Search

There is growing interest in the use of micro-sized hydrogels, including bioactive signals, as efficient platforms for tissue regeneration because they are able to mimic cell niche structure and selected functionalities. Herein, it is proposed to optimize bioactive composite microgels via electrohydrodynamic atomization (EHDA) to regenerate the dentin-pulp complex. The addition of disodium phosphate (Na 2 HPO 4 ) salts as mineral precursors triggered an in situ reaction with divalent ions in solution, thus promoting the encapsulation of different amounts of apatite-like phases. Morphological analysis via image analysis of optical images confirmed a narrow distribution of perfectly rounded particles, with an average diameter ranging from 223 ± 18 μm to 502 ± 64 μm as a function of mineral content and process parameters used. FTIR, TEM, and EDAX analyses confirmed the formation of calcium phosphates with a characteristic Ca/P ratio close to 1.67 and a needle-like crystal shape. In vitro studies-using dental pulp stem cells (DPSCs) in crown sections of natural teeth slices-showed an increase in cell viability until 14 days, recording a decay of proliferation at 21 days, independent on the mineral amount, suggesting that differentiation is started, as confirmed by the increase of ALP activity at 14 days. In this view, mineralized microgels could be successfully used to support in vitro osteogenesis, working as an interesting model to study dental tissue regeneration.

Huntington's disease (HD) is a monogenetic neurodegenerative disorder that is caused by the expansion of a polyglutamine region within the huntingtin (HTT) protein, but there is still an incomplete understanding of the molecular mechanisms that drive pathology. Expression of the mutant form of HTT is a key aspect of diseased tissues, and the most promising therapeutic approaches aim to lower expanded HTT levels. Consequently, the investigation of HTT expression in time and in multiple tissues, with assays that accurately quantify expanded and non-expanded HTT, are required to delineate HTT homeostasis and to best design and interpret pharmacodynamic readouts for HTT lowering therapeutics. Here we evaluate mutant polyglutamine-expanded (mHTT) and polyglutamine-independent HTT specific immunoassays for validation in human HD and control fibroblasts and use to elucidate the CSF/brain and peripheral tissue expression of HTT in preclinical HD models.

In the last two decades, alginate scaffolds have been variously studied as extracellular matrix analogs for tissue engineering. However, relevant evidence is still lacking concerning their ability to mimic the microenvironment of hierarchical tissues such as bone. Hence, an increasing amount of attention has recently been devoted to the fabrication of macro/microporous sponges with pore anisotropy able to more accurately replicate the cell niche structure as a trigger for bioactive functionalities. This paper presents an in vivo study of alginate sponges with anisotropic microporous domains (MAS) formed by ionic crosslinking in the presence of different fractions (30 or 50% v) of hydroxyapatite (HA). In comparison with unloaded sponges (MAS0), we demonstrated that HA confers peculiar physical and biological properties to the sponge, depending upon the inorganic fraction used, enabling the sponge to bio-mimetically support the regeneration of newly formed bone. Scanning electron microscopy analysis showed a preferential orientation of pores, ascribable to the physical constraints exerted by HA particles during the pore network formation. Energy dispersive spectroscopy (EDS) and X-Ray diffraction (XRD) confirmed a chemical affinity of HA with the native mineral phase of the bone. In vitro studies via WST-1 assay showed good adhesion and proliferation of human Dental Pulp-Mesenchymal Stem Cells (hDP-MSC) that increased in the presence of the bioactive HA signals. Moreover, in vivo studies via micro-CT and histological analyses of a bone model (e.g., a rat calvaria defect) confirmed that the maximum osteogenic response after 90 days was achieved with MAS30, which supported good regeneration of the calvaria defect without any evidence of inflammatory reaction. Hence, all of the results suggested that MAS is a promising scaffold for supporting the regeneration of hard tissues in different body compartments., (Copyright © 2020 Serrano-Bello, Cruz-Maya, Suaste-Olmos, González-Alva, Altobelli, Ambrosio, Medina, Guarino and Alvarez-Perez.)

In recent years, different processing technologies have been engineered to fabricate capsules or particles with peculiar properties (e.g., swelling, pH-sensitive response) at the micro and sub-micrometric size scale, to be used as carriers for controlled drug and molecular release. Herein, the development of cellulose acetate (CA) micro-carriers with mono- (MC) or bi-phasic (BC) composition is proposed, fabricated via electrohydrodynamic atomization (EHDA)-an electro-dropping technology able to micro-size polymer solution by the application of high voltage electrostatic forces. Image analysis allows identification of the process parameters to optimize morphology, in terms of size distribution and shape. Meanwhile, an accurate rheological study has enabled investigating the interface between CA solutions with different viscosities to optimize BC systems. Release tests have confirmed that BC carriers can retain the drug more efficiently in acidic conditions, also providing a more gradual and sustained release until six days, with respect to MC carriers. Hence, all these results have proven that biphasic architecture significantly improves the capability of CA microcarriers to release ketoprofen lysinate, thus suggesting a new route to design core/shell systems for the retarded oral administration of anti-inflammatory drugs.

Herein, we propose the fabrication of a new carrier with core/shell structure-inner core of cellulose acetate (CA) coated by a micrometric layer of chitosan (CS)-fabricated through an integrated process, which combines Electro Dynamic Atomization (EDA) and layer-by-layer (LbL) technique. We demonstrate that CA based microspheres possess a unique capability to relevantly retain the drugs-that is, Ketoprofen Lysinate (KL)-along the gastric tract, while providing a massive release along the intestine. CS shell slightly influences the morphology and water retention under different pH conditions, improving drug encapsulation without compromising drug release kinetics. In vitro studies in simulated gastric and intestine fluids (SGF, SIF) with physiological enzymes, show a moderate release of LSK during the first 2 h (ca. 20% at pH 2), followed by a sustained release during the next 6 h (ca. 80% at pH 7). The obtained results demonstrate that CA-based microspheres hold strong potential to be used as carriers for a delayed oral administration of anti-inflammatory drugs. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 2636-2644, 2018., (© 2018 Wiley Periodicals, Inc.)

Autophagy is a cellular mechanism that can generate energy for cells or clear misfolded or aggregated proteins, and upregulating this process has been proposed as a therapeutic approach for neurodegenerative diseases. Here we describe a novel set of LC3B-II and p62 time-resolved fluorescence resonance energy transfer (TR-FRET) assays that can detect changes in autophagy in the absence of exogenous labels. Lipidated LC3 is a marker of autophagosomes, while p62 is a substrate of autophagy. These assays can be employed in high-throughput screens to identify novel autophagy upregulators, and can measure autophagy changes in cultured cells or tissues after genetic or pharmacological interventions. We also demonstrate that different cells exhibit varying autophagic responses to pharmacological interventions. Overall, it is clear that a battery of readouts is required to make conclusions about changes in autophagy.

The main limitation of conventional antibiotic therapies concerns the low efficacy to fight bacteria attacks during long treatment times. In this context, the integrated use of electrofluidodynamics (EFDs)-basically electrospinning and electrospraying-may represent an interesting route for designing nanostructured platforms with controlled release to prevent the formation of bacterial biofilms in oral implant sites. They allow for the deposition of nanofibres and nanoparticles by different modes-i.e. sequential, simultaneous-for the fabrication of more efficacious systems in terms of degradation protection, pharmacokinetic control and drug distribution to the surrounding tissues. Herein, we will investigate EFDs processing modes and conditions to decorate polycaprolactone nanofibres surfaces by chitosan nano-reservoirs for the administration of Amoxicillin Trihydrate as an innovative antibacterial treatment of the periodontal pocket.

The removal of dye and toxic ionic pollutants from water is an extremely important issue that requires systematic and efficient adsorbent preparation strategies. To address this challenge, we developed composite chitosan (CS)-based hydrogels containing hyper-crosslinked polymer (HCP) particles to be used as broad-spectrum adsorbents. The goal is to efficiently combine the dye adsorption ability of chitosan and the capacity of the porous particles of trapping pollutant molecules. The HCP particles are well distributed and firmly embedded into the chitosan matrix and the composite hydrogels exhibit improved mechanical properties. Adsorption experiments reveal a synergistic effect between CS and HCP particles, and the samples are able to remove both anionic and cationic dyes (indigo carmine, rhodamine 6G and sunset yellow) from water. The maximum dye uptake is higher than that of comparable biosorbents. Moreover, the mechanical properties of the composite hydrogels are enhanced respect to pure CS, and the samples can be regenerated and reused keeping their adsorption ability unaltered over successive cycles of adsorption, desorption, and washing., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The sequence of events which leads to the interfacial crowding of plate-like nanoparticles in co-continuous polymer blends is investigated through a combination of morphological and rheological analyses. Very low amounts (∼0.2 vol%) of organo-modified clay are sufficient to suppress phase coarsening in a co-continuous polystyrene/poly(methyl methacrylate) blend, while lower particle loading allows for a tuning of the characteristic size of the polymer phases at the μm-scale. In any case, an interfacial network of nanoparticles eventually forms, which is driven by the preferred polymer-polymer interface. The elastic features and stress-bearing ability of this peculiar nanoparticle assembly are studied in detail by means of a descriptive two-phase viscoelastic model, which allows isolation of the contribution of the filler network. The role of the co-continuous matrix in driving the space arrangement of the nanoparticles is emphasized by means of comparative analysis with systems based on the same polymers and nanoparticles, but in which the matrix is either a pure polymer or a blend with drop-in-matrix morphology. The relaxation dynamics of the interfacial network was found not to depend on the matrix microstructure, which instead substantially affects the assembly of the nanoplatelets. When the host medium is co-continuous, the particles align along the preferred polymer-polymer interface, percolating at a very low amount (∼0.17 vol%) and prevalently interacting edge-to-edge. The stress bearing ability of such a network is much higher than that in the case of matrix based on a homogeneous polymer or a drop-in-matrix blend, but its elasticity shows low sensitivity to the filler content.

Electrofluidodynamics techniques (EFDTs) are emerging methodologies based on liquid atomization induced by electrical forces to obtain a fine suspension of particles from hundreds of micrometers down to nanometer size. As a function of the characteristic size, these particles are interesting for a wide variety of applications, due to the high scalability of chemical and physical properties in comparison to the bulk form. Here, we propose the optimization of EFDT techniques to design chitosan systems in the form of microgels or nanoparticles for several biomedical applications. Different microscopy techniques (Optical, SEM, TEM) have been used to investigate the morphology of chitosan systems at multiple size scale. The proposed study confirms the high versatility and feasibility of EFDTs for creating micro and nano-sized carriers for cells and drug species.

A large variety of processes and tools has been investigated to acquire better knowledge on the natural evolution of healthy or pathological tissues in 3D scaffolds to discover new solutions for tissue engineering and cancer therapy. Among them, electrodynamic techniques allow revisiting old scaffold manufacturing approach by utilizing electrostatic forces as the driving force to assemble fibers and/or particles from an electrically charged solution. By carefully selecting materials and processing conditions, they allow to fine control of characteristic shapes and sizes from micro to sub-micrometric scale and incorporate biopolymers/molecules (e.g., proteins, growth factors) for time- and space-controlled release for use in drug delivery and passive/active targeting. This review focuses on current advances to design micro or nanostructured polymer platforms by electrodynamic techniques, to be used as innovative scaffolds for tissue engineering or as 3D models for preclinical in vitro studies of in vivo tumor growth.

Background: CD30 expression can be considered a marker of Th2 cells. We investigated the presence of CD30+ cells in the lesional skin of children with atopic dermatitis (AD). We also analyzed the possible relationship between CD30+ cells and serum soluble CD 30 (sCD30) levels, and IgE, soluble interleukin-2 (IL-2) receptor (sIL-2R) or soluble CD23 (sCD23) levels in the blood, and clinical score., Methods: Ten eczematous children (4 males, 6 females; median age: 4 years and 5 months; range: 11 months to 14 years), 9 sex- and age-matched control children and an adult control group were studied. A clinical score (SCORAD index), was given to eczematous lesions. Blood was taken for the determination of IgE, sCD30, sIL-2R and sCD23 levels. Punch biopsies of lesional skin were stained with hematoxylin and eosin or incubated with anti-CD30 monoclonal antibodies. Skin prick tests (SPTs) were also performed., Results: In the biopsy specimens, CD30 expression was observed in high proportions of infiltrating cells. In children with AD, total serum IgE, sCD30, sIL-2R, sCD23 and eosinophils were significantly elevated compared to controls. CD30+ cells were not associated with serum IgE, sCD30, sIL-2R, sCD23, or SPT results, score of inflammatory cells in lesional skin or clinical score. Children with AD who had high total IgE and specific IgE antibodies did not differ from those with normal total IgE and negative specific IgE in respect of age, clinical score, number of CD30+ cells, sCD30, sIL-2R and sCD23 levels, score of inflammatory cells in skin or clinical score., Conclusion: Our results showed remarkable numbers of CD30+ cells in the lesional skin and high sCD30 in the serum of children with AD. CD30+ cells did not correlate with systemic markers of IgE reaction., (Copyright 2000 S. Karger AG, Basel)

On a sample of 35 healthy newborns the authors have determined the IgE in the blood of the umbilical cord, using the PRIST method. A geometric average of 1.152 +/- 0.117 was obtained. No statistically significant difference appeared between the two groups, which contained individuals with positive and negative allergic family anamnesis, respectively. Thus, the possibility of predicting neonatal atopy by IgE determination appears to be no better than can be deduced through an accurate family anamnesis. The observed differences in the IgE levels of the funicular blood might then be explained by physiological, paraphysiological and pathological motivations.

Two groups of children from 6 to 14 years old have been compared to explain correlation between haematic eosinophils and allergic respiratory diseases related with the environment where they live. First group was formed by 350 pupils of Sesto S. Giovanni, and industrialized town, the second group by 345 pupils of Cogliate, a country place. The screening was been preceded by a set of questions self-managed and it consisted in the determinations percentage of peripheral blood eosinophils. Patients whose eosinophils were more than 4% have been considered eosinophilia carrier. The results have been elaborated with statistical method "chi square". Results proved the greatest prevalence of allergic respiratory diseases in urban population with a greater eosinophilia spreading (33% in comparison with 26%; p less than 0.05). Eosinophilia appeared connected with recurrent rhinitis (p less than 0.001) relapsing bronchitis (p less than 0.001) relapsing bronchitis (p less than 0.001) asthmatic bronchitis (p less than 0.001) positive results of skin tests to 4 allergens (p less than 0.001) and in rural district also with male sex (p less than 0.05). In industrialized areas, in special way there is a closed relation between respiratory diseases and peripheral blood eosinophils increased. Eosinophilia will be find in relapsing respiratory pathology and it will apply to allergology controls. It could represent an unfavourable element of prognosis foresee the possibility that episodic bronchus-spasms will be frequent in patients with asthmatic syndrome.

This paper present the case of a 5 year old girl affected by ichthyosis vulgaris, atopic diathesis (asthma - eczema), hair anomalies as pili torti, trichorrhexis invaginata. Such data are compared with the most frequent symptoms Netherton's syndrome. Although some difference be extant, the authors deem it appropriate that this case be included within the framework of this syndrome. The problem is therefore posed once again of the boundaries of Netherton's syndrome. Also considered are the most interesting hypotheses recently set forth by different authors with the aim of better defining this disease whose biological mechanism is still unknown.