Speaker
Description
Background:
The global rise of skin infections, exacerbated by antibiotic-resistant bacteria, is nowadays a great challenge to healthcare. Traditional treatments are increasingly losing their potential, leading to severe impacts on patient quality of life and survival rates. Consequently, many efforts are made to develop innovative antibacterial therapies. To test this new strategies, conventional methods, such as 2D cell culture and planktonic bacteria culture, often fail to replicate the complexity of infection environments, highlighting the potential of 3D in vitro skin models as a superior alternative.
Methods:
The initial focus of the study has been the development of a 3D in vitro skin model. The dermis compartment was biofabricated by embedding human fibroblasts in a gelatin methacryloyl matrix, with human keratinocytes seeded on top the dermis to form the epidermis. The models were cultured for 31 days in an air-liquid interface configuration. The model demonstrated robust barrier formation and extracellular matrix remodelling. A 3 mm biopsy puncher was used to simulate controlled wounding, followed by inoculation of Staphylococcus aureus (Gram-positive) or Escherichia coli (Gram-negative) to mimic infection. Antibiotic treatment was applied after 24 hours. The samples were subsequently analysed for bacterial survival and skin model responses.
Results:
Results were remarkable: differences between wounded and unwounded samples underlined the barrier effect: indeed, the bacterial proliferation was lower in unwounded samples, for both bacterial strains, meaning that the developed epidermis was able to partially stop the bacterial proliferation. Moreover, the 3D skin model was able to react to both wound and infection in a complete and complex way in terms of extracellular matrix deposition and remodeling, inflammatory response, antimicrobial peptides production and change in cellular behaviors, from epithelial to mesenchymal and from fibroblasts to myofibroblasts. Another important outcome was the change in skin response during infection, showing the ability of both bacteria, in different ways, to impair the model immune response. Also, the antibiotic interacted with the model, modulating some markers, giving some evidences of the release of endotoxins with Escherichia coli death.
Conclusions:
In conclusion the developed 3D in vitro skin model has the potential to become a future landmark as platform for infection investigation and novel therapies testing. Indeed, it demonstrated to be able to behave in a complex and complete way despite being easily producible and low cost. These properties put it in a prominent position for future standardization of platforms to test antibacterial therapies and strategies.
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