Loricrin and Cytokeratin Disorganisation in Severe Forms of Periodontitis.

OBJECTIVE: The aim of this research was to investigate the role of the cornified epithelium, the outermost layer of the oral mucosa, engineered to prevent water loss and microorganism invasion, in severe forms of periodontitis (stage III or IV, grade C). METHODS: Porphyromonas gingivalis, a major periodontal disease pathogen, can affect cornified epithelial protein expression through chronic activation of signal transducer and activator of transcription 6 (Stat6). We used a mouse model, Stat6VT, that mimics this to determine the effects of barrier defect on P gingivalis-induced inflammation, bone loss, and cornified epithelial protein expression, and compared histologic and immunohistologic findings with tissues obtained from human controls and patients with stage III and IV, grade C disease. Alveolar bone loss in mice was assessed using micro-computerised tomography, and soft tissue morphology was qualitatively and semi-quantitatively assessed by histologic examination for several proteins, including loricrin, filaggrin, cytokeratin 1, cytokeratin 14, a proliferation marker, a pan-leukocyte marker, as well as morphologic signs of inflammation. Relative cytokine levels were measured in mouse plasma by cytokine array. RESULTS: In the tissues from patients with periodontal disease, there were greater signs of inflammation (rete pegs, clear cells, inflammatory infiltrates) and a decrease and broadening of expression of loricrin and cytokeratin 1. Cytokeratin 14 expression was also broader and decreased in stage IV. P gingivalis-infected Stat6VT mice showed greater alveolar bone loss in 9 out of 16 examined sites, and similar patterns of disruption to human patients in expression of loricrin and cytokeratins 1 and 14. There were also increased numbers of leukocytes, decreased proliferation, and greater signs of inflammation compared with P gingivalis-infected control mice. CONCLUSIONS: Our study provides evidence that changes in epithelial organisation can exacerbate the effects of P gingivalis infection, with similarities to the most severe forms of human periodontitis.

A 2-plane micro-computed tomographic alveolar bone measurement approach in mice.

PURPOSE: This study introduces a standardized 2-plane approach using 8 landmarks to assess alveolar bone levels in mice using micro-computed tomography. MATERIALS AND METHODS: Bone level differences were described as distance from the cemento-enamel junction (CEJ) to alveolar bone crest (ABC) and as percentages of vertical bone height and vertical bone loss, comparing mice infected with Porphyromonas gingivalis (Pg) to controls. Eight measurements were obtained per tooth: 2 in the sagittal plane (mesial and distal) and 6 in the coronal plane (mesiobuccal, middle-buccal, distobuccal, mesiolingual, middle-lingual, and distolingual). RESULTS: Significant differences in the CEJ-to-ABC distance between Pg-infected mice and controls were found in the coronal plane (middle-lingual, mesiobuccal, and distolingual for the first molar; and mesiobuccal, middle-buccal, and distolingual for the second molar). In the sagittal plane, the distal measurement of the second molar was different. The middle-buccal, mesiobuccal, and distolingual sites of the first and second molars showed vertical bone loss relative to controls; the second molar middle-lingual site was also different. In the sagittal plane, the mesial sites of the first and second molars and the distal site of the second molar showed loss. Significantly different vertical bone height percentages were found for the mesial and distal sites of the second molar (sagittal plane) and the middle-lingual and distolingual sites of the first molar(coronal plane). CONCLUSION: A reliable, standardized technique for linear periodontal assessments in mice is described. Alveolar bone loss occurred mostly on the lingual surface of the coronal plane, which is often omitted in studies.