RESUMEN
Extracellular matrix (ECM) regulates cellular responses through mechanotransduction. The standard approach of in vitro culturing on plastic surfaces overlooks this phenomenon, so there is a need for biocompatible materials that exhibit adjustable mechanical and structural properties, promote cell adhesion and proliferation at low cost and for use in 2D or 3D cell cultures. This study presents a new tunable hydrogel system prepared from high-molecular hyaluronic acid (HA), Bovine serum albumin (BSA), and gelatin cross-linked using EDC/NHS. Hydrogels with Young's moduli (E) ranging from subunit to units of kilopascals were prepared by gradually increasing HA and BSA concentrations. Concentrated high-molecular HA network led to stiffer hydrogel with lower cluster size and swelling capacity. Medium and oxygen diffusion capability of all hydrogels showed they are suitable for 3D cell cultures. Mechanical and structural changes of mouse embryonic fibroblasts (MEFs) on hydrogels were compared with cells on standard cultivation surfaces. Experiments showed that hydrogels have suitable mechanical and cell adhesion capabilities, resulting in structural changes of actin filaments. Lastly, applying hydrogel for a more complex HL-1 cell line revealed improved mechanical and electrophysiological contractile properties.
Asunto(s)
Ácido Hialurónico , Hidrogeles , Animales , Ratones , Hidrogeles/farmacología , Ácido Hialurónico/farmacología , Ácido Hialurónico/química , Mecanotransducción Celular , Fibroblastos , Materiales BiocompatiblesRESUMEN
Nanoporous surfaces are promising for label-free electrochemical biosensing. We formed nanopores directly on the electrode surface by means of assembling a dense layer of nonconductive nanoparticles. In our model affinity biosensor, covalent attachment of albumin protein on top of 40 nm polystyrene nanoparticles represented a capture of an analyte, resulting in blockage of the nanopores. Different bulk concentrations of the ferro/ferricyanide redox pair were probed by Faradaic electrochemical impedance spectroscopy and fast chronoamperometry. The character of the redox probe permeation towards the electrode surface differed in dependence on its concentration. These data were compared with the theoretical behavior of the free diffusion according to the Cottrell equation. Both the bulk concentration of the redox probe and the timescale of the experiment affected the performance of the electrochemical detection, demonstrating the importance of controlling these parameters in immunosensing applications.
Asunto(s)
Técnicas Biosensibles , Nanoporos , Técnicas Biosensibles/métodos , Espectroscopía Dieléctrica/métodos , Electrodos , Oxidación-Reducción , Técnicas Electroquímicas , Oro/químicaRESUMEN
Four ferrocene derivatives - ferrocenecarboxylic acid, ferrocenium salt, ferroceneboronic acid, and aminoferrocene - were characterized electrochemically, and their cytotoxicity was probed using cancer cells (line MG-63). We related the observed cytotoxicity with the determined redox potentials of these four ferrocenes - aminoferrocene with its lowest redox potential exhibited the highest cytotoxicity. Thus, we synthesized four derivatives consisting of aminoferrocene and phenylboronic acid residue with the intent to employ them as ROS-activated prodrugs (ROS - reactive oxygen species). We characterized them and studied their time-dependent stability in aqueous environments. Then, we performed electrochemical measurements at oxidative conditions to confirm ROS-responsivity of the synthesized molecules. Finally, the cytotoxicity of the synthesized molecules was tested using cancer MG-63 cells and noncancerous NIH-3T3 cells. The experiments revealed sought behaviour, especially for para-regioisomers of synthesized ferrocenyliminoboronates.
Asunto(s)
Compuestos Ferrosos/química , Metalocenos/química , Profármacos/química , Especies Reactivas de Oxígeno/metabolismo , Animales , Ácidos Borónicos/química , Línea Celular Tumoral , Supervivencia Celular/efectos de los fármacos , Compuestos Ferrosos/farmacología , Humanos , Metalocenos/farmacología , Ratones , Estructura Molecular , Oxidación-Reducción , Profármacos/farmacologíaRESUMEN
In this paper, the d(GCGAAGC) heptamer and the closely related d(GCGAGC) hexamer are examined via electrochemical (cyclic voltammetry) and spectroscopic (circular dichroism) methods. Dramatic changes in the CD spectroscopic and CV electrochemical properties, induced by the loss of only one single nucleotide (A), are detected. The CD spectra and native polyacrylamide gel electrophoresis (PAGE) confirmed structural changes taking place in the relevant chain-like oligodeoxynucleotide assemblies. Dedicated studies suggest that the heptamer (Hp) possesses a hairpin structure, whereas the hexamer (Hx) appears to be rather a duplex. Both of the structures exhibited completely different adsorption behavior at the hanging mercury drop electrode, and this factor was readily confirmed by means of elimination voltammetry with linear scan (EVLS). We established that the Hp hairpin (~-1300 mV), compared to the Hx duplex (~-1360 mV), is the thermodynamically favored electron acceptor. The adsorption isotherms were constructed based on the voltammetric peak height values, reflecting the reduction of the adenine (A) and cytosine (C) moieties as well as the oxidation of the 7,8-dihydroguanine (7,8-DHG) moieties. Finally, as revealed by the spectroscopic and electrochemical results, Hx forms a bimolecular antiparallel homo-duplex carrying both Watson-Crick base pairs (CG or GC) and mismatched edge-to-edge base pairs (GA or AG).
Asunto(s)
ADN/química , ADN/genética , Polimorfismo de Nucleótido Simple , Secuencia de Bases , Dicroismo Circular , ADN/metabolismo , Electroquímica , Guanina/análogos & derivados , Guanina/metabolismo , Modelos Moleculares , Conformación MolecularRESUMEN
Blockage of a nanopore by an analyte molecule has emerged as a promising concept for electrochemical biosensing. Nanoporous structures can be formed on the electrode surface simply by packing spherical nanoparticles in a dense planar arrangement. Modification of the nanoparticles with human serum albumin (HSA) and its interaction with the corresponding antibody (anti-HSA) can induce nanopore-blockage which significantly hinders permeation of the redox probe ([Fe(CN6)]4-/3-). Interfaces of different parameters were studied using Electrochemical Impedance Spectroscopy (EIS), and counterintuitively, the influence of charge of the nanoparticles and other immobilized entities played a substantial role in the measurement. Our study reveals dominant effects including the presence of mixed output signal and resolves corresponding EIS biosensing-related challenges. Consequently, blocking the nanopores was introduced as an efficient technique which enables the application of EIS-based biosensing to real-world analytical issues.