A Prussian blue-based electrode array (PBEA) constituted by eight stencil-printed electrodes on a flexible PET (polyethylene terephthalate) substrate is proposed for in-situ HeLa cell culturing and real-time detection of the released H2O2. The array was suitably interfaced with a poly- (methyl methacrylate) (PMMA) well-containing holders resulting in a low cost multichambered chip. PBEA fabrication was carried out employing a xurography-based cost-effective benchtop microfabrication technology using just a desktop cutting plotter and office grade thermal-laminator. The hydrophobicity of the PET isolating layer allows to constrain cell-containing drops directly on top of the electrochemical cells. HeLa cells growth in the very close vicinity of the working electrode ensures in-situ cell seeding, incubation, and further electrochemical detection of the H2O2 released, enabling high-throughput analysis. Selective and sensitive electrochemical sensing of hydrogen peroxide was carried out at −100 mV vs Ag|AgCl; the resulting LOD was 1.9 μM. Remarkably, the analytical exploitability of the approach was demonstrated by detection of the hydrogen peroxide released from HeLa cells stimulated with N-Formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP) and after pretreatment of the cells with cocoa polyphenols, that induced a decreased oxidative stress levels. These data make our approach a promising tool for oxidative stress evaluation in cell cultures and biological systems.

Oxidative stress on-chip: Prussian blue-based electrode array for in situ detection of H2O2 from cell populations

Della Pelle F.;Del Carlo M.;Compagnone D.
;
2020-01-01

Abstract

A Prussian blue-based electrode array (PBEA) constituted by eight stencil-printed electrodes on a flexible PET (polyethylene terephthalate) substrate is proposed for in-situ HeLa cell culturing and real-time detection of the released H2O2. The array was suitably interfaced with a poly- (methyl methacrylate) (PMMA) well-containing holders resulting in a low cost multichambered chip. PBEA fabrication was carried out employing a xurography-based cost-effective benchtop microfabrication technology using just a desktop cutting plotter and office grade thermal-laminator. The hydrophobicity of the PET isolating layer allows to constrain cell-containing drops directly on top of the electrochemical cells. HeLa cells growth in the very close vicinity of the working electrode ensures in-situ cell seeding, incubation, and further electrochemical detection of the H2O2 released, enabling high-throughput analysis. Selective and sensitive electrochemical sensing of hydrogen peroxide was carried out at −100 mV vs Ag|AgCl; the resulting LOD was 1.9 μM. Remarkably, the analytical exploitability of the approach was demonstrated by detection of the hydrogen peroxide released from HeLa cells stimulated with N-Formyl-L-methionyl-L-leucyl-L-phenylalanine (fMLP) and after pretreatment of the cells with cocoa polyphenols, that induced a decreased oxidative stress levels. These data make our approach a promising tool for oxidative stress evaluation in cell cultures and biological systems.
File in questo prodotto:
Non ci sono file associati a questo prodotto.

I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.

Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11575/111050
Citazioni
  • ???jsp.display-item.citation.pmc??? 3
  • Scopus 23
  • ???jsp.display-item.citation.isi??? ND
social impact