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Selective persistence of Propionibacterium species FMA5 following sealing of infected dentinal matrix

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Abstract

The objective of the present study was to test the hypothesis that nutrient deprivation by effective isolation should inactivate causative saccharolytic bacteria occupying carious lesions. Vital maxillary third molar teeth were prepared by removing only the superficial necrotic material, leaving behind infected dentinal matrix, before the cavity was sealed with glass ionomer cement (GIC). Before sealing, lesions were biopsied to provide reference bacterial DNA for microbial analysis. After an interval of 10–12 months, the teeth were extracted and, after careful removal of GIC restoration, the underlying dentine was biopsied again for post-treatment microbial analysis. Microbial diversity for nine taxa in 45 carious lesions, before and after minimal intervention therapy, was quantified by real-time polymerase chain reaction (PCR). Except for Propionibacterium sp. FMA5, Fusobacterium nucleatum and Pseudoramibacter alactolyticus, representation of all other taxa showed reduction in the post-restoration biopsy samples. However, Propionibacterium sp. FMA5 was the only species predominantly detected in 80% of the pre-intervention, 82% of the post-restoration and 73% of the paired pre- and post-restoration biopsy samples. The median bacterial load for Propionibacterium sp. FMA5, lactobacilli and bacteria from the family Coriobacteriaceae was higher than the median bacterial load for the remaining six taxa. Significant reduction in the median bacterial load for lactobacilli was evident in post-restoration biopsy samples, implying effective control by GIC after minimal intervention. However, the median bacterial load for Propionibacterium sp. FMA5 increased in post-restoration biopsy samples. Incorporation of antimicrobial agents effective against Propionibacterium species FMA5 could add to more effective conservative management of advanced carious lesions.

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Acknowledgements

Kerstin Angner was a recipient of a Bela Schwartz Fellowship. Ky-Anh Nguyen and Shanika Nanayakkara from the Institute of Dental Research, Westmead Hospital and Faculty of Dentistry, The University of Sydney, are thanked for the statistical analysis of the data.

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    Authors and Affiliations

    1. Institute of Dental Research, Westmead Centre for Oral Health and The Westmead Institute for Medical Research, The University of Sydney, Westmead, Australia

      M. A. Nadkarni, K. Angner & N. Hunter

    2. Faculty of Dentistry, The University of Sydney, Sydney, New South Wales, Australia

      M. A. Nadkarni, K. Angner & N. Hunter

    Authors
    1. M. A. Nadkarni
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    2. K. Angner
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    3. N. Hunter
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    Correspondence to M. A. Nadkarni.

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    Funding

    This study was supported by a National Health and Medical Research Council grant (512524).

    Conflict of interest

    The authors declare that they have no conflict of interest.

    Ethical approval

    The Human Research Ethics Committee, Sydney West Area Health Service, ref no. HS/TG HREC2005/8/4.21(2169).

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    Written consent was obtained from participating subjects.

    Additional information

    M. A. Nadkarni and K. Angner contributed equally as first authors.

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    Fig. S1

    Schematic representation of pre-restoration and post-restoration, end-point bur biopsies. After removing the denatured dentine at the lesion surface of a decayed wisdom tooth, two bur biopsies were taken at two separate lesion sites (A). After the 10–12 months’ time interval, the tooth is extracted, the temporary restoration is removed and a single bur biopsy is taken from the underlying dentine (B) (GIF 126 kb)

    High-resolution image (TIF 91 kb)

    Fig. S2

    Fluorescence in-situ hybridisation (FISH) analysis of post-extraction FISH on 2-μm sections of bisected tooth. Universal probe labelled with Alexa 594 (red), Propionibacterium sp. FMA5 labelled with Alexa 488 (green) (GIF 176 kb)

    High-resolution image (TIF 199 kb)

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    Nadkarni, M.A., Angner, K. & Hunter, N. Selective persistence of Propionibacterium species FMA5 following sealing of infected dentinal matrix. Eur J Clin Microbiol Infect Dis 36, 869–878 (2017). https://doi.org/10.1007/s10096-016-2875-6

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    • DOI: https://doi.org/10.1007/s10096-016-2875-6

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