RESUMO
There is a widely recognized need to reduce human activity's impact on the environment. Many industries of the leather and textile sector (LTI), being aware of producing a significant amount of residues (Keßler et al. 2021; Liu et al. 2021), are adopting measures to reduce the impact of their processes on the environment, starting with a more comprehensive characterization of the chemical risk associated with the substances commonly used in LTI. The present work contributes to these efforts by compiling and toxicologically annotating the substances used in LTI, supporting a continuous learning strategy for characterizing their chemical safety. This strategy combines data collection from public sources, experimental methods and in silico predictions for characterizing four different endpoints: CMR, ED, PBT, and vPvB. We present the results of a prospective validation exercise in which we confirm that in silico methods can produce reasonably good hazard estimations and fill knowledge gaps in the LTI chemical space. The proposed protocol can speed the process and optimize the use of resources including the lives of experimental animals, contributing to identifying potentially harmful substances and their possible replacement by safer alternatives, thus reducing the environmental footprint and impact on human health.
Assuntos
Segurança Química , Indústria Têxtil , Animais , Humanos , IndústriasRESUMO
Aerogels from natural polymers are endowed with attractive textural and biological properties for biomedical applications due to their high open mesoporosity, low density, and reduced toxicity. Nevertheless, the lack of macroporosity in the aerogel structure and of a sterilization method suitable for these materials restrict their use for regenerative medicine purposes and prompt the research on getting ready-to-implant dual (macro + meso)porous aerogels. In this work, zein, a family of proteins present in materials for tissue engineering, was evaluated as a sacrificial porogen to obtain macroporous starch aerogels. This approach was particularly advantageous since it could be integrated in the conventional aerogel processing method without extra leaching steps. Physicochemical, morphological, and mechanical characterization were performed to study the effect of porogen zein at various proportions (0:1, 1:2, and 1:1 zein:starch weight ratio) on the properties of the obtained starch-based aerogels. From a forward-looking perspective for its clinical application, a supercritical CO2 sterilization treatment was implemented for these aerogels. The sterilization efficacy and the influence of the treatment on the aerogel final properties were evaluated mainly in terms of absence of microbial growth, cytocompatibility, as well as physicochemical, structural, and mechanical modifications.
Assuntos
Géis/química , Porosidade , Amido/química , Engenharia Tecidual , Dióxido de Carbono/química , Humanos , Polímeros/química , Próteses e ImplantesRESUMO
Microbial laccases are powerful enzymes capable of degrading lignin and other recalcitrant compounds including endocrine disrupting chemicals (EDCs). Efficient EDC removal on an industrial scale requires robust, stable, easy to handle and cost-effective immobilized biocatalysts. In this direction, magnetic biocatalysts are attractive due to their easy separation through an external magnetic field. Recently, a bioinspired immobilization technique that mimics the natural biomineralization reactions in diatoms has emerged as a fast and versatile tool for generating robust, cheap, and highly stable (nano) biocatalysts. In this work, bioinspired formation of a biotitania matrix is triggered on the surface of magnetic particles in the presence of laccase in order to produce laccase-biotitania (lac-bioTiO2 ) biocatalysts suitable for environmental applications using a novel, fast and versatile enzyme entrapment technique. Highly active lac-bioTiO2 particles have been produced and the effect of different parameters (enzyme loading, titania precursor concentration, pH, duration of the biotitania formation, and laccase adsorption steps) on the apparent activity yield of these biocatalysts were evaluated, the concentration of the titania precursor being the most influential. The lac-bioTiO2 particles were able to catalyze the removal of bisphenol A, 17α-ethinylestradiol and diclofenac in a mixture of six model EDCs and retained 90% of activity after five reaction cycles and 60% after 10 cycles.
Assuntos
Disruptores Endócrinos/isolamento & purificação , Enzimas Imobilizadas/metabolismo , Lacase/metabolismo , Magnetismo , Titânio/metabolismo , Poluentes da Água/isolamento & purificação , Compostos Benzidrílicos/isolamento & purificação , Diclofenaco/isolamento & purificação , Etinilestradiol/isolamento & purificação , Fenóis/isolamento & purificação , Purificação da Água/métodosRESUMO
Additive manufacturing (AM) is widely recognized as a versatile tool for achieving complex geometries and customized functionalities in designed materials. However, the challenge lies in selecting an appropriate AM method that simultaneously realizes desired microstructures and macroscopic geometrical designs in a single sample. This study presents a direct ink writing method for 3D printing intricate, high-fidelity macroscopic cellulose aerogel forms. The resulting aerogels exhibit tunable anisotropic mechanical and thermal characteristics by incorporating fibers of different length scales into the hydrogel inks. The alignment of nanofibers significantly enhances mechanical strength and thermal resistance, leading to higher thermal conductivities in the longitudinal direction (65 mW m-1 K-1) compared to the transverse direction (24 mW m-1 K-1). Moreover, the rehydration of printed cellulose aerogels for biomedical applications preserves their high surface area (≈300 m2 g-1) while significantly improving mechanical properties in the transverse direction. These printed cellulose aerogels demonstrate excellent cellular viability (>90% for NIH/3T3 fibroblasts) and exhibit robust antibacterial activity through in situ-grown silver nanoparticles.
Assuntos
Celulose , Impressão Tridimensional , Celulose/química , Camundongos , Animais , Células NIH 3T3 , Géis/química , Nanofibras/química , Prata/química , Antibacterianos/farmacologia , Antibacterianos/química , Nanopartículas Metálicas/químicaRESUMO
Gold nanoparticles (AuNPs) are attractive materials for the immobilization of enzymes due to several advantages such as high enzyme loading, absence of internal diffusion limitations, and Brownian motion in solution, compared to the conventional immobilization onto porous macroscopic supports. The affinity of AuNPs to different groups present at the protein surface enables direct enzyme binding to the nanoparticle without the need of any coupling agent. Enzyme activity and stability appear to be improved when the biocatalyst is immobilized onto AuNPs. Rhamnulose-1-phosphate aldolase (RhuA) was selected as model enzyme for the immobilization onto AuNPs. The enzyme loading was characterized by four different techniques: surface plasmon resonance (SPR) shift and intensity, dynamic light scattering (DLS), and transmission electron microscopy (TEM). AuNPs-RhuA complexes were further applied as biocatalyst of the aldol addition reaction between dihydroxyacetone phosphate (DHAP) and (S)-Cbz-alaninal during two reaction cycles. In these conditions, an improved reaction yield and selectivity, together with a fourfold activity enhancement were observed, as compared to soluble RhuA.
Assuntos
Aldeído Liases/química , Aldeído Liases/metabolismo , Aldeídos/química , Biocatálise , Enzimas Imobilizadas/química , Enzimas Imobilizadas/metabolismo , Nanoconjugados/química , Alanina/análogos & derivados , Alanina/metabolismo , Fosfato de Di-Hidroxiacetona/metabolismo , Estabilidade Enzimática , Escherichia coli/enzimologia , Ouro/química , Nanopartículas Metálicas/química , Ramnose/análogos & derivados , Ramnose/metabolismoRESUMO
Demand of scaffolds for hard tissue repair increases due to a higher incidence of fractures related to accidents and bone-diseases that are linked to the ageing of the population. Namely, scaffolds loaded with bioactive agents can facilitate the bone repair by favoring the bone integration and avoiding post-grafting complications. Supercritical (sc-)foaming technology emerges as a unique solvent-free approach for the processing of drug-loadenu7d scaffolds at high incorporation yields. In this work, medicated poly(ε-caprolactone) (PCL) scaffolds were prepared by sc-foaming coupled with a leaching process to overcome problems of pore size tuning of the sc-foaming technique. The removal of the solid porogen (BA, ammonium bicarbonate) was carried out by a thermal leaching taking place at 37 °C and in the absence of solvents for the first time. Macroporous scaffolds with dual porosity (50-100 µm and 200-400 µm ranges) were obtained and with a porous structure directly dependent on the porogen content used. The processing of ketoprofen-loaded scaffolds using BA porogen resulted in drug loading yields close to 100% and influenced its release profile from the PCL matrix to a relevant clinical scenario. A novel solvent-free strategy has been set to integrate the incorporation of solid porogens in the sc-foaming of medicated scaffolds.
RESUMO
One of the main challenges of cardiovascular tissue engineering is the development of bioresorbable and compliant small-diameter vascular grafts (SDVG) for patients where autologous grafts are not an option. In this work, electrospun bilayered bioresorbable SDVG based on blends of poly(L-lactic acid) (PLLA) and segmented polyurethane (PHD) were prepared and evaluated. The inner layer of these SDVG was surface-modified with heparin, following a methodology involving PHD urethane functional groups. Heparin was selected as anticoagulant agent, and also due to its ability to promote human umbilical vein endothelial cells (HUVECs) growth and to inhibit smooth muscle cells over-proliferation, main cause of neointimal hyperplasia and restenosis. Immobilized heparin was quantified and changes in SDVG microstructure were investigated through SEM. Tensile properties of the heparin-functionalized SDVG resembled those of saphenous vein. Vascular grafts were seeded with HUVECs and cultured on a flow-perfusion bioreactor to analyze the effect of heparin on graft endothelization under simulated physiological-like conditions. The analysis of endothelial cells attachment and gene expression (Real-Time PCR) pointed out that the surface functionalization with heparin successfully promoted a stable and functional endothelial cell layer.
Assuntos
Anticoagulantes/metabolismo , Bioprótese , Prótese Vascular , Heparina/metabolismo , Células Endoteliais da Veia Umbilical Humana/citologia , Engenharia Tecidual/métodos , Humanos , Poliésteres/química , Poliuretanos/químicaRESUMO
Biopolymer-based aerogels can be obtained by supercritical drying of wet gels and endowed with outstanding properties for biomedical applications. Namely, polysaccharide-based aerogels in the form of microparticles are of special interest for wound treatment and can also be loaded with bioactive agents to improve the healing process. However, the production of the precursor gel may be limited by the viscosity of the polysaccharide initial solution. The jet cutting technique is regarded as a suitable processing technique to overcome this problem. In this work, the technological combination of jet cutting and supercritical drying of gels was assessed to produce chitosan aerogel microparticles loaded with vancomycin HCl (antimicrobial agent) for wound healing purposes. The resulting aerogel formulation was evaluated in terms of morphology, textural properties, drug loading, and release profile. Aerogels were also tested for wound application in terms of exudate sorption capacity, antimicrobial activity, hemocompatibility, and cytocompatibility. Overall, the microparticles had excellent textural properties, absorbed high amounts of exudate, and controlled the release of vancomycin HCl, providing sustained antimicrobial activity.
RESUMO
Chronic wounds are a prevailing cause of decreased quality of life, being microbial burden a factor hindering the normal wound healing process. Aerogels are nanostructured materials with large surface area (>250 m2/g) and high porosity (>96%). In this work, vancomycin-loaded chitosan aerogel beads were tested as a potential formulation to treat and prevent infections at the wound site. Processing of chitosan in the form of aerogels endowed this polysaccharide with enhanced water sorption capacity and air permeability. The morphological and textural properties of the particles were studied by image and N2 adsorption-desorption analysis and scanning electron microscopy. Vancomycin content and release profiles from aerogel carriers showed a fast drug release that permitted to efficiently achieve local therapeutic levels. Cell studies with fibroblasts and antimicrobial tests against S. aureus showed that the vancomycin-loaded aerogel particles were cytocompatible and effective in preventing high bacterial loads at the wound site.
Assuntos
Antibacterianos/farmacologia , Quitosana/química , Portadores de Fármacos/química , Hidrogéis/química , Vancomicina/farmacologia , Animais , Antibacterianos/química , Antibacterianos/toxicidade , Linhagem Celular , Quitosana/toxicidade , Colagenases/metabolismo , Portadores de Fármacos/toxicidade , Liberação Controlada de Fármacos , Fibroblastos/efeitos dos fármacos , Hidrogéis/toxicidade , Camundongos , Porosidade , Staphylococcus aureus/efeitos dos fármacos , Vancomicina/química , Vancomicina/toxicidadeRESUMO
Smart electroactive biomaterials are sought to allow the direct delivery of electrical, electrochemical and electromechanical signals to biological tissues. Specifically, poly-(3,4-ethylenedioxythiophene) (PEDOT) is a polymer of special interest attending to its biocompatibility, tuneable electrical conductivity and processing versatility. In this work, nanostructured PEDOT was synthesized using starch/κ-carrageenan aerogels as templates. κ-carrageenan biopolymer acted as doping agent of the conductive polymer to enhance the biocompatibility and the electrical response. The physicochemical, morphological, mechanical and electrical properties of the nanostructured PEDOT and templates were characterized. The incorporation of κ-carrageenan to the nanostructured materials resulted in an increase in the compressive strength of ca. 40% and a decrease in the electrical impedance of one order-of-magnitude. The synergistic combination of the inherent electrical properties of the PEDOT, the advantageous features of κ-carrageenan as doping agent and the porous morphology of the aerogel template resulted in electroactive PEDOT nanostructures with relevant properties for biomedical applications.
Assuntos
Compostos Bicíclicos Heterocíclicos com Pontes/química , Carragenina/química , Polímeros/química , Amido/química , Nanoestruturas/química , Tiofenos/químicaRESUMO
In recent years, there has been an increase in efforts to improve wastewater treatment as the concentration of dangerous pollutants, such as endocrine disrupting chemicals, in wastewater increases. These compounds, which mimic the effect of hormones, have a negative impact on human health and are not easily removed from water. One way to effectively eliminate these pollutants is to use enzymatically activated materials. In this study, we report on the use of laccase from the white rot fungus Trametes versicolor immobilized onto polyamide 6/chitosan (PA6/CHIT) nanofibers modified using two different spacers (bovine serum albumin and hexamethylenediamine). We then tested the ability of the PA6/CHIT-laccase biocatalysts to eliminate a mixture containing 50µM of two endocrine disrupting chemicals: bisphenol A and 17α-ethinylestradiol. The PA6/CHIT nanofiber matrix used in this study not only proved to be a suitable carrier for immobilized and modified laccase but was also efficient in the removal of a mixture of endocrine disrupting chemicals in three treatment cycles.
Assuntos
Disruptores Endócrinos/metabolismo , Proteínas Fúngicas/metabolismo , Lacase/metabolismo , Trametes/enzimologia , Biodegradação Ambiental , Caprolactama/análogos & derivados , Caprolactama/química , Quitosana/química , Enzimas Imobilizadas/metabolismo , Humanos , Nanofibras/química , Nanofibras/ultraestrutura , Polímeros/químicaRESUMO
: In vitro multienzymatic bioreaction systems are attracting increasing attention for the development of bioproduction systems. The de-coupling of the biocatalytic pathway from the cellular machinery aimed at growth and survival allows achievement of high product yields and thus reduces byproduct or waste generation. Additionally, the use of several enzymes allows the realization of much more complex synthetic schemes, thus expanding the chemical diversity of synthetic compounds with new chemical properties or bioactivities. This chapter provides a survey of in vitro multienzymatic bioreaction systems used for biosynthesis, discusses process design aspects for the technical realization of multienzymatic bioreactions, highlights advantages and recent developments in using multienzyme microreactors, and finally reviews examples and strategies of the co-immobilization of multienzymes in nano/microsized materials.
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Biocatálise , Complexos MultienzimáticosRESUMO
In vitro enzymatic activity highly depends on the reaction medium. One of the most important parameters is the buffer used to keep the pH stable. The buffering compound prevents a severe pH-change and therefore a possible denaturation of the enzyme. However buffer agents can also have negative effects on the enzymatic activity, such as competitive substrate inhibition. We assess this effect with a computational approach based on a protein-ligand docking method and the HYDE scoring function. Our method predicts competitive binding of the buffer compound to the active site of the enzyme. Using data from literature and new experimental data, the procedure is evaluated on nine different enzymatic reactions. The method predicts buffer-enzyme interactions and is able to score these interactions with the correct trend of enzymatic activities. Using the new method, possible buffers can be selected or discarded prior to laboratory experiments.