Molecular analyses of two bacterial sampling methods in ligature‐induced periodontitis in rats

Abstract

The prevalence profile of periodontal pathogens in dental plaque can vary as a function of the detection method; however, the sampling technique may also play a role in determining dental plaque microbial profiles. We sought to determine the bacterial composition comparing two sampling methods, one well stablished and a new one proposed here. In this study, a ligature‐induced periodontitis model was used in 30 rats. Twenty‐seven days later, ligatures were removed and microbiological samples were obtained directly from the ligatures as well as from the periodontal pockets using absorbent paper points. Microbial analysis was performed using DNA probes to a panel of 40 periodontal species in the checkerboard assay. The bacterial composition patterns were similar for both sampling methods. However, detection levels for all species were markedly higher for ligatures compared with paper points. Ligature samples provided more bacterial counts than paper points, suggesting that the technique for induction of periodontitis could also be applied for sampling in rats. Our findings may be helpful in designing studies of induced periodontal disease‐associated microbiota.

Macrophage immunomodulation in chronic osteolytic diseases—the case of periodontitis

“…One approach to simulate human PD in vertebrate animals is to generate subgingival biofilm retentive milieus by placing silk ligatures inside gingival sulci of molars, which facilitate the onset of inflammation mediated alveolar bone loss . Changes in ligature‐associated biofilms show similar microbial shifts to humans with PD compared to health . The ligature model has been used in rodents and larger vertebrates including dogs or nonhuman primates such as rhesus monkeys, cynomolgus monkeys, and baboons due to similarities with oral human structures that facilitate naturally occurring dental plaque, calculus, putative human periodontal pathogens, and PD .…”

“…43 Changes in ligature-associated biofilms show similar microbial shifts to humans with PD compared to health. 44,45 The ligature model has been used in rodents and larger vertebrates including dogs or nonhuman primates such as rhesus monkeys, cynomolgus monkeys, and baboons due to similarities with oral human structures that facilitate naturally occurring dental plaque, calculus, putative human periodontal pathogens, and PD. 46,47 However, larger vertebrates are less frequently used due to ethical and economic constraints, and experimental PD in rodents using ligatures or pathogen inoculation do not fully reproduce human PD.…”

Comparison of Periodontopathic Bacterial Profiles of Different Periodontal Disease Severity Using Multiplex Real-Time Polymerase Chain Reaction

“…Belibasakis and colleagues [ 27 ] reported similar profiles of ‘Red complex’ both from paperpoint and from curette samples. In another study, ligature that induced experimental periodontitis and paperpoint subsequently inserted in the same sites after ligature removal were compared with checkerboard DNA-DNA hybridization [ 28 ]. Considering that similar bacterial profiles between the ligature and paperpoint, it is suggested that a ligature on a tooth may also be used as a sampling method.…”