Human skin as an ex vivo model for maintaining 'Mycobacterium leprae' and leprosy studies

The in vitro cultivation of 'M. leprae' has not been possible since it was described as causing leprosy, and the limitation of animal models for clinical aspects makes studies on leprosy and bacteria-human host interaction a challenge. Our aim was to standardize the ex vivo skin model (hOS...

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Bibliographic Details
Published in:Tropical medicine and infectious disease 2024-06, Vol.9 (6), p.1-14
Main Authors: de Paula, Natália Aparecida, Leite, Marcel Nani, de Faria Bertoluci, Daniele Ferreira, Soares, Cleverson Teixeira, Rosa, Patrícia Sammarco, Frade, Marco Andrey Cipriani
Format: Article
Language:English
Subjects:
Analysis
animal-use alternatives
Antibiotics
Evaluation
ex vivo skin culture
hOSEC
Immunology
Laboratories
Leprosy
Mycobacterium leprae
Patients
Penicillin
Skin
Technique
Tissue culture
tissue culture techniques
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Summary:The in vitro cultivation of 'M. leprae' has not been possible since it was described as causing leprosy, and the limitation of animal models for clinical aspects makes studies on leprosy and bacteria-human host interaction a challenge. Our aim was to standardize the ex vivo skin model (hOSEC) to maintenance and study of 'M. leprae' as an alternative animal model. Bacillary suspensions were inoculated into human skin explants and sustained in DMEM medium for 60 days. Explants were evaluated by RT-PCR-16SrRNA and cytokine gene expression. The viability and infectivity of bacilli recovered from explants (D28 and D60) were evaluated using the Shepard's model. All explants were RT-PCR-16SrRNA positive. The viability and infectivity of recovered bacilli from explants, analyzed after 5 months of inoculation in mice, showed an average positivity of 31%, with the highest positivity in the D28 groups (80%). Furthermore, our work showed different patterns in cytokine gene expression (TGF-beta, IL-10, IL-8, and TNF-alpha) in the presence of alive or dead bacilli. Although changes can be made to improve future experiments, our results have demonstrated that it is possible to use the hOSEC to maintain 'M. leprae' for 60 days, interacting with the host system, an important step in the development of experimental models for studies on the biology of the bacillus, its interactions, and drug susceptibility.
ISSN:2414-6366
2414-6366
DOI:10.3390/tropicalmed9060135