In Vitro Electrochemical Detection of Hydrogen Peroxide in Activated Macrophages via a Platinum Microelectrode Array
Oxidative stress, an excess of endogenous or exogenous reactive oxygen species (ROS) in the human body, is closely aligned with inflammatory responses. ROS such as hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>), superoxide, and radical hydroxyl ions serve essential functions...
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MDPI AG
2021-08-01
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| Series: | Sensors |
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| Online Access: | https://www.mdpi.com/1424-8220/21/16/5607 |
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| author | Victor M. Carriere Jolin P. Rodrigues Chao Tan Prabhu Arumugam Scott Poh |
| author_facet | Victor M. Carriere Jolin P. Rodrigues Chao Tan Prabhu Arumugam Scott Poh |
| author_sort | Victor M. Carriere |
| collection | DOAJ |
| description | Oxidative stress, an excess of endogenous or exogenous reactive oxygen species (ROS) in the human body, is closely aligned with inflammatory responses. ROS such as hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>), superoxide, and radical hydroxyl ions serve essential functions in fighting infection; however, chronic elevation of these species irreversibly damages cellular components. Given the central role of inflammation in a variety of diseases, including Alzheimer’s disease and rheumatoid arthritis, a low-cost, extracellular, non-invasive assay of H<sub>2</sub>O<sub>2</sub> measurement is needed. This work reports the use of a platinum microelectrode array (Pt MEA)-based ceramic probe to detect time- and concentration-dependent variations in H<sub>2</sub>O<sub>2</sub> production by activated RAW 264.7 macrophages. First, these cells were activated by lipopolysaccharide (LPS) to induce oxidative stress. Chronoamperometry was then employed to detect the quantity of H<sub>2</sub>O<sub>2</sub> released by cells at various time intervals up to 48 h. The most stimulatory concentration of LPS was identified. Further experiments assessed the anti-inflammatory effect of dexamethasone (Dex), a commonly prescribed steroid medication. As expected, the probe detected significantly increased H<sub>2</sub>O<sub>2</sub> production by LPS-doped macrophages, subsequently diminishing the pro-inflammatory effect in LPS-doped cells treated with Dex. These results strongly support the use of this probe as a non-invasive, robust, point-of-care test of inflammation, with a high potential for multiplexing in further studies. |
| first_indexed | 2024-03-10T08:23:30Z |
| format | Article |
| id | doaj.art-7e43798b7c194defa1aafcedf7df45a1 |
| institution | Directory Open Access Journal |
| issn | 1424-8220 |
| language | English |
| last_indexed | 2024-03-10T08:23:30Z |
| publishDate | 2021-08-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Sensors |
| spelling | doaj.art-7e43798b7c194defa1aafcedf7df45a12023-11-22T09:42:43ZengMDPI AGSensors1424-82202021-08-012116560710.3390/s21165607In Vitro Electrochemical Detection of Hydrogen Peroxide in Activated Macrophages via a Platinum Microelectrode ArrayVictor M. Carriere0Jolin P. Rodrigues1Chao Tan2Prabhu Arumugam3Scott Poh4Biomedical Engineering Department, Louisiana Tech University, Ruston, LA 71272, USABiomedical Engineering Department, Louisiana Tech University, Ruston, LA 71272, USAInstitute for Micromanufacturing, Louisiana Tech University, Ruston, LA 71272, USAInstitute for Micromanufacturing, Louisiana Tech University, Ruston, LA 71272, USACollege of Engineering and Science-Chemistry, Louisiana Tech University, Ruston, LA 71272, USAOxidative stress, an excess of endogenous or exogenous reactive oxygen species (ROS) in the human body, is closely aligned with inflammatory responses. ROS such as hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>), superoxide, and radical hydroxyl ions serve essential functions in fighting infection; however, chronic elevation of these species irreversibly damages cellular components. Given the central role of inflammation in a variety of diseases, including Alzheimer’s disease and rheumatoid arthritis, a low-cost, extracellular, non-invasive assay of H<sub>2</sub>O<sub>2</sub> measurement is needed. This work reports the use of a platinum microelectrode array (Pt MEA)-based ceramic probe to detect time- and concentration-dependent variations in H<sub>2</sub>O<sub>2</sub> production by activated RAW 264.7 macrophages. First, these cells were activated by lipopolysaccharide (LPS) to induce oxidative stress. Chronoamperometry was then employed to detect the quantity of H<sub>2</sub>O<sub>2</sub> released by cells at various time intervals up to 48 h. The most stimulatory concentration of LPS was identified. Further experiments assessed the anti-inflammatory effect of dexamethasone (Dex), a commonly prescribed steroid medication. As expected, the probe detected significantly increased H<sub>2</sub>O<sub>2</sub> production by LPS-doped macrophages, subsequently diminishing the pro-inflammatory effect in LPS-doped cells treated with Dex. These results strongly support the use of this probe as a non-invasive, robust, point-of-care test of inflammation, with a high potential for multiplexing in further studies.https://www.mdpi.com/1424-8220/21/16/5607reactive oxygen species (ROS)inflammationRAW 264.7macrophageschronoamperometryplatinum microelectrode array |
| spellingShingle | Victor M. Carriere Jolin P. Rodrigues Chao Tan Prabhu Arumugam Scott Poh In Vitro Electrochemical Detection of Hydrogen Peroxide in Activated Macrophages via a Platinum Microelectrode Array Sensors reactive oxygen species (ROS) inflammation RAW 264.7 macrophages chronoamperometry platinum microelectrode array |
| title | In Vitro Electrochemical Detection of Hydrogen Peroxide in Activated Macrophages via a Platinum Microelectrode Array |
| title_full | In Vitro Electrochemical Detection of Hydrogen Peroxide in Activated Macrophages via a Platinum Microelectrode Array |
| title_fullStr | In Vitro Electrochemical Detection of Hydrogen Peroxide in Activated Macrophages via a Platinum Microelectrode Array |
| title_full_unstemmed | In Vitro Electrochemical Detection of Hydrogen Peroxide in Activated Macrophages via a Platinum Microelectrode Array |
| title_short | In Vitro Electrochemical Detection of Hydrogen Peroxide in Activated Macrophages via a Platinum Microelectrode Array |
| title_sort | in vitro electrochemical detection of hydrogen peroxide in activated macrophages via a platinum microelectrode array |
| topic | reactive oxygen species (ROS) inflammation RAW 264.7 macrophages chronoamperometry platinum microelectrode array |
| url | https://www.mdpi.com/1424-8220/21/16/5607 |
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