Shear stress enhances human endothelial cell wound closure in vitro

2000 ◽  
Vol 279 (1) ◽  
pp. H293-H302 ◽  
Author(s):  
Maria Luiza C. Albuquerque ◽  
Christopher M. Waters ◽  
Ushma Savla ◽  
H. William Schnaper ◽  
Annette S. Flozak
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Endothelial Cell ◽  
Wound Closure ◽  
Type I Collagen ◽  
Umbilical Vein ◽  
Type I ◽  
Laminar Shear Stress ◽  
Human Coronary Artery ◽  
Closure Rate

Repair of the endothelium occurs in the presence of continued blood flow, yet the mechanisms by which shear forces affect endothelial wound closure remain elusive. Therefore, we tested the hypothesis that shear stress enhances endothelial cell wound closure. Human umbilical vein endothelial cells (HUVEC) or human coronary artery endothelial cells (HCAEC) were cultured on type I collagen-coated coverslips. Cell monolayers were sheared for 18 h in a parallel-plate flow chamber at 12 dyn/cm2 to attain cellular alignment and then wounded by scraping with a metal spatula. Subsequently, the monolayers were exposed to a laminar shear stress of 3, 12, or 20 dyn/cm2 under shear-wound-shear (S-W-sH) or shear-wound-static (S-W-sT) conditions for 6 h. Wound closure was measured as a percentage of original wound width. Cell area, centroid-to-centroid distance, and cell velocity were also measured. HUVEC wounds in the S-W-sH group exposed to 3, 12, or 20 dyn/cm2 closed to 21, 39, or 50%, respectively, compared with only 59% in the S-W-sT cells. Similarly, HCAEC wounds closed to 29, 49, or 33% (S-W-sH) compared with 58% in the S-W-sT cells. Cell spreading and migration, but not proliferation, were the major mechanisms accounting for the increases in wound closure rate. These results suggest that physiological levels of shear stress enhance endothelial repair.

scholarly journals Attachment of Human Endothelial Cells to Polyester Vascular Grafts: Pre-Coating With Adhesive Protein Assemblies and Resistance to Short-Term Shear Stress

2014 ◽  
pp. 167-177 ◽  
Author(s):  
J. CHLUPÁČ ◽  
E. FILOVÁ ◽  
T. RIEDEL ◽  
M. HOUSKA ◽  
E. BRYNDA ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Type I Collagen ◽  
Type I ◽  
Laminar Shear Stress ◽  
Protein Assemblies ◽  
Vascular Prostheses ◽  
Thrombotic Risk ◽  
Adhesive Protein ◽  
Fibrin Network

Cardiovascular prosthetic bypass grafts do not endothelialize spontaneously in humans, and so they pose a thrombotic risk. Seeding with cells improves their performance, particularly in small-caliber applications. Knitted tubular polyethylene-terephthalate (PET) vascular prostheses (6 mm) with commercial type I collagen (PET/Co) were modified in the lumen by the adsorption of laminin (LM), by coating with a fibrin network (Fb) or a combination of Fb and fibronectin (Fb/FN). Primary human saphenous vein endothelial cells were seeded (1.50 × 105/cm2), cultured for 72 h and exposed to laminar shear stress 15 dyn/cm2 for 40 and 120 min. The control static grafts were excluded from shearing. The cell adherence after 4 h on PET/Co, PET/Co +LM, PET/Co +Fb and PET/Co +Fb/FN was 22 %, 30 %, 19 % and 27 % of seeding, respectively. Compared to the static grafts, the cell density on PET/Co and PET/Co +LM dropped to 61 % and 50 %, respectively, after 120 min of flow. The cells on PET/Co +Fb and PET/Co +Fb/FN did not show any detachment during 2 h of shear stress. Pre-coating the clinically-used PET/Co vascular prosthesis with LM or Fb/FN adhesive protein assemblies promotes the adherence of endothelium. Cell retention under flow is improved particularly on fibrin-containing (Fb and Fb/FN) surfaces.

Abstract 418: Co-Culture of Endothelial Cells, Smooth Muscle Cells and Monocytes in Collagen Scaffold: A Novel i n vitro Model of Atherosclerosis

2017 ◽  
Vol 37 (suppl_1) ◽  
Author(s):  
Martin Liu ◽  
Angelos Karagiannis ◽  
Matthew Sis ◽  
Srivatsan Kidambi ◽  
Yiannis Chatzizisis
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Type I Collagen ◽  
Smooth Muscle Actin ◽  
Muscle Cells ◽  
Type I ◽  
Collagen Gels ◽  
Human Coronary Artery

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.

scholarly journals Investigating potential wound healing properties of polysaccharides extracted from Grewia mollis Juss. and Hoheria populnea A. Cunn. (Malvaceae)

2020 ◽  
Author(s):  
N Pearman ◽  
SR Moxon ◽  
Susan Carnachan ◽  
ME Cooke ◽  
EI Nep ◽  
...  
Keyword(s):  
Wound Healing ◽  
Traditional Medicine ◽  
Quantitative Pcr ◽  
Wound Closure ◽  
Type I Collagen ◽  
Type I ◽  
Closure Rate ◽  
Scratch Assay ◽  
Grewia Gum ◽  
Positive Effect

© 2019 Elsevier Ltd The Malvaceae family is a group of flowering plants that include approximately 244 genera, and 4225 species. Grewia mollis, and Hoheria populnea (lacebark), are examples of the Malvaceae family that are used in traditional medicine. For this study polysaccharide samples were extracted from the inner bark of Grewia mollis (unmodified (GG) and destarched grewia gum (GGDS)) and from the leaves of Hoheria populnea (lacebark polysaccharide (LB)). Wound healing properties of grewia gum and lacebark polysaccharides were investigated using 3T3 fibroblast cells cultured in supplemented DMEM. Deposition of collagen using van Gieson's stain, expression of the COL1A1 gene which encodes type I collagen using quantitative PCR, and chemotaxis using a scratch plate assay were analysed following treatment of cells with the test polysaccharides. Quantitative PCR results indicated that all three polysaccharides increased the levels of COL1A1 mRNA, with GG showing the greatest fold change. Histological staining also indicated that the fibroblasts treated with GG deposited more collagen than control cells. Additionally, scratch assay data indicated that simulated cell ‘wounds’ treated with each polysaccharide showed increased wound closure rate over a 36 h period post treatment, with GG exhibiting the greatest effect on wound closure. Analysis of the Malvaceae derived polysaccharides indicates that they could have a positive effect on mechanisms that are integral to wound healing, potentially providing greater scientific understanding behind their use in traditional medicine.

scholarly journals Primary cilia of human endothelial cells disassemble under laminar shear stress

2004 ◽  
Vol 164 (6) ◽  
pp. 811-817 ◽  
Author(s):  
Carlo Iomini ◽  
Karla Tejada ◽  
Wenjun Mo ◽  
Heikki Vaananen ◽  
Gianni Piperno
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Primary Cilia ◽  
Umbilical Vein ◽  
Intraflagellar Transport ◽  
Mechanical Stimuli ◽  
Laminar Shear Stress ◽  
Human Endothelial Cells ◽  
Umbilical Vein Endothelial Cells ◽  
Laminar Shear

We identified primary cilia and centrosomes in cultured human umbilical vein endothelial cells (HUVEC) by antibodies to acetyl-α-tubulin and capillary morphogenesis gene-1 product (CMG-1), a human homologue of the intraflagellar transport (IFT) protein IFT-71 in Chlamydomonas. CMG-1 was present in particles along primary cilia of HUVEC at interphase and around the oldest basal body/centriole at interphase and mitosis. To study the response of primary cilia and centrosomes to mechanical stimuli, we exposed cultured HUVEC to laminar shear stress (LSS). Under LSS, all primary cilia disassembled, and centrosomes were deprived of CMG-1. We conclude that the exposure to LSS ends the IFT in cultured endothelial cells.

Effects of Fluid Shear Stress on Endothelial Cell Invasion Into Three-Dimensional Matrices

2007 ◽  
Author(s):  
Hojin Kang ◽  
Kayla J. Bayless ◽  
Roland Kaunas
Keyword(s):  
Endothelial Cells ◽  
Shear Stress ◽  
Endothelial Cell ◽  
Cell Invasion ◽  
Fluid Shear Stress ◽  
Vascular Endothelial Cells ◽  
Umbilical Vein ◽  
Shear Stresses ◽  
Collagen Gels ◽  
Fluid Shear

We have previously developed a cell culture model to study the effects of angiogenic factors, such as sphingosine-1-phosphate (S1P), on the invasion of endothelial cells into the underlying extracellular matrix. In addition to biochemical stimuli, vascular endothelial cells are subjected to fluid shear stress due to blood flow. The present study is aimed at determining the effects of fluid shear stress on endothelial cell invasion into collagen gels. A device was constructed to apply well-defined fluid shear stresses to confluent human umbilical vein endothelial cells (HUVECs) seeded on collagen gels. Fluid shear stress induced significant increases in cell invasion with a maximal induction at ∼5 dyn/cm2. These results provide evidence that fluid shear stress is a significant stimulus for endothelial cell invasion and may play a role in regulating angiogenesis.

Endothelial Cell Seeding on Crosslinked Collagen: Effects of Crosslinking on Endothelial Cell Proliferation and Functional Parameters

2000 ◽  
Vol 84 (08) ◽  
pp. 325-331 ◽  
Author(s):  
M. J. B. Wissink ◽  
M. J. A. van Luyn ◽  
R. Beernink ◽  
F. Dijk ◽  
A. A. Poot ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Plasminogen Activator ◽  
Type I Collagen ◽  
Vascular Grafts ◽  
Small Diameter ◽  
Endothelial Cell Proliferation ◽  
Type I ◽  
Cell Seeding ◽  
Endothelial Cell Seeding

SummaryEndothelial cell seeding, a promising method to improve the performance of small-diameter vascular grafts, requires a suitable substrate, such as crosslinked collagen. Commonly used crosslinking agents such as glutaraldehyde and formaldehyde cause, however, cytotoxic reactions and thereby hamper endothelialization of currently available collagen-coated vascular graft materials.The aim of this study was to investigate the effects of an alternative method for crosslinking of collagen, using N-(3-dimethylaminopropyl)-N’-ethylcarbodiimide (EDC) in combination with N-hydroxysuccinimide (NHS), on various cellular functions of human umbilical vein endothelial cells (HUVECs) in vitro. Compared to non-crosslinked type I collagen, proliferation of seeded endothelial cells was significantly increased on EDC/NHS-crosslinked collagen. Furthermore, higher cell numbers were found with increasing crosslink densities. Neither the morphology of the cells nor the secretion of prostacyclin (PGI2), von Willebrand factor (vWF), tissue plasminogen activator (t-PA) and plasminogen activator inhibitor (PAI-1) was affected by the crosslink density of the collagen substrate. Therefore, EDC/NHScrosslinked collagen is candidate substrate for in vivo application such as endothelial cell seeding of collagen-coated vascular grafts.

scholarly journals IL-8 reduces VCAM-1 secretion of smooth muscle cells by increasing p-ERK expression when 3-D co-cultured with vascular endothelial cells

2011 ◽  
Vol 34 (3) ◽  
pp. 138 ◽  
Author(s):  
Zhi Zhang ◽  
Guang Chu ◽  
Hong-Xian Wu ◽  
Ni Zou ◽  
Bao-Gui Sun ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Smooth Muscle ◽  
Smooth Muscle Cells ◽  
Type I Collagen ◽  
Vascular Endothelial Cells ◽  
Umbilical Vein ◽  
Muscle Cells ◽  
Type I ◽  
Vascular Endothelial ◽  
Culture Model

Objective: The goal of this study was to investigate the crosstalk between vascular endothelial cells (ECs) and smooth muscle cells (SMCs) using a three-dimensional (3-D) co-culture model. In addition, the role of IL-8 in this crosstalk was investigated. Methods: A 3-D co-culture model was constructed using a Transwell chamber system and type I collagen gel. Human umbilical artery smooth muscle cells (HUASMCs) were suspended in the gel and added to the upper compartment of the Transwell. Human umbilical vein endothelial cells (HUVECs) were then grown on the surface of the gel. The growth of HUASMCs was tested with a CFDA SE cell proliferation kit. IL-8 and other bioactive substances were investigated by ELISA and real-time PCR. The alteration of p-ERK expression related to the change in IL-8 levels was also examined by Western blot analysis. Results: The proliferation rate of HUASMCs in the 3-D co-culture model was 0.679 ± 0.057. Secretion and transcription of VEGF, t-PA, NO and VCAM-1 in the 3-D co-culture model were different than in single (2-D) culture. When 3-D co-cultured, IL-8 released by HUVECs was significantly increased (2.35 ± 0.16 fold) (P﹤0.05) and the expression of VCAM-1 from HUASMCs was reduced accordingly (0.55±0.09 fold). In addition, increasing or decreasing the level of IL-8 changed the level of p-ERK and VCAM-1 expression. The reduction of VCAM-1, resulting from increased IL-8, could be blocked by the MEK inhibitor, PD98059. Conclusion: Crosstalk between HUVECs and HUASMCs occurred and was probably mediated by IL-8 in this 3-D co-culture model.

scholarly journals Effects of Oral Anticoagulant Therapy on Gene Expression in Crosstalk between Osteogenic Progenitor Cells and Endothelial Cells

2019 ◽  
Vol 8 (3) ◽  
pp. 329 ◽  
Author(s):  
Luca Dalle Carbonare ◽  
Monica Mottes ◽  
Anna Brunelli ◽  
Michela Deiana ◽  
Samuele Cheri ◽  
...  
Keyword(s):  
Gene Expression ◽  
Bone Marrow ◽  
Endothelial Cells ◽  
Anticoagulant Therapy ◽  
Type I Collagen ◽  
Umbilical Vein ◽  
Oral Anticoagulant Therapy ◽  
Marker Gene ◽  
Marker Genes ◽  
Type I

Direct oral anti-coagulants (DOACs) are employed in clinical practice for the prevention and treatment of recurrent venous thromboembolism and for the prevention of stroke in non-valvular atrial fibrillation. DOACs directly and reversibly inhibit activated factor X or thrombin and can interfere with other pathophysiological processes such as inflammation, lipid metabolism, and bone turnover. We aimed to evaluate the possible effects of DOACs on osteogenesis and angiogenesis. We treated 34 patients affected by cardiovascular disorders with DOACs; biochemical and molecular analyses were performed before and after three months of treatment. Circulating progenitors (CPs; CD34−, CD45−, CD14−, CD73+, CD105+), which share typical bone marrow stem cell (MSCs) features, were harvested from peripheral blood of the study subjects to monitor the expression of osteogenesis-related genes RUNX2 and SPARC. Human umbilical vein endothelial cells (HUVECs) were used to probe angiogenesis-related VEGF, CD31, and CD105 gene expression. We performed co-culture experiments using a commercial human mesenchymal stem cells line (hMSCs) obtained from bone marrow and HUVECs. Clinical parameters related to bone metabolism, coagulation, renal and liver function, and the lipid profile were evaluated. Values of the C-terminal telopeptide type I collagen (CTX) increased after the treatment. We found a significant increase in osteogenesis marker gene expression in CPs after three months of anticoagulant therapy. An increase in the RUNX2 expression determinant alone was detected instead in hMSCs co-cultured with HUVECs in the presence of treated patients’ sera. The VEGF, CD31, and CD105 marker genes appeared to be significantly upregulated in HUVECs co-cultured with hMSCs in the presence of treated patients’ sera. Under these conditions, new vessel formation increased as well. Our results highlight an unexpected influence of DOAC therapy on osteogenic commitment and vascular endothelial function promotion.

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