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1.
Mater Sci Eng C Mater Biol Appl ; 115: 110931, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32600679

RESUMO

Alkaline phosphatase (ALP) is an essential biomarker of osteoblastic activity. Currently, ALP activity has been used to study bone mineralization mechanisms and osteoactive biomaterials among others. The ALP quantification is usually performed by destructive methods either on growing cells or cells lysate in which the osteoconductive biomaterial is being assessed. This work addresses the evaluation of a non-destructive colorimetric approach for the determination of ALP activity on osteoblast-derived exosomes from culture supernatants. The efficiency of the method was evaluated on osteoconductive electrospun scaffolds of PCL compounded with ZnO as a reference biomaterial. The results demonstrated that the osteoblast cell line mineralization induced by osteoconductive scaffolds can be monitorized over time by the non-destructive measurement of ALP activity on osteoblast derived exosomes. Consequently, this non-destructive approach suggested to be a reliable alternative technique for the quantification of biomaterials osteoconductivity or even evaluation of osteoblastic response at stem cells.


Assuntos
Fosfatase Alcalina/análise , Meios de Cultura/análise , Exossomos/metabolismo , Osteoblastos/citologia , Desenvolvimento Ósseo , Regeneração Óssea , Calorimetria , Técnicas de Cultura de Células , Linhagem Celular , Humanos , Osteoblastos/metabolismo
2.
Mater Sci Eng C Mater Biol Appl ; 109: 110575, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-32228910

RESUMO

We report a straightforward route for the preparation of flexible, electrochemically stable and easily functionalizable poly(3,4-ethylenedioxythiophene) (PEDOT) composite films deposited on PET foils as biosensing platforms. For this purpose, poly(allylamine) hydrochloride (PAH) was blended with PEDOT to provide amine-bearing sites for further biofunctionalization as well as to improve the mechanical properties of the films. The conducting PEDOT-PAH composite films were characterized by cyclic voltammetry, UV-vis and Raman spectroscopies. An exhaustive stability study was carried out from the mechanical, morphological and electrochemical viewpoint. Subsequent sugar functionalization of the available amine groups from PAH allowed for the specific recognition of lectins and the subsequent self-assembly of glycoenzymes (glucose oxidase and horseradish peroxidase) concomitant with the prevention of non-specific protein fouling. The platforms presented good bioelectrochemical performance (glucose oxidation and hydrogen peroxide reduction) in the presence of redox mediators. The developed composite films constitute a promising option for the construction of all-polymer biosensing platforms with great potential owing to their flexibility, high transmittance, electrochemical stability and the possibility of glycosylation, which provides a simple route for specific biofunctionalization as well as an effective antifouling strategy.


Assuntos
Aspergillus niger/enzimologia , Técnicas Biossensoriais , Compostos Bicíclicos Heterocíclicos com Pontes/química , Técnicas Eletroquímicas , Proteínas Fúngicas/química , Glucose Oxidase/química , Glucose/análise , Membranas Artificiais , Poliaminas/química , Polímeros/química , Peroxidase do Rábano Silvestre/química
3.
Glob Chall ; 4(2): 1900076, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-32042446

RESUMO

Due to its deleterious effects on health, development of new methods for detection and removal of pesticide residues in primary and derived agricultural products is a research topic of great importance. Among them, imazalil (IMZ) is a widely used post-harvest fungicide with good performances in general, and is particularly applied to prevent green mold in citrus fruits. In this work, a composite film for the impedimetric sensing of IMZ built from metal-organic framework nanocrystallites homogeneously distributed on a conductive poly(3,4-ethylene dioxythiophene) (PEDOT) layer is presented. The as-synthetized thin films are produced via spin-coating over poly(ethylene terephtalate (PET) substrate following a straightforward, cost-effective, single-step procedure. By means of impedance spectroscopy, electric transport properties of the films are studied, and high sensitivity towards IMZ concentration in the range of 15 ppb to 1 ppm is demonstrated (featuring 1.6 and 4.2 ppb limit of detection, when using signal modulus and phase, respectively). The sensing platform hereby presented could be used for the construction of portable, miniaturized, and ultrasensitive devices, suitable for pesticide detection in food, wastewater effluents, or the assessment of drinking-water quality.

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