Silvana Fiorito

Nanosurfaces and nanostructures for artificial orthopedic implants

Nanomaterials and structures, such as nanoparticles, nanofibers, nanosurfaces, nanocoatings, nanoscaffolds and nanocomposites, are considered for various applications in orthopedics and traumatology. This review looks at proposed nanotechnology inspired applications for implants from the perspective of the orthopedic industry. Investigations support consistently the theory that most nanomaterials in various physical forms are able to enhance the cell response selectively for biological tissue integration or increase the strength and wear resistance of current orthopedic materials. At this stage, most of the studies are at the laboratory scale or in early in vivo testing. Significant basic and applied research and development is needed to realize their full clinical potential and biological, manufacturing, economic and regulatory issues have to be addressed. Nevertheless, a crucial factor for success is well-coordinated multimethod and multidiscipline teamwork with profound industrial and medical expertise.

Proinflammatory effects of diesel exhaust nanoparticles on scleroderma skin cells.

Autoimmune diseases are complex disorders of unknown etiology thought to result from interactions between genetic and environmental factors. We aimed to verify whether environmental pollution from diesel engine exhaust nanoparticulate (DEP) of actually operating vehicles could play a role in the development of a rare immune-mediated disease, systemic sclerosis (SSc), in which the pathogenetic role of environment has been highlighted. The effects of carbon-based nanoparticulate collected at the exhaust of newer (Euro 5) and older (Euro 4) diesel engines on SSc skin keratinocytes and fibroblasts were evaluated in vitro by assessing the mRNA expression of inflammatory cytokines (IL-1α, IL-6, IL-8, and TNF-α) and fibroblast chemical mediators (metalloproteases 2, 3, 7, 9, and 12; collagen types I and III; VEGF). DEP was shown to stimulate cytokine gene expression at a higher extent in SSc keratinocytes versus normal cells. Moreover, the mRNA gene expression of all MMPs, collagen types, and VEGF genes was significantly higher in untreated SSc fibroblasts versus controls. Euro 5 particle exposure increased the mRNA expression of MMP-2, -7, and -9 in SSc fibroblasts in a dose dependent manner and only at the highest concentration in normal cells. We suggest that environmental DEP could trigger the development of SSc acting on genetically hyperreactive cell systems.

Supplementing π-systems: eumelanin and graphene-like integration towards highly conductive materials for the mammalian cell culture bio-interface

Organic (bio)electronics appears to be the first target for competitive exploitation in the materials science of eumelanins, black insoluble photoprotective human biopolymers. Nonetheless, the low conductivity of these pigments is limiting the implementation of eumelanin-based devices. Here we present a novel organic/organic hybrid material (EUGL) by integration of conductive graphene-like (GL) layers within the EUmelanin pigment (EU). GL layers were obtained by a two-step oxidation/reduction of carbon black. The stability of GL layers over a wide pH range and the self-assembling tendency place this material in a leading position for the fabrication of hybrid materials in aqueous media. EUGL was obtained by inducing the polymerization of eumelanin precursors (5,6-dihydroxyindole, DHI and 5,6-dihydroxyindole-2 carboxylic acid, DHICA) in aqueous media containing GL layers. The new material featured promising biocompatibility and an increased conductivity with respect to eumelanin by four orders of magnitude.

Inhibition of microbial growth by carbon nanotube networks.

In the last years carbon nanotubes have attracted increasing attention for their potential applications in the biomedical field as diagnostic and therapeutic nano tools. Here we investigate the antimicrobial activity of different fully characterized carbon nanotube types (single walled, double walled and multi walled) on representative pathogen species: Gram-positive Staphylococcus aureus, Gram-negative Pseudomonas aeruginosa and the opportunistic fungus Candida albicans. Our results show that all the carbon nanotube types possess a highly significant antimicrobial capacity, even though they have a colony forming unit capacity and induction of oxidative stress in all the microbial species to a different extent. Moreover, scanning electron microscopy analysis revealed that the microbial cells were wrapped or entrapped by carbon nanotube networks. Our data taken together suggest that the reduced capacity of microbial cells to forming colonies and their oxidative response could be related to the cellular stress induced by the interactions of pathogens with the CNT network.

Antimitochondrial autoantibodies in myocardial hypertrophy: Comparison between hypertrophic cardiomyopathy, hypertensive heart disease, and athlete's heart☆

Antimitochondrial autoantibodies (AMA) were tested by indirect immunofluorescence in three groups of subjects with different types of myocardial hypertrophy: 35 patients affected with hypertrophic cardiomyopathy (HC), 20 patients with cardiac hypertrophy secondary to essential hypertension, and 35 active endurance athletes with exercise-induced left ventricular hypertrophy. Forty-two healthy subjects served as a control group. Left ventricular hypertrophy was considered a left ventricular mass (LVM) echocardiographically calculated (Devereux formula), exceeding 244 gm or a LVM index exceeding 122 gm/m2 (greater than 2 SD from a previously studied normal population). AMA were found in 15 of 35 (43%) patients with HC and in 6 of 20 (30%) patients with hypertensive heart disease (p less than 0.01); in contrast, AMA were not present in the sera of athletes or in the sera of controls. Although the significance of AMA in subjects with pathologic myocardial hypertrophy has not yet been established, their absence in the sera of athletes strengthens the opinion that cellular changes, as a compensatory response of the myocardium to a work overload, have a physiologic fashion in these cases. Moreover, identification of AMA in the sera of athletes with disproportionate severe left ventricular hypertrophy of uncertain origin may be helpful to ensure a single diagnosis.

Antimicrobial properties of graphene-like nanoparticles: coating effect on Staphylococcus aureus

The exploitation of nanomaterials with antimicrobial properties has attracted an ever-growing interest in the recent years. Carbon-based materials, such as graphene and graphene family materials (GFMs), have gained most of the attention for application in many biomedical fields. Here, we describe the antimicrobial activity of graphene-like (GL) layers derived from the chemical demolition of carbon black, against the planktonic growth of Staphylococcus aureus cells, primary cause of hospital and community-acquired infections, often leading to bacteremia and sepsis. The inhibitory capabilities of GL layers on the formation of S. aureus biofilm are also assessed. The antimicrobial properties seem based mainly on the interaction between GL layers and bacteria surfaces. FESEM and AFM analyses suggest that the GL layers coat the cells as soon as they get in contact with them, as also indicated by the wettability of the GLs.

Highly electroconductive multiwalled carbon nanotubes as potentially useful tools for modulating calcium balancing in biological environments

Aiming to explore the mechanisms modulating cell-carbon nanotube interactions, we investigated whether Ca(2+) ion balancing between intra- and extracellular environments could be affected by multiwalled carbon nanotubes (MWCNTs). We analyzed the effects induced by two different kinds of MWCNTs (as prepared and annealed at 2400°C) on the intracellular Ca(2+) ion levels in rat electrically sensitive cells and on the intercellular junction integrity of rat adenocarcinoma colon cells and platelet aggregation ability, which depend on the Ca(2+) concentration in the medium. MWCNTs, purified by annealing and more electroconductive as compared to nonannealed MWCNTs, affected Ca(2+) ion balancing between extra- and intracellular environments and induced changes on Ca(2+) ion-dependent cellular junctions and platelet aggregation, behaving as the calcium chelator ethylene glycol tetraacetic acid. This could be due to the sorption of cationic Ca(2+) ions on CNTs surface because of the excess of negatively charged electrons on the aromatic units formed on MWCNTs after annealing. From the ClinicAL Editor: The authors investigated whether Ca(2+) ion balance between intra- and extracellular space can be modulated by multiwalled carbon nanotubes (MWCNTs). Annealed nanotubes induced changes on Ca(2+) dependent cellular junctions and platelet aggregation, behaving similary to ethylene glycol tetraacetic acid, an established calcium chelator.

Evaluation of the efficacy of carbon nanotubes for delivering peptides into mitochondria

Mitochondrial (mt) diseases are devastating neurodegenerative pathologies due to mutations in nuclear or mt genes. Among mtDNA pathogenic mutations, more than one half have been identified in transfer RNA (tRNA) genes. These are responsible for a wide range of pathologies including myopathies, encephalopathies, cardiomyopathies and deafness for which no effective treatment is available at present. Therefore, new strategies to suppress their damaging effects are required to envisage therapeutic approaches for these diseases. Here we report data for carbon nanotube (CNT) derivatives showing that the conjugates bearing a specific peptide sequence are able to target the mitochondria in yeast and human monocyte cells while the control derivative without the peptide diffuses into the cytoplasm. Moreover the compounds do not affect cellular viability and cytotoxicity both in vitro and in vivo. Toxicity of the constructs is also assessed on the simple pluricellular model Caenorhabditis elegans.

Diesel exhaust particles induce autophagy and citrullination in Normal Human Bronchial Epithelial cells

A variety of environmental agents has been found to influence the development of autoimmune diseases; in particular, the studies investigating the potential association of systemic autoimmune rheumatic diseases with environmental micro and nano-particulate matter are very few and contradictory. In this study, the role of diesel exhaust particles (DEPs), one of the most important components of environment particulate matter, emitted from Euro 4 and Euro 5 engines in altering the Normal Human Bronchial Epithelial (NHBE) cell biological activity was evaluated. NHBE cells were exposed in vitro to Euro 4 and Euro 5 particle carbon core, sampled upstream of the typical emission after-treatment systems (diesel oxidation catalyst and diesel particulate filter), whose surfaces have been washed from well-assessed harmful species, as polycyclic aromatic hydrocarbons (PAHs) to: (1) investigate their specific capacity to affect cell viability (flow cytometry); (2) stimulate the production of the pro-inflammatory cytokine IL-18 (Enzyme-Linked ImmunoSorbent Assay -ELISA-); (3) verify their specific ability to induce autophagy and elicit protein citrullination and peptidyl arginine deiminase (PAD) activity (confocal laser scanning microscopy, immunoprecipitation, Sodium Dodecyl Sulphate-PolyAcrylamide Gel Electrophoresis -SDS-PAGE- and Western blot, ELISA). In this study we demonstrated, for the first time, that both Euro 4 and Euro 5 carbon particles, deprived of PAHs possibly adsorbed on the soot surface, were able to: (1) significantly affect cell viability, inducing autophagy, apoptosis and necrosis; (2) stimulate the release of the pro-inflammatory cytokine IL-18; (3) elicit protein citrullination and PAD activity in NHBE cells. In particular, Euro 5 DEPs seem to have a more marked effect with respect to Euro 4 DEPs.