scholarly journals Altered shear stress on endothelial cells leads to remodeling of extracellular matrix and induction of angiogenesis

PLoS ONE ◽  
2020 ◽  
Vol 15 (11) ◽  
pp. e0241040
Author(s):  
T. A. Russo ◽  
A. M. M. Banuth ◽  
H. B. Nader ◽  
J. L. Dreyfuss
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Shear Stress ◽  
Pathological Condition ◽  
Cell Interaction ◽  
Tube Formation ◽  
Mechanical Forces ◽  
Matrix Remodeling ◽  
Collagen Iii ◽  
Remodeling Of Extracellular Matrix

Endothelial cells (ECs) are subjected to physical forces such as shear stress (SS) induced by blood flow that leads to significant changes in morphology, physiology and gene expression. The abnormal mechanical forces applied in the cardiovascular system can influence the development of conditions and diseases such as thrombosis, hypertension and atherosclerosis. This study investigated the expression of glycosaminoglycans (GAGs), proteoglycans and extracellular matrix molecules in ECs exposed to normal and altered SS. ECs were exposed to SS of 12 dyn/cm2 (artery physiological condition) and 4 dyn/cm2 (artery pathological condition). Subsequently, ECs were subjected to immunofluorescence, qPCR, GAG biosynthesis analyses and cell-based assays. SS induced changes in ECs morphology. There were other pathological consequences of altered SS, including inhibited adhesion, stimulation of migration and capillary-like tube formation, as well as increases of GAG synthesis. We observed higher expression of syndecan-4, perlecan, decorin, fibronectin and collagen III α1 and growth factors, including VEGF-A and TGFβ-1. ECs exposed to SS displayed extracellular matrix remodeling as well as expression of cell-matrix and cell-cell interaction molecules. This study contributes to the understanding of how vascular biology is affected by mechanical forces and how these molecules can be affected in cardiovascular diseases.

Naturally Produced Extracellular Matrix Is an Excellent Substrate for Canine Endothelial Cell Proliferation and Resistance to Shear Stress on PTFE Vascular Grafts

1997 ◽  
Vol 78 (05) ◽  
pp. 1392-1398 ◽  
Author(s):  
A Schneider ◽  
M Chandra ◽  
G Lazarovici ◽  
I Vlodavsky ◽  
G Merin ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Shear Stress ◽  
Endothelial Cell ◽  
Cell Attachment ◽  
Thrombus Formation ◽  
Endothelial Cell Proliferation ◽  
Ptfe Graft ◽  
Vascular Prostheses ◽  
Endothelial Cell Attachment

SummaryPurpose: Successful development of a vascular prosthesis lined with endothelial cells (EC) may depend on the ability of the attached cells to resist shear forces after implantation. The present study was designed to investigate EC detachment from extracellular matrix (ECM) precoated vascular prostheses, caused by shear stress in vitro and to test the performance of these grafts in vivo. Methods: Bovine aortic endothelial cells were seeded inside untreated polytetrafluoro-ethylene (PTFE) vascular graft (10 X 0.6 cm), PTFE graft precoated with fibronectin (FN), or PTFE precoated with FN and a naturally produced ECM (106 cells/graft). Sixteen hours after seeding the medium was replaced and unattached cells counted. The strength of endothelial cell attachment was evaluated by subjecting the grafts to a physiologic shear stress of 15 dynes/cm2 for 1 h. The detached cells were collected and quantitated. PTFE or EC preseeded ECM coated grafts were implanted in the common carotid arteries of dogs. Results: While little or no differences were found in the extent of endothelial cell attachment to the various grafts (79%, 87% and 94% of the cells attached to PTFE, FN precoated PTFE, or FN+ECM precoated PTFE, respectively), the number of cells retained after a shear stress was significanly increased on ECM coated PTFE (20%, 54% and 85% on PTFE, FN coated PTFE, and FN+ECM coated PTFE, respectively, p <0.01). Implantation experiments in dogs revealed a significant increase in EC coverage and a reduced incidence of thrombus formation on ECM coated grafts that were seeded with autologous saphenous vein endothelial cells prior to implantation. Conclusion: ECM coating significantly increased the strength of endothelial cell attachment to vascular prostheses subjected to shear stress. The presence of adhesive macromolecules and potent endothelial cell growth promoting factors may render the ECM a promising substrate for vascular prostheses.

scholarly journals Cardiac Extracellular Vesicles (EVs) Released in the Presence or Absence of Inflammatory Cues Support Angiogenesis in Different Manners

2019 ◽  
Vol 20 (24) ◽  
pp. 6363 ◽  
Author(s):  
Christien Madlen Beez ◽  
Maria Schneider ◽  
Marion Haag ◽  
Kathleen Pappritz ◽  
Sophie Van Linthout ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Conditioned Medium ◽  
Extracellular Vesicles ◽  
Quantitative Polymerase Chain Reaction ◽  
Cell Interaction ◽  
Tube Formation ◽  
Vascular Endothelial ◽  
Human Proteins ◽  
Intracellular Detection ◽  
Endothelial Growth Factor

Cells release extracellular vesicles (EVs) to communicate in a paracrine manner with other cells, and thereby influence processes, such as angiogenesis. The conditioned medium of human cardiac-derived adherent proliferating (CardAP) cells was recently shown to enhance angiogenesis. To elucidate whether their released EVs are involved, we isolated them by differential centrifugation from the conditioned medium derived either in the presence or absence of a pro-inflammatory cytokine cocktail. Murine recipient cells internalized CardAP-EVs as determined by an intracellular detection of human proteins, such as CD63, by a novel flow cytometry method for studying EV–cell interaction. Moreover, endothelial cells treated for 24 h with either unstimulated or cytokine stimulated CardAP-EVs exhibited a higher tube formation capability on Matrigel. Interestingly, unstimulated CardAP-EVs caused endothelial cells to release significantly more vascular endothelial growth factor and interleukin (IL)-6, while cytokine stimulated CardAP-EVs significantly enhanced the release of IL-6 and IL-8. By nCounter® miRNA expression assay (NanoString Technologies) we identified microRNA 302d-3p to be enhanced in unstimulated CardAP-EVs compared to their cytokine stimulated counterparts, which was verified by quantitative polymerase chain reaction. This study demonstrates that both CardAP-EVs are pro-angiogenic by inducing different factors from endothelial cells. This would allow to select potent targets for a safe and efficient therapeutic application.

scholarly journals Metabolic Disorder of Extracellular Matrix Mediated by Decorin Upregulation Is Associated With Brain Arteriovenous Malformation Diffuseness

2020 ◽  
Vol 12 ◽  
Author(s):  
Maogui Li ◽  
Qingyuan Liu ◽  
Junhua Yang ◽  
Pengjun Jiang ◽  
Yi Yang ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Signaling Pathway ◽  
Transforming Growth Factor Beta ◽  
Proteomic Analysis ◽  
Metabolic Disorder ◽  
Transforming Growth Factor ◽  
Tube Formation ◽  
Normal Brain ◽  
Altered Expression

Background and ObjectiveDiffuse brain arteriovenous malformations (BAVMs) are mixed up with normal brain parenchyma and therefore increase the difficulty of surgical resection, leading to poor surgical prognosis. Since the mechanism underlying BAVM diffuseness remains unknown, a quantitative proteomic analysis was performed to investigate the altered expression of proteins in diffuse BAVMs compared to compact ones.MethodsWe performed proteomic analysis on five diffuse BAVMs and five compact BAVMs. Bioinformatics analysis was conducted to identify potential signals related to BAVM diffuseness. Candidate proteins were then investigated in BAVM specimens using immunofluorescence and Western blot analysis. Tube formation assays were used to investigate the effects of candidate proteins on the angiogenesis of human umbilical endothelial cells (HUVECs). Finally, Masson, Sirius red staining, and immunofluorescence were used to evaluate the characteristics of extracellular matrix (ECM) in BAVM tissues.ResultsA total of 58 proteins were found to be differentially expressed between diffuse and compact BAVMs via proteomic analysis. TGF-β (transforming growth factor-beta) signaling pathway, ECM–receptor pathway, relaxin signaling pathway, and several other pathways were associated with BAVM diffuseness. The TGF-β signaling pathway is associated with angiogenesis; the role of this pathway in the formation of diffuse BAVMs was investigated, and the decorin (DCN) upregulation played an important role in this process. Immunofluorescence showed that DCN was significantly upregulated within and around the malformed vessels of diffuse BAVMs. Functional assays showed that exogenous DCN could promote the tube formation ability of HUVECs through inhibiting the TGF-β signaling pathway and overproducing ECM. Histological staining demonstrated the overproduction of ECM in diffuse BAVMs.ConclusionTGF-β signaling pathway inhibited by DCN in vascular endothelial cells is related to BAVM diffuseness. The metabolic disorder of ECM caused by DCN upregulation may significantly contribute to the formation of diffuse BAVMs.

Regulation of Extracellular Matrix Remodeling and MMP-2 activation in Cultured Rat Adrenal Medullary Endothelial Cells

Endothelium ◽  
2001 ◽  
Vol 8 (3) ◽  
pp. 181-194 ◽  
Author(s):  
E. Papadimitriou ◽  
C. R. Waters ◽  
V. G. Manolopoulos ◽  
B. R. Unsworth ◽  
M. E. Maragoudakis ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Extracellular Matrix Remodeling ◽  
Matrix Remodeling

Abstract P016: GYY4137, a Hydrogen Sulfide Donor, Mitigates Ang II-induced Extracellular Matrix Remodeling in Glomerular Endothelial and Mesangial Cells

Hypertension ◽  
2015 ◽  
Vol 66 (suppl_1) ◽  
Author(s):  
Matthew Amin ◽  
Sathnur Pushpakumar ◽  
Sourav Kundu ◽  
Geetansh Tyagi ◽  
Aaron Tyagi ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Hydrogen Sulfide ◽  
Mesangial Cells ◽  
Collagen Iv ◽  
Tissue Inhibitors Of Metalloproteinases ◽  
Matrix Remodeling ◽  
Ang Ii ◽  
Hypertensive Nephropathy ◽  
Glomerular Endothelial Cells

Hypertensive nephropathy is associated with progressive alteration of extracellular matrix (ECM) components. Both mesangial and glomerular endothelial cells have the ability to synthesize and degrade ECM proteins such as collagens by changes in the activity of matrix metalloproteinases (MMPs) and their inhibitors, tissue inhibitors of metalloproteinases (TIMPs). Endo180 is an extracellular fibronectin type II domain involved in lysosomal degradation of collagen which has been shown to mitigate renal fibrosis. More recently, hydrogen sulfide (H2S) has also been shown to mitigate hypertensive renal remodeling, however, its mechanism remains unclear. In this study, our aim was to investigate whether Angiotensin-II (Ang II) treatment alters the expression of Endo180, MMPs and TIMPs leading to dysregulation of collagen metabolism and whether GYY4137 (H2S donor) restores their levels to achieve homeostasis. Mesangial and mouse glomerular endothelial cells (MCs and MGECs respectively) were treated without or with Ang II (200 nM) and GYY4137 (250 μM) for 48hrs. Cell lysates were analyzed for MMP-13, -14, TIMP-1, Endo180, and collagen IV by Western blot, RT-PCR, and immunohistochemistry. In MGECs, Ang II treatment compared to its control decreased MMP-13/TIMP-1 ratio (0.75±0.44 vs. 2.48 ±0.73), and upregulated MMP-14/TIMP-1 ratio (0.64±2.10 vs. 0.96±1.47), and collagen IV (0.77±0.07 vs. 0.58±0.06). GYY4137 treatment mitigated these changes. In contrast, Ang II treatment in MCs decreased Endo180 compared to control (0.72±0.04 vs. 1.07±0.23), but did not alter the expression of MMP-13/TIMP-1, MMP-14/TIMP-1 ratios, and collagen IV level compared to control or MGECs. Similarly, immunostaining showed downregulation of MMP-13 and Endo180 in Ang II treated MGECs which was normalized following GYY4137 treatment. Endo180 was also normalized in MCs following GYY4137 treatment however, there was no change in MMP-14/TIMP-1 ratio or collagen IV level. We conclude that Ang II treatment causes adverse ECM remodeling in MGECs via downregulation of Endo180 and MMP-13 and upregulation of MMP-14 and TIMP-1 and in MCs by decreasing Endo180, and GYY4137 mitigates these changes in part, by modulating Endo180/MMP/TIMP pathway.

Construction of multi-functional extracellular matrix proteins that promote tube formation of endothelial cells

Biomaterials ◽  
2008 ◽  
Vol 29 (20) ◽  
pp. 2977-2986 ◽  
Author(s):  
Makiko Nakamura ◽  
Masayasu Mie ◽  
Hisakazu Mihara ◽  
Makoto Nakamura ◽  
Eiry Kobatake
Keyword(s):  
Extracellular Matrix ◽  
Endothelial Cells ◽  
Extracellular Matrix Proteins ◽  
Tube Formation ◽  
Matrix Proteins

scholarly journals An In Vitro Study on Extracellular Vesicles From Adipose-Derived Mesenchymal Stem Cells in Protecting Stress Urinary Incontinence Through MicroRNA-93/F3 Axis

2021 ◽  
Vol 12 ◽  
Author(s):  
Lu Wang ◽  
Yali Wang ◽  
Yuancui Xiang ◽  
Jinping Ma ◽  
Hui Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Extracellular Matrix ◽  
Urinary Incontinence ◽  
Stress Urinary Incontinence ◽  
Extracellular Vesicles ◽  
Satellite Cells ◽  
Extracellular Matrix Remodeling ◽  
Matrix Remodeling ◽  
Collagen Iii ◽  
Primary Fibroblasts

Since the potential roles of extracellular vesicles secreted by adipose-derived mesenchymal stem cells (ADSCs) are not well understood in collagen metabolism, the purpose of this research was to evaluate the effects of ADSCs-extracellular vesicles in stress urinary incontinence and the regulatory mechanism of delivered microRNA-93 (miR-93). ADSCs were isolated and cultured, and ADSCs-extracellular vesicles were extracted and identified. Stress urinary incontinence primary fibroblasts or satellite cells were treated with ADSCs-extracellular vesicles to detect the expression of Elastin, Collagen I, and Collagen III in fibroblasts and Pax7 and MyoD in satellite cells. After transfecting ADSCs with miR-93 mimics or inhibitors, extracellular vesicles were isolated and treated with stress urinary incontinence primary fibroblasts or satellite cells to observe cell function changes. The online prediction and luciferase activity assay confirmed the targeting relationship between miR-93 and coagulation factor III (F3). The rescue experiment verified the role of ADSCs-extracellular vesicles carrying miR-93 in stress urinary incontinence primary fibroblasts and satellite cells by targeting F3. ADSCs-extracellular vesicles treatment upregulated expression of Elastin, Collagen I, and Collagen III in stress urinary incontinence primary fibroblasts and expression of Pax7 and MyoD in stress urinary incontinence primary satellite cells. miR-93 expression was increased in stress urinary incontinence primary fibroblasts or satellite cells treated with ADSCs-extracellular vesicles. Extracellular vesicles secreted by ADSCs could deliver miR-93 to fibroblasts and then negatively regulate F3 expression; ADSCs-extracellular vesicles could reverse the effect of F3 on extracellular matrix remodeling in stress urinary incontinence fibroblasts. miR-93 expression was also increased in stress urinary incontinence primary satellite cells treated by ADSCs-extracellular vesicles. Extracellular vesicles secreted by ADSCs were delivered to satellite cells through miR-93, which directly targets F3 expression and upregulates Pax7 and MyoD expression in satellite cells. Our study indicates that miR-93 delivered by ADSCs-extracellular vesicles could regulate extracellular matrix remodeling of stress urinary incontinence fibroblasts and promote activation of stress urinary incontinence satellite cells through targeting F3.

scholarly journals Coordinated interactions between endothelial cells and macrophages in the islet microenvironment promote β cell regeneration

2020 ◽  
Author(s):  
Diane C. Saunders ◽  
Kristie I. Aamodt ◽  
Tiffany M. Richardson ◽  
Alec Hopkirk ◽  
Radhika Aramandla ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Cell Proliferation ◽  
Endothelial Cells ◽  
Cell Loss ◽  
Cell Regeneration ◽  
Matrix Remodeling ◽  
Cell Recovery ◽  
Β Cells ◽  
Β Cell ◽  
Elevated Expression

ABSTRACTEndogenous β cell regeneration could alleviate diabetes, but proliferative stimuli within the islet microenvironment are incompletely understood. We previously found that β cell recovery following hypervascularization-induced β cell loss involves interactions with endothelial cells (ECs) and macrophages (MΦs). Here we show that proliferative ECs modulate MΦ infiltration and phenotype during β cell loss, and recruited MΦs are essential for β cell recovery. Furthermore, VEGFR2 inactivation in quiescent ECs accelerates islet vascular regression during β cell recovery and leads to increased β cell proliferation without changes in MΦ phenotype or number. Transcriptome analysis of β cells, ECs, and MΦs reveals that β cell proliferation coincides with elevated expression of extracellular matrix remodeling molecules and growth factors likely driving activation of proliferative signaling pathways in β cells. Collectively, these findings suggest a new β cell regeneration paradigm whereby coordinated interactions between intra-islet MΦs, ECs, and extracellular matrix mediate β cell self-renewal.

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