red S staining and calcium content assay, associated with down-regulation of osteogenic markers including type I collagen (COL I), RUNX2, BMP2 and osteocalcin (OCN). These results were further confirmed in aortic rings ex vivo. Moreover, DMY ameliorated vascular calcification in rats with CKD. Additionally, we found that AKT signaling was activated during vascular calcification, whereas significantly inhibited by DMY administration. DMY treatment significantly reversed AKT activator-induced vascular calcification. Furthermore, inhibition of AKT signaling efficiently attenuated calcification, which was similar to that after treatment with DMY alone, and DMY had a better inhibitory effect on calcification as compared to AKT inhibitor. The present study demonstrated that DMY has a potent inhibitory role in vascular calcification partially by inhibiting AKT activation, suggesting that DMY may act as a promising therapeutic candidate for patients suffering from vascular calcification.

scholarly journals Regulation of neutrophil gelatinase-associated lipocalin in aortic valve stenosis

2021 ◽  
Vol 42 (Supplement_1) ◽  
Author(s):  
E Jover ◽  
L Matilla ◽  
M Garaikoetxea ◽  
A Fernandez-Celis ◽  
R Sabada ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Aortic Valve ◽  
Type I ◽  
Serum Samples ◽  
Alizarin Red ◽  
Neutrophil Gelatinase Associated Lipocalin ◽  
Wide Range ◽  
Neutrophil Gelatinase ◽  
And Oxidative Stress

Abstract Background Aortic valve (AV) stenosis is the commonest form of adult valvular heart disease (VHD) and affects 4.5% of the general population aged over 60 years. Owing to multifactorial and complex molecular events, the valve interstitial cell (VIC) undergoes myofibroblast and osteoblast differentiation. Neutrophil gelatinase-associated lipocalin (NGAL) is a pleiotropic glycoprotein belonging to the lipocalin family and it is expressed in a wide range of tissues and cell types. It is deregulated in several diseases with both detrimental and beneficial effects. NGAL mainly signals towards 24p3R. Purpose We aimed to confirm the expression of NGAL in human AV stenosis and its association with inflammation, oxidative stress, fibrosis/remodeling and calcification, and its regulation in calcifying VICs. Methods Surgical AV leftovers were harvested from patients undergoing elective surgical valve replacement with no kidney disease. Serum samples were collected within the 24h before the surgery. AV were histologically assessed by hematoxylin-eosin, Movat, Alizarin Red and Alcian blue/Sirius Red staining and immunohistochemistry. Western blotting, ELISA and zymography were used in molecular biology studies. VICs were challenged for 2, 4 and 8 days with hyperphosphate (2.6mM, HP) media ± rhNGAL for in vitro validation studies. Results NGAL was quantified in AVs and serum samples from 126 donors (57.4% men). Circulating NGAL was associated with circulating inflammation (Tumor Necrosis Factor-α, Interleukin (IL)-6 and ICAM-1) and oxidative stress (Myeloperoxidase (MPO) and 8OHdG) markers. Likewise, tissue NGAL was correlated with inflammation (IL-6, RANTES and Galectin-3), oxidative stress (MPO, Endothelial Nitric Oxide Synthase, Malondialdehyde (MDA) and Carboxy Methyl Lysine (CML)) and fibrosis (Collagen type I). Osteoblast markers, metalloproteinase (MMP)-9 expression or its activity were not associated with NGAL. NGAL was greater expressed in men than women (217.70±23.41 vs. 119.5±11.31, p=0.0098). In vitro, intracellular NGAL and 24p3R were strongly down-regulated in calcifying men-derived VICs (n=6) whereas secretion of NGAL was enhanced from day 4 (0.55±0.15, p=0.0283; 0.32±0.09 p<0.0001; 8.00±2.32, p=0.0053 fold changes, respectively). This may reflect reduced 24p3R-dependent signalling in osteoblast-like VICs. Such effects were overall negated in women-derived VICs (n=3). RhNGAL endowed calcifying VICs with increased necrosis (52KDa-cPARP1), apoptosis (cCaspase 3) and oxidative stress (CML, MDA, nitrotyrosine and pNF-kB) at day 8. Conclusions NGAL is associated with inflammation, oxidative stress and fibrosis in AV stenosis, and promotes pro-apoptotic and necrotic phenotypes in vitro. Our results suggest that NGAL signaling may drive sex-dependent differences clinically relevant to the VHD pathogenesis. The challenge is now to understand how NGAL signals in men/women-derived VICs. FUNDunding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Instituto de Salud Carlos III

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