Search results for " Immobilized"
showing 10 items of 25 documents
Development of enzymatically-active bacterial cellulose membranes through stable immobilization of an engineered beta-galactosidase
2018
Enzymatically-active bacterial cellulose (BC) was prepared by non-covalent immobilization of a hybrid enzyme composed by a β-galactosidase from Thermotoga maritima (TmLac) and a carbohydrate binding module (CBM2) from Pyrococcus furiosus. TmLac-CBM2 protein was bound to BC, with higher affinity at pH 6.5 than at pH 8.5 and with high specificity compared to the non-engineered enzyme. Both hydrated (HBC) and freeze-dried (DBC) bacterial cellulose showed equivalent enzyme binding efficiencies. Initial reaction rate of HBC-bound enzyme was higher than DBC-bound and both of them were lower than the free enzyme. However, enzyme performance was similar in all three cases for the hydrolysis of 5% l…
Comparative study of the production of soluble factors in human placenta-derived mesenchymal stromal/stem cells grown in adherent conditions or as ag…
2019
Abstract Different approaches have been studied in both preclinical and clinical settings to develop cell-based therapies and/or engineered cell-based therapies to better integrate grafts with the host. In these techniques, much attention is addressed to the use of adult stem cells such as mesenchymal stem cells (MSCs), but identifying and obtaining sufficient numbers of therapeutic cells, and the right route of administration, is often a challenge. In this study, we tested the feasibility of encapsulating human amnion-derived MSCs (hAMSCs) in a semipermeable and biocompatible fiber as a new approach for regenerative medicine. Our data showed that hAMSCs aggregated in the device constitutes…
Lubricin is expressed in chondrocytes derived from osteoarthritic cartilage encapsulated in poly (ethylene glycol) diacrylate scaffold
2011
Osteoarthritis (OA) is characterized by degenerative changes within joints that involved quantitative and/or qualitative alterations of cartilage and synovial fluid lubricin, a mucinous glycoprotein secreted by synovial fibroblasts and chondrocytes. Modern therapeutic methods, including tissue-engineering techniques, have been used to treat mechanical damage of the articular cartilage but to date there is no specific and effective treatment. This study aimed at investigating lubricin immunohistochemical expression in cartilage explant from normal and OA patients and in cartilage constructions formed by Poly (ethylene glycol) (PEG) based hydrogels (PEG-DA) encapsulated OA chondrocytes. The e…
Delivering Inorganic and Organic Reagents and Enzymes from Zein and Developing Optical Sensors.
2018
Nowadays, interest in using environmentally friendly materials is increasing in many fields. However, the rational design of sensors with biodegradable materials is a challenge. The main aim of this work is to show the possibility of using zein, a protein from corn, as a biodegradable and low-cost material for immobilizing, stabilizing, and delivering different kind of reagents for developing optical sensors. Enzymes, metallic salts, and aromatic and small organic compounds were tested. In addition, different techniques of immobilization, entrapment and adsorption, were used, and different formats, such as solid devices and also multiwell platforms, were proposed. The capacity of zein for i…
Multiple-scan voltammetry and OCP: Archaeometric tools for dating archaeological bronzes
2021
[EN] The application of a multiple-scan strategy to nanosamples taken from 18 cross-sections of Bronze Age arms and armour, as well as two Roman coins using two solid-state electrochemical techniques, the voltammetry of immobilized microparticles (VIMP) and open circuit potential measurements (OCP) is described. The voltammetric responses in contact with aqueous acetate buffer can be attributed to the reduction of cuprite with variable degree of compaction and crystallinity revealing significant differences in the gradient of such properties with depth. Such differences are also revealed by "dry" OCP measurements connecting points in the cross section near and separated from the corrosion l…
Response of human chondrocytes to a non-uniform distribution of hydrophilic domains on poly (ethyl acrylate-co-hydroxyethyl methacrylate) copolymers.
2005
A series of polymer and copolymer networks with varying hydrophilicity and distribution of the hydrophilic groups was synthesized and biologically tested with monolayer culture of human chondrocytes in vitro. Cell viability (MTT), proliferation (BrdU incorporation) and aggrecan expression (PG ELISA) were quantified at 7 and 14 days from seeding. Both assays (MTT and BrdU) showed complementary results that are consistent with positive cellular adhesion on the material. When human chondrocytes were cultured on polymer substrates in which the hydrophilic groups were homogeneously distributed, hydrophobic substrates showed higher values in all the biological parameters analysed. Adhesion, proli…
Enzyme-Controlled Nanodevice for Acetylcholine-Triggered Cargo Delivery Based on Janus Au–Mesoporous Silica Nanoparticles
2017
[EN] This work reports a new gated nanodevice for acetylcholine-triggered cargo delivery. We prepared and characterized Janus Au-mesoporous silica nanoparticles functionalized with acetylcholinesterase on the Au face and with supramolecular b-cyclodextrin: benzimidazole inclusion complexes as caps on the mesoporous silica face. The nanodevice is able to selectively deliver the cargo in the presence of acetylcholine via enzyme-mediated acetylcholine hydrolysis, locally lowering the pH and opening the supramolecular gate. Given the key role played by ACh and its relation with Parkinson's disease and other nervous system diseases, we believe that these findings could help design new therapeuti…
2′,5′-oligoadenylate synthetase from a lower invertebrate, the marine sponge Geodia cydonium, does not need dsRNA for its enzymatic activity
2002
AbstractRecently, the presence of 2′,5′-linked oligoadenylates and a high 2′,5′-oligoadenylate synthetase activity were discovered in a lower invertebrate, the marine sponge Geodia cydonium. It has been demonstrated that mammalian 2–5A synthetase isozymes require a dsRNA cofactor for their enzymatic activity. Our results show that, unlike mammalian 2–5A synthetases, the 2–5A synthetase from the sponge acts in a dsRNA-independent manner in vitro. A prolonged incubation of the G. cydonium extract with a high concentration of a micrococcal nuclease had no effect on the activity of the 2–5A synthetase. At the same time, the micrococcal nuclease was effective within 30 min in degrading dsRNA nee…
A Protein-Interaction Array Inside a Living Cell
2013
Cell phenotype is determined by protein network states that are maintained by the dynamics of multiple protein interactions.1 Fluorescence microscopy approaches that measure protein interactions in individual cells, such as by Forster resonant energy transfer (FRET), are limited by the spectral separation of fluorophores and thus are most suitable to analyze a single protein interaction in a given cell. However, analysis of correlations between multiple protein interactions is required to uncover the interdependence of protein reactions in dynamic signal networks. Available protein-array technologies enable the parallel analysis of interacting proteins from cell extracts, however, they can …
Characterizing archaeological bronze corrosion products intersecting electrochemical impedance measurements with voltammetry of immobilized particles
2017
Application of electrochemical impedance measurements to microparticulate deposits of copper corrosion products attached to graphite electrodes in contact with 0.10 M aqueous HClO4 electrolyte is described. The impedance measurements were sensitive to the applied potential and the amount of solid sample and were modeled taking into account the contribution of the uncovered base electrode. Several pairs of circuit elements provide monotonic variations which are able to characterize different corrosion compounds regardless the amount of microparticulate solid on the electrode. Application to a set of archaeological samples from the archaeological Roman site of Gadara (Jordan, 4th century AD) …