Establishment of a Human Immunocompetent 3D Tissue Model to Enable the Long-Term Examination of Biofilm–Tissue Interactions

Author:

Murkar Rasika1ORCID,von Heckel Charlotte1,Walles Heike1ORCID,Moch Theresia Barbara1,Arens Christoph2ORCID,Davaris Nikolaos2ORCID,Weber André3,Zuschratter Werner4,Baumann Sönke5,Reinhardt Jörg6,Kopp Sascha1ORCID

Affiliation:

1. Core Facility Tissue Engineering, Otto-von-Guericke University Magdeburg, Universitätsplatz 2, 39106 Magdeburg, Germany

2. Department of Otorhinolaryngology, Head and Neck Surgery, University Clinic Giessen, 35392 Giessen, Germany

3. Photonscore GmbH, Brenneckestr. 20, 39118 Magdeburg, Germany

4. Leibniz Institute for Neurobiology, 39118 Magdeburg, Germany

5. Omicron-Laserage® Laserprodukte GmbH, Raiffeisenstr. 5e, 63110 Rodgau, Germany

6. MedFact Engineering GmbH, Hammerstrasse 3, 79540 Lörrach, Germany

Abstract

Different studies suggest an impact of biofilms on carcinogenic lesion formation in varying human tissues. However, the mechanisms of cancer formation are difficult to examine in vivo as well as in vitro. Cell culture approaches, in most cases, are unable to keep a bacterial steady state without any overgrowth. In our approach, we aimed to develop an immunocompetent 3D tissue model which can mitigate bacterial outgrowth. We established a three-dimensional (3D) co-culture of human primary fibroblasts with pre-differentiated THP-1-derived macrophages on an SIS-muc scaffold which was derived by decellularisation of a porcine intestine. After establishment, we exposed the tissue models to define the biofilms of the Pseudomonas spec. and Staphylococcus spec. cultivated on implant mesh material. After 3 days of incubation, the cell culture medium in models with M0 and M2 pre-differentiated macrophages presented a noticeable turbidity, while models with M1 macrophages presented no noticeable bacterial growth. These results were validated by optical density measurements and a streak test. Immunohistology and immunofluorescent staining of the tissue presented a positive impact of the M1 macrophages on the structural integrity of the tissue model. Furthermore, multiplex ELISA highlighted the increased release of inflammatory cytokines for all the three model types, suggesting the immunocompetence of the developed model. Overall, in this proof-of-principle study, we were able to mitigate bacterial overgrowth and prepared a first step for the development of more complex 3D tissue models to understand the impact of biofilms on carcinogenic lesion formation.

Funder

Bundesministerium für Bildung und Forschung

Otto-von-Guericke University

Publisher

MDPI AG

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