Chiono, Valeria

Ciardelli, Gianluca

Idrees, Ayesha

Salber, Jochen

Viebahn, Richard

English

Ayesha Idrees

Richard Viebahn

Gianluca Ciardelli

Valeria Chiono

Jochen Salber

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

04 August 2020POLITECNICO DI TORINORepository ISTITUZIONALES. aureus Colonized Human Skin Equivalent - In Vitro Bio-evaluation Tool for Antibacterial Polymers / Idrees, Ayesha;Viebahn, Richard; Ciardelli, Gianluca; Chiono, Valeria; Salber, Jochen. - (2018). ((Intervento presentato al convegnoDrug-free antibacterial technology for medical applications tenutosi a The Hauser Forum, Cambridge nel 14 December,2018.OriginalS. aureus Colonized Human Skin Equivalent - In Vitro Bio-evaluation Tool for Antibacterial PolymersPublisher: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: 11583/2722662 since: 2019-03-06T15:06:25ZNot applicableHyMedPoly has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement number 643050.www.hymedpoly.euDrug-Free Antibacterial Hybrid Biopolymers for Medical ApplicationsAyesha Idrees1, 2, Richard Viebahn2, Gianluca Ciardelli1, Valeria Chiono1, Jochen Salber21Department of Mechanical and Aerospace Engineering (DIMEAS), Politecnico di Torino, Italy 2 UMC Knappschaftskrankenhaus GmbH, Clinic of Surgery, Hospital of the Ruhr-University, Bochum, GermanyA bacterial colonized human skin equivalent (c-HSE)The aim of this study was the development of a human skin model and human skin wound infection model for the bio-evaluation ofantimicrobial biomaterials intended for wound healing purposes. The three-dimensional in vitro models will represent advanced and complexsystems to perform more reliable preclinical studies. These models will be employed for in vitro screening of both antibacterial activity as wellas cytocompatibility of new biomaterials and wound dressings to optimize their in vivo performances. In the study, the 3D systems weredeveloped and their structure was characterized. The antibacterial activity and cytotoxicity of a model wound dressing releasing Ag+ wasanalyzed in the models.020406080100Percent Viability of Skin Equivalent  (%)Decreasing concentration of Ag+ 020406080100Ag+ MBC Ag+ wounddressingPercent Reduction of S. aureus in Liquid Broth CFU assay (%)77.47.88.28.699.49.8Ag+ MBC Ag+wounddressingUntreatedLog CFU per S. aureus Colonized Skin equivalent (Log) 3D Dermal Fibroblast Model020406080100Ag+ wounddressingWounddressingcontrol(Without Ag+)UntreatedcontrolPercent Viability of Skin Equivalent  (%)S. aureus Colonized Human Skin Equivalent - In Vitro Bio-evaluation Tool for Antibacterial Polymers: ESR14Optimizing the dermal part of human skin: Z-stacked imagingrevealed the filopodia like morphology and a uniform distributionof human fibroblasts at different planes inside a Col-I matrix.Fluorescent microscopic images show cell nuclei stained withDAPI and cytoskeletal F-actin stained with Phalloidin. Scalebar=100 µmH&E stained cross section of in vitro HSE model:- Epidermal layer (1)- Dermal layer (2)The HSE epidermis has a characteristic structure: Stratum corneum, granulosum, spinosum and basale.Histological Analysis of the HSE Immunohistochemistry of the 3D Human Skin Model Ultrastructure Analysis   S. aureus-colonized HSE    TEM image:- Collagen-I fibres (Col I)- Epidermal-Dermal Junction (DEJ).- DEJ presents lamina lucida (LL) and lamina densa (LD).Antibacterial Analysis Cytocompatibility Analysis Inoculated bacteria adhere to the dermal surface, colonize, and replicate to make large structures of  biofilm.Big arrows: Bacteria located within a biofilm matrix inside the dermis. Small arrows: Bacteria surrounding keratinocytes in epidermis.100 µmDAPI/K10bDAPI/Ki67/βTubulin100 µmcDAPI/Lam 550 µmd100 µmDAPI/K14aESR14 Email: ayesha.idrees@polito.it; ayeshaidrees19@gmail.comSupervisors’ Emails: gianluca.ciardelli@polito.it; valeria.chiono@polito.it; jochen.salber@kk-bochum.de; jochen.salber@hotmail.comAffiliation: Department of Mechanical and Aerospace Engineering (DIMEAS), Politecnico di Torino, Italy; UMC Knappschaftskrankenhaus GmbH, Clinic of Surgery, Hospital of the Ruhr-University, Bochum, GermanyCell viability measuring in the 3D system. The 3D skin model was exposed to a range ofsilver ion concentrations (Ag+) for a period of 24 hours.A commercially available Ag+ releasing wound dressing served the purpose of a modelmaterial and was tested in a 3D system along with its control material (without Ag+).The graph demonstrates the treatment of infected skin equivalents with acommercially available Ag+ releasing wound dressing. Skin equivalents were infectedwith S.aureus and thereafter, Ag+ releasing wound dressing or Ag+ in PBS wasapplied onto the skin equivalents.Conclusion: Development of colonized human skin equivalent (c-HSE); Risk assessment platform for cytocompatibility evaluation; Efficacy assessment of antibacterial materials; Comparison of 2D vs. 3D systems; Understanding “Host-Pathogen Interaction”; Development of complex skin models.DEJLL Col ILD(a) (b) (a) (b)IHC verification of the HumanSkin Equivalent (HSE):(a) Keratin 14 (K14) red;(b) Keratin 10 (K10) green;(c) Ki67 (arrows) green;(d) Laminin 5 (Lam5) green;Cell nuclei are shown in blue byusing DAPI staining. Laminin 5is used as a marker of dermal-epidermal junction (DEJ) andappeared as a thin line.12

S. aureus Colonized Human Skin Equivalent - In Vitro Bio-evaluation Tool for Antibacterial Polymers

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S. aureus Colonized Human Skin Equivalent - In Vitro Bio-evaluation Tool for Antibacterial Polymers

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