Irina Titorencu

Circulating microparticles and endothelial progenitor cells in atherosclerosis: pharmacological effects of irbesartan

Summary.  Aims: This study aimed to (i) employ our newly designed model, the hypertensive–hypercholesterolemic hamster (HH), in order to find out whether a correlation exists between circulating microparticles (MPs), endothelial progenitor cells (EPCs) and their contribution to vascular dysfunction and (ii) to assess the effect of irbesartan treatment on HH animals (HHI).Methods and Results: The results showed that compared with the control (C) group, HH displayed: (i) a significant increase in plasma cholesterol and triglyceride concentration, and an augmentation of systolic and diastolic arterial blood pressure, and of heart rate; (ii) a marked elevation of MPs and a significant decrease in EPCs; (iii) structural modifications of the arterial wall correlated with altered protein expression of MMP2, MMP9, MMP12, TIMP1, TIMP2 and collagen type I and III; (iv) a considerably altered reactivity of the arterial wall closely correlated with MPs and EPC adherence; and (v) an inflammatory process characterized by augmented expression of P‐Selectin, E‐Selectin, von Willebrand factor, tissue factor, IL‐6, MCP‐1 and RANTES. Additionally, the experiments showed the potential of irbesartan to correct all altered parameters in HH and to mobilize EPCs by NO, chemokines and adhesion molecule‐dependent mechanisms.Conclusions: Hypertension associated with hypercholesterolemia is accompanied by structural modifications and expression of pro‐inflammatory molecules by the vessel wall, the alteration of vascular tone, enhanced release of MPs and reduced EPCs; the ratio between the latter two may be considered as a marker of vascular dysfunction. Irbesartan, which exhibits a pharmacological control on the levels of MPs and EPCs, has the potential to restore homeostasis of the arterial wall.

The two step nanotube formation on TiZr as scaffolds for cell growth

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.

Osteoblast ontogeny and implications for bone pathology: an overview.

Osteoblasts are specialized mesenchyme-derived cells accountable for bone synthesis, remodelling and healing. Differentiation of osteoblasts from mesenchymal stem cells (MSC) towards osteocytes is a multi-step process strictly controlled by various genes, transcription factors and signalling proteins. The aim of this review is to provide an update on the nature of bone-forming osteoblastic cells, highlighting recent data on MSC—osteoblast—osteocyte transformation from a molecular perspective and to discuss osteoblast malfunctions in various bone diseases. We present here the consecutive stages occurring in the differentiation of osteoblasts from MSC, the transcription factors involved and the role of miRNAs in the process. Recent data concerning the pathogenic mechanisms underlying the loss of bone mass and architecture caused by malfunctions in the synthetic activity and metabolism of osteoblasts in osteoporosis, osteogenesis imperfecta, osteoarthritis and rheumatoid arthritis are discussed. The newly acquired knowledge of the ontogeny of osteoblasts will assist in unravelling the abnormalities taking place during their differentiation and will facilitate the prevention and/or treatment of bone diseases by therapy directed against altered molecules and mechanisms.

The promise of EPC-based therapies on vascular dysfunction in diabetes

Diabetes mellitus is one of the most common metabolic diseases in the world and the vascular dysfunction represents a challenging clinical problem. In diabetes, endothelial cells (ECs), lining the inner wall of blood vessels, do not function properly and contribute to impaired vascular function. Circulating endothelial progenitor cells (EPCs), the precursor of mature EC, actively participate in endothelial repair, by moving to the vascular injury site to form mature EC and new blood vessels. Knowing that the therapeutic interventions can improve only a part of EC dysfunction in diabetes, this review addresses recent findings on the use of EPCs for cell therapy. The strategies proposed in review are based on in vivo and in vitro studies and, thus, their physiological relevance is confirmed. EPC therapy shows great promise for the prevention and cure of diabetes-induced vascular dysfunction.

Irbesartan administration therapeutically influences circulating endothelial progenitor cell and microparticle mobilization by involvement of pro-inflammatory cytokines

Circulating microparticles (MPs) and endothelial progenitor cells (EPCs) correlate with endothelial dysfunction and contribute to the pathogenesis of atherosclerosis. In this context, we explored whether the angiotensin II type I receptor antagonist, irbesartan, exerts a pharmacological control in the atherosclerotic process by the improvement of EPC mobilization and inhibitory effects on MP release and VEGF and SDF-1α levels in the hypertensive-hypercholesterolemic (HH) hamster model. The HH hamsters were treated with irbesartan (50mg/kg b.w/day administered by gavage) for 4 month (HHI). We analyzed MP/EPC infiltration in vascular wall before and after irbesartan administration as well as the endothelial function and expression of VEGF/SDF-1α in plasma and tissue and of molecular pathways activated by them. The results showed that treatment with irbesartan significantly increased EPC infiltration and decreased MP infiltration. The mechanisms underlying this response include the reduction/increase of a number of specific membrane receptors exposed by MPs (TF, P-Selectin, E-Selectin, PSGL-1, Rantes), respectively, by EPCs (β2-Integrins, α4β1-integrin), the augmentation of endothelium-mediated vasodilation and the reduction of protein expression of VEGF/SDF-1α followed by: (1) the diminishment of pro-inflammatory endothelial cytokines: VEGFR1, VEGFR2, CXCR4, Tie2, PIGF with role in EPC homing to sites of damaged endothelium; and (2) the increase of protein expression of COX-2, PGI2 synthase molecules with role in the improvement of arterial wall vasodilatation. In conclusion, the study underlines that irbesartan administration therapeutically improves/reduces EPC, respectively, MP mobilization and this action may be of salutary relevance contributing to its beneficial cardiovascular effects.

Collagen Wound Dressings with Anti-Inflammatory Activity

The aim of this study was to develop modern wound dressings such as controlled drug delivery systems. These systems consist in collagen as release support and niflumic acid as drug. The scaffolds were prepared by lyophilization in order to obtain porous structures and were evaluated by release profile of niflumic acid, water absorption, collagenase degradation and biocompatibility with fibroblast cells. The collagen scaffold with 0.75% niflumic acid solved in laurel oil was optimal in terms of biodegradability, absorbability and fibroblast cells biocompatibility. Thus, the obtained collagen scaffolds could be used as wound dressings with absorbent, antibacterial and anti-inflammatory properties.

Mechanical properties and biocompatibility of the sputtered Ti doped hydroxyapatite

The hydroxyapatite enriched with Ti were prepared as possible candidates for biomedical applications especially for implantable devices that are in direct contact to the bone. The hydroxyapatites with different Ti content were prepared by RF magnetron sputtering on Ti-6Al-4V alloy using pure hydroxyapatite and TiO2 targets. The content of Ti was modified by changing the RF power fed on TiO2 target. The XPS and FTIR analyses revealed the presence of hydroxyapatite structure. The hardness and elastic modulus of the hydroxyapatite were increased by Ti addition. After 5 days of culture, the cell viability of the Ti-6Al-4V was enhanced by depositing with undoped or doped hydroxyapatite. The Ti additions led to an increase in cell viability of hydroxyapatite, after 5 days of culture. The electron microscopy showed the presence of more cells on the surface of Ti-enriched hydroxyapatite than those observed on the surface of the uncoated alloys or undoped hydroxyapatite.

In Vitro Biocompatibility of Human Endothelial Cells with Collagen-Doxycycline Matrices

The aim of this study was to test the biocompatibility between collagen-doxycycline matrices with various porosities and human endothelial cells. Collagen matrices were prepared by freeze-drying process. The crosslinking degree of collagen matrices was evaluated by FT-IR spectroscopy, the matrices morphology by SEM microscopy. The biocompatibility of collagen matrices with endothelial cells was monitored byfluorescence and transmission electron microscopy. We found that the three-dimensional structures of matrix with 1.2% collagen, 0.2% doxycycline crosslinked with 0.25% glutaraldehyde was optimal in terms of biodegradability, morphology and endothelial cells biocompatibility indicating the use of these scaffolds in tissueengineering.

Natural Polymer-Cell Bioconstructs for Bone Tissue Engineering

The major goal of bone tissue engineering is to develop bioconstructs which substitute the functionality of damaged natural bone structures as much as possible if critical-sized defects occur. Scaffolds that mimic the structure and composition of bone tissue and cells play a pivotal role in bone tissue engineering applications. First, composition, properties and in vivo synthesis of bone tissue are presented for the understanding of bone formation. Second, potential sources of osteoprogenitor cells have been investigated for their capacity to induce bone repair and regeneration. Third, taking into account that the main property to qualify one scaffold as a future bioconstruct for bone tissue engineering is the biocompatibility, the assessments which prove it are reviewed in this paper. Forth, various types of natural polymer- based scaffolds consisting in proteins, polysaccharides, minerals, growth factors etc, are discussed, and interaction between scaffolds and cells which proved bone tissue engineering concept are highlighted. Finally, the future perspectives of natural polymer-based scaffolds for bone tissue engineering are considered.

Enoxaparin reduces adrenergic contraction of resistance arterioles in aging and in aging associated with diabetes via engagement of MAP kinase pathway

The arterial endothelial dysfunction in aging and diabetes remains a clinical problem. We questioned the effect of the low-molecular-weight heparin, enoxaparin, on arterioles contractility in aging and in aging associated with diabetes, and investigated the involvement of the mitogen-activated protein (MAP) kinases pathway in the enoxaparin-mediated effect. The experiments were performed on the isolated resistance arteries of young (4 months old), aged (16 months old), and aged-diabetic hamsters (16 months old and 5 months since streptozotocin injection). The techniques used were myography, molecular biology, and immunoblotting. The results showed that 60 μg/ml enoxaparin has favorable effects on the arteriole reactivity in aged and aged-diabetic conditions, reducing the contractile response to 10−8–10−6 mol/l noradrenaline. The diminishment of contractility is exerted via MAP kinase pathway, and involves reduction of c-fos gene expression and of transcription factor AP-1 protein expression. These results suggest that enoxaparin preserves the arterial endothelial function in a mechanism independent of its anticoagulant activity. Understanding the signal transduction mechanisms involved in the altered contractility of vascular wall could provide useful information on the development of specific MAP kinase inhibitors with therapeutic benefits and reduced side effects.