Chiono, V.

Ciardelli, G.

Gatermann, S. G.

Greiser, U.

Idrees, Ayesha

Marlinghaus, L.

Pacharra, S.

Salber, J.

Vasquez, J. M.

Viebahn, R.

Wang, W.

English

IDREES, AYESHA

S. Pacharra

L. Marlinghaus

V. Chiono

G. Ciardelli

J. M. Vasquez

U. Greiser

W. Wang

R. Viebahn

S. G. Gatermann

J. Salber

PORTO@iris (Publications Open Repository TOrino - Politecnico di Torino)

04 August 2020POLITECNICO DI TORINORepository ISTITUZIONALEAntimicrobial and Cytotoxicity Evaluations of Hydrogen peroxide - Towards Clinical Application of AntimicrobialBiomaterials for Wound Dressings / Idrees, Ayesha; Pacharra, S.; Marlinghaus, L.; Chiono, V.; Ciardelli, G.; Vasquez, J.M.; Greiser, U.; Wang, W.; Viebahn, R.; Gatermann, S. G.; Salber, J.. - (2017). ((Intervento presentato al convegnoAnnual Meeting of the German Society for Biomaterials 2017 – Würzburg, November 09 – 11 tenutosi a Würzburg,Germany nel November 09 – 11, 2017.OriginalAntimicrobial and Cytotoxicity Evaluations of Hydrogen peroxide - Towards Clinical Application ofAntimicrobial Biomaterials for Wound DressingsPublisher:PublishedDOI:Terms of use:openAccessPublisher copyright(Article begins on next page)This article is made available under terms and conditions as specified in the  corresponding bibliographic description inthe repositoryAvailability:This version is available at: 11583/2691360 since: 2019-01-11T19:02:29ZBiomedical Engineering / Biomedizinische TechnikA Idrees1,2,3, S Pacharra3, L Marlinghaus4, V Chiono1, G Ciardelli1, JM Vasquez1,5, U Greiser5, W Wang5, R Viebahn2, SG Gatermann4, J Salber2,31Department of Mechanical and Aerospace Engineering, Politecnico di Torino, Corso Duca Degli Abruzzi 24, 10129 Turin, Italy.2Clinic of Surgery, Knappschaftskrankenhaus Bochum, Hospital of the RUHR-University, In der Schornau 23-25, 44892 Bochum, Germany.3Clinical Research Center, RUHR University Bochum, Universitätsstraße 150, 44801 Bochum4Department of Medical Microbiology, Institute for Hygiene and Microbiology, Ruhr-University Bochum, Universitätsstraße 150, 44801, Germany.5Vornia Biomaterials Ltd, Dublin.Acknowledgements: HyMedPoly received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska‐Curie grant agreement No643050. The authors thank all the HyMedPoly Partners in this HyMedPoly project.Antimicrobial and Cytotoxicity Evaluations of Hydrogen peroxide-Towards Clinical Application of Antimicrobial Biomaterials for Wound DressingsFigure 1: Evaluation of cytotoxic effects of H2O2 on L929 fibroblastsFigure 1: Cell viability data shows that:1. H2O2 did not favor cell viability at 20 mM, 10 mM, and 1 mMconcentrations in our experimental set-up (L929 cell culture).2. H2O2 concentration of 10 µM did not alter cell viability with respectto control conditions (cells only) and further that H2O2 might bestimulating cell growth at the concentration of 10 µM.1,23. Cytotoxic effect started decreasing below 100 µM H2O2concentration and was no more relevant at further lowerconcentrations (10, 1, 0.1 µM). H2O2 concentration of 10 µM did notaffect cell behavior.Introduction: Chronic wound infections and emerging drug resistance are serious problems in the present world causing a considerablemorbidity and a high healthcare costs. For this purpose, investigation on novel antimicrobial strategies is of great interest. Use of honey fromancient times is reputed for its wound-healing and antibacterial properties. It has been reported that the major antibacterial factor in honey is therelease of hydrogen peroxide (H2O2) produced by glucose oxidase. This approach can be exploited to prepare novel polymer-based antimicrobialbiomaterials for wound healing purposes. In this study, the inhibitory effect of H2O2 on the growth of numerous bacteria of clinical significancewas investigated. To determine the “safe” antimicrobial concentration of H2O2, cytocompatibility analysis was also performed for H2O2 inducedcellular cytotoxicity.Methods: The effect of externally added H2O2 was performed by exposing L929 fibroblasts to various H2O2 concentrations. At different timepoints after exposure with H2O2, cell viability was assessed by measuring cell metabolic activity, cell membrane integrity and cell morphology.Antimicrobial efficacy was evaluated against a wide range of gram-positive and gram-negative bacteria that are involved in chronic wounds namelyS. aureus, S. epidermidis, S. lugdunensis, E. faecalis, E. coli, P. aeruginosa, K. pneumoniae, and A. baumanii. Antimicrobial tests were performedusing broth microdilution method for the determination of Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration(MBC). MIC is the lowest concentration of antimicrobial agent that inhibits bacterial growth, while the MBC is the lowest level of antimicrobialagent that kills the bacteria (by reducing the viability of the initial bacterial inoculum by ≥99.9%).Figure 2: H2O2concentrations of 1 mMand 100 µM did not showimmediate cytotoxic effect,rather cytotoxic effectincreased with time. Thisalso correlated with theaffected morphologicalchanges that appeared overtime.H2O2 at the concentrationof 20 mM and 10 mM,caused cell membranedamage within 4 hour ofexposure.10 µM concentration didnot show cytotoxicity until24 hours of exposure.Figure 2: Evaluation of cytotoxic effects of H2O2 on L929 fibroblasts over timeFigure 3: Antimicrobial activity of H2O2 against different clinically significant  bacterial speciesFigure 4: MIC of H2O2 against different bacterial species Figure 5: MBC of H2O2 against different bacterial species References: 1Wiese AG, Pacifici RE, Davies KJ. Transient adaptation to oxidative stress in mammalian cells. Archives of biochemistry and biophysics. 1995;318(1):231-40.2Burdon RH. Superoxide and hydrogen peroxide in relation to mammalian cell proliferation. Free Radical Biology and Medicine. 1995;18(4):775-94.Results & Conclusion: Results (Figure 3, 4 and 5) showed different MIC and MBC values of H2O2 for different bacterial species indicating theirdifferences in susceptibility to treatment. There was no clear preference between Gram positive and Gram negative bacteria. Among the tested bacteria, S.aureus showed 99.9% bacterial reduction at the concentration of 0.5 mM, while E. faecalis showed the highest MIC value of 5 mM. H2O2 was found mosteffective against S. aureus (one of the most pathogenic bacteria) and less effective against E. faecalis. Acinetobacter baumanii being one of the mostpathogenic bacteria involved in serious skin wound infections also showed the same MIC value (0.5 mM) as S. aureus.Cytotoxicity results (figure 1 and 2) showed two distinct patterns in our experimental set-up: the highest concentrations rapidly induced cell deathcharacterized by morphological evidence and plasma membrane damage as compared to the concentrations of 1 mM and 100 µM where the cytotoxiceffect only gradually increased with time. Results showed that 10 µM concentration did not show cytotoxicity. This data also indicated the concentrationdependent distinct pathways of H2O2 -induced cytotoxicity. Also, we speculated that the cytotoxic effects would differ depending on “at once” H2O2exposure or exposure to “gradually” produced H2O2 by glucose oxidase and glucose entrapped into a matrix. Different H2O2 administration influences theability of th cells to elimin te and detoxify H2O2 nd nee s further investigation.S.aureus S.epidermidisS.lugdunensisE.faecalisE.coliP.aeruginosaK.pneumoniaeA.baumanii0.02.55.0Hydrogen peroxide (mM)

Antimicrobial and Cytotoxicity Evaluations of Hydrogen peroxide - Towards Clinical Application of Antimicrobial Biomaterials for Wound Dressings

https://iris.polito.it/retrieve/handle/11583/2691360/225155/DGBM%20Poster%20Presentation_Ayesha%20Idrees_SJV.pdf

Antimicrobial and Cytotoxicity Evaluations of Hydrogen peroxide - Towards Clinical Application of Antimicrobial Biomaterials for Wound Dressings

Abstract

Similar works

Full text

Available Versions

PORTO@iris (Publications Open Repository TOrino - Politecnico di Torino)