Chiono, Valeria

Ciardelli, Gianluca

Idrees, Ayesha

Pacharra, Sandra

Salber, Jochen

English

Ayesha Idrees

Sandra Pacharra

Gianluca Ciardelli

Valeria Chiono

Jochen Salber

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

04 August 2020POLITECNICO DI TORINORepository ISTITUZIONALEBio-Analyses of Antimicrobial Biomaterials for Wound Dressings – A Step Forward to Prevent Microbial InfectionDiseases / Idrees, Ayesha; Pacharra, Sandra; Ciardelli, Gianluca; Chiono, Valeria; Salber, Jochen. - (2016). ((Interventopresentato al convegno PhD Day tenutosi a Politecnico di Torino, Italy nel October 2016.OriginalBio-Analyses of Antimicrobial Biomaterials for Wound Dressings – A Step Forward to Prevent MicrobialInfection DiseasesPublisher: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: 11583/2714991 since: 2019-03-06T15:08:57ZNot applicableA. Idrees1Academic Tutor: V. Chiono1Co-Authors: J. Salber2,3, S. Pacharra2, G. Ciardelli11Department of Mechanical and Aerospace Engineering, Politecnico di Torino, Corso Duca degli Abruzzi 24, 10129 Torino, 2ClinicalResearch Center, RUHR University Bochum, Universitätsstraße 150, 44801 Bochum, 3Universitätsklinikum KnappschaftskrankenhausBochum GmbH, In der Schornau 23-25, D-44892 BochumAcknowledgements:The funding was provided by the European Union’s Horizon 2020 research and innovation programme under the grantagreement No. 643050 (HyMedPoly ). The authors thank all the HyMedPoly Partners in this HyMedPoly project.Bio-Analyses of Antimicrobial Biomaterials for Wound Dressings –A Step Forward to Prevent Microbial Infection DiseasesIntroduction: The aim of this study is the bio-evaluation of antimicrobialbiomaterials intended for wound dressings. As a wound dressing will be indirect contact with the wound and therefore the skin cells, it should not onlybe antibacterial to prevent infection at wound site but also be non-toxic to thesurrounding tissue. For this purpose, testing of the newly developedbiomaterials will be performed using in vitro assays based on 2D cell culture(according to ISO 10993-5) & 3D cell culture systems as well as testing againstmicrobial biofilm formation.Therefore the first part of my PhD thesis is focused on theestablishment of a suitable 3D skin model and, even more importantly, on thedevelopment of cyto-compatibility assay protocols suitable for the testing ofmaterial samples in this 3D system. In this study, we established a dermallayer 3D model using L929 fibroblasts. We used this model for trials andoptimization of different methods of cytocompatibility analysis for example,Live/Dead staining as well as quantitative assays to measure the amount ofviable and dead cells.0100002000030000400005000060000Day 5 Day 7 Day 10 Day 12 Day 14 Day 16 Day 18 Day 20Fluorescence 560/590nm (RFU)Cell Viability CytoToxicityLive/Dead stainingBright-fieldL929 fibroblast cells inside Collagen I MatrixL929 fibroblast cells on TCPSScheme of the construction of 3D skin model:A collagen I gel (mimicking the extracellularmatrix) is embedded with human dermalfibroblasts. After dermal fibroblasts contractand remodel the collagen matrix, keratinocytesare then seeded onto it. Tissues are then raisedto an air-liquid interface to initiate skin-analogue tissue formation.Effect of Repeated Exposure of CellTiter Reagent on Cells in 3DCell Viability and Cell Death of 3D FibroblastsSchools/Workshops/Industrial trainings/Courses/Publications:-HYMEDPOLY WINTER SCHOOL 1 and workshop: 8th–10th February 2016, “Biodegradable polymers synthesis and functionalisation” Politechnico di Torino,Turin, Italy-HYMEDPOLY Meeting and Workshop: 19th—20th July, 2016, “Biomaterials and cells interaction – new concepts of drug-free antibacterial therapies, University of Westminster, London, UK-Workshop of the project Smart Injectable Drug-Delivery systems for Parkinson’s and Alzheimer’s Disease Treatment (PAD-INJ): 19th -20th November, 2015, “Engineered Biomaterials and BiomedicalDevices in the Regenerative Medicine of the Nervous System”, Doctoral School of Politecnico di Torino, Turin, Italy (Prof. Valeria Chiono and Prof. Maria Grazia Spillantini)-BioPmed Seminar: 26th January 2016, “Biological evaluation of Medical Device: ISO 10993” , Bioindustry Park, Ivrea (Vicent Legay)-Course: 11-13th November 2015, “Advancement in the knowledge and the treatment of neurodegenerative diseases” ,Doctoral School of Politecnico di Torino, Turin, Italy, (Prof. Maria Grazia Spillantini)-Course: 23rd November 2015 , Enterpreneurship, Doctoral School of Politecnico di Torino, Turin, Italy, (Professor Erasmo Carrera and Dr. Marco Petrolo)-Anticipated publication of the review article: “Drug-free antibacterial polymers for biomedical applications” , Journal: Biomedical Science and Engineering Page Press, (in progress)PhD Day- PhD Cycle 31- 4th October, 2016-Politechnico di Torino, ItalyBoth assays (CellTiter-Blue and CytoTox-ONE; www.promega.com) are adaptable to 3D system. Cells were viable for at least 20 days in matrix which is a prerequisite for such experiments.Repeated exposure of cells to reagent for up to 18 days has no lasting toxiceffect. Therefore this assay can be used as a method to monitor cell viabilityover an extended time frame.Cells grown on conventional 2D surface have flattenedmorphology, while the cells in the 3D matrix show filopodia (thincell membrane protrusions), and this morphology resembles thein vivo situation.This evidence shows that Live/Dead staining is also applicable in3D skin model, indicating that cells can directly be observed toassess morphological changes and sudden cell death as a resultof interaction with biomaterials.H & E stained dermal layer showing fibroblasts with dendritic extensions in the matrixFuture Activity Plan: The knowledge deriving from this 3D model will be used to develop a human primary cell based in-vitro model (withhuman dermal fibroblasts and epidermal keratinocytes) able to mimic human skin. Then the development and optimization of bio-assays in thissystem will be performed to evaluate the cell compatibility of biomaterials.Biofilm development and anti bacterial testing will be performed to evaluate the antibacterial properties. Work will start after meeting at theDepartment for Medical Microbiology RUB (also National Reference Center for gram-negative bacteria) on the 19th of October.0100002000030000400005000060000Day 0 Day 1 Day 3 Day 5 Day 7 Day 10 Day 12 Day 14 Day 16 Day 18Fluorescence 560/590nm (RFU)Cell Viability (cells never exposed to cellTiter reagent before)Cell Viability (cells repeatedly exposed to cellTiter reagent for several days)Cyto Toxicity (cells never exposed to cellTiter reagent before)Cyto Toxicity (cells repeatedly exposed to cellTiter reagent for several days)Development and Optimization of Quantitative Cytocompatibility Assays in 3D3D skin model DevelopmentVisualization of Cells in 3D

Bio-Analyses of Antimicrobial Biomaterials for Wound Dressings – A Step Forward to Prevent Microbial Infection Diseases

https://iris.polito.it/retrieve/handle/11583/2714991/225165/PhD%20day_Poster%20Presentation_Ayesha%20Idrees_04.10.2016_SP_JS_gc_VC.pdf

Bio-Analyses of Antimicrobial Biomaterials for Wound Dressings – A Step Forward to Prevent Microbial Infection Diseases

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