P. Di Nardo

Brain Natriuretic Peptide (BNP) regulates the production of inflammatory mediators in human THP-1 macrophages.

Brain Natriuretic Peptide (BNP), besides retaining vasodilatory, diuretic and natriuretic properties, is a vasoactive hormone that it is also involved in several cardiac diseases as well as severe sepsis and septic shock. All these conditions are characterized by an ongoing inflammatory response consisting in a complex interaction of pleiotropic mediators derived from plasma or cells, including monocytes and macrophages. However, the relationship between this hormone and inflammation remains to be elucidated. Therefore, the aim of the present study was to evaluate a possible BNP immunomodulatory activity on macrophages. Our results demonstrate that BNP regulates the production of major inflammatory molecules, such as reactive oxygen- and nitrogen species (ROS and RNS), leukotriene B(4) (LTB(4)), prostaglandin E(2) (PGE(2)); modulates the cytokines (TNF-alpha, IL-12 and IL-10) profile, and affects cell motility. These results furnish novel and brand-new proofs on BNP ability of modulating the production of inflammatory mediators in macrophages whose role has broad implications in inflammatory states where increased BNP levels have been reported.

Role of atrial natriuretic peptide in the suppression of lysophosphatydic acid-induced rat aortic smooth muscle (RASM) cell growth

Lysophosphatidic acid (LPA) is a lipid mediator with multiple biological functions. In the present study we investigated the possible role of atrial natriuretic peptide (ANP), a hormone affecting cardiovascular homeostasis and inducing antimitogenic effects in different cell types, on LPA-induced cell growth and reactive oxygen species (ROS) production in rat aortic smooth muscle (RASM) cells. Both LPA effects on cell growth and levels of ROS were totally abrogated by physiological concentrations of ANP, without modifying the overexpression of LPA-receptors. These effects were also affected by cell pretreatment with wortmannin, an inhibitor of phosphatidylinositol 3-kinase (PI3K). Moreover, the LPA-induced activation of Akt, a downstream target of PI3K, was completely inhibited by physiological concentrations of ANP, which were also able to inhibit p42/p44 phosphorylation. Taken together, our data suggest that PI3K may represent an important step in the LPA signal transduction pathway responsible for ROS generation and DNA synthesis in RASM cells. At same time, the enzyme could also represent an essential target for the antiproliferative effects of ANP.

Tuning hierarchical architecture of 3D polymeric scaffolds for cardiac tissue engineering

Tissue engineering combines the fields of engineering, chemistry, biology, and medicine to fabricate replacement tissues able to restore, maintain, or improve structurally and functionally damaged organs. The approach of regenerative medicine is of paramount importance for treating patients with severe cardiac diseases. For successful exploitation, the challenge for cardiac regenerative medicine is to identify the suitable combination between the best cell source for cardiac repair and the design of the optimal scaffold as a template for tissue replacement. Adult stem cells have the potential to improve regenerative medicine with their peculiar feature to self-renew and differentiate into various phenotypes. Insights into the stem cell field lead to the identification of the suitable scaffold features that enhance the ex vivo proliferation and differentiation of stem cells. Scaffolds composed of natural and/or synthetic polymers can organise stem cells into complex architectures that mimic native tissues. To achieve this, a proper design of the chemical, mechanical, and morphological characteristics of the scaffold at different length scales is needed to reproduce the tissue complexity at the cell-scaffold interface. Hierarchical porosities are needed in a single construct, at the millimetre scale to help nutrition and vascularisation, at the micrometer scale to accommodate cells, and at the nanometre scale to favour the expression of extra-cellular matrix components. The present study has been undertaken to setup strategies to integrate stem cells and tailored scaffolds, as a tool to control cardiac tissue regeneration. Among the many available techniques for scaffold fabrication, porogen leaching, phase separation, and electrospinning were selected as low-cost and user-friendly technologies to fabricate tuneable, hierarchically porous matrices that mimic aspects of the cell native surroundings. The biological validation of these scaffolds was performed by implanting adult stem cells.

Activatory Properties of Lysophosphatidic Acid on Human THP-1 Cells

Excessive leukocyte proliferation and proinflammatory mediators release represent common phenomena in several chronic inflammatory diseases. Multiple evidences identify lysophosphatidic acid (LPA), a small lipid endowed with pleiotropic activities, as an important modulator of both proliferation and activation of different cell types involved in several inflammation-associated pathologies. However, its possible role on monocyte proinflammatory activation is not fully understood yet. Aim of the present study was to investigate LPA effects on THP-1 cells in terms of proliferation, reactive oxygen intermediates (ROI) production and release of arachidonic acid-derived inflammatory mediators. Actually, LPA significantly increased both DNA synthesis and ROI production as well as prostaglandin E2 release and the upregulation of LPA3 receptor expression. These findings identified LPA as both a growth factor and a triggering mediator of proinflammatory response in THP-1 cells.

Oxidant-induced pHi/ca2+ changes in rat aortic smooth muscle cells. The role of atrial natriuretic peptide

The aim of this study was to investigate the effects of oxidative stress on PLD activity, [Ca2+]i and pHi levels and the possible relationship among them. Moreover, since atrial natriuretic peptide (ANP) protects against oxidant-induced injury, we investigated the potential protective role of the hormone in Rat Aortic Smooth Muscle (RASM) cells exposed to oxidative stress. Water-soluble 2,2′-Azobis (2hyphen;amidinopropane) dihydrochloride (AAPH) was used as free radical generating system, since it generates peroxyl radicals with defined reaction and the half time of peroxyl radicals is longer than other ROS. A significant increase of PLD activity was related to a significant decrease in pHi, while [Ca2+]i levels showed an increase followed by a decrease after cell exposure to AAPH. [Ca2+]i changes and pHi fall induced by AAPH were prevented by cadmium which inhibits a plasma membrane Ca2+ ATPase coupled to Ca2+/H+ exchanger, that operates the efflux of Ca2+ coupled to H+ influx. The involvement of PLD in pHi and [Ca2+]i changes was confirmed by calphostin-c treatment, a potent inhibitor of PLD, which abolished all AAPH-induced effects. Pretreatment of RASM cells with pharmacological concentrations of ANP attenuated the AAPH effects on PLD activity as well as [Ca2+]i and pHi changes, while no effects were observed with physiological ANP concentrations, suggesting a possible role of the hormone as defensive effector against early events of the oxidative stress.

Atrial natriuretic factor inhibits mitogen-induced growth in aortic smooth muscle cells

Atrial natriuretic factor (ANF) is a polypeptide able to affect cardiovascular homeostasis exhibiting diuretic, natriuretic, and vasorelaxant activities. ANF shows antimitogenic effects in different cell types acting through R2 receptor. Excessive proliferation of smooth muscle cells is a common phenomenon in diseases such as atherosclerosis, but the role of growth factors in the mechanism which modulate this process has yet to be clarified. The potential antimitogenic role of ANF on the cell growth induced by growth factors appears very intriguing. Aim of the present study was to investigate the possible involvement of ANF on rat aortic smooth muscle (RASM) cells proliferation induced by known mitogens and the mechanism involved. Our data show that ANF, at physiological concentration range, inhibits RASM cell proliferation induced by known mitogens such as PDGF and insulin, and the effect seems to be elicited through the modulation of phosphatidic acid (PA) production and MAP kinases involvement. J. Cell. Physiol. 193: 103–109, 2002.

Selective changes in DNA binding activity of transcription factors in UM-X7. 1 cardiomyopathic hamsters

UM-X7.1 hamsters (CH) are considered a representative model for human cardiomyopathy. CH display the loss of the cytoskeletal delta-sarcoglycan protein, associated with myocardium remodeling and fatal reduction of heart functional efficiency. Even though altered redox balance and calcium homeostasis have already been reported to affect cardiomyocyte function, the molecular mechanisms underlying this pathology are largely unknown. We found no significant differences in DNA binding activity of redox-related (NF-kappaB, Sp1, AP-1 and AP-2) transcription factors in heart ventricles of 90 day-old CH, compared to normal animals. On the other hand, DNA binding activity of calcium-dependent transcription factors NF-AT3 and CREB were increased and decreased respectively in CH vs. normal ventricles. Western blot experiments confirmed the down regulation of CREB levels and suggest a novel regulation mechanism for this transcription factor in the heart. Our results are consistent with recent studies on NF-AT3, GATA4 and CREB transgenic mice, and provide clues for the comprehension of pathogenetic mechanisms of hamster hereditary cardiomyopathy.

Atrial natriuretic peptide induces cell death in human hepatoblastoma (HepG2) through the involvement of NADPH oxidase

Atrial natriuretic peptide induces cell death in human hepatoblastoma (HepG2) through the involvement of NADPH oxidase

Influence of ceria nanoparticles on chemical structure and properties of segmented polyesters.

In this work, we present new nanocomposite materials derived from segmented copolyesters, comprising ethylene terephthalate (PET) segments and dimerized linoleic acid (DLA), and nanometric cerium oxide particles (CeO2). Nanoparticles were incorporated in situ during polycondensation in various concentrations, from 0.1 up to 0.6 wt.%. It was found that preparation of nanocomposites in situ, during polycondensation, had no significant influence on changes in segmental composition as determined from (1)H and (13)C, as well as 2D NMR. Thermal analysis and calculated degree of crystallinity showed that increasing concentration of ceria nanoparticles lead to an increase in mass content of PET crystallites in hard segments. The XRD investigations also showed an increased intensity of characteristic signals with increasing ceria concentration. Simultaneously, the incorporation of CeO2 led to an increase in tensile strength and elongation at break, indicating a reinforcing and plasticizing effect of ceria nanoparticles. However, the modulus at 10% strain decreased with increasing amount of nanoparticles. The in vitro culture of human cardiac progenitor cells (hCPCs) on the new materials indicated a homogenous cell displacement across the samples after 5 days with no signs of cytotoxicity, indicating good biocompatibility in vitro of CeO2-based nanocomposites and a potential for biomedical applications.

Application of Evolutionary Game Theory on stem cells interaction in bio-active scaffolds

Cardiac diseases represent one of the biggest death causes in modern society. Current therapies, despite their importance, do not give an efficient solution to the problem. However, stem cells research could represent a future hope for a more efficient solution. In particular, there is currently a great interest in myocardial tissue generation from stem cells cultures in 3D bio-active scaffolds. Those polymeric structures intend to mimic the environment inside the heart where cardiac cells, cardiomyocytes, are generated. Well designed scaffolds along with chemical substances contribute to the differentiation process of the stem cell. How this happens, and how a stem cells population becomes a myocardial tissue? There are many models that describe the differentiation process, but they consider the stem cell acting alone, subjected only to the environment. However, a cell lives surrounded by other cells, some stem and some differentiating, and they will necessarily interact exchanging signals. We studied a solution using Game Theory (GT) and in particular Evolutionary Game Theory (EGT). This theory has been widely applied to biology to describe animals behaviour. We set a very simple model to describe the basic concepts of EGT and present some early results.