Microbiome analysis of feline odontoclastic resorptive lesion (FORL) and feline oral health.

Introduction. Feline odontoclastic resorptive lesion (FORL) is one of the most common and painful oral diseases of the cat. It is characterised by tooth resorption due to destructive activity of odontoclasts. FORL can result in tooth loss. While the aetiology of FORL is not clearly understood, it is thought to be multifactorial and bacteria are likely to play a major role.Hypothesis. Dysbiosis of the normal feline oral microbiota leads to an alteration in commensal bacteria populations, which results in the development of FORL.Aim. The purpose of the current study was to determine the composition of the microbiomes associated with feline oral health and FORL.Methodology. Supragingival plaque was collected from 25 cats with a healthy oral cavity and 40 cats with FORL. DNA was extracted from each sample, the V4 region of the 16S rRNA gene amplified by polymerase chain reaction and amplicons sequenced. Diversity and species richness analyses were performed, principal component analysis was used to explore differences between the oral microbiomes of healthy cats and those with FORL, and linear discriminant analysis effect size was used to assess differences between the groups.Results. The six most abundant bacterial genera identified were Bergeyella, Capnocytophaga, Lampropedia, Morexella, Porphyromonas and Treponema. Two-step cluster analysis of the data identified two FORL sub-groups (FORL-1, FORL-2). The FORL-2 sub-group was very similar to the healthy group, whilst the FORL-1 sub-group was clearly different from both the FORL-2 sub-group and the healthy groups. In this analysis, Capnocytophaga (P <0.001) and Lampropedia (P <0.01) were found at significantly lower levels and Porphyromonas at a slightly higher level in the FORL-1 sub-group compared to the healthy and FORL-2 sub-groups. Microbial diversity was found to be less in the FORL-1 sub-group than in the healthy group. Lampropedia sp., a phosphate-accumulating oral commensal species, was significantly lower in the FORL-1 sub-group.Conclusion. The oral microbiota associated with the FORL-1 sub-group is distinct from that found in the healthy group and FORL-2 sub-group. Lampropedia species may influence the local calcium-phosphate ratio, which could be a factor in tooth and bone resorption observed in FORL.

Anti-Periodontitis Effect of Ethanol Extracts of Alpinia Katsumadai Seeds.

Oral microbes are intimately associated with many oral and systemic diseases. Ongoing research is seeking to elucidate drugs that prevent and treat microbial diseases. Various functions of Alpinia Katsumadai seed extracts have been reported such as their anti-viral, anti-oxidant, anti-inflammatory, anti-puritic, anti-emetic, and cytoprotective effects. Here, we investigated the anti-periodontitis effect of an ethanol extract of Alpinia Katsumadai seeds (EEAKSs) on dental plaque bacteria (DPB)-induced inflammation and bone resorption. DPB and Porphyromonas gingivalis (P. gingivalis) were cultured and lipopolysaccharide (LPS) was extracted. Prostaglandin E2 (PGE2) and cyclooxygenase 2 (COX-2) levels were estimated using ELISA. Cytotoxicity was also verified. Proteases were screened using a protease antibody array method. Osteoclastic bone resorption was also investigated. EEAKSs suppressed P. gingivalis growth on agar plates. LPS prepared from dental plaque bacteria (DPB-LPS) and P. gingivalis (PG-LPS) significantly increased PGE2 and COX2 levels in immortalized gingival fibroblasts (IGFs), immortalized human oral keratinocytes (IHOKs), and RAW264.7 macrophage cells. However, DPB-LPS and PG-LPS-induced PGE2 and COX-2 increases were effectively abolished by EEAKS treatment at non-cytotoxic concentrations. In the protease antibody array, matrix metalloproteinase (MMP)-2, MMP-3, MMP-7, kallikrein 10, cathepsin D, and cathepsin V levels were increased by PG-LPS stimulation. However, increases in protease levels except for cathepsin D were suppressed by EEAKS treatment. In addition, RANKL-induced osteoclast differentiation was significantly inhibited by EEAKS treatment, leading to reductions in resorption pit formation. These results suggest that EEAKSs exerted a beneficial oral health effect to help prevent DPB-mediated periodontal disease.

Isolation of Treponema and Tannerella spp. from equine odontoclastic tooth resorption and hypercementosis related periodontal disease.

REASONS FOR PERFORMING STUDY: Red complex bacteria, i.e. Porphyromonas gingivalis, Treponema denticola and Tannerella forsythia, are involved in the onset and progression of periodontal disease in man, yet seldom inhabit the oral cavity of healthy individuals. Periodontal disease is also encountered in horses, with equine odontoclastic tooth resorption and hypercementosis (EOTRH) constituting a particular form of disease. However, only little is known about the oral microbiome of healthy and periodontitis-affected equids. OBJECTIVE: We aimed to test the hypothesis that red complex bacteria are also associated with EOTRH-related periodontal disease. STUDY DESIGN: Controlled cross-sectional study. METHODS: We screened DNA purified from crevicular fluid derived from 23 EOTRH-affected and 21 disease-free horses for the presence of Treponema spp., Tannerella spp. and Porphyromonas gingivalis DNA by polymerase chain reaction. Subsequently, amplified DNA was bidirectionally sequenced and identified via BLAST analysis. RESULTS: Treponema and/or Tannerella DNA was detected in 100% of periodontitis-related samples and in 52.2% of DNA derived from healthy horses. Twenty-six amplicon sequences were 98-100% homologous to published bacterial sequences, which mostly corresponded to Treponema pectinovorum, oral Treponema clones JU025 and OMZ 840, and Tannerella forsythia. P. gingivalis DNA was only found in 3 EOTRH-related samples. Forty-three amplicon sequences revealed weaker homologies ranging between 80% and 97% to known Treponema or Tannerella strains, partly because of their heterogeneity, partly because they obviously represented so far unknown types. CONCLUSIONS: This is the first report in which known and novel Treponema and Tannerella spp. were isolated in association with EOTRH-related periodontal disease.

Management of tooth resorption.

A correct diagnosis and an understanding of the aetiology and dynamics of the processes involved in tooth resorption is critical to effective management. Tooth resorptions can be classified as: (1) trauma induced; (2) infection induced; or (3) hyperplastic invasive. Some transient trauma induced resorptions require no treatment but must be carefully monitored to check that there are no complicating issues such as infection. In cases of trauma induced replacement resorption, a multidisciplinary approach is usually necessary to ensure an optimal long-term solution. Infection induced tooth resorptions require the removal of the invading micro-organisms by endodontic therapy including intra-canal medication which can also facilitate repair of the resorbed tooth structure. The hyperplastic invasive tooth resorptions pose considerable challenges in management due to the complexity and aggressive nature of the resorptive process. With careful case selection and complete inactivation of resorptive tissue successful management can be achieved.

Histopathologic study of physiological and pathological resorptions in human primary teeth.

This study presents a histological analysis through optical microscopy of primary teeth with physiological and pathological resorptions to outline the histological profile of resorptions. Sixty teeth were examined: 19 primary teeth with physiological resorption and 41 primary teeth with pathological resorption. To analyze the histological conditions of the pulp, periradicular tissue, and the resorption areas, and to investigate the presence, intensity, and location of bacteria, slides were prepared using the hematoxylin-eosin and the Brown-Brenn techniques. For the teeth with physiological resorption, normal pulps and no evidence of bacteria were found. For the teeth with pathological resorption, pulpal alterations, atypical resorption, and bacteria were observed.

In vivo killing of Porphyromonas gingivalis by toluidine blue-mediated photosensitization in an animal model.

Porphyromonas gingivalis is one of the major causative organisms of periodontitis and has been shown to be susceptible to toluidine blue-mediated photosensitization in vitro. The aims of the present study were to determine whether this technique could be used to kill the organism in the oral cavities of rats and whether this would result in a reduction in the alveolar bone loss characteristic of periodontitis. The maxillary molars of rats were inoculated with P. gingivalis and exposed to up to 48 J of 630-nm laser light in the presence of toluidine blue. The number of surviving bacteria was then determined, and the periodontal structures were examined for evidence of any damage. When toluidine blue was used together with laser light there was a significant reduction in the number of viable P. gingivalis organisms. No viable bacteria could be detected when 1 mg of toluidine blue per ml was used in conjunction with all light doses used. On histological examination, no adverse effect of photosensitization on the adjacent tissues was observed. In a further group of animals, after time was allowed for the disease to develop in controls, the rats were killed and the level of maxillary molar alveolar bone was assessed. The bone loss in the animals treated with light and toluidine blue was found to be significantly less than that in the control groups. The results of this study show that toluidine blue-mediated lethal photosensitization of P. gingivalis is possible in vivo and that this results in decreased bone loss. These findings suggest that photodynamic therapy may be useful as an alternative approach for the antimicrobial treatment of periodontitis.