Adhesion and spreading of corneal endothelial cells on collagens type I and IV in vitro: a model to study mechanisms of endothelial repair

Objectives: To develop and validate a 3D in-vitro model of atherosclerosis that enables direct interaction between various cell types and/or extracellular matrix. Methods and Results: Type I collagen (0.75 mg/mL) was mixed with human artery smooth muscle cells (SMCs; 6x10 5 cells/mL), medium, and water. Human coronary artery endothelial cells (HCAECs; 10 5 /cm 2 ) were plated on top of the collagen gels and activated with oxidized low density lipoprotein cholesterol (LDL-C). Monocytes (THP-1 cells; 10 5 /cm 2 ) were then added on top of the HCAECs. Immunofluorescence showed the expression of VE-cadherin by HCAECs (A, B) and α-smooth muscle actin by SMCs (A). Green-labelled LDL-C particles were accumulated in the subendothelial space, as well as in the cytoplasm of HCAECs and SMCs (C). Activated monocytes were attached to HCAECs and found in the subendothelial area (G-I). Both HCAECs and SMCs released IL-1β, IL-6, IL-8, PDGF-BB, TGF-ß1, and VEGF. Scanning and transmission electron microscopy showed the HCAECs monolayer forming gap junctions and the SMCs (D-F) and transmigrating monocytes within the collagen matrix (G-I). Conclusions: In this work, we presented a novel, easily reproducible and functional in-vitro experimental model of atherosclerosis that has the potential to enable in-vitro sophisticated molecular and drug development studies.

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Disparate cytokine regulation of ICAM-1 in rat alveolar epithelial cells and pulmonary endothelial cells in vitro

AJP Lung Cellular and Molecular Physiology ◽

10.1152/ajplung.1995.269.1.l127 ◽

1995 ◽

Vol 269 (1) ◽

pp. L127-L135 ◽

Cited By ~ 5

Author(s):

W. W. Barton ◽

S. Wilcoxen ◽

P. J. Christensen ◽

R. Paine

Keyword(s):

Endothelial Cells ◽

Epithelial Cells ◽

Inflammatory Cytokines ◽

Alveolar Epithelial Cells ◽

Normal Lung ◽

Type I ◽

Type Ii Cells ◽

Type Ii ◽

Alveolar Epithelial

Intercellular adhesion molecule-1 (ICAM-1) is expressed at high levels on type I alveolar epithelial cells in the normal lung and is induced in vitro as type II cells spread in primary culture. In contrast, in most nonhematopoetic cells ICAM-1 expression is induced in response to inflammatory cytokines. We have formed the hypothesis that the signals that control ICAM-1 expression in alveolar epithelial cells are fundamentally different from those controlling expression in most other cells. To test this hypothesis, we have investigated the influence of inflammatory cytokines on ICAM-1 expression in isolated type II cells that have spread in culture and compared this response to that of rat pulmonary artery endothelial cells (RPAEC). ICAM-1 protein, determined both by a cell-based enzyme-linked immunosorbent assay and by Western blot analysis, and mRNA were minimally expressed in unstimulated RPAEC but were significantly induced in a time- and dose-dependent manner by treatment with tumor necrosis factor-alpha, interleukin-1 beta, or interferon-gamma. In contrast, these cytokines did not influence the constitutive high level ICAM-1 protein expression in alveolar epithelial cells and only minimally affected steady-state mRNA levels. ICAM-1 mRNA half-life, measured in the presence of actinomycin D, was relatively long at 7 h in alveolar epithelial cells and 4 h in RPAEC. The striking lack of response of ICAM-1 expression by alveolar epithelial cells to inflammatory cytokines is in contrast to virtually all other epithelial cells studied to date and supports the hypothesis that ICAM-1 expression by these cells is a function of cellular differentiation.(ABSTRACT TRUNCATED AT 250 WORDS)

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Uvaol Improves the Functioning of Fibroblasts and Endothelial Cells and Accelerates the Healing of Cutaneous Wounds in Mice

Molecules ◽

10.3390/molecules25214982 ◽

2020 ◽

Vol 25 (21) ◽

pp. 4982

Author(s):

Julianderson Carmo ◽

Polliane Cavalcante-Araújo ◽

Juliane Silva ◽

Jamylle Ferro ◽

Ana Carolina Correia ◽

...

Keyword(s):

Endothelial Cells ◽

Healing Process ◽

Virgin Olive Oil ◽

Mapk Signaling ◽

Control Treatment ◽

Type I ◽

Pentacyclic Triterpene ◽

Cutaneous Wounds ◽

Positive Effects

Uvaol is a natural pentacyclic triterpene that is widely found in olives and virgin olive oil, exerting various pharmacological properties. However, information remains limited about how it affects fibroblasts and endothelial cells in events associated with wound healing. Here, we report the effect of uvaol in the in vitro and in vivo healing process. We show the positive effects of uvaol on migration of fibroblasts and endothelial cells in the scratch assay. Protein synthesis of fibronectin and laminin (but not collagen type I) was improved in uvaol-treated fibroblasts. In comparison, tube formation by endothelial cells was enhanced after uvaol treatment. Mechanistically, the effects of uvaol on cell migration involved the PKA and p38-MAPK signaling pathway in endothelial cells but not in fibroblasts. Thus, the uvaol-induced migratory response was dependent on the PKA pathway. Finally, topical treatment with uvaol caused wounds to close faster than in the control treatment using experimental cutaneous wounds model in mice. In conclusion, uvaol positively affects the behavior of fibroblasts and endothelial cells, potentially promoting cutaneous healing.

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Equid Herpesvirus 1 Targets the Sensitization and Induction Steps To Inhibit the Type I Interferon Response in Equine Endothelial Cells

Journal of Virology ◽

10.1128/jvi.01342-19 ◽

2019 ◽

Vol 93 (23) ◽

Cited By ~ 2

Author(s):

Fatai S. Oladunni ◽

Sanjay Sarkar ◽

Stephanie Reedy ◽

Udeni B. R. Balasuriya ◽

David W. Horohov ◽

...

Keyword(s):

Endothelial Cells ◽

Cellular Level ◽

Immune Defense ◽

Interferon Response ◽

Type I ◽

Type I Ifn ◽

Viral Pathogen ◽

Equid Herpesvirus ◽

Herpesvirus 1

ABSTRACT Equid herpesvirus 1 (EHV-1) is a viral pathogen of horse populations worldwide spread by the respiratory route and is known for causing outbreaks of neurologic syndromes and abortion storms. Previously, we demonstrated that an EHV-1 strain of the neuropathogenic genotype, T953, downregulates the beta interferon (IFN-β) response in vitro in equine endothelial cells (EECs) at 12 h postinfection (hpi). In the present study, we explored the molecular correlates of this inhibition as clues toward an understanding of the mechanism. Data from our study revealed that EHV-1 infection of EECs significantly reduced both Toll-like receptor 3 (TLR3) and TLR4 mRNA expression at 6 hpi and 12 hpi. While EHV-1 was able to significantly reduce IRF9 mRNA at both 6 hpi and 12 hpi, the virus significantly reduced IFN regulatory factor 7 (IRF7) mRNA only at 12 hpi. EHV-1 did not alter the cellular level of Janus-activated kinase 1 (JAK1) at any time point. However, EHV-1 reduced the cellular level of expression of tyrosine kinase 2 (TYK2) at 12 hpi. Downstream of JAK1-TYK2 signaling, EHV-1 blocked the phosphorylation and activation of signal transducer and activator of transcription 2 (STAT2) when coincubated with exogenous IFN, at 12 hpi, although not at 3 or 6 hpi. Immunofluorescence staining revealed that the virus prevented the nuclear translocation of STAT2 molecules, confirming the virus-mediated inhibition of STAT2 activation. The pattern of suppression of phosphorylation of STAT2 by EHV-1 implicated viral late gene expression. These data help illuminate how EHV-1 strategically inhibits the host innate immune defense by limiting steps required for type I IFN sensitization and induction. IMPORTANCE To date, no commercial vaccine label has a claim to be fully protective against the diseases caused by equid herpesvirus 1 (EHV-1), especially the neurologic form. The interferon (IFN) system, of which type I IFN is of great importance, still remains a viable immunotherapeutic option against EHV-1 infection. The type I IFN system has been exploited successfully to treat other viral infections, such as chronic hepatitis B and C in humans. The current state of research on how EHV-1 interferes with the protective effect of type I IFN has indicated transient induction of type I IFN production followed by a rapid shutdown in vitro in equine endothelial cells (EECs). The significance of our study is the identification of certain steps in the type I IFN signaling pathway targeted for inhibition by EHV-1. Understanding this pathogen-host relationship is essential for the long-term goal of developing effective immunotherapy against EHV-1.

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Corneal endothelial cell dysfunction: etiologies and management

Therapeutic Advances in Ophthalmology ◽

10.1177/2515841418815802 ◽

2018 ◽

Vol 10 ◽

pp. 251584141881580 ◽

Cited By ~ 13

Author(s):

Sepehr Feizi

Keyword(s):

Endothelial Cells ◽

Endothelial Cell ◽

Endothelial Dysfunction ◽

Corneal Endothelial Cell ◽

Endothelial Cell Dysfunction ◽

Endothelial Keratoplasty ◽

Corneal Endothelial Cells ◽

Cell Dysfunction ◽

Corneal Endothelial Dysfunction

A transparent cornea is essential for the formation of a clear image on the retina. The human cornea is arranged into well-organized layers, and each layer plays a significant role in maintaining the transparency and viability of the tissue. The endothelium has both barrier and pump functions, which are important for the maintenance of corneal clarity. Many etiologies, including Fuchs’ endothelial corneal dystrophy, surgical trauma, and congenital hereditary endothelial dystrophy, lead to endothelial cell dysfunction. The main treatment for corneal decompensation is replacement of the abnormal corneal layers with normal donor tissue. Nowadays, the trend is to perform selective endothelial keratoplasty, including Descemet stripping automated endothelial keratoplasty and Descemet’s membrane endothelial keratoplasty, to manage corneal endothelial dysfunction. This selective approach has several advantages over penetrating keratoplasty, including rapid recovery of visual acuity, less likelihood of graft rejection, and better patient satisfaction. However, the global limitation in the supply of donor corneas is becoming an increasing challenge, necessitating alternatives to reduce this demand. Consequently, in vitro expansion of human corneal endothelial cells is evolving as a sustainable choice. This method is intended to prepare corneal endothelial cells in vitro that can be transferred to the eye. Herein, we describe the etiologies and manifestations of human corneal endothelial cell dysfunction. We also summarize the available options for as well as recent developments in the management of corneal endothelial dysfunction.

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MT1-MMP–dependent neovessel formation within the confines of the three-dimensional extracellular matrix

The Journal of Cell Biology ◽

10.1083/jcb.200405001 ◽

2004 ◽

Vol 167 (4) ◽

pp. 757-767 ◽

Cited By ~ 223

Author(s):

Tae-Hwa Chun ◽

Farideh Sabeh ◽

Ichiro Ota ◽

Hedwig Murphy ◽

Kevin T. McDonagh ◽

...

Keyword(s):

Extracellular Matrix ◽

Endothelial Cells ◽

Cell Surface ◽

Type I Collagen ◽

Ex Vivo ◽

Three Dimensional ◽

Collagenolytic Activity ◽

Type I ◽

Ex Vivo Model

During angiogenesis, endothelial cells initiate a tissue-invasive program within an interstitial matrix comprised largely of type I collagen. Extracellular matrix–degradative enzymes, including the matrix metalloproteinases (MMPs) MMP-2 and MMP-9, are thought to play key roles in angiogenesis by binding to docking sites on the cell surface after activation by plasmin- and/or membrane-type (MT) 1-MMP–dependent processes. To identify proteinases critical to neovessel formation, an ex vivo model of angiogenesis has been established wherein tissue explants from gene-targeted mice are embedded within a three-dimensional, type I collagen matrix. Unexpectedly, neither MMP-2, MMP-9, their cognate cell-surface receptors (i.e., β3 integrin and CD44), nor plasminogen are essential for collagenolytic activity, endothelial cell invasion, or neovessel formation. Instead, the membrane-anchored MMP, MT1-MMP, confers endothelial cells with the ability to express invasive and tubulogenic activity in a collagen-rich milieu, in vitro or in vivo, where it plays an indispensable role in driving neovessel formation.

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Activation of a procollagenase by low-molecular-weight angiogenesis factor

Bioscience Reports ◽

10.1007/bf01121948 ◽

1983 ◽

Vol 3 (2) ◽

pp. 171-177 ◽

Cited By ~ 33

Author(s):

Jacqueline B. Weiss ◽

C. R. Hill ◽

R. J. Davis ◽

B. McLaughlin ◽

K. A. Sedowofia ◽

...

Keyword(s):

Molecular Weight ◽

Endothelial Cells ◽

Endothelial Cell ◽

Cell Growth ◽

Low Molecular Weight ◽

Type I ◽

Angiogenesis Factor ◽

The Matrix ◽

Endothelial Cell Growth

Avascular tumours have the ability to establish a blood supply for themselves by secreting a humoral factor which stimulates their host's endothelial cells to proliferate and to migrate towards the tumour source. The mechanism of action of such a humoral anglo-genesis factor is more than that of an endothelial-cell growth factor since it requires an oriented migration of cells towards the tumour. We report here the activation of pure skin-fibroblast procollagenase by a low-molecular-weight angiogenesis factor capable of stimulating endothelial-cell growth in vitro. The activation was observed when either Type I or III collagen was used as substrate. It is suggested that at least one function of angiogenesis factor is to promote limited degradation of the connective tissue through which it passes causing channeling in the matrix along which stimulated endothelial cells may

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Transplantation of Adult Human or Porcine Corneal Endothelial Cells onto Human Recipients In Vitro. Part I

Cornea ◽

10.1097/00003226-199903000-00010 ◽

1999 ◽

Vol 18 (2) ◽

pp. 199 ◽

Cited By ~ 50

Author(s):

Katrin Engelmann ◽

Dagmar Drexler ◽

Matthias Böhnke

Keyword(s):

Endothelial Cells ◽

Corneal Endothelial Cells ◽

Adult Human

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