Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy

Bucharest, Romania

Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy

Bucharest, Romania

Time filter

Source Type

Preda M.B.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Preda M.B.,University of Oslo | Ronningen T.,University of Oslo | Burlacu A.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | And 4 more authors.
Stem Cells | Year: 2014

Cardioprotection can be evoked through extracardiac approaches. This prompted us to investigate whether remote transplantation of stem cells confers protection of the heart against ischemic injury. The cardioprotective effect of subcutaneous transplantation of naïve versus heme oxygenase-1 (HMOX-1)-overexpressing mouse mesenchymal stem cells (MSC) to mice was investigated in hearts subjected to ischemia-reperfusion in a Langendorff perfusion system. Mice were transplanted into the interscapular region with naïve or HMOX-1 transfected MSC isolated from transgenic luciferase reporter mice and compared to sham-treated animals. The fate of transplanted cells was followed by in vivo bioluminescence imaging, revealing that MSC proliferated, but did not migrate detectably from the injection site. Ex vivo analysis of the hearts showed that remote transplantation of mouse adipose-derived MSC (mASC) resulted in smaller infarcts and improved cardiac function after ischemia-reperfusion compared to sham-treated mice. Although HMOX-1 overexpression conferred cytoprotective effects on mASC against oxidative stress in vitro, no additive beneficial effect of HMOX-1 transfection was noted on the ischemic heart. Subcutaneous transplantation of MSC also improved left ventricular function when transplanted in vivo after myocardial infarction. Plasma analysis and gene expression profile of naïve- and HMOX-1-mASC after transplantation pointed toward pentraxin 3 as a possible factor involved in the remote cardioprotective effect of mASC. These results have significant implications for understanding the behavior of stem cells after transplantation and development of safe and noninvasive cellular therapies with clinical applications. Remote transplantation of MSC can be considered as an alternative procedure to induce cardioprotection. © 2014 AlphaMed Press.


Manea A.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Manea A.,Petru Poni Institute of Macromolecular Chemistry | Manea S.-A.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Todirita A.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | And 4 more authors.
Cell and Tissue Research | Year: 2015

High glucose induces vascular smooth muscle cell (SMC) dysfunction by generating oxidative stress attributable, in part, to the up-regulated NADPH oxidases (Nox). We have attempted to elucidate the high-glucose-generated molecular signals that mediate this effect and hypothesize that products of high-glucose-induced lipid peroxidation regulate Nox by activating peroxisome proliferator-activated receptors (PPARs). Human aortic SMCs were exposed to glucose (5.5–25 mM) or 4-hydroxynonenal (1–25 μM, 4-HNE). Lucigenin assay, real-time polymerase chain reaction, western blot, and promoter analyses were employed to investigate Nox. We found that high glucose generated an increase in Nox activity and expression. It also promoted oxidative stress that consequently induced lipid peroxidation, which resulted in the production of 4-HNE. Pharmacological inhibition of Nox activity significantly reduced the formation of high-glucose-induced 4-HNE. Exposure of SMCs to non-cytotoxic concentrations (1–10 μM) of 4-HNE alone mimicked the effect of high glucose incubation, whereas scavenging of 4-HNE by N-acetyl L-cysteine completely abolished both the effects of high glucose and 4-HNE. The latter exerted its effect by activating PPARα and PPARβ/δ, but not PPARγ, as assessed pharmacologically by the inhibitory effect of selective antagonists and following the silencing of the expression of these receptors. These new data indicate that 4-HNE, generated following Nox activation, functions as an endogenous activator of PPARα and PPARβ/δ. The newly discovered “lipid peroxidation products–PPARs–Nox axis” represents a novel mechanism of Nox regulation and an additional therapeutic target for oxidative stress in diabetes. © 2015, Springer-Verlag Berlin Heidelberg.


Butoi E.D.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Gan A.M.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Manduteanu I.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Stan D.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | And 6 more authors.
Biochimica et Biophysica Acta - Molecular Cell Research | Year: 2011

Objective: In atherosclerotic lesions, fractalkine (CX3CL1) and its receptor (CX3CR1) expressed by smooth muscle cells (SMC) and monocytes/macrophages, mediate the heterotypic anchorage and chemotaxis of these cells. We questioned whether, during the close interaction of monocytes with SMC, the CX3CL1/CX3CR1 pair modulates the expression of pro-atherogenic molecules in these cells. Methods and results: SMC were co-cultured with monocytes or LPS-activated monocytes (18. h) and then the cells were separated and individually investigated for the gene and protein expression of TNFα, IL-1β, IL-6, CX3CR1 and metalloproteinases (MMP-2, MMP-9). We found that SMC-monocyte interaction induced, in each cell type, an increased mRNA and protein expression of TNFα, IL-1β, IL-6, CX3CR1, MMP-2 and MMP-9. Blocking the binding of fractalkine to CX3CR1 (by pre-incubation of monocytes with anti-CX3CR1 or by CX3CR1 siRNA transfection) before cell co-culture decreased the production of TNFα, CX3CR1 and MMP-9. Monocyte-SMC interaction induced the phosphorylation of p38MAPK and activation of AP-1 transcription factor. Silencing the p65 (NF-kB subunit) inhibited the IL-1β and IL-6 and silencing c-jun inhibited the TNFα, CX3CR1 and MMP-9 induced by SMC-monocyte interaction. Conclusions: The cross-talk between SMC and monocytes augments the inflammatory response in both cell types as revealed by the increased expression of TNFα, IL-1β, IL-6, CX3CR1 and MMPs. Up-regulation of TNFα, CX3CR1 and MMP-9 is further increased upon interaction of SMC with activated monocytes and is dependent on fractalkine/CXRCR1 pair. These data imply that the fractalkine/CX3RCR1 axis may represent a therapeutic target to impede the inflammatory process associated with atherosclerosis. © 2011 Elsevier B.V.


Pirvulescu M.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Manduteanu I.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Gan A.M.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Stan D.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | And 5 more authors.
Biochemical and Biophysical Research Communications | Year: 2012

Resistin is a significant local and systemic regulatory cytokine involved in inflammation. Suppressors of cytokine signaling (SOCS) proteins are intracellular regulators of receptor signal transduction induced by several cytokines in a cytokine and cell specific manner. Resistin up-regulates SOCS3 expression in mice adipocytes but it is not known whether this is a common occurrence in other cells. We questioned whether resistin-induces SOCS3 in human endothelial cells and if signal transducer and activator of transcription (STAT) proteins are involved in the process. The Real-Time PCR and Western blot analysis showed that in resistin-activated HEC the gene and protein expression of SOCS3 were significantly increased. Furthermore, resistin induced activation of STAT3 as characterized by increased tyrosine phosphorylation. Resistin-induced SOCS3 expression was blocked by specific inhibitors of STAT3 signaling and by the transfection of siRNA specific for STAT3. Silencing of SOCS3 gene expression by transfection with SOCS3 siRNA reduced the expression of resistin induced-P-selectin and fractalkine in HEC. Together, our results demonstrate that in HEC (1) resistin up-regulates SOCS3 expression and activates STAT3 transcription factor; (2) the increase in SOCS3 mRNA and protein expression as well as STAT3 activation have a long-lasting effect (up to 18. h); (3) inhibition of SOCS3 function prevents resistin-induced expression of cell adhesion molecules P-selectin and fractalkine and thus activation of endothelial cells. The data uncover a new resistin-mediated mechanism in human endothelial cells and designate SOCS3 as a novel therapeutic target to modulate resistin-dependent inflammation in vessel wall diseases. © 2012 Elsevier Inc.


Alexandru N.,Petru Poni Institute of Macromolecular Chemistry | Alexandru N.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Popov D.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Georgescu A.,Petru Poni Institute of Macromolecular Chemistry
Trends in Cardiovascular Medicine | Year: 2010

This article provides an overview of the current knowledge on intraplatelet oxidative/nitrative stress, an abnormality associated with platelet activation and hyper-reactivity. The first issue discussed is related to induction of platelet endogenous stress by the molecules present within the circulating (extracellular) milieu that bathes these cells. The second issue concerns the intraplatelet oxidative/nitrative stress associated with specific pathologies or clinical procedures and action of particular molecules and platelet agonists as well as of the specialized intraplatelet milieu and its redox system; the biomarkers of endogenous oxidative/nitrative stress are also briefly outlined. Next, the association between intraplatelet oxidative/nitrative stress and the risk factors of the metabolic syndrome is presented. Then, the most recent strategies aimed at the control/regulation of platelet endogenous oxidative/nitrative stress, such as exploitation of circulating extracellular reactive oxygen species scavengers, manipulation of platelet molecules, and the use of antioxidants, are discussed. Finally, the results of studies on platelet-dependent redox mechanisms, which deserve immediate attention for potential clinical exploitation, are illustrated. © 2010 Elsevier Inc..


Preda M.B.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Burlacu A.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Burlacu A.,Petru Poni Institute of Macromolecular Chemistry | Simionescu M.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy
Tissue and Cell | Year: 2013

The pluripotent nature of embryonic stem (ES) cells makes them powerful tools in cell replacement therapy for severe degenerative diseases, such as heart failure. However, the development of strategies to increase the efficiency of cardiomyocyte (CMC) differentiation is still needed to produce a sufficient amount of cells for clinical applications. This paper evaluates the impact of the size and the aggregation of embryoid bodies (EBs) on the efficiency of ES cell differentiation into CMCs. ES cells were generated from RAP inbred mice. These cells expressed pluripotency markers and induced teratomas when injected into syngeneic mice, which made them suitable for differentiation into CMCs. We found that the EBs that were formed as a result of in vitro ES cell aggregation generated contractile tissue in direct correlation with the initial number of ES cells. Furthermore, the presence of knock-out serum replacement (KO-SR) during ES cell aggregation resulted in less compacted EBs and increased cell differentiation into CMCs compared to the presence of foetal bovine serum. In conclusion, cardiac differentiation of ES cells is dependent on the size and the degree of compaction of EBs, and the presence of KO-SR during initiation of EBs may lead to improved cardiogenic differentiation of ES cells. © 2012 Elsevier Ltd.


Grigorescu S.,Polytechnic University of Bucharest | Pruna V.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Titorencu I.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Jinga V.V.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | And 4 more authors.
Bioelectrochemistry | Year: 2014

Various TiO2 nanotubes on Ti50Zr alloy have been fabricated via a two step anodization method in glycol with 15vol.% H2O and 0.2M NH4F under anodization controlled voltages of 15, 30 and 45V. A new sonication treatment in deionized water with three steps and total sonication time as 1min was performed after the first anodization step in order to remove the oxide layer grown during 2h. The second step of anodization was for 1h and took place at the same conditions. The role of removed layer as a nano-prepatterned surface was evidenced in the formation of highly ordered nanotubular structures and morphological features were analyzed by SEM, AFM and surface wettability. The voltage-controlled anodization leads to various nanoarhitectures, with diameters in between 20 and 80nm. As biological assay, cell culture tests with MG63 cell line originally derived from a human osteosarcoma were performed. A correlation between nanostructure morphological properties as a result of voltage-controlled anodization and cell response was established. © 2014 Elsevier B.V.


Stancu C.S.,Institute of Cellular Biology and Pathology nicolae Simionescu of the Romanian Academy | Toma L.,Institute of Cellular Biology and Pathology nicolae Simionescu of the Romanian Academy | Sima A.V.,Institute of Cellular Biology and Pathology nicolae Simionescu of the Romanian Academy
Cell and Tissue Research | Year: 2012

The endothelium is a key constituent of the vascular wall, being actively involved in maintaining the structural integrity and proper functioning of blood vessels. Hyperlipidemia, diabetes, hypertension, smoking and aging are important risk factors for the dysfunction of endothelial cells (EC). Circulating lipoproteins (Lp) synthesized and secreted from the intestine or liver have an important role in supplying peripheral tissues with fatty acids from triglyceride rich lipoproteins (TGRLp) for energy production or storage, and cholesterol from low density lipoproteins (LDL) or high density lipoproteins (HDL) for the synthesis of cellular membranes and steroid hormones. Under pathological conditions, Lp may suffer alterations in concentration and composition and become aggressors for EC. Modified LDL, remnant Lp, TGRLp lipolysis products, dysfunctional HDL are involved in the changes induced in EC morphology (reduced glycocalyx, overdeveloped endoplasmic reticulum, Golgi apparatus and basement membrane), loose intercellular junctions, increased oxidative and inflammatory stress, nitric oxide/redox imbalance, excess Lp transport and storage, as well as loss of anti-thrombotic properties, all of these being characteristics of endothelial dysfunction. Normal HDL are able to counteract the harmful effects of atherogenic Lp in EC but under persistent pathological conditions they lose the protective properties and become pro-atherogenic. This review summarises recent advances in understanding the role of Lp in the induction of endothelial dysfunction and the initiation and progression of atherosclerotic lesions. Its main focus is the antagonistic role of atherogenic Lp (LDL, VLDL, dysfunctional HDL) versus anti-atherogenic Lp (HDL), also pointing out the potential targets for arresting or reversing this process. © Springer-Verlag 2012.


Manea S.-A.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Todirita A.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Raicu M.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Manea A.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy
Journal of Cellular and Molecular Medicine | Year: 2014

In atherosclerosis, oxidative stress-induced vascular smooth muscle cells (SMCs) dysfunction is partially mediated by up-regulated NADPH oxidase (Nox); the mechanisms of enzyme regulation are not entirely defined. CCAAT/enhancer-binding proteins (C/EBP) regulate cellular proliferation and differentiation, and the expression of many inflammatory and immune genes. We aimed at elucidating the role of C/EBP in the regulation of Nox in SMCs exposed to pro-inflammatory conditions. Human aortic SMCs were treated with interferon-γ (IFN-γ) for up to 24 hrs. Lucigenin-enhanced chemiluminescence, real-time PCR, Western blot, promoter-luciferase reporter analysis and chromatin immunoprecipitation assays were employed to investigate Nox regulation. IFN-γ dose-dependently induced Nox activity and expression, nuclear translocation and up-regulation of C/EBPα, C/EBPβ and C/EBPδ protein expression levels. Silencing of C/EBPα, C/EBPβ or C/EBPδ reduced significantly but differentially the IFN-γ-induced up-regulation of Nox activity, gene and protein expression. In silico analysis indicated the existence of typical C/EBP sites within Nox1, Nox4 and Nox5 promoters. Transient overexpression of C/EBPα, C/EBPβ or C/EBPδ enhanced the luciferase level directed by the promoters of the Nox subtypes. Chromatin immunoprecipitation demonstrated the physical interaction of C/EBPα, C/EBPβ and C/EBPδ proteins with the Nox1/4/5 promoters. C/EBP transcription factors are important regulators of Nox enzymes in IFN-γ-exposed SMCs. Activation of C/EBP may induce excessive Nox-derived reactive oxygen species formation, further contributing to SMCs dysfunction and atherosclerotic plaque development. Pharmacological targeting of C/EBP-related signalling pathways may be used to counteract the adverse effects of oxidative stress. © 2014 The Authors.


Stancu C.S.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Sanda G.M.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Deleanu M.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy | Sima A.V.,Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy
Molecular Nutrition and Food Research | Year: 2014

Scope: Hyperlipidemia, hyperglycemia, and the oxidative stress are among the known risk factors of atherosclerosis. Our aim was to assess the hypolipidemic and antioxidant effects of a probiotic mix (Lactobacillus acidophilus and Bifidobacterium animalis) in hyperlipidemic hamsters (HL). Methods and results: Male Golden Syrian hamsters developed hyperlipidemia after 21 weeks of fat diet. For the last 5 weeks of experiment, ten HL were treated with the probiotic mix (HLP), ten received water (HL). Ten animals received standard chow (N). Increase of plasma total cholesterol (TC), triglycerides (TG), phospholipids (PL), oxidized LDL, glucose, of 4-hydroxynonenal (4-HNE) in plasma, liver, and myocardium, and of intestinal Niemann Pick C1 like 1 (NPC1L1) and microsomal TG transfer protein (MTTP) expression was observed in HL versus N. The probiotic mix decreased plasma TC, TG, PL, oxidized LDL, 4-HNE, and glucose levels and increased paraoxonase-1 activity, decreased NPC1L1 and MTTP protein expression compared to HL. In HLP liver, a significant reduction of TC, TG, and fatty acids was observed. PL increased and 4-HNE levels decreased in the liver and myocardium of HLP versus HL. Conclusion: Our data support the administration of probiotics to humans because of their hypolipidemic (through decreasing intestinal NPC1L1 and MTTP) and antioxidant effects (stimulating HDL-associated paraoxonase-1). © 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Loading Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy collaborators
Loading Institute of Cellular Biology and Pathology Nicolae Simionescu of the Romanian Academy collaborators