Graphene-based nanomaterials for tissue engineering in the dental field

The world of dentistry is approaching graphene-based nanomaterials as substitutes for tissue engineering. Apart from its exceptional mechanical strength, electrical conductivity and thermal stability, graphene and its derivatives can be functionalized with several bioactive molecules. They can also be incorporated into different scaffolds used in regenerative dentistry, generating nanocomposites with improved characteristics. This review presents the state of the art of graphene-based nanomaterial applications in the dental field. We first discuss the interactions between cells and graphene, summarizing the available in vitro and in vivo studies concerning graphene biocompatibility and cytotoxicity. We then highlight the role of graphene-based nanomaterials in stem cell control, in terms of adhesion, proliferation and differentiation. Particular attention will be given to stem cells of dental origin, such as those isolated from dental pulp, periodontal ligament or dental follicle. The review then discusses the interactions between graphene-based nanomaterials with cells of the immune system; we also focus on the antibacterial activity of graphene nanomaterials. In the last section, we offer our perspectives on the various opportunities facing the use of graphene and its derivatives in associations with titanium dental implants, membranes for bone regeneration, resins, cements and adhesives as well as for tooth-whitening procedure

Genetical stability and osteogenic ability of mesenchimal stem cells on demineralized bone matrices

Journal of Osseointegration Volume 7, Issue 1, 1 March 2015, Pages 2-7 Open Access Genetical stability and osteogenic ability of mesenchimal stem cells on demineralized bone matrices (Article) Pozzuoli, A.a, Gardin, C.b, Aldegheri, R.a, Bressan, E.c, Isola, M.d, Calvo-Guirado, J.L.e, Biz, C.a, Arrigoni, P.a, Feroni, L.b, Zavan, B.b a Department of Surgical,Oncological and Gastroenterological Sciences, University of Padua, Padua, Italy b Department of Biomedical Sciences, University of Padua, Padua, Italy c Department of Neurosciences, University of Padua, Padua, Italy d Department of Animal Medicine, Production and Health (MAPS), Italy e Department of General Dentistry, Faculty of Medicine and Dentistry, University of Murcia, Murcia, Spain Hide additional affiliations View references (44) Abstract Aim: Tissue engineering is a rapidly expanding field with regard to the use of biomaterials and stem cells in the orthopedic surgery. Many experimental studies have been done to understand the best characteristics of cells, materials and laboratory methods for safe clinical applications. The aim of this study was to compare the ability of 2 different human demineralized bone matrices (DBMs), the one enriched and the other not enriched with hyaluronic acid, to stimulate in vitro the proliferation and the osteogenic differentiation of human adipose-derived stem cells (ADSCs) seeded onto an osteoconductive scaffold. Materials and Methods: ADSCs were isolated, by enzymatic digestion, from abdominal adipose tissue of 5 patients undergoing cosmetic lipoaspiration surgery. ADSCs were then seeded onto a 3D scaffold in the presence of the two different osteoinductive matrices of human demineralized bone and evaluated for proliferation and osteogenic differentiation. The safety of the methods was verified using array-Comparative Genomic Hybridization (array-CGH). Results: ADSCs were able to differentiate in osteogenic sense. Both DBMs showed the ability to induce osteogenic differentiation of the cells. Conclusion: array-CGH showed no changes at genome level, thus confirming the safety of materials and method

Traditional landscape and rural development: comparative study in three terraced areas in northern, central and southern Italy to evaluate the efficacy of GAEC standard 4.4 of cross compliance

The recent National Strategic Plan 2007-2013 has introduced landscape as a strategic objective of the rural sector. This represents a minor revolution in the way of visualizing the role of the landscape, together with that of agriculture and the rural territory as a whole, and demonstrates the importance of treating the landscape with a systematic point of view. As part of the Efficond project, three sample areas have been identified, each of about 800-1000 hectares, in zones with important historical - cultural landscapes that are included in the National Catalogue of Historical Rural Landscapes. For each sample area a methodology has been applied, defined Historical Cultural Evaluation Approach, developed as part of a project for the monitoring of the Tuscan landscape that we have simplified and adapted. This methodology is based on the consideration that the landscape is the result of the centuries-old interaction between man and the environment, and so to define an element of the landscape as characteristic it is necessary to evaluate the land use dynamics and landscape changes that took place in the past, identifying those that have persisted for a long time, are slowly evolving or stabilized. The study of the historical landscape, which in the proposed methodology refers to the 50’s, has been done through the interpretation and analysis of aerial photographs taken on the GAI flight in 1954, and has allowed the characteristic, traditional and historical elements of that landscape to be identified and an insight to be gained into the cultural identity of the area. Through the creation of specific indices of density and intensity of the terracing obtained by photo-interpretation, field surveys and GIS elaborations, it was possible to classify the sample areas for this specific and important landscape element, compare the results in two periods and evaluate their frequency in the territory. Multi-temporal comparative analysis is being used increasingly often, especially for the study of territories of value, and in our case has been accompanied both by mapping of the landscape dynamics, which identifies the areas subject to transformations in the considered period, and by tables and figures that allow the evolution of a unit of land use to be followed, observing how this has evolved over time. The evaluation of these evolutionary dynamics has then been integrated with a set of indices, in part borrowed from landscape ecology, and in part specifically developed for areas historically shaped by man, which demonstrate that the landscape has become less fragmented and that the layout of fields has been adapted to a different agricultural model that has profoundly changed the structure of the traditional landscape. The efficacy of the laws protecting the characteristic elements of the landscape is strictly linked to the maintenance of its diversity and typicality and conservation of the complexity of the landscape mosaic. Its evaluation necessitates a historical analysis of the evolutionary dynamics conducted at a purely local level

Controlling Exchange Pathways in Dynamic Supramolecular Polymers by Controlling Defects

Supramolecular fibers composed of monomers that self-assemble directionally via noncovalent interactions are ubiquitous in nature, and of great interest in chemistry. In these structures, the constitutive monomers continuously exchange in-and-out the assembly according to a well-defined supramolecular equilibrium. However, unraveling the exchange pathways and their molecular determinants constitutes a nontrivial challenge. Here, we combine coarse-grained modeling, enhanced sampling, and machine learning to investigate the key factors controlling the monomer exchange pathways in synthetic supramolecular polymers having an intrinsic dynamic behavior. We demonstrate how the competition of directional vs. nondirectional interactions between the monomers controls the creation/annihilation of defects in the supramolecular polymers, from where monomers exchange proceeds. This competition determines the exchange pathway, dictating whether a fiber statistically swaps monomers from the tips or from all along its length. Finally, thanks to their generality, our models allow the investigation of molecular approaches to control the exchange pathways in these dynamic assemblies

Avaliação econômica dos sistemas de produção de morango: convencional, integrado e orgânico.

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Hyperbaric oxygen therapy improves the osteogenic and vasculogenic properties of mesenchymal stem cells in the presence of inflammation in vitro

Hyperbaric oxygen (HBO) therapy has been reported to be beneficial for treating many conditions of inflammation-associated bone loss. The aim of this work was to in vitro investigate the effect of HBO in the course of osteogenesis of human Mesenchymal Stem Cells (MSCs) grown in a simulated pro-inflammatory environment. Cells were cultured with osteogenic differentiation factors in the presence or not of the pro-inflammatory cytokine Tumor Necrosis Factor-α (TNF-α), and simultaneously exposed daily for 60 min, and up to 21 days, at 2,4 atmosphere absolute (ATA) and 100% O2. To elucidate osteogenic differentiation-dependent effects, cells were additionally pre-committed prior to treatments. Cell metabolic activity was evaluated by means of the MTT assay and DNA content quantification, whereas osteogenic and vasculogenic differentiation was assessed by quantification of extracellular calcium deposition and gene expression analysis. Metabolic activity and osteogenic properties of cells did not differ between HBO, high pressure (HB) alone, or high oxygen (HO) alone and control if cells were pre-differentiated to the osteogenic lineage. In contrast, when treatments started contextually to the osteogenic differentiation of the cells, a significant reduction in cell metabolic activity first, and in mineral deposition at later time points, were observed in the HBO-treated group. Interestingly, TNF-α supplementation determined a significant improvement in the osteogenic capacity of cells subjected to HBO, which was not observed in TNF-α-treated cells exposed to HB or HO alone. This study suggests that exposure of osteogenic-differentiating MSCs to HBO under in vitro simulated inflammatory conditions enhances differentiation towards the osteogenic phenotype, providing evidence of the potential application of HBO in all those processes requiring bone regeneration

774-5 Effect of Cardiac Translation on Measurement of Left Ventricular Wall Velocities: Implications for Doppler Imaging of Myocardium

Doppler imaging of the myocardium is a new application which has the potential to record myocardial velocities. These recorded velocities, however, include cardiac motion independent of ventricular contraction. A measured myocardial velocity, therefore, represents the net vector of contraction, translation, and rotation. To determine the effects of cardiac translation on myocardial velocities, 2-dimensional (2D) and M-mode echocardiographic recordings were obtained in 10 normal subjects. The average anteroseptal (AS) and posterior wall (PW) velocities were measured by 2D echo directed M-mode in the centerline of the parasternal short-axis view. Translation was measured from 2D echo cine-loop display as the displacement of the epicardial junction of the right ventricular free wall and interventricular septum during systole. The average translational velocity is reported as the component of the displacement vector parallel to the M-mode beam (+=toward transducer). The AS and PW velocities (cm/sec) displayed in the table represent net measured velocities, which include the translational vector.ResultsASPWTranslationMean±SD3.2±0.54.5±1.1+1.3±0.6Range2.4 to 4.03.4 to 6.9-l.4 to+2.4In 8/10 subjects the velocity vector was positive. The mean percent error in the M-mode derived velocities due to translation was 41% for the AS wall and 31% for the PW.Conclusions1) As measured by 2D echocardiography, the magnitude of the translational vector is significant when compared to the M-mode derived myocardial velocities. 2) The relative error demonstrated in the measured velocities may be further modified when applied in two dimensions, due to the angle of incidence of the Doppler beam. 3) New techniques for measuring myocardial velocities, such as Doppler imaging of the myocardium, should incorporate algorithms which correct for the translational vector

Doppler color flow “proximal isovelocity surface area” method for estimating volume flow rate: Effects of orifice shape and machine factors

AbstractPreviously described Doppler color How mapping methods for estimating the severity of valvular regurgitation have focused on the distal jet. In this study, a newer Doppler color flow technique, focusing on the flow proximal to an orifice, was used. This method identifies a proximal isovelocity surface area (PISA) by displaying an aliasing interface. Volume flow rate (cm3/s) can be calculated as PISA (cm2) × aliasing velocity (cm/s). For planar circular orifices, a hemi-elliptic model accurately approximated the shape of PISA.Clinically, however, orifice shapes may be noncircular. In vitro flow experiments (n = 226) using orifices of various shapes (ellipse, square, triangle, star, rectangle) were performed. Volume flow rate calculated using a hemi-elliptic model for PISA was accurate, with average percent differences from actual flow rate = +4.3% for a square, −4.2% for a triangle, −4.7% for a star, −4.5% for an ellipse and −2.8% for a rectangle. However, average percent differences for calculated volume flow rates using a hemispheric model for PISA shape ranged from −11.6% (square) to −34.8% (rectangle).In addition, to evaluate whether PISA is influenced by machine factors, in vitro studies (n = 83) were performed. For a volume flow rate of 13 liters/min, the color aliasing radius was not affected by: 1) system gain(radius = 9.9 mm at −20 dB versus 10.4 mm at +20 dB); 2) wall filter(10.8 mm at high versus 11.2 mm at tow); 3) frame rate(11.2 mm at 6/s versus 10.6 mm at 22/s); 4) transmit power(10.3 mm at high versus 10.1 mm at low); and 5) packet size(10.6 mm at 4 samples/line versus 10.5 mm at 8 samples/line). The aliasing radius was lower at higher aliasing velocities. However, the calculated volume flow rate was not affected by changes in aliasing velocity.It is concluded that differences in planar orifice shape do not affect volume flow rate calculated using a hemi-elliptic model. Furthermore, changes in machine factors do not alter volume flow rate calculated using the PISA method. The PISA method may have advantages over previous Doppler color flow methods in calculating volume flow rate and may be useful clinically in estimating valvular regurgitant or shunt volume

Citrate Mediates Crosstalk between Mitochondria and the Nucleus to Promote Human Mesenchymal Stem Cell In Vitro Osteogenesis

Citrate, generated in the mitochondria, is a key metabolite that might link metabolism with signaling, chromatin structure and transcription to orchestrate mesenchymal stem cells (MSCs) fate determination. Based on a detailed morphological analysis of 3D reconstruction of mitochondria and nuclei in single cells, we identified contact sites between these organelles that drastically increase in volume and number during the early stage of mesenchymal stem cell differentiation. These contact sites create a microdomain that facilitates exchange of signals from mitochondria to the nucleus. Interestingly, we found that the citrate derived from mitochondria is necessary for osteogenic lineage determination. Indeed, inhibition of the citrate transporter system dramatically affected osteogenesis, reduced citrate levels that could be converted in α-ketoglutarate, and consequently affected epigenetic marker H3K9me3 associated with the osteogenesis differentiation process. These findings highlight that mitochondrial metabolites play key regulatory roles in the MSCs differentiation process. Further in-depth investigation is needed to provide novel therapeutic strategies in the field of regenerative medicine

Mineral-doped poly(L-lactide) acid scaffolds enriched with exosomes improve osteogenic commitment of human adipose-derived mesenchymal stem cells

Exosomes derived from mesenchymal stem cells are extracellular vesicles released to facilitate cell communication and function. Recently, polylactic acid (PLA), calcium silicates (CaSi), and dicalcium phosphate dihydrate (DCPD) have been used to produce bioresorbable functional mineral-doped porous scaffolds-through thermally induced phase separation technique, as materials for bone regeneration. The aim of this study was to investigate the effect of mineral-doped PLA-based porous scaffolds enriched with exosome vesicles (EVs) on osteogenic commitment of human adipose mesenchymal stem cells (hAD-MSCs). Two different mineral-doped scaffolds were produced: PLA-10CaSi-10DCPD and PLA-5CaSi-5DCPD. Scaffolds surface micromorphology was investigated by ESEM-EDX before and after 28 days immersion in simulated body fluid (HBSS). Exosomes were deposited on the surface of the scaffolds and the effect of exosome-enriched scaffolds on osteogenic commitment of hAD-MSCs cultured in proximity of the scaffolds has been evaluated by real time PCR. In addition, the biocompatibility was evaluated by direct-contact seeding hAD-MSCs on scaffolds surface-using MTT viability test. In both formulations, ESEM showed pores similar in shape (circular and elliptic) and size (from 10\u201330 \ub5m diameter). The porosity of the scaffolds decreased after 28 days immersion in simulated body fluid. Mineral-doped scaffolds showed a dynamic surface and created a suitable bone-forming microenvironment. The presence of the mineral fillers increased the osteogenic commitment of hAD-MSCs. Exosomes were easily entrapped on the surface of the scaffolds and their presence improved gene expression of major markers of osteogenesis such as collagen type I, osteopontin, osteonectin, osteocalcin. The experimental scaffolds enriched with exosomes, in particular PLA-10CaSi-10DCPD, increased the osteogenic commitment of MSCs. In conclusion, the enrichment of bioresorbable functional scaffolds with exosomes is confirmed as a potential strategy to improve bone regeneration procedures