Organotypic 3D Co-Culture of Human Pleura as a Novel In Vitro Model of Staphylococcus aureus Infection and Biofilm
Olga Kurow1, Rima Nuwayhid1, Peggy Stock2
1Department of Orthopedic, Trauma and Plastic Surgery, University Hospital of Leipzig, 04103 Leipzig, Germany.
Abstract:
Bacterial pleural infections are associated with high mortality. Treatment is complicated due to biofilm formation. A common causative pathogen is Staphylococcus aureus (S. aureus). Since it is distinctly human-specific, rodent models do not provide adequate conditions for research. The purpose of this study was to examine the effects of S. aureus infection on human pleural mesothelial cells using a recently established 3D organotypic co-culture model of pleura derived from human specimens. After infection of our model with S. aureus, samples were harvested at defined time points. Histological analysis and immunostaining for tight junction proteins (c-Jun, VE-cadherin, and ZO-1) were performed, demonstrating changes comparable to in vivo empyema. The measurement of secreted cytokine levels (TNF-α, MCP-1, and IL-1β) proved host-pathogen interactions in our model. Similarly, mesothelial cells produced VEGF on in vivo levels. These findings were contrasted by vital, unimpaired cells in a sterile control model. We were able to establish a 3D organotypic in vitro co-culture model of human pleura infected with S. aureus resulting in the formation of biofilm, including host-pathogen interactions. This novel model could be a useful microenvironment tool for in vitro studies on biofilm in pleural empyema.
Insights
This study developed a novel 3D human pleural co-culture model to study Staphylococcus aureus infections. The model effectively replicates biofilm formation and host-pathogen interactions seen in empyema.
Area of Science:
- Infectious Diseases
- Cell Biology
- Biomaterials
Background:
- Bacterial pleural infections, particularly those caused by Staphylococcus aureus, lead to high mortality.
- Biofilm formation complicates treatment strategies for these infections.
- Existing rodent models are inadequate for studying human-specific pathogens like S. aureus.
Purpose of the Study:
- To establish and validate a 3D organotypic co-culture model of human pleura.
- To investigate the effects of Staphylococcus aureus infection on human pleural mesothelial cells within this model.
- To analyze host-pathogen interactions and biofilm formation in vitro.
Main Methods:
- Development of a 3D organotypic co-culture model using human pleural specimens.
- Infection of the model with Staphylococcus aureus and sample harvesting at defined time points.
- Histological analysis, immunostaining for tight junction proteins (c-Jun, VE-cadherin, ZO-1), and cytokine level measurements (TNF-α, MCP-1, IL-1β).
Main Results:
- The model demonstrated histological changes comparable to in vivo empyema.
- Evidence of host-pathogen interactions, including cytokine production (TNF-α, MCP-1, IL-1β) and VEGF secretion by mesothelial cells.
- Successful induction of Staphylococcus aureus biofilm formation within the co-culture system.
Conclusions:
- A novel 3D organotypic in vitro co-culture model of human pleura infected with Staphylococcus aureus was successfully established.
- This model mimics key aspects of pleural empyema, including biofilm formation and host-pathogen interactions.
- The model serves as a valuable tool for in vitro research on pleural empyema and biofilm-related infections.
