Shuli Chen (Hannover, DE), Dr. Carina Mikolai (Hannover, DE), Diana Strauch (Hannover, DE), Henning Hartwig (Hannover, DE), Jennifer Ulrich (Hannover, DE), Dr. Katharina Nikutta-Doll (Hannover, DE), Dr. Andreas Winkel (Hannover, DE), Dr. Muhammad Imran Rahim (Hannover, DE), Dr. Daniela Paasch (Hannover, DE), Prof. Dr. Nico Lachmann (Hannover, DE), Prof. Dr. Dagmar Wirth (Braunschweig, DE), Prof. Dr. Henning Menzel (Braunschweig, DE), Prof. Dr. Meike Stiesch (Hannover, DE)
Abstract text (incl. figure legends and references)
Introduction
Recently, a 3D model consisting of artificial human mucosa, implant material and oral biofilm was established to study host-microbe interactions at peri-implant sites in vitro. However, immune cells are so far not represented in the model despite the decisive impact they have on the course of inflammatory processes in patients.
Objectives
The aim of the study was to integrate macrophages into the existing 3D peri-implant oral mucosa-biofilm model (INTERbACT) and to validate macrophage presence after model build-up.
Material and Methods
First, the compatibility and performance of the involved cell types (gingival fibroblasts, oral keratinocytes, macrophages) in various media combinations had to be tested. Using adapted culture conditions, M0 macrophages were included in different proportions during model setup. The maturated mucosa was harvested and the content and viability of macrophages was evaluated using microscopy and FACS analyses.
Results
Mixtures of culture media used for growth cultures as well as in the build-up of the 3D model had no negative effect on the viability of the involved cells. Under these culture conditions it was possible to include different proportions of M0 macrophages (1 up to 16x105 MΦs) in the models. No significant differences in the morphology of the mucosa could be observed over a period of 23 days.
The presence and viability of immune cells in the maturated 3D model could be documented. The comparison of CD14- versus CD33-positive cells proved the incorporation of M0. Different amounts of incorporated macrophages are detectable using FACS analyses.
Conclusion
Macrophages could be integrated into the INTERbACT model by adapting the culture medium. By adjusting the number of involved cells, a gradual increase in macrophage proportion could be obtained, suggesting that the model has the potential to mimic different stages of inflammation.