Membrane Microparticles from Patients with Steroid-Induced Avascular Necrosis of Femoral Head Increase the Expression of Fas in Endothelial Cells In Vitro.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 3903-3903
Author(s):  
Xi-sheng Weng ◽  
Zhi-hong Wu ◽  
Yi Wei ◽  
Gui-xing Qiu
Keyword(s):  
Endothelial Cells ◽  
Femoral Head ◽  
Avascular Necrosis ◽  
Umbilical Vein ◽  
Cell Types ◽  
High Dose ◽  
Necrosis Of Femoral Head ◽  
Membrane Microparticles

Abstract High-dose or long-term usage of steroid have been suggested to be at great risk of developing avascular necrosis of femoral head (ANFH). A theory proposed to decipher the mechanism behind the development of steroid-induced ANFH involves vascular compromise and cell death. Membrane microparticles (MMPs) are fragments shed from plasma membrane blebs of virtually all cell types when submitted to a number of stress conditions, including apoptosis. It has been reported that high-dose dexamethasone cause dysfunction and apoptosis of endothelial cells; and MMPs isolated from the plasma of patients with myocardial infarction of preclampsia were found to cause damage in isolated arteries in vitro. We hypothesize that MMPs generated after high-dose or long-term administration of steroid facilitates the apoptosis of endothelial cells initiated by steroid, which contributes to the development of ANFH. MMPs were isolated from the blood of 4 healthy individuals, and 5 patients demonstrated to have ANFH by magnetic resonance imaging (MRI) and X-ray. The mean duration and accumulated dose of steroid administration were 8 months and 2000 mg respectively. The 3rd generation of human umbilical vein endothelial cells (HUVEC) was exposed to MMPs (corresponding to 0.2-fold circulating plasma level) prepared above. After 48 hours, part of cells was lysed to make total cell lysate. And the rest were used to prepare mRNA. The expression level of Fas was detected with reverse-transcript PCR and Western Blot. Our results shown that treatment of HUVEC with MMPs from ANFH patient’s blood significantly increases the transcription and expression of Fas, indicating that MMPs derived from patients with steroid-induced ANFH exacerbates high-dose steroid-induced apoptosis of endothelial cells by enhancing the expression of Fas.

scholarly journals Hydrogel-Coated Textile Scaffolds as Three-Dimensional Growth Support for Human Umbilical Vein Endothelial Cells (HUVECs): Possibilities as Coculture System in Liver Tissue Engineering

2002 ◽  
Vol 11 (4) ◽  
pp. 369-377 ◽  
Author(s):  
Makarand V. Risbud ◽  
Erdal Karamuk ◽  
René Moser ◽  
Joerg Mayer
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Three Dimensional ◽  
Mesh Size ◽  
Cell Types ◽  
Cell Number ◽  
Human Umbilical Vein ◽  
Hydrogel Matrix ◽  
Umbilical Vein Endothelial Cells

Three-dimensional (3-D) scaffolds offer an exciting possibility to develop cocultures of various cell types. Here we report chitosan–collagen hydrogel-coated fabric scaffolds with defined mesh size and fiber diameter for 3-D culture of human umbilical vein endothelial cells (HUVECs). These scaffolds did not require pre-coating with fibronectin and they supported proper HUVEC attachment and growth. Scaffolds preserved endothelial cell-specific cobblestone morphology and cells were growing in compartments defined by the textile mesh. HUVECs on the scaffold maintained the property of contact inhibition and did not exhibit overgrowth until the end of in vitro culture (day 6). MTT assay showed that cells had preserved mitochondrial functionality. It was also noted that cell number on the chitosan-coated scaffold was lower than that of collagen-coated scaffolds. Calcein AM and ethidium homodimer (EtD-1) dual staining demonstrated presence of viable and metabolically active cells, indicating growth supportive properties of the scaffolds. Actin labeling revealed absence of actin stress fibers and uniform distribution of F-actin in the cells, indicating their proper attachment to the scaffold matrix. Confocal microscopic studies showed that HUVECs growing on the scaffold had preserved functionality as seen by expression of von Willebrand (vW) factor. Observations also revealed that functional HUVECs were growing at various depths in the hydrogel matrix, thus demonstrating the potential of these scaffolds to support 3-D growth of cells. We foresee the application of this scaffold system in the design of liver bioreactors wherein hepatocytes could be cocultured in parallel with endothelial cells to enhance and preserve liver-specific functions.

scholarly journals AICAR Protects Vascular Endothelial Cells from Oxidative Injury Induced by the Long-Term Palmitate Excess

2021 ◽  
Vol 23 (1) ◽  
pp. 211
Author(s):  
Mikhail V. Samsonov ◽  
Nikita V. Podkuychenko ◽  
Asker Y. Khapchaev ◽  
Eugene E. Efremov ◽  
Elena V. Yanushevskaya ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Insulin Signaling ◽  
Umbilical Vein ◽  
Blood Levels ◽  
Ros Generation ◽  
No Production ◽  
Vascular Endothelial ◽  
Long Term Effects

Hyperlipidemia manifested by high blood levels of free fatty acids (FFA) and lipoprotein triglycerides is critical for the progression of type 2 diabetes (T2D) and its cardiovascular complications via vascular endothelial dysfunction. However, attempts to assess high FFA effects in endothelial culture often result in early cell apoptosis that poorly recapitulates a much slower pace of vascular deterioration in vivo and does not provide for the longer-term studies of endothelial lipotoxicity in vitro. Here, we report that palmitate (PA), a typical FFA, does not impair, by itself, endothelial barrier and insulin signaling in human umbilical vein endothelial cells (HUVEC), but increases NO release, reactive oxygen species (ROS) generation, and protein labeling by malondialdehyde (MDA) hallmarking oxidative stress and increased lipid peroxidation. This PA-induced stress eventually resulted in the loss of cell viability coincident with loss of insulin signaling. Supplementation with 5-aminoimidazole-4-carboxamide-riboside (AICAR) increased endothelial AMP-activated protein kinase (AMPK) activity, supported insulin signaling, and prevented the PA-induced increases in NO, ROS, and MDA, thus allowing to maintain HUVEC viability and barrier, and providing the means to study the long-term effects of high FFA levels in endothelial cultures. An upgraded cell-based model reproduces FFA-induced insulin resistance by demonstrating decreased NO production by vascular endothelium.

HUMAN ENDOTHELIAL CELLS MODIFICATIONS DURING LONG TERM CULTURE

1987 ◽  
Author(s):  
M P Wautier ◽  
J L Wautier
Keyword(s):  
Endothelial Cells ◽  
Doubling Time ◽  
Umbilical Vein ◽  
Light And Electron Microscopy ◽  
Major Change ◽  
Culture Conditions ◽  
Human Endothelial Cells ◽  
Long Term Culture

The culture of human endothelial cells is largely used for vascular research. The possibility of developping long term culture of human endothelial cells (EC) raised the question regarding the identity after several passages. To further investigate this aspect we have cultured human umbilical vein EC until the 12th passage on fibronectin coated dishes supplemented with ECGF. We have studied the EC morphology by light and electron microscopy, the reactivity with 51Cr labelled platelets, and prostacyclin synthesis. Until the 6th passage no major change could be noted, except the occurence of rare large EC and a reduction in the doubling time between 2nd and 5th passage. After the 7th passage up to the 10th EC became more elongated and did not grow in strict monolayer. The number of vacuoles and mitochondria increased as well as the doubling time. After the 12th passage the EC were still viable but proliferated very slowly. The adhesion of radiolabelled platelets dramatically increased (150%) and PGI2 production significantly decreased (6 Keto PGF1α : 1st passage 13±2.5 ng; 6th passage 0.33±0.27 ng/106 EC). In our culture conditions EC kept most of their original characteristics up to the 6th passage but then lost some of them. At any passage EC contained Weibel Palade bodies and von Willebrand factor. We can conclude that after the 7th passage EC in culture are different from the original cells and could possibly represent an in vitro model of EC ageing.

A proteasome inhibitor reduces concurrent, sequential, and long-term IL-1β- and TNF-α-induced ECAM expression and adhesion

2003 ◽  
Vol 285 (4) ◽  
pp. C813-C822 ◽  
Author(s):  
Nilesh M. Dagia ◽  
Douglas J. Goetz
Keyword(s):  
Endothelial Cells ◽  
Cell Adhesion ◽  
Proteasome Inhibitor ◽  
Umbilical Vein ◽  
Hl60 Cell ◽  
Induced Expression ◽  
Tnf Α ◽  
Short Period

A promising approach for reducing aberrant leukocyte-endothelial adhesion during pathological inflammation is to inhibit endothelial cell adhesion molecule (ECAM) expression at the transcription level. Several compounds have been shown to decrease cytokine-induced upregulation of ECAMs primarily by modulating the activity of transcription factors [e.g., nuclear factor-κB (NF-κB)]. The majority of the in vitro studies have focused on the effect of transcription inhibitors on endothelial cells exposed to a single cytokine [primarily tumor necrosis factor-α (TNF-α)] for a relatively short period of time (primarily 4-6 h). However, in the in vivo setting, multiple cytokines [e.g., interleukin-1β (IL-1β) and TNF-α] may be present for extended periods of time. Thus we studied the effects of a transcription inhibitor, the proteasome inhibitor lactacystin, on ECAM expression and myeloid (HL60) cell adhesion to human umbilical vein endothelial cells (HUVEC) activated by concurrent, sequential, and long-term (24 h) treatment with IL-1β and TNF-α. We show, for the first time, that lactacystin inhibits 1) 4-h concurrent IL-1β- and TNF-α-induced expression of E-selectin, VCAM-1, ICAM-1, and HL60 cell adhesion to HUVEC; 2) 4-h TNF-α-induced expression of E-selectin, VCAM-1, and HL60 cell adhesion to HUVEC that have become desensitized to IL-1β activation; 3) 24-h TNF-α-induced expression of E-selectin and VCAM-1 but not ICAM-1; and 4) 24-h TNF-α-induced HL60 cell adhesion to HUVEC. Combined, our results demonstrate that a proteasome inhibitor can reduce concurrent, sequential, and long-term IL-1β- and TNF-α-induced ECAM expression and myeloid cell adhesion.

scholarly journals Poloxamer 407-induced atherosclerosis in mice appears to be due to lipid derangements and not due to its direct effects on endothelial cells and macrophages

2003 ◽  
Vol 12 (3) ◽  
pp. 147-155 ◽  
Author(s):  
Thomas P. Johnston ◽  
Yuai Li ◽  
Ahmed S. Jamal ◽  
Daniel J. Stechschulte ◽  
Kottarappat N. Dileepan
Keyword(s):  
Endothelial Cells ◽  
Mouse Model ◽  
Umbilical Vein ◽  
Density Lipoprotein ◽  
Poloxamer 407 ◽  
Direct Effects ◽  
Direct Stimulation

Coronary heart disease secondary to atherosclerosis is still the leading cause of death in the US. Animal models used for elucidating the pathogenesis of this disease primarily involve rabbits and pigs. Previous studies from this laboratory have demonstrated intraperitoneal injections of poloxamer 407 (P-407) in both male and female mice will lead to hyperlipidemia and atherosclerosis, suggesting the use of this polymer to develop a mouse model of atherosclerosis. In order to understand the mechanism of P-407-induced hyperlipidemia and vascular lesion formation, we evaluated the direct effects of P-407 on endothelial cell and macrophage functionsin vitro, and itsin vivoeffects on the oxidation of circulating lipids following long-term (4 month) administration. Our results demonstrated that incubation of P-407 with human umbilical vein endothelial cells in culture did not influence either cell proliferation or interleukin-6 and interleukin-8 production over a concentration range of 0-40 μM. In addition, nitric oxide production by macrophages was not affected by P-407 over a concentration range of 0-20 μM. Finally, we demonstrated that while P-407 could not induce the oxidation of LDL-Cin vitro, long-term (4 month) administration of P-407 in mice resulted in elevated levels of oxidized lipids in the plasma. Thus, it is suggested that the formation of atherosclerotic lesions in this mouse model of atherosclerosis does not result from either direct stimulation of endothelial cells or macrophage activation by P-407. Instead, these data would support the premise that oxidation of lipids (perhaps low-density lipoprotein cholesterol) by an indirect mechanism following injection of P-407 may represent one of the mechanisms responsible for atheroma formation.

Allicin inhibits osteoblast apoptosis and steroid-induced necrosis of femoral head progression by activating the PI3K/AKT pathway

2020 ◽  
Vol 11 (9) ◽  
pp. 7830-7841 ◽  
Author(s):  
Jingdi Zhan ◽  
Zijian Yan ◽  
Mengyao Zhao ◽  
Weihui Qi ◽  
Jian Lin ◽  
...  
Keyword(s):  
Femoral Head ◽  
Avascular Necrosis ◽  
Major Complication ◽  
Akt Pathway ◽  
Clinical Use ◽  
Osteoblast Apoptosis ◽  
Necrosis Of Femoral Head

Steroid-induced avascular necrosis of the femoral head (SANFH) is a major complication of long-term or excessive clinical use of glucocorticoids.

Upregulation of microRNA-320 decreases the risk of developing steroid-induced avascular necrosis of femoral head by inhibiting CYP1A2 both in vivo and in vitro

Gene ◽  
2018 ◽  
Vol 660 ◽  
pp. 136-144 ◽  
Author(s):  
Ji-Hua Wei ◽  
Qun-Qiang Luo ◽  
Yu-Jin Tang ◽  
Ji-Xia Chen ◽  
Chun-Lan Huang ◽  
...  
Keyword(s):  
Femoral Head ◽  
Avascular Necrosis ◽  
Necrosis Of Femoral Head

scholarly journals Up-regulation of CD81 inhibits cytotrophoblast invasion and mediates maternal endothelial cell dysfunction in preeclampsia

2017 ◽  
Vol 114 (8) ◽  
pp. 1940-1945 ◽  
Author(s):  
Li Shen ◽  
Zhenyu Diao ◽  
Hai-Xiang Sun ◽  
Gui-Jun Yan ◽  
Zhiqun Wang ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Endothelial Cell ◽  
Cell Activation ◽  
Umbilical Vein ◽  
Normal Pregnancy ◽  
Uterine Wall ◽  
High Dose ◽  
Endothelial Cell Activation ◽  
Umbilical Vein Endothelial Cells

Preeclampsia (PE) is initiated by abnormal placentation in the early stages of pregnancy, followed by systemic activation of endothelial cells of the maternal small arterioles in the late second or third trimester (TM) of pregnancy. During normal pregnancy, placental cytotrophoblasts (CTBs) invade the maternal uterine wall and spiral arteries, whereas this process is interrupted in PE. However, it is not known how the malformed placenta triggers maternal endothelial crisis and the associated manifestations. Here, we have focused on the association of CD81 with PE. CD81, a member of the tetraspanin superfamily, plays significant roles in cell growth, adhesion, and motility. The function of CD81 in human placentation and its association with pregnancy complications are currently unknown. In the present study, we have demonstrated that CD81 was preferentially expressed in normal first TM placentas and progressively down-regulated with gestation advance. In patients with early-onset severe PE (sPE), CD81 expression was significantly up-regulated in syncytiotrophoblasts (STBs), CTBs and the cells in the villous core. In addition, high levels of CD81 were observed in the maternal sera of patients with sPE. Overexpressing CD81 in CTBs significantly decreased CTB invasion, and culturing primary human umbilical vein endothelial cells (HUVECs) in the presence of a high dose of exogenous CD81 resulted in interrupted angiogenesis and endothelial cell activation in vitro. Importantly, the phenotype of human PE was mimicked in the CD81-induced rat model.

scholarly journals Endothelial Jak3 expression enhances pro-hematopoietic angiocrine function in mice

2021 ◽  
Vol 4 (1) ◽  
Author(s):  
José Gabriel Barcia Durán ◽  
Tyler Lu ◽  
Sean Houghton ◽  
Fuqiang Geng ◽  
Ryan Schreiner ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Endothelial Cells ◽  
Cell Types ◽  
Bone Marrow Niche ◽  
Hematopoietic Stem ◽  
Sinusoidal Endothelium ◽  
Bone Marrow Vascular Niche ◽  
Different Cell Types

AbstractJak3 is the only non-promiscuous member of the Jak family of secondary messengers. Studies to date have focused on understanding and targeting the cell-autonomous role of Jak3 in immunity, while functional Jak3 expression outside the hematopoietic system remains largely unreported. We show that Jak3 is expressed in endothelial cells across hematopoietic and non-hematopoietic organs, with heightened expression in the bone marrow. The bone marrow niche is understood as a network of different cell types that regulate hematopoietic function. We show that the Jak3–/– bone marrow niche is deleterious for the maintenance of long-term repopulating hematopoietic stem cells (LT-HSCs) and that JAK3-overexpressing endothelial cells have increased potential to expand LT-HSCs in vitro. This work may serve to identify a novel function for a highly specific tyrosine kinase in the bone marrow vascular niche and to further characterize the LT-HSC function of sinusoidal endothelium.

Ultrastructural observations on the in vitro cytoadherence of Plasmodium falciparum infected red blood cells

1990 ◽  
Vol 48 (3) ◽  
pp. 914-915
Author(s):  
D.J.P. Ferguson ◽  
A.R. Berendt ◽  
J. Tansey ◽  
K. Marsh ◽  
C.I. Newbold
Keyword(s):  
Plasmodium Falciparum ◽  
Red Blood Cells ◽  
Blood Cells ◽  
Umbilical Vein ◽  
Cell Types ◽  
Amelanotic Melanoma ◽  
Cytoplasmic Process ◽  
Umbilical Vein Endothelial Cells

In human malaria, the most serious clinical manifestation is cerebral malaria (CM) due to infection with Plasmodium falciparum. The pathology of CM is thought to relate to the fact that red blood cells containing mature forms of the parasite (PRBC) cytoadhere or sequester to post capillary venules of various tissues including the brain. This in vivo phenomenon has been studied in vitro by examining the cytoadherence of PRBCs to various cell types and purified proteins. To date, three Ijiost receptor molecules have been identified; CD36, ICAM-1 and thrombospondin. The specific changes in the PRBC membrane which mediate cytoadherence are less well understood, but they include the sub-membranous deposition of electron-dense material resulting in surface deformations called knobs. Knobs were thought to be essential for cytoadherence, lput recent work has shown that certain knob-negative (K-) lines can cytoadhere. In the present study, we have used electron microscopy to re-examine the interactions between K+ PRBCs and both C32 amelanotic melanoma cells and human umbilical vein endothelial cells (HUVEC).We confirm previous data demonstrating that C32 cells possess numerous microvilli which adhere to the PRBC, mainly via the knobs (Fig. 1). In contrast, the HUVEC were relatively smooth and the PRBCs appeared partially flattened onto the cell surface (Fig. 2). Furthermore, many of the PRBCs exhibited an invagination of the limiting membrane in the attachment zone, often containing a cytoplasmic process from the endothelial cell (Fig. 2).

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