<i>Ex Vivo</i> Murine Skin Model for <i>B. burgdorferi</i> Biofilm
<i>Borrelia burgdorferi</i>, the causative agent of Lyme disease, has been recently shown to form biofilm structures in vitro and <i>in vivo</i>. Biofilms are tightly clustered microbes characterized as resistant aggregations that allow bacteria to withstand harsh environment...
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| Main Authors: | , , , , , |
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| Format: | Book |
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MDPI AG,
2020-08-01T00:00:00Z.
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| Summary: | <i>Borrelia burgdorferi</i>, the causative agent of Lyme disease, has been recently shown to form biofilm structures in vitro and <i>in vivo</i>. Biofilms are tightly clustered microbes characterized as resistant aggregations that allow bacteria to withstand harsh environmental conditions, including the administration of antibiotics. Novel antibiotic combinations have recently been identified for <i>B. burgdorferi in vitro</i>, however, due to prohibiting costs, those agents have not been tested in an environment that can mimic the host tissue. Therefore, researchers cannot evaluate their true effectiveness against <i>B. burgdorferi</i>, especially its biofilm form. A skin ex vivo model system could be ideal for these types of experiments due to its cost effectiveness, reproducibility, and ability to investigate host-microbial interactions. Therefore, the main goal of this study was the establishment of a novel ex vivo murine skin biopsy model for <i>B. burgdorferi</i> biofilm research. Murine skin biopsies were inoculated with <i>B. burgdorferi</i> at various concentrations and cultured in different culture media. Two weeks post-infection, murine skin biopsies were analyzed utilizing immunohistochemical (IHC), reverse transcription PCR (RT-PCR), and various microscopy methods to determine <i>B. burgdorferi</i> presence and forms adopted as well as whether it remained live in the skin tissue explants. Our results showed that murine skin biopsies inoculated with 1 × 10<sup>7</sup> cells of <i>B. burgdorferi</i> and cultured in BSK-H + 6% rabbit serum media for two weeks yielded not just significant amounts of live <i>B. burgdorferi</i> spirochetes but biofilm forms as well. IHC combined with confocal and atomic force microscopy techniques identified specific biofilm markers and spatial distribution of <i>B. burgdorferi</i> aggregates in the infected skin tissues, confirming that they are indeed biofilms. In the future, this ex vivo skin model can be used to study development and antibiotic susceptibility of <i>B. burgdorferi</i> biofilms in efforts to treat Lyme disease effectively. |
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| Item Description: | 10.3390/antibiotics9090528 2079-6382 |