scholarly journals Oligonucleotide and Parylene Surface Coating of Polystyrene and ePTFE for Improved Endothelial Cell Attachment and Hemocompatibility

2012 ◽  
Vol 2012 ◽  
pp. 1-14 ◽  
Author(s):  
Martina Schleicher ◽  
Jan Hansmann ◽  
Bentsian Elkin ◽  
Petra J. Kluger ◽  
Simone Liebscher ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Human Blood ◽  
Cell Attachment ◽  
Culture Conditions ◽  
Dna Oligonucleotides ◽  
Para Xylene ◽  
Coated Surfaces ◽  
Blood Vessel Substitutes ◽  
Cardiovascular Implants

In vivoself-endothelialization by endothelial cell adhesion on cardiovascular implants is highly desirable. DNA-oligonucleotides are an intriguing coating material with nonimmunogenic characteristics and the feasibility of easy and rapid chemical fabrication. The objective of this study was the creation of cell adhesive DNA-oligonucleotide coatings on vascular implant surfaces. DNA-oligonucleotides immobilized by adsorption on parylene (poly(monoaminomethyl-para-xylene)) coated polystyrene and ePTFE were resistant to high shear stress (9.5 N/m2) and human blood serum for up to 96 h. Adhesion of murine endothelial progenitor cells, HUVECs and endothelial cells from human adult saphenous veins as well as viability over a period of 14 days of HUVECs on oligonucleotide coated samples under dynamic culture conditions was significantly enhanced (P<0.05). Oligonucleotide-coated surfaces revealed low thrombogenicity and excellent hemocompatibility after incubation with human blood. These properties suggest the suitability of immobilization of DNA-oligonucleotides for biofunctionalization of blood vessel substitutes for improvedin vivoendothelialization.

scholarly journals 3D culture conditions support Kaposi’s sarcoma herpesvirus (KSHV) maintenance and viral spread in endothelial cells

2021 ◽  
Vol 99 (3) ◽  
pp. 425-438
Author(s):  
Tatyana Dubich ◽  
Anne Dittrich ◽  
Kristine Bousset ◽  
Robert Geffers ◽  
Guntram Büsche ◽  
...  
Keyword(s):  
Cell Culture ◽  
Endothelial Cells ◽  
Endothelial Cell ◽  
Kaposi's Sarcoma ◽  
3D Cell Culture ◽  
Kaposi’S Sarcoma ◽  
Culture Conditions ◽  
List Type ◽  
Infection Frequency

Abstract Kaposi’s sarcoma–associated herpesvirus (KSHV) is a human tumorigenic virus and the etiological agent of an endothelial tumor (Kaposi’s sarcoma) and two B cell proliferative diseases (primary effusion lymphoma and multicentric Castleman’s disease). While in patients with late stage of Kaposi’s sarcoma the majority of spindle cells are KSHV-infected, viral copies are rapidly lost in vitro, both upon culture of tumor-derived cells or from newly infected endothelial cells. We addressed this discrepancy by investigating a KSHV-infected endothelial cell line in various culture conditions and in tumors of xenografted mice. We show that, in contrast to two-dimensional endothelial cell cultures, KSHV genomes are maintained under 3D cell culture conditions and in vivo. Additionally, an increased rate of newly infected cells was detected in 3D cell culture. Furthermore, we show that the PI3K/Akt/mTOR and ATM/γH2AX pathways are modulated and support an improved KSHV persistence in 3D cell culture. These mechanisms may contribute to the persistence of KSHV in tumor tissue in vivo and provide a novel target for KS specific therapeutic interventions. Key messages In vivo maintenance of episomal KSHV can be mimicked in 3D spheroid cultures 3D maintenance of KSHV is associated with an increased de novo infection frequency PI3K/Akt/mTOR and ATM/ γH2AX pathways contribute to viral maintenance

Enhanced re-endothelialization of acellular kidney scaffolds for whole organ engineering via antibody conjugation of vasculatures

TECHNOLOGY ◽  
2014 ◽  
Vol 02 (03) ◽  
pp. 243-253 ◽  
Author(s):  
In Kap Ko ◽  
Mehran Abolbashari ◽  
Jennifer Huling ◽  
Cheil Kim ◽  
Sayed-Hadi Mirmalek-Sani ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Cell Attachment ◽  
Great Promise ◽  
Cell Seeding ◽  
Seeding Method ◽  
Vascular Patency ◽  
Antibody Conjugation ◽  
Endothelial Cell Seeding ◽  
Whole Organ Engineering

Decellularization of whole organs, such as the kidney hold great promise in addressing donor shortage for transplantation. However, successful implantation of engineered whole kidney constructs has been challenged by the inability to maintain endothelial cell coverage of the vasculature matrix, resulting in excessive blood clots, loss of vascular patency, and cell death within the construct. In this study, we describe an endothelial cell seeding approach that permits effective coating of the vascular matrix of the decellularized porcine kidney scaffold using a combination of static and ramping perfusion cell seeding. Furthermore, conjugation of CD31 antibodies to the vascular matrix improved endothelial cell retention on the vasculatures, which enhanced vascular patency of the implanted scaffold. These results demonstrate that our endothelial cell seeding method combined with antibody conjugation improves endothelial cell attachment and retention leading to vascular patency of tissue-engineered whole kidney in vivo.

scholarly journals Plasma-Mediated Immobilization of Antibody with PEG as Spacer for Enhanced Endothelial Cell Adhesion and Proliferation

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Yuan Yuan ◽  
Jing Zhang ◽  
Min Yin ◽  
Changsheng Liu
Keyword(s):  
Endothelial Cell ◽  
Cold Plasma ◽  
Cell Attachment ◽  
Specific Antigen ◽  
Argon Plasma ◽  
Ethylene Vinyl Acetate ◽  
Efficiency Increase ◽  
Spacer Arm ◽  
Cardiovascular Implants ◽  
Acetate Copolymer

Immobilization of anti-CD34 antibody is proven to be an effective strategy to accelerate reendothelialization and thereby lower the thrombosis of blood contacting grafts. To realize highly efficient immobilization of anti-CD34 antibody, an argon cold plasma-mediated graft process was developed with PEG as spacer arm in this study. In this process, the 316L stainless steel (316LSS) model substrate was first coated with ethylene vinyl acetate copolymer (EVA) followed by argon plasma treatment and PEG400 modification (EVA-PEG). The EVA-PEG was further ignited by argon plasma and then the anti-CD34 antibody was immobilized. XPS measurement indicated the successful immobilization of the EVA and the anti-CD34 antibody molecules. Compared with the anti-CD34 antibody anchored without PEG, the immobilized EVA-PEG-anti-CD34 antibody exhibited better capturing efficiency (increase about 1-fold) of specific antigen. Consequently, the endothelial cell attachment (before 12 h) and proliferation (1~4 days) were significantly improved. Further study showed that this EVA-PEG-anti-CD34 coating could reduce blood coagulation. Therefore, this cold plasma-mediated graft process with PEG spacer arm developed here is a promising strategy to immobilize antibody with higher bioactivity for rapid reendothelialization of the cardiovascular implants.

Hydroxyapatite Coatings of Varying Crystallinity

1993 ◽  
Vol 331 ◽  
Author(s):  
Suzanne H. Maxian ◽  
Janice B. Liesch ◽  
Moti L. Tiku ◽  
Joseph P. Zawadsky ◽  
Michael G. Dunn
Keyword(s):  
Tissue Culture ◽  
Alkaline Phosphatase ◽  
Cell Attachment ◽  
Fibroblast Cell ◽  
Bone Cell ◽  
Titanium Implants ◽  
Interface Failure ◽  
Hydroxyapatite Coatings ◽  
Coated Surfaces

AbstractQuestions remain as to the integrity of hydroxyapatite (HA) coatings on orthopaedic implants and the possibility of failure at the coating/implant interface in vivo. Previous investigations of resorbable HA coated implants showed bone bonding strengths equal to non-resorbable HA coated surfaces in interference fit surgical model. If the concept of a resorbing coating will alleviate concerns of interface failure, further investigation as to the mode of coating resorption and the cellular events at the interface are required. The effect of coating these implants with Matrigel, as a means to enhance bone cell attachment and growth was studied. Fibroblast cell behavior on resorbable, non-resorbable HA coated and uncoated rough titanium implants were studied at 4 and 24 hours for attachment and spreading. Postnatal rat calvarial cell attachment, growth and morphology studies were also performed on these three implants and on tissue culture plates with and without Matrigel at 2, 7, and 10 days. Resorbable HA coatings showed greater surface activity and cell spreading, but not greater cell attachment at 4 and 24 hours. Greater alkaline phosphatase production per cell was seen on Matrigel coated tissue culture plates than uncoated wells. Cytoplasmic processes similar to osteocytic canalicular networks were seen. Matrigel coating on the three types of implants showed slightly greater alkaline phosphatase production of cells than without Matrigel coating.

Ultrastructural Study of a differentiating human colon carcinoma cell line

1990 ◽  
Vol 48 (3) ◽  
pp. 208-209
Author(s):  
Sylvie Polak-Charcon ◽  
Mehrdad Hekmati ◽  
Yehuda Ben Shaul
Keyword(s):  
Cell Line ◽  
Morphological Changes ◽  
Secretory Granules ◽  
Human Colon ◽  
Cell Types ◽  
Culture Conditions ◽  
Morphological Evidence ◽  
Human Colon Carcinoma Cell ◽  
Colon Cell

The epithelium of normal human colon mucosa “in vivo” exhibits a gradual pattern of differentiation as undifferentiated stem cells from the base of the crypt of “lieberkuhn” rapidly divide, differentiate and migrate toward the free surface. The major differentiated cell type of the intestine observed are: absorptive cells displaying brush border, goblet cells containing mucous granules, Paneth and endocrine cells containing dense secretory granules. These different cell types are also found in the intestine of the 13-14 week old embryo.We present here morphological evidence showing that HT29, an adenocarcinoma of the human colon cell line, can differentiate into various cell types by changing the growth and culture conditions and mimic morphological changes found during development of the intestine in the human embryo.HT29 cells grown in tissue-culture dishes in DMEM and 10% FCS form at late confluence a multilayer of morphologically undifferentiated cell culture covered with irregular microvilli, and devoid of tight junctions (Figs 1-3).

Endogenous hydrogen sulfide sulfhydrates IKKβ at cysteine 179 to control pulmonary artery endothelial cell inflammation

2019 ◽  
Vol 133 (20) ◽  
pp. 2045-2059 ◽  
Author(s):  
Da Zhang ◽  
Xiuli Wang ◽  
Siyao Chen ◽  
Selena Chen ◽  
Wen Yu ◽  
...  
Keyword(s):  
Hydrogen Sulfide ◽  
Endothelial Cell ◽  
Pulmonary Artery ◽  
Cell Model ◽  
Site Directed Mutagenesis ◽  
Critical Event ◽  
Hypertensive Rats ◽  
Pulmonary Artery Endothelial Cell

Abstract Background: Pulmonary artery endothelial cell (PAEC) inflammation is a critical event in the development of pulmonary arterial hypertension (PAH). However, the pathogenesis of PAEC inflammation remains unclear. Methods: Purified recombinant human inhibitor of κB kinase subunit β (IKKβ) protein, human PAECs and monocrotaline-induced pulmonary hypertensive rats were employed in the study. Site-directed mutagenesis, gene knockdown or overexpression were conducted to manipulate the expression or activity of a target protein. Results: We showed that hydrogen sulfide (H2S) inhibited IKKβ activation in the cell model of human PAEC inflammation induced by monocrotaline pyrrole-stimulation or knockdown of cystathionine γ-lyase (CSE), an H2S generating enzyme. Mechanistically, H2S was proved to inhibit IKKβ activity directly via sulfhydrating IKKβ at cysteinyl residue 179 (C179) in purified recombinant IKKβ protein in vitro, whereas thiol reductant dithiothreitol (DTT) reversed H2S-induced IKKβ inactivation. Furthermore, to demonstrate the significance of IKKβ sulfhydration by H2S in the development of PAEC inflammation, we mutated C179 to serine (C179S) in IKKβ. In purified IKKβ protein, C179S mutation of IKKβ abolished H2S-induced IKKβ sulfhydration and the subsequent IKKβ inactivation. In human PAECs, C179S mutation of IKKβ blocked H2S-inhibited IKKβ activation and PAEC inflammatory response. In pulmonary hypertensive rats, C179S mutation of IKKβ abolished the inhibitory effect of H2S on IKKβ activation and pulmonary vascular inflammation and remodeling. Conclusion: Collectively, our in vivo and in vitro findings demonstrated, for the first time, that endogenous H2S directly inactivated IKKβ via sulfhydrating IKKβ at Cys179 to inhibit nuclear factor-κB (NF-κB) pathway activation and thereby control PAEC inflammation in PAH.

In Vivo Assessment of Synovial Microcirculation and Leukocyte–Endothelial Cell Interaction in Mouse Antigen-Induced Arthritis

1999 ◽  
Vol 6 (4) ◽  
pp. 281-290 ◽  
Author(s):  
A N D R E A S VEIHELMANN ◽  
ANTHONY G U S T A V E HARRIS ◽  
F R I T Z KROMBACH ◽  
E L K E SCHÜTZE ◽  
HANS JÜRGEN REFIOR ◽  
...  
Keyword(s):  
Endothelial Cell ◽  
Cell Interaction ◽  
Endothelial Cell Interaction ◽  
In Vivo Assessment

An Electron Microscopic Study of Platelet Thrombus Formation in the Rabbit with Particular Regard to 5-Hydroxytryptamine Release

1967 ◽  
Vol 18 (03/04) ◽  
pp. 592-604 ◽  
Author(s):  
H. R Baumgartner ◽  
J. P Tranzer ◽  
A Studer
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Vessel Wall ◽  
Thrombus Formation ◽  
Endothelial Damage ◽  
Electron Microscopic ◽  
Rabbit Ear ◽  
Basement Lamina ◽  
Platelet Thrombus

SummaryElectron microscopic and histologic examination of rabbit ear vein segments 4 and 30 min after slight endothelial damage have yielded the following findings :1. Platelets do not adhere to damaged endothelial cells.2. If the vessel wall is denuded of the whole endothelial cell, platelets adhere to the intimai basement lamina as do endothelial cells.3. The distance between adherent platelets as well as endothelial cells and intimai basement lamina measures 10 to 20 mµ, whereas the distance between aggregated platelets is 30 to 60 mµ.4. 5-hydroxytryptamine (5-HT) is released from platelets during viscous metamorphosis at least in part as 5-HT organelles.It should be noted that the presence of collagen fibers is not necessary for platelet thrombus formation in vivo.

Glycoprotein Ib Can Mediate Endothelial Cell Attachment to a von Willebrand Factor Substratum

1995 ◽  
Vol 73 (02) ◽  
pp. 309-317 ◽  
Author(s):  
Dorothy A Beacham ◽  
Miguel A Cruz ◽  
Robert I Handin
Keyword(s):  
Endothelial Cell ◽  
Von Willebrand Factor ◽  
Cell Attachment ◽  
Umbilical Vein ◽  
Single Amino Acid ◽  
Cell Surface Expression ◽  
Platelet Glycoprotein ◽  
Glycoprotein Ib ◽  
Von Willebrand ◽  
Willebrand Factor

SummaryIntroduction of single amino acid substitutions into the C-terminal Arg-Gly-Asp-Ser (RGDS) site of von Willebrand Factor, referred to as RGD mutant vWF, selectively abrogated vWF binding to platelet glycoprotein IIb/IIIa (GpIIb/IIIa, αIIbβ3 and abolished human umbilical vein endothelial cell (HUVEC) spreading, but not attachment, to RGD mutant vWF (Beacham, D. A., Wise, R. J., Turci, S. M. and Handin, R. I. 1992. J. Biol. Chem. 167, 3409-3415). These results suggested that in addition to the vitronectin receptor (VNR, αvβ3), a second endothelial membrane glycoprotein can mediate HUVEC adhesion to vWF. HUVEC attachment to wild-type (WT) and RGD-mutant vWF was reduced by two proteins known to block the vWF-platelet glycoprotein Ib/IX (GpIb/IX) interaction, the monoclonal antibody AS-7 and the recombinant polypeptide, vWF-A1. The addition of cytochalasin B or DNase I to disrupt potential GPIbα-cytoskeletal interactions enhanced the immunoprecipitation of endothelial GPIbα, caused HUVEC to round up, and increased HUVEC adhesion to RGD mutant vWF. These results indicate that while the VNR is the primary adhesion receptor for vWF, endothelial GPIbα can mediate HUVEC attachment to vWF. GpIb-dependent attachment could contribute to HUVEC adhesion under conditions when cell surface expression of the VNR is downregulated, and VNR-dependent adhesion is reduced.

Organische Nitrate und Thrombozytenfunktion

1988 ◽  
Vol 08 (02) ◽  
pp. 90-99 ◽  
Author(s):  
H. Schröder ◽  
K. Schrör
Keyword(s):  
Endothelial Cell ◽  
Angina Pectoris ◽  
Ex Vivo

ZusammenfassungOrganische Nitrate unterschiedlicher chemischer Struktur sowie Nitroprussidnatrium und Molsidomin (bzw. ihre biologisch aktiven Metaboliten) können die (primäre) Aggregation und Sekretion von Humanthrombozyten in vitro und ex vivo hemmen. Eine solche Wirkung wird für Molsidomin (SIN-1) und Nitroprussidnatrium in vitro in Konzentrationen beobachtet, die in der gleichen Größenordnung liegen wie die vasodilatierenden Effekte der Substanzen. Dagegen sind für eine direkte Antiplättchenwirkung organischer Nitrate (Glyzeryltrinitrat, Isosorbiddinitr at, Isosorbidmononitrate, Teopranitol) in vitro Konzentrationen erforderlich, die ca. 100- bis 1000fach höher sind als die Plasmaspiegel der Substanzen nach therapeutischer Dosierung bzw. die Konzentrationen, die isolierte Gefäßstreifen relaxieren. Als gemeinsamer Wirkungsmechanismus der direkten thrombozy-tenfunktionshemmenden und gefäßerweiternden Wirkung all dieser Substanzen kann heute eine Stickoxid-(NO)-vermittelte Stimulation der cGMP-Bildung angenommen werden, das aus organischen Nitraten als »Pro-drug« entsteht. Die Freisetzung von NO, eines »endothelial cell-derived relaxing factors« (EDRF) aus Nitroprussidnatrium und SIN-1 erfolgt spontan. Dagegen erfordert die Freisetzung von NO aus organischen Nitraten einen enzymatischen Stoffwechselweg, der in isolierten Thrombozyten nicht vorhanden ist. Eine Antiplättchenwirkung organischer Nitrate in vivo bzw. ex vivo wird daher über die Stimulation eines endothelialen, thrombozyteninhibitorischen Faktors erklärt. Hierbei sind Prostazyklin sowie ein bisher unbekannter Endothel-zellfaktor neben einer synergistischen Wirkung organischer Nitrate mit endogenem Prostazyklin in Diskussion. Eine thrombozytenfunktionshemmen-de Wirkung organischer Nitrate könnte in Kombination mit ihren hämody-namischen Effekten auch für die an-tianginöse Wirkung in der Klinik bedeutsam sein, insbesondere zur Verhinderung vasospastischer Zustände bei der instabilen Angina pectoris.

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