Gene Expression of Adhesion Molecules in Endothelial Cells from Patients with Peripheral Arterial Disease Is Reduced after Surgical Revascularization and Pharmacological Treatment

Atherosclerosis is an inflammatory disease characterized by immunological activity, in which endothelial dysfunction represents an early event leading to subsequent inflammatory vascular damage. We investigated gene expression of the adhesion molecules (AMs) ICAM-1, VCAM-1, andβ1-integrin in endothelial cells (ECs) isolated from venous blood (circulating EC, cEC) and purified from femoral plaques (pEC) obtained from 9 patients with peripheral artery disease (PAD) submitted to femoral artery thrombendarterectomy (FEA). In addition, in peripheral blood mononuclear cells (PBMCs) of the same subjects, we investigated gene expression of IFN-γ, IL-4, TGF-β, and IL-10. Patients were longitudinally evaluated 1 month before surgery, when statin treatment was established, at the time of surgery, and after 2 and 5 months. All AM mRNA levels, measured by means of real-time PCR, in cEC diminished during the study, up to 41–50% of initial levels at followup. AM mRNA expression was significantly higher in pEC than in cEC. During the study, in PBMCs, TGF-βand IL-10 mRNA levels remained unchanged while IFN-γand IL-4 levels increased; however, the ratio IFN-γ/IL-4 showed no significant modification. In PAD patients, FEA and statin treatment induce a profound reduction of AM expression in cEC and affect cytokine mRNA expression in PBMCs.

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Human mononuclear cells express 12-LX: coordinated mRNA regulation with 5-LX and FLAP genes

Blood ◽

10.1182/blood.v87.1.331.bloodjournal871331 ◽

1996 ◽

Vol 87 (1) ◽

pp. 331-340

Author(s):

WE Kaminski ◽

E Jendraschak ◽

K Baumann ◽

R Kiefl ◽

S Fischer ◽

...

Keyword(s):

Gene Expression ◽

Mrna Expression ◽

Time Course ◽

Cell Activation ◽

Mononuclear Cells ◽

Constitutive Expression ◽

Ionophore A23187 ◽

Mrna Levels ◽

Rt Pcr ◽

Human Mononuclear Cells

Lipoxygenases (LXs) catalyze formation of leukotrienes and hydroxy-eicosatetraenoic acids (HETEs), proinflammatory, and spasmogenic autacoids that are critical for host defense systems. We studied the expression and regulation of LX genes (12-LX, 5-LX, and 15-LX) and the 5-lipoxygenase activating protein (FLAP) in human mononuclear cells (MNC) and granulocytes using a quantitative reverse transcription polymerase chain reaction (RT-PCR) technique. We show that 12-LX mRNA is constitutively expressed in resting platelet-free MNC. 12-LX gene expression was upregulated by activation with lipopolysaccharide (LPS). The formation of 12-HETE was inducible with ionophore in MNC, as assessed by high-performance liquid chromatography (HPLC) and gas chromatography, and increased after LPS pretreatment. In addition to 12- LX, resting MNC expressed the genes for 5-LX and FLAP constitutively. Quantitative time course analyses of 12-LX, 5-LX, and FLAP gene expression suggested coregulation of 12-LX and FLAP mRNAs, and reciprocal regulation of 5-LX and FLAP mRNAs. During cell stimulation with LPS 5-LX mRNA levels remained unchanged, whereas FLAP gene expression increased. No 15-LX mRNA expression or 15-HETE formation was detectable in unstimulated and activated MNC. In contrast to MNC, quantitative RT-PCR mRNA analysis showed intermittent intraindividual expression of the 5-LX and FLAP genes in resting granulocytes. mRNAs for 12-LX and 15-LX were not expressed. On stimulation of granulocytes ex vivo, mRNA expression of 5-LX and FLAP was upregulated. Stimulation by LPS differed from that by ionophore A23187. Neither LPS nor ionophore induced gene expression of 12-LX or 15-LX in granulocytes. Our data indicate that resting human MNC and granulocytes express LX and FLAP genes in a cell-specific manner. Cell activation induces coordinated upregulation of 12-LX and FLAP genes in MNC, and 5-LX and FLAP genes in granulocytes, respectively. The constitutive expression of 12-LX mRNA, its upregulation on cell activation, and the formation of 12-HETE clearly indicate the presence of a functional 12-LX in human MNC.

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Human mononuclear cells express 12-LX: coordinated mRNA regulation with 5-LX and FLAP genes

Blood ◽

10.1182/blood.v87.1.331.331 ◽

1996 ◽

Vol 87 (1) ◽

pp. 331-340 ◽

Cited By ~ 12

Author(s):

WE Kaminski ◽

E Jendraschak ◽

K Baumann ◽

R Kiefl ◽

S Fischer ◽

...

Keyword(s):

Gene Expression ◽

Mrna Expression ◽

Time Course ◽

Cell Activation ◽

Mononuclear Cells ◽

Constitutive Expression ◽

Ionophore A23187 ◽

Mrna Levels ◽

Rt Pcr ◽

Human Mononuclear Cells

Abstract Lipoxygenases (LXs) catalyze formation of leukotrienes and hydroxy-eicosatetraenoic acids (HETEs), proinflammatory, and spasmogenic autacoids that are critical for host defense systems. We studied the expression and regulation of LX genes (12-LX, 5-LX, and 15-LX) and the 5-lipoxygenase activating protein (FLAP) in human mononuclear cells (MNC) and granulocytes using a quantitative reverse transcription polymerase chain reaction (RT-PCR) technique. We show that 12-LX mRNA is constitutively expressed in resting platelet-free MNC. 12-LX gene expression was upregulated by activation with lipopolysaccharide (LPS). The formation of 12-HETE was inducible with ionophore in MNC, as assessed by high-performance liquid chromatography (HPLC) and gas chromatography, and increased after LPS pretreatment. In addition to 12- LX, resting MNC expressed the genes for 5-LX and FLAP constitutively. Quantitative time course analyses of 12-LX, 5-LX, and FLAP gene expression suggested coregulation of 12-LX and FLAP mRNAs, and reciprocal regulation of 5-LX and FLAP mRNAs. During cell stimulation with LPS 5-LX mRNA levels remained unchanged, whereas FLAP gene expression increased. No 15-LX mRNA expression or 15-HETE formation was detectable in unstimulated and activated MNC. In contrast to MNC, quantitative RT-PCR mRNA analysis showed intermittent intraindividual expression of the 5-LX and FLAP genes in resting granulocytes. mRNAs for 12-LX and 15-LX were not expressed. On stimulation of granulocytes ex vivo, mRNA expression of 5-LX and FLAP was upregulated. Stimulation by LPS differed from that by ionophore A23187. Neither LPS nor ionophore induced gene expression of 12-LX or 15-LX in granulocytes. Our data indicate that resting human MNC and granulocytes express LX and FLAP genes in a cell-specific manner. Cell activation induces coordinated upregulation of 12-LX and FLAP genes in MNC, and 5-LX and FLAP genes in granulocytes, respectively. The constitutive expression of 12-LX mRNA, its upregulation on cell activation, and the formation of 12-HETE clearly indicate the presence of a functional 12-LX in human MNC.

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Serum Interferon (IFN)-Neutralizing Antibodies and Bioactivities of IFNs in Patients with Severe Type II Essential Mixed Cryoglobulinemia

Clinical and Diagnostic Laboratory Immunology ◽

10.1128/cdli.10.1.70-77.2003 ◽

2003 ◽

Vol 10 (1) ◽

pp. 70-77 ◽

Cited By ~ 6

Author(s):

Carolina Scagnolari ◽

Milvia Casato ◽

Francesca Bellomi ◽

Francesca De Pisa ◽

Ombretta Turriziani ◽

...

Keyword(s):

Mrna Expression ◽

Clinical Response ◽

Neutralizing Antibodies ◽

Mononuclear Cells ◽

Mixed Cryoglobulinemia ◽

Mrna Levels ◽

Type Ii ◽

Essential Mixed Cryoglobulinemia ◽

Peripheral Blood Mononuclear ◽

Severe Type

ABSTRACT The efficacy of alpha interferon (IFN-α) in the treatment of severe type II essential mixed cryoglobulinemia (EMC) has been reported previously. In some patients, the development of neutralizing antibodies to recombinant IFN-α (rIFN-α) can affect the clinical response achieved with rIFN-α; a second treatment with natural IFN-α preparations may reinduce the clinical response. In the present study the ability of leukocyte IFN (LeIFN) to restore the response was investigated from a pharmacodynamic viewpoint. Specifically, the pharmacodynamic profiles of different IFN-α preparations were studied by measuring the serum neopterin levels and the levels of expression of protein MxA mRNA in in vivo peripheral blood mononuclear cells in two patients with EMC whose resistance to rIFN-α2a treatment increased concomitantly with the development of neutralizing antibodies. These markers were measured before injection and at 24 and 48 h after a single injection of rIFN-α2a, consensus IFN [(C)IFN], or LeIFN. No increase or only a slight increase in MxA mRNA levels was detectable after administration of rIFN-α2a or (C)IFN, whereas a significant increase (≥10-fold) in MxA mRNA expression was recorded following administration of LeIFN. The neutralizing antibodies to rIFN-α2a cross-react with (C)IFN. Sera from these patients neutralized most but not all of the subtypes present in the natural IFN-α (LeIFN) mixture, and no significant increase in neopterin levels was observed after these patients were switched to LeIFN treatment. In summary, the data demonstrate that the problem of neutralizing antibodies still exists and that LeIFN may induce an increase in the level of MxA mRNA expression but not an increase in neopterin levels in patients who are resistant to treatment with rIFN-α2a or (C)IFN.

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Elevated Expression of the NLRP3 Inflammasome and Its Correlation with Disease Activity in Adult-onset Still Disease

The Journal of Rheumatology ◽

10.3899/jrheum.161354 ◽

2017 ◽

Vol 44 (8) ◽

pp. 1142-1150 ◽

Cited By ~ 17

Author(s):

Chia-Wei Hsieh ◽

Yi-Ming Chen ◽

Chi-Chen Lin ◽

Kuo-Tung Tang ◽

Hsin-Hua Chen ◽

...

Keyword(s):

Disease Activity ◽

Mrna Expression ◽

Nlrp3 Inflammasome ◽

Mononuclear Cells ◽

Mrna Levels ◽

Adult Onset ◽

Peripheral Blood Mononuclear ◽

Protein Expressions ◽

Still Disease ◽

Stimulation Of

Objective.The dysregulation of the NLRP3 (NLR containing a pyrin domain) inflammasome is involved in autoinflammatory diseases. Adult-onset Still disease (AOSD) is regarded as an autoinflammatory disease. However, the pathogenic involvement of NLRP3 inflammasome in AOSD remains unclear and NLRP3 activators in AOSD are currently unknown.Methods.The mRNA expression of NLRP3 inflammasome signaling in peripheral blood mononuclear cells (PBMC) from 34 patients with AOSD and 14 healthy subjects was determined using quantitative-PCR (qPCR). The changes in mRNA and protein levels of NLRP3 inflammasome signaling in PBMC treated with the potential activator [imiquimod (IMQ)] or inhibitor of NLRP3 were evaluated using qPCR and immunoblotting, respectively. The supernatant levels of interleukin (IL)-1β and IL-18 were determined by ELISA.Results.Significantly higher mRNA levels of NLRP3 inflammasome signaling were observed in patients with AOSD compared with healthy controls. NLRP3 expressions were positively correlated with disease activity in patients with AOSD. IMQ (an effective Toll-like receptor 7 ligand; 10 µg/ml and 25 µg/ml) stimulation of PBMC from patients with AOSD induced dose-dependent increases of mRNA expression of NLRP3 (mean ± standard error of the mean, 2.06 ± 0.46 and 6.05 ± 1.84, respectively), caspase-1 (1.81 ± 0.23 and 4.25 ± 0.48), IL-1β (5.68 ± 1.51 and 12.13 ± 3.71), and IL-18 (2.32 ± 0.37 and 4.81 ± 0.51) compared with controls (all p < 0.005). IMQ stimulation of PBMC from patients similarly induced greater increases in protein expressions of NLRP3 inflammasome compared with controls. The protein expressions of NLRP3, IL-1β, and IL-18 on PBMC significantly decreased after treatment with NLRP3 inhibitor in patients with AOSD.Conclusion.Increased expression of NLRP3 inflammasome and its positive correlation with disease activity in AOSD suggest its involvement in disease pathogenesis. IMQ upregulated expressions of NLRP3 inflammasome signaling, and IMQ might be an activator of NLRP3 inflammasome in AOSD.

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Expression of the antiviral protein Mx in peripheral blood mononuclear cells of pregnant and bred, non-pregnant ewes

Journal of Endocrinology ◽

10.1677/joe.0.170r007 ◽

2001 ◽

Vol 170 (2) ◽

pp. R7-11 ◽

Cited By ~ 54

Author(s):

SJ Yankey ◽

BA Hicks ◽

KG Carnahan ◽

AM Assiri ◽

SJ Sinor ◽

...

Keyword(s):

Gene Expression ◽

Immune System ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Fold Increase ◽

Mrna Levels ◽

Antiviral Protein ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

Interferon-tau (IFN tau) acts locally on the endometrium to suppress estrogen and oxytocin receptor expression and block luteolysis in ruminants. Systemic administration of conceptus homogenates or recombinant ovine IFN tau does not block luteolysis or enhance pregnancy rates in sheep or cattle, respectively. However, IFN tau up-regulates expression of the antiviral protein Mx throughout the entire uterine wall during early pregnancy. These studies determined if conceptus-derived IFN tau also up-regulates Mx expression in components of the circulating immune system that migrate through the endometrial wall. In experiment one, peripheral blood mononuclear cells (PBMC) were isolated from ewes at D26 post-artificial insemination (AI) and Mx mRNA levels examined by Northern and slot-blot hybridization. Pregnancy resulted in a two-fold increase in Mx mRNA levels compared to bred, non-pregnant ewes at D26. In experiment two, PBMC were isolated from ewes at AI, and every three days from D9 to D30. Results showed a four-fold increase in Mx mRNA levels in PBMC from pregnant versus bred, non-pregnant ewes at D15. Increased Mx mRNA, which remained elevated through D30, was accompanied by increased levels of Mx protein. These results show that pregnancy recognition signaling rapidly induces Mx gene expression in PBMC, and are the first to suggest that IFN tau activates gene expression in components of the circulating immune system.

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Integrating the lncRNA and mRNA expression profiles of TLR4-primed mesenchymal stem cells in ankylosing spondylitis

10.21203/rs.3.rs-45893/v1 ◽

2020 ◽

Author(s):

Yuxi Li ◽

Ming Li ◽

Jiajun Huang ◽

Yuwei Liang ◽

Junshen Huang ◽

...

Keyword(s):

Ankylosing Spondylitis ◽

Mrna Expression ◽

High Throughput Sequencing ◽

Mononuclear Cells ◽

Expression Profiles ◽

Functional Networks ◽

Control Group ◽

Mrna Levels ◽

Peripheral Blood Mononuclear ◽

Qrt Pcr

Abstract Background Our previous study found that the toll-like receptor 4 (TLR4) expression of ankylosing spondylitis (AS) patients was significantly different from that of healthy donors. The goals of this study were to explore the expression profiles and functional networks of lncRNAs and mRNAs in TLR4-primed mesenchymal stromal cells from AS patients (AS-MSCs) and to clarify the mechanisms by which TLR4-primed MSCs exert immunoregulatory effects in AS. Methods Firstly, the immunoregulatory effects of MSCs were determined after TLR4 activation. Then, the differentially expressed (DE) lncRNAs and mRNAs between the control group (AS-MSCs without stimulation) and experimental group (AS-MSCs stimulated with lipopolysaccharide) were identified through high-throughput sequencing followed by qRT-PCR confirmation. Finally, bioinformatic analyses were performed to identify the critical biological functions, signalling pathways and associated functional networks involved in the TLR4-primed immunoregulatory function of AS-MSCs. Results TLR4-primed AS-MSCs showed a strong ability to inhibit the proliferation of peripheral blood mononuclear cells (PBMCs) with 1 µg/ml LPS stimulation for 4 hours. A total of 147 DE lncRNAs and 698 DE mRNAs were identified between TLR4-primed AS-MSCs and unstimulated AS-MSCs. Significant fold changes in lncRNA and mRNA levels were confirmed by qRT-PCR. GO and KEGG analysis demonstrated that the DE mRNAs and lncRNAs were highly associated with the inflammatory response. Cis-regulation prediction revealed 9 novel lncRNAs while trans-regulation prediction revealed 15 lncRNAs, respectively. Conclusions Our research describes the lncRNA and mRNA expression profiles and functional networks in TLR4-primed AS-MSCs, which is supposed to enhance the understanding of the pathogenesis of AS-MSC immunoregulatory dysfunction.

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PO-050 Effects of Exercise on Gene Expression Correlative to Bone Metabolism in Peripheral Blood Mononuclear Cells

Exercise Biochemistry Review ◽

10.14428/ebr.v1i3.10153 ◽

2018 ◽

Vol 1 (3) ◽

Author(s):

Yongjie Yang ◽

Li Gao

Keyword(s):

Gene Expression ◽

Cell Adhesion ◽

Bone Metabolism ◽

Adhesion Molecules ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Cell Adhesion Molecules ◽

Mononuclear Cells ◽

Peripheral Blood Mononuclear ◽

Blood Mononuclear Cells

Objective To examine the gene expression pattern of Peripheral Blood Mononuclear Cells （PBMCs）and to explore exercise-sensitive genes correlative to bone metabolism in PBMCs. Thus to provide a theoretical basis for exercise intervention to prevent and treat osteoporosis. Methods Uphill (+8° ) and downhill (-8° ) training were used for the exercise loading in two-month-old male Sprague-Dawley rats. The exercise method performed at 25 m/min, training 50 min/d, 5 d/wk, for 12 wk, respectively. Bone mineral density of distal femurs was measured using dual X-ray absorptiometry, and the expressed gene profile of PBMCs was examined using Gene Chip IVT Labeling Kit (Affymetrix). Results Compared with control (CON) group, the BMD of the femur in the downhill (DOWN) group was significantly increased. Compared with the uphill （UP）group, the BMD and BMC of the femur in the DOWN group were significantly increased. There were 38 genes detected differentially expressed between two exercise groups together with CON group. The expression of genes modified by running involved in immunity, cell proliferation，Rheumatoid arthritis，Cell adhesion molecules and Tnf signaling pathway. There were 105 differently expressed genes between the DOWN group and the UP group which were mainly enriched in biological processes and pathways such as response to hydrogen peroxide，lipopolysaccharide，cell factor and mechanical stimulus，Cell adhesion molecules，cell migration，collagen biosynthetic process and Tnf signaling pathway. Tnf, Cxcl2, Ccl2, Jun and Mmp9 as the key nodes of protein interaction network were identified as candidate genes related to bone metabolism and sensitive to exercise. Conclusions With weight gaining，age increasing and training time prolonging, long-term and high-intensity exercise will be harm for bone. At the same time and same running speed, downhill running conduces to increase bone density more than uphill running. It may be associated with differential expression of exercise-sensitive genes involved in bone metabolism in PBMCs.

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Gene expression of chemokines CX3CL1 and CXCL16 and their receptors, CX3CR1 and CXCR6, in peripheral blood mononuclear cells of patients with relapsing-remitting multiple sclerosis - a pilot study

Vojnosanitetski pregled ◽

10.2298/vsp180717035s ◽

2020 ◽

Vol 77 (9) ◽

pp. 967-973

Author(s):

Ljiljana Stojkovic ◽

Aleksandra Stankovic ◽

Ivan Zivotic ◽

Evica Dincic ◽

Dragan Alavantic ◽

...

Keyword(s):

Gene Expression ◽

Multiple Sclerosis ◽

Peripheral Blood Mononuclear Cells ◽

Peripheral Blood ◽

Mononuclear Cells ◽

Fold Change ◽

Mrna Levels ◽

Peripheral Blood Mononuclear ◽

Relapsing Remitting ◽

Blood Mononuclear Cells

Background/Aim. In vitro and in vivo studies show that CX3CL1 and CXCL16 chemokines and their specific receptors, CX3CR1 and CXCR6, respectively, mediate mechanism of neuroinflammation during the pathogenesis of multiple sclerosis (MS). The aim of this study was to investigate relative messenger ribonucleic acid (mRNA) levels of CX3CL1, CXCL16, CX3CR1 and CXCR6 in peripheral blood mononuclear cells, as potential molecular markers of relapsing-remitting (RR) MS. Methods. The study included 43 unrelated RR MS patients, 20 of them with clinically active disease (relapse) and 23 with clinically stable disease (remission), and 28 unrelated healthy subjects as controls. Real-time polymerase chain reactions (PCR) were performed using TaqMan? gene expression assays. Relative expression (mRNA) level of each target gene in each sample of peripheral blood mononuclear cells was calculated as the mean normalized expression. Results. The levels of CX3CR1 mRNA were significantly higher in clinically active RR MS patients compared to controls [fold change = 1.38, p (Mann-Whitney U test) = 0.009], and significantly lower in clinically stable vs active RR MS patients [fold change = - 1.43, p (t-test) = 0.03]. Stable RR MS patients had significantly higher CXCL16 mRNA levels than controls [fold change = 1.33, p (Mann-Whitney U test) = 0.006]. A trend of increased CXCR6 gene expression was found in active RR MS patients compared to controls [fold change = 1.23, p (Mann-Whitney U test) = 0.08]. In either active or stable RR MS patients there were no significant correlations of the clinical parameters with expression levels of the target genes. Conclusion. The current results show that increased CX3CR1 mRNA levels in peripheral blood mononuclear cells could represent a proinflammatory molecular marker of clinically active RR MS.

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Efficacy of β-D-Mannuronic Acid [M2000] on the Pro-Apoptotic Process and Inflammatory-Related Molecules NFκB, IL-8 and Cd49d using Healthy Donor PBMC

Current Drug Discovery Technologies ◽

10.2174/1570163815666181109165837 ◽

2020 ◽

Vol 17 (2) ◽

pp. 225-232

Author(s):

Atousa Khalatbari ◽

Mehdi Mahdavi ◽

Fahimeh Jafarnezhad ◽

Sanaz Afraei ◽

Farzaneh Tofighi Zavareh ◽

...

Keyword(s):

Gene Expression ◽

Healthy Volunteers ◽

Illicit Drugs ◽

Mononuclear Cells ◽

Venous Blood ◽

Untreated Group ◽

High Dose ◽

Peripheral Blood Mononuclear ◽

Expression Levels ◽

Mannuronic Acid

Objective: This investigation evaluates the pro-apoptotic and anti-inflammatory effects of β-D-mannuronic acid [M2000] compared to diclofenac, based on gene expression involved in apoptosis and inflammation process [including Bcl2, NFκB, IL-8 and Cd49d] in Peripheral Blood Mononuclear Cells [PBMCs] of healthy donors under exvivo conditions. Material: The venous blood samples of twelve healthy volunteers with aged 25-60 years were collected in heparinized tubes. The healthy volunteers were selected from no smoking group and without using illicit drugs and suffering from diabetes. The PBMCs were separated and divided into untreated and treated groups. Methods: The PBMCs of each sample were cultured in 5 wells of culture plate, so that the first well consisted of 2×106 cells exposed by LPS-EB [1μg/ml] to stimulate PBMCs and absence of M2000 [untreated well]. The second, third, fourth and fifth wells containing 2×106 cells/well and LPS-EB, after 4 hours incubation at 37ºC, received 5, 25 and 50 μg/well of M2000 and 5 μg/well of diclofenac, respectively as treated group. Results: The PBMCs were separated and RNAs were then extracted and cDNAs synthesized and gene expression levels were assessed by qRT-PCR. Furthermore, we studied whether M2000 is able to facilitate apoptosis in PBMCs. Our findings represent that the high dose of M2000 could significantly decrease the expression level of NFκB gene compared to untreated group (p < 0.0002). On the other hand, no significant change was observed in treated cells with diclofenac. All doses of M2000 could significantly augment apoptosis compared to untreated group [p < 0.0001]. Additionally, we observed the same apoptotic effects between the medium dose of M2000 and diclofenac. Besides, no significant reduction was shown in expression levels of IL8, Bcl2 and Cd49d genes in all doses of M2000 and diclofenac compared to untreated group. This experiment demonstrates M2000 as a new effective NSAID with immunosuppressive characteristics capable of stimulating apoptosis through lowering expression levels of NFκB gene, which might be probably considered as an appropriate drug for reducing the risk of developing inflammatory diseases and cancer.

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Abstract 17236: Regulating Optimal Angiogenesis and Leukocyte Adhesion of Human Endothelial Cells by Gradient Nanopore Substrate

Circulation ◽

10.1161/circ.138.suppl_1.17236 ◽

2018 ◽

Vol 138 (Suppl_1) ◽

Author(s):

Soon Jun Hong ◽

Li-Hua Huang

Keyword(s):

Endothelial Cells ◽

Cellular Response ◽

Mononuclear Cells ◽

Leukocyte Adhesion ◽

Umbilical Vein ◽

Mrna Levels ◽

Human Endothelial Cells ◽

Peripheral Blood Mononuclear ◽

Angiogenic Potential ◽

Cellular Effects

Introduction: Understanding signals in the microenvironment that regulate endothelial cell behavior are important in tissue engineering. Hypothesis: Although many studies have examined the cellular effects of nanotopography, no study has investigated the functional regulation of human endothelial cells grown on nano-sized gradient pore substrate. Methods: We examined the cellular response of human umbilical vein endothelial cells (HUVECs) by using a gradient nanopore substrate (GPS) with three different types of hole nanopore patterns (HP): which diameters were described in HP1, 120-200 nm; HP2, 200-280 nm; HP3, 280-360 nm. Results: Our study revealed that: (1) HUVECs showed highest proliferation and attachment capacity on HP2 (200-280 nm) GPS, (2) HP2 GPS was determined to be the optimal diameter for extending greater angiogenic potential to HUVECs and (3) both HP2 and HP3 GPS significantly decrease leukocyte adhesion compared to Flat. Moreover, expressions of VCL, TLN1, PXN, ITGA2, ITGA6, ITGAV, ITGB3 and ROCK1 in HUVECs cultured on HP2 GPS were significantly greater than that of Flat. Conversely, TLN1, PXN, ROCK 1 and ROCK2 mRNA levels decreased on HP3 GPS compared to those of Flat. Mature tube structures at 24 hours were found in HUVECs stimulated by HP2 GPS. Expressions of angiogenesis related markers (ANGPT1, TIE2 and CXCR4) and endothelial-related marker (vWF) were significantly increased in HUVECs grown on HP2 GPS. The adhesion of peripheral blood mononuclear cells (PBMNCs) to HUVECs stimulated by HP2 and HP3 GPS was significantly reduced compared to those of Flat at 3, 6 and 24 hours after PBMNC seeding. Conclusions: Optimal functional regulations of HUVECs were achieved by nanopore substrate with 200-280 nm-sized pores. Our results provide the possibility of applying nanopore interface-based implant devices in the field of cardiovascular regenerative therapy.

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