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Vascular tissue engineering endeavors to design, fabricate, and validate biodegradable and bioabsorbable small-diameter vascular scaffolds engineered with bioactive molecules, capable of meeting the challenges imposed by commercial vascular prostheses. A comprehensive investigation of these engineered scaffolds in bioreactor is deemed essential as a prerequisite before any in vivo experimentation in order to get information regarding their behavior under physiological conditions and predict the biological activities they will possess. This study focuses on an innovative electrospun scaffold made of poly(caprolactone) and poly(glycerol sebacate), integrating quercetin, able to modulate inflammation, and gelatin, necessary to reduce permeability. A custom-made bioreactor was used to assess the performances of the scaffolds maintained under different pressure regimes, covering the human physiological pressure range. As results, the 3D microfibrous architecture was notably influenced by the release of bioactives, maintaining the adequate properties needed for the in vivo regeneration and scaffolds showed mechanical properties similar to human native artery. Release of gelatin was adequate to avoid blood leakage and useful to make the material porous during the testing period, whereas the amount of released quercetin was useful to counteract the post-surgery inflammation. This study showcases the successful validation of an engineered scaffold in a bioreactor, enabling to consider it as a promising candidate for vascular substitutes in in vivo applications. Our approach represents a significant leap forward in the field of vascular tissue engineering, offering a multifaceted solution to the complex challenges associated with small-diameter vascular prostheses. .
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Introduction: Coastal seawater pollution poses a public health risk due to the potential ingestion of contaminated water during recreational activities. Wastewater-based epidemiology has revealed the abundant presence of SARS-CoV-2 in seawater emitted from wastewater outlets. The objective of this research was to investigate the impact of seawater on SARS-CoV-2 infectivity to assess the safety of recreational activities in seawater. Methods: Wild SARS-CoV-2 was collected from oral swabs of COVID-19 affected patients and incubated for up to 90 min using the following solutions: (a) standard physiological solution (control), (b) reconstructed seawater (3.5% NaCl), and (c) authentic seawater (3.8%). Samples were then exposed to two different host systems: (a) Vero E6 cells expressing the ACE2 SARS-CoV-2 receptor and (b) 3D multi-tissue organoids reconstructing the human intestine. The presence of intracellular virus inside the host systems was determined using plaque assay, quantitative real-time PCR (qPCR), and transmission electron microscopy. Results: Ultrastructural examination of Vero E6 cells revealed the presence of virus particles at the cell surface and in replicative compartments inside cells treated with seawater and/or reconstituted water only for samples incubated up to 2 min. After a 90-min incubation, the presence of the virus and its infectivity in Vero E6 cells was reduced by 90%. Ultrastructural analysis performed in 3D epi-intestinal tissue did not reveal intact viral particles or infection signs, despite the presence of viral nucleic acid detected by qPCR. Indeed, viral genes (Orf1ab and N) were found in the intestinal luminal epithelium but not in the enteric capillaries. These findings suggest that the intestinal tissue is not a preferential entry site for SARS-CoV-2 in the human body. Additionally, the presence of hypertonic saline solution did not increase the susceptibility of the intestinal epithelium to virus penetration; rather, it neutralized its infectivity. Conclusion: Our results indicate that engaging in recreational activities in a seawater environment does not pose a significant risk for COVID-19 infection, despite the possible presence of viral nucleic acid deriving from degraded and fragmented viruses.
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COVID-19 , Ácidos Nucleicos , Humanos , SARS-CoV-2 , Saúde Pública , Água do Mar , Água , PermeabilidadeRESUMO
The Sauter mean diameter, d32, is a representative parameter in emulsions that indicates the average size of the oil droplets once the emulsion becomes stable. Several mathematical and physical approaches have been employed in the literature to seek expressions for d32 under different conditions. The present work sheds light on this rich literature and emphasizes that the characterization of emulsions is still a fertile field for investigation. In this paper, a new Π-theorem-based model to predict the normalized Sauter mean diameter for the specific case of rotor-stator emulsification is sought by applying a multiple regression analysis on experimental data of oil-in-water (O-W) emulsions produced using three different oils: paraffin, soybean oil, and isopropyl myristate, at different oil-to-water (O/W) ratios and rotor speeds. The proposed model quantifies the roles of the viscous, inertial, and interfacial tension forces, besides the O/W ratio, in the emulsification process within the turbulent inertial subrange. The developed empirical correlation is then contrasted with relevant literature models for reliability assessment; predictions of the present explicit model are proven to be more accurate for the fluid properties and the experimental conditions under study.
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Considering the global increase in fish consumption, the growing side-streams coming from the fish supply chain (e.g., skin, fins, tail, heads ), also including undersized or "unwanted catches", have been recently proposed as source of high-value bioactive compounds (e.g., peptides and fatty acids). In this case study, hydrolyzed collagen peptides (HCPs) were extracted from different parts of Mugil cephalus L. using environmentally friendly techniques such as ultrasounds and enzymatic treatments. Both a mixed biomass derived from the skin, fins, and tail, and a whole fish, were considered as starting biomass, simulating the unsorted processing side-streams and an undersized/unwanted catch, respectively. The extracted HCPs were purified in fractions (<3 KDa and >3 KDa) whose yields (about 5% and 0.04-0.3%, respectively) demonstrated the efficiency of the hydrolysis process. The extraction protocol proposed allowed us to also isolate the intermediate products, namely the lipids (about 8-10%) and the non-collagenous proteins (NCs, 16-23%), whose exploitation could be considered. Each sample was characterized using Sircol, UltraViolet-Spectra, and hydroxyproline assay, and the viability of their collagen fractions was tested on human endothelial cells. Significant effects were obtained at a fraction of <3 KDa, in particular at a concentration of 0.13 µg/mL. The T-scratch test was also performed, with positive results in all fractions tested.
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Colágeno , Células Endoteliais , Animais , Humanos , Células Endoteliais/metabolismo , Colágeno/química , Pele/metabolismo , Antioxidantes/química , Peixes/metabolismo , Peptídeos/químicaRESUMO
This study presents for the first time a scalable process for the extraction of valuable proteins starting from samples of unsorted mixed tuna scraps which were previously dehydrated by an industrial patented process. The aims of this work were both to avoid the onerous sorting step of tuna leftovers, which generally consists of isolating skin and bones for collagen/gelatin extraction, and to improve the logistic of managing highly perishable biomass thanks to the reduction in its volume and to its microbiological stabilization. In view of a zero-waste economy, all the protein fractions (namely, non-collagenous proteins NCs and ALKs, gelatin, and hydrolyzed gelatin peptides, HGPs) isolated in the proposed single cascade flowchart were stabilized and preliminarily characterized. The extraction flowchart proposed allows one to obtain the following most promising compounds: 1.7 g of gelatin, 3.2 g of HGPs, and 14.6 g of NCs per 100 g of dehydrated starting material. A focus on oven-dried gelatin was reported in terms of proximate analysis, amino acid composition, color parameters, FT-IR spectrum, pH, and viscoelastic properties (5 mPa·s of viscosity and 14.3 °C of gelling temperature). All the obtained extracts are intended to be exploited in food supplements, feed, fertilizers/plant bio-stimulants, packaging, and the cosmetic industry.
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Polyphenolic compounds constitute a diverse group of natural components commonly occurring in various plant species, known for their potential to exert both beneficial and detrimental effects. Additionally, these polyphenols have also been implicated as endocrine-disrupting (ED) chemicals, raising concerns about their widespread use in the cosmetics industry. In this comprehensive review, we focus on the body of literature pertaining to the estrogenic properties of ED chemicals, with a particular emphasis on the interaction of isoflavones with estrogen receptors. Within this review, we aim to elucidate the multifaceted roles and effects of polyphenols on the skin, exploring their potential benefits as well as their capacity to act as ED agents. By delving into this intricate subject matter, we intend to provoke thoughtful consideration, effectively opening a Pandora's box of questions for the reader to ponder. Ultimately, we invite the reader to contemplate whether polyphenols should be regarded as friends or foes in the realm of skincare and endocrine disruption.
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Many different amphibian skin peptides have been characterized and proven to exert various biological actions, such as wound-healing, immunomodulatory, anti-oxidant, anti-inflammatory and anti-diabetic effects. In this work, the possible anti-steatotic effect of macrotympanain A1 (MA1) (FLPGLECVW), a skin peptide isolated from the Chinese odorous frog Odorrana macrotympana, was investigated. We used a well-established in vitro model of hepatic steatosis, consisting of lipid-loaded rat hepatoma FaO cells. In this model, a 24 h treatment with 10 µg/mL MA1 exerted a significant anti-steatotic action, being able to reduce intracellular triglyceride content. Accordingly, the number and diameter of cytosolic lipid droplets (LDs) were reduced by peptide treatment. The expression of key genes of hepatic lipid metabolism, such as PPARs and PLINs, was measured by real-time qPCR. MA1 counteracted the fatty acid-induced upregulation of PPARγ expression and increased PLIN3 expression, suggesting a role in promoting lipophagy. The present data demonstrate for the first time a direct anti-steatotic effect of a peptide from amphibian skin secretion and pave the way to further studies on the use of amphibian peptides for beneficial actions against metabolic diseases.
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Fígado Gorduroso , Ratos , Animais , Fígado Gorduroso/metabolismo , Ranidae/metabolismo , Pele/metabolismo , Peptídeos/farmacologia , Peptídeos/química , PPAR gama/metabolismoRESUMO
Atherosclerosis represents one of the main causes of death in the Western world. It is a multifactorial pathology characterized by lesions that reduce the lumen of the vessels causing serious clinical events. The extra-domain B of fibronectin is overexpressed during angiogenesis and in tissues undergoing growth and extensive remodeling, i.e., atherosclerotic plaque. Bevacizumab is a recombinant humanized monoclonal antibody that can play a central role against angiogenesis reducing the risk associated with this process in atherosclerosis. In this work, an innovative nanosystem for the targeted delivery of bevacizumab to the atherosclerotic lesion is proposed. A production protocol for poly(lactic-co-glycolic acid)-polyethylene glycol nanoparticles loaded with bevacizumab and functionalized with immunouteroglobin-1 was designed. Once functionalized nanoparticles with immunouteroglobin-1 were produced, they were characterized in terms of morphology, mean diameter, ζ-potential, association and conjugation efficiencies, bevacizumab release profile, both in phosphate buffered saline and in serum, bevacizumab stability after release, cytocompatibility, and hemocompatibility. Nanoparticle mean diameter was in the range of 217-265 nm, their surface charge was between -22 and -8 mV, and the association and conjugation efficiencies of about 76 and 59 %, respectively. Fourier transform infrared spectroscopy analysis confirmed the functionalization of their surface with immunouteroglobin-1. In vitro assays showed that the studied nanoparticles were cytocompatible, once in contact with human endothelial and murine macrophage cell lines up to 72 h, and hemocompatible, once in contact with red blood cells, at different concentrations of encapsulated bevacizumab (0.1, 1, 10, and 100 µg/mL).
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Aterosclerose , Nanopartículas , Humanos , Camundongos , Animais , Bevacizumab/farmacologia , Copolímero de Ácido Poliláctico e Ácido Poliglicólico/química , Nanopartículas/química , Polietilenoglicóis , Neovascularização Patológica , Tamanho da PartículaRESUMO
Aluminum is an element found in nature and in cosmetic products. It can interfere with the metabolism of other cations, thus inducing gastrointestinal disorder. In cosmetics, aluminum is used in antiperspirants, lipsticks, and toothpastes. The aim of this work is to investigate aluminum bioavailability after accidental oral ingestion derived from the use of a toothpaste containing a greater amount of aluminum hydroxide than advised by the Scientific Committee on Consumer Safety (SCCS). To simulate in vitro toothpaste accidental ingestion, the INFOGEST model was employed, and the amount of aluminum was measured through the ICP-AES analysis. Tissue barrier integrity was analyzed by measuring transepithelial electric resistance, and the tissue architecture was checked through light microscopy. The margin of safety was also calculated. Overall, our results indicate that the acute exposure to aluminum accidentally ingested from toothpastes is safe for the final user, even in amounts higher than SCCS indications.
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Alumínio , Cosméticos , Disponibilidade Biológica , Qualidade de Produtos para o Consumidor , Cosméticos/toxicidade , Cremes DentaisRESUMO
This work is a comparative study among three different biocompatible and biodegradable polymers, poly(lactic-co-glycolic acid), poly(ε-caprolactone), and poly(lactic acid), used to produce microparticles for the encapsulation of bevacizumab for drug delivery purposes. All the formulations were produced using the double emulsion water-oil-water evaporation method and characterized in terms of particle mean diameter, particle size distribution, and bevacizumab entrapment efficiency. Bevacizumab cumulative release was taken into consideration to study the dissolution kinetics from the three different polymeric delivery platforms for a period of 50 days at 37 °C in phosphate buffered saline and mathematical models of the drug release kinetic were attempted in order to describe the release phenomena from the different types of the studied microparticles. Finally, cell viability on human endothelial cell line EA.hy926 was studied to define the maximum cytocompatible concentration for each microsystem, registering the mitochondrial functionality through MTS assay.
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Cardiovascular diseases are the leading cause of mortality in the Western world. Among them, atherosclerosis represents one of the most common diseases in the modern society due to a common sedentary lifestyle, high-fat diet, and smoking. In the near future, a new approach could potentially improve the therapy of vascular pathologies, where to date the non-specific treatments present several limitations, such as poor biodistribution, quick elimination from the body, and undesired side-effects. In this field, nanotechnology has a great potential for the therapy and diagnosis of atherosclerosis with more and more recent and innovative publications. This review is a critical analysis of the results reported in the literature regarding the different and possible new approaches for the therapy and diagnosis of atherosclerosis.
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Anti-Inflamatórios/uso terapêutico , Aterosclerose/tratamento farmacológico , Sistemas de Liberação de Medicamentos , Nanopartículas/química , Nanotecnologia , Anti-Inflamatórios/química , Aterosclerose/patologia , HumanosRESUMO
Over the previous years, the design, development, and potential application of nanocarriers in the medical field have been intensively studied for their ability to preserve drug properties, especially their pharmacological activity, and to improve their bioavailability. This work is a comparative study between two different types of nanocarriers, poly (lactic-co-glycolic acid)-based nanoparticles and phosphatidylcholine-based nanoliposomes, both prepared for the encapsulation of bovine serum albumin as a model protein. Polymeric nanoparticles were produced using the double emulsion water-oil-water evaporation method, whereas nanoliposomes were obtained by the thin-film hydration method. Both nanocarriers were characterized by morphological analysis, particle mean size, particle size distribution, and protein entrapment efficiency. Invitro release studies were performed for 12 days at 37 °C. In order to explore a possible application of these nanocarriers for a targeted therapy in the cardiovascular field, hemolytic activity and biocompatibility, in terms of cell viability, were performed by using human red blood cells and EA.hy926 human endothelial cell line, respectively.