Abstract
Objective: In this study, we investigated the effects of a flavonoid-rich fraction of Bergamot juice (BJe) in rats subjected to experimental periodontitis induced by a single intragingival injection of lipopolysaccharides (LPS).Main Methods: Periodontitis was induced by a single intragingival injection of 1 μl LPS (10 μg/μl) derived from Salmonella typhimurium in sterile saline solution. The injection was made in the mesolateral side at the interdental papilla between the first and the second molar. Fourteen days after LPS injection, we performed radiographic analyses and then we surgically removed the gingivomucosal tissue surrounding the mandibular first molar for histological, immunohistochemical and molecular analysis.Results: LPS significantly induced oedema, tissue damage and increased neutrophil infiltration. At molecular level, we found increased NF-κB translocation as well as raised both TNF-α and IL-1β expression, other than modulation of apoptosis-associated proteins. Moreover, the increased myeloperoxidase activity was associated with up-regulation of adhesion molecules. Immunohistochemical analysis for nitrotyrosine and poly ADP-ribose displayed an intense staining in the gingivomucosal tissue. Oral administration of BJe for 14 consecutive days reduced tissue injury and several markers of gingival inflammation including nuclear NF-κB translocation, cytokines expression, myeloperoxidase activity and the expression of some adhesion molecules such as ICAM and P-selectin. BJe also decreased both nitrosative stress and PARP positive staining. Moreover, it caused down-regulation of Bax and up-regulation of Bcl-2 expression.Conclusion: Our findings demonstrate that BJe improves LPS-induced periodontitis in rats by reducing the typical markers of inflammation, thus suggesting its potential in the treatment of periodontal diseases.
Highlights
Periodontal disease is a major health problem that affects a great portion of the world population (Thomas and Puleo, 2011)
Before starting this study, we have repeated the quali-quantitative analysis of flavonoids present in BJe by high performance liquid chromatography system (HPLC; Figure 1), following the procedure described by Gattuso and coworkers (Gattuso et al, 2007)
Fourteen days after the beginning of experiments, we evaluated the dental alveolar bone level defined as the distance from the cemento-enamel junction (CEJ) to the maximum coronal level of the alveolar bone crest (CEJ–bone distance) using IMAGE J processing software (Image J software, National Institutes of Health, Bethesda, MD, United States) (Gugliandolo et al, 2018)
Summary
Periodontal disease is a major health problem that affects a great portion of the world population (up to 90%) (Thomas and Puleo, 2011). Periodontitis is an inflammatory disease involving the supporting structures of the teeth, and is caused by an accumulation of periodontopathic bacteria at the gingival sulcus. The etiopathogenesis of periodontal disease is influenced by host response, presence of pathogenic bacteria and risk factors (Kinane et al, 2017). The latter include external (such as smoking) and systemic influences (such as diabetes mellitus), intrinsic and local factors like gender, race, oral hygiene, age, socioeconomic status, systemic health status and use of medications. The inflammatory process involves the stimulation of fibroblasts by interleukin-1β (IL-1β), the increase of collagen breakdown and the rise of osteoclast activity by tumor necrosis factor-α (TNF-α), resulting all together in bone resumption (Kinney et al, 2007)
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