Direct real-time measurements of superoxide release from skeletal muscles in rat limbs and human blood platelets using an implantable Cytochrome C microbiosensor.

Autor: Deshpande AS; Department of Chemistry and Biomolecular Science, Clarkson University, 8 Clarkson Avenue, Potsdam, NY, 13699-5810, USA., Muraoka W; U.S. Army Institute of Surgical Research, Blood and Shock Resuscitation, Fort Sam Houston, TX, 78234, USA., Wait J; Department of Physics, Clarkson University, 8 Clarkson Avenue, Potsdam, NY, 13699, USA., Çolak A; Department of Physics, Clarkson University, 8 Clarkson Avenue, Potsdam, NY, 13699, USA., Andreescu S; Department of Chemistry and Biomolecular Science, Clarkson University, 8 Clarkson Avenue, Potsdam, NY, 13699-5810, USA. Electronic address: eandrees@clarkson.edu.
Jazyk: angličtina
Zdroj: Biosensors & bioelectronics [Biosens Bioelectron] 2023 Nov 15; Vol. 240, pp. 115664. Date of Electronic Publication: 2023 Sep 02.
DOI: 10.1016/j.bios.2023.115664
Abstrakt: Oxidative stress and excessive accumulation of the superoxide (O 2 .- ) anion are at the genesis of many pathological conditions and the onset of several diseases. The real time monitoring of (O 2 .- ) release is important to assess the extent of oxidative stress in these conditions. Herein, we present the design, fabrication and characterization of a robust (O 2 .- ) biosensor using a simple and straightforward procedure involving deposition of a uniform layer of L-Cysteine on a gold wire electrode to which Cytochrome C (Cyt c) was conjugated. The immobilized layers, studied using conductive Atomic Force Microscopy (c-AFM) revealed a stable and uniformly distributed redox protein on the gold surface, visualized as conductivity and surface topographical plots. The biosensor enabled detection of (O 2 .- ) at an applied potential of 0.15 V with a sensitivity of 42.4 nA/μM and a detection limit of 2.4 nM. Utility of the biosensor was demonstrated in measurements of real time (O 2 .- ) release in activated human blood platelets and skeletal rat limb muscles following ischemia reperfusion injury (IRI), confirming the biosensor's stability and robustness for measurements in complex biological systems. The results demonstrate the ability of these biosensors to monitor real time release of (O 2 .- ) and estimate the extent of oxidative injury in models that could easily be translated to human pathologies.
Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.
(Copyright © 2023 Elsevier B.V. All rights reserved.)
Databáze: MEDLINE
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