Toshihiro Sanbe

Effects of Ethanol Consumption on Periodontal Inflammation in Rats

Studies suggest a correlation between ethanol consumption and periodontal disease. We hypothesized that elevated levels of blood reactive oxygen species following ethanol consumption may increase inflammation in periodontal tissue. Rats were divided into 4 groups (6–7 rats/group). Two groups were fed an ethanol-containing liquid diet, and 2 groups were fed a pair-fed control diet. In one of each dietary group, periodontitis was ligature-induced, while the other group was left unligated. Chronic ethanol feeding alone decreased the ratio of reduced/oxidized glutathione and increased 8-hydroxydeoxy-guanosine and tumor necrosis factor (TNF)-α levels in the gingiva. Blood hydroperoxides were also increased. In ligature-induced periodontitis lesions, ethanol feeding enhanced polymorpho-nuclear leukocyte infiltration and TNF-α expression. The results suggest that chronic alcohol consumption increased periodontal inflammation, oxidative damage, and TNF-α production and had an additive effect on polymorphonuclear leukocyte infiltration and gingival oxidative damage, increasing the severity of periodontal inflammation in the ligature model.

Effects of vitamin C intake on gingival oxidative stress in rat periodontitis.

Increased levels of oxidative stress due to excessive production of reactive oxygen species are involved in the pathogenesis of periodontitis. Studies suggest a negative association between plasma vitamin C level and the severity of periodontitis. We hypothesized that increases in plasma vitamin C levels after vitamin C intake might clinically reduce gingival oxidative stress in a rat periodontitis model. A ligature was placed around rat mandibular molars for 4 weeks to induce periodontitis, and the rats were then given drinking water with or without 1 g/L vitamin C for 2 weeks after the ligature was removed. The periodontitis-induced rats showed a 149% increase in 8-hydroxydeoxyguanosine level and a 40% decrease in reduced:oxidized glutathione ratio in gingival tissue. Vitamin C intake induced a 175% increase in plasma vitamin C level, resulting in an improvement in the gingival 8-hydroxydeoxyguanosine level (decreased) and in the reduced:oxidized glutathione ratio (increased). Furthermore, in ligature-induced periodontitis lesions, gene expression encoding inflammation, including interleukin-1 alpha and interleukin-1 beta, was more than twofold down-regulated by vitamin C intake. The results suggest that systemic administration of vitamin C could be clinically beneficial in improving periodontitis-induced oxidative stress by down-regulating inflammatory gene expression.

Oxidative damage of periodontal tissue in the rat periodontitis model: effects of a high-cholesterol diet.

Studies suggest an association between consumption of a high‐cholesterol diet and periodontitis. We addressed the mechanism by which high dietary cholesterol could be detrimental to periodontal health in a rat model. Feeding a high‐cholesterol diet augmented the effects of bacterial pathogens and their products (e.g., lipopolysaccharide and proteases) on production of pro‐inflammatory cytokines in fibroblasts. High dietary cholesterol also increased mitochondrial 8‐hydroxydeoxyguanosine in the periodontal tissues. These results suggest that excessive tissue oxidative damage induced by high dietary cholesterol could potentiate pro‐inflammatory cytokine production by fibroblasts stimulated with bacterial pathogens.

Preventive effects of a cocoa-enriched diet on gingival oxidative stress in experimental periodontitis.

BACKGROUND Oxidative stress affects the progression of periodontitis. Cocoa is a rich source of flavonoids with antioxidant properties, which could suppress gingival oxidative stress in periodontal lesions. The purpose of the present study was to investigate the effects of a cocoa-enriched diet on gingival oxidative stress in a rat-periodontitis model. METHODS In this 4-week study, rats were divided into three groups (n = 8/group): a control group (fed a regular diet) and two periodontitis groups (fed a regular diet or cocoa-enriched diet [10% of food intake]). Periodontitis was induced by ligature placement around the mandibular first molars. Serum levels for reactive oxygen metabolites were measured at baseline and 2 and 4 weeks. At 4 weeks, the levels of 8-hydroxydeoxyguanosine and reduced/oxidized glutathione ratio were determined to evaluate gingival oxidative damage and antioxidant status, respectively. RESULTS Rats with experimental periodontitis that were fed a regular diet showed an increase in the level of serum reactive oxygen metabolites in a time-dependent manner. These rats also had an increased 8-hydroxydeoxyguanosine level and decreased reduced/oxidized glutathione ratio in the gingival tissue, inducing alveolar bone loss and polymorphonuclear leukocyte infiltration. Although experimental periodontitis was induced in the rats fed a cocoa-enriched diet, they did not show impairments in serum reactive oxygen metabolite level and gingival levels for 8-hydroxydeoxyguanosine and reduced/oxidized glutathione ratio. Alveolar bone loss and polymorphonuclear leukocyte infiltration after ligature placement were also inhibited by cocoa intake. CONCLUSION Consuming a cocoa-enriched diet could diminish periodontitis-induced oxidative stress, which, in turn, might suppress the progression of periodontitis.

Vitamin C intake attenuates the degree of experimental atherosclerosis induced by periodontitis in the rat by decreasing oxidative stress.

OBJECTIVE Periodontitis has been causally linked to cardiovascular disease, which is mediated through the oxidative stress induced by periodontitis. Since vitamin C has been suggested to limit oxidative damage, we hypothesized that vitamin C intake may reduce endothelial oxidative stress induced by periodontitis in the aorta. The aim of this study was to investigate the effects of vitamin C intake on the initiation of atherosclerosis in a ligature-induced rat periodontitis model. DESIGN Eighteen 8-week-old-male Wistar rats were divided into three groups of six rats and all rats received daily fresh water and powdered food through out the 6-week study. In the vitamin C and periodontitis groups, periodontitis was ligature-induced for the first 4 weeks. In the vitamin C group, rats were given distilled water containing 1 g/L vitamin C for the 2 weeks after removing the ligature. RESULTS In the periodontitis group, there was lipid deposition in the descending aorta and significant increases of serum level of hexanoyl-lysine (HEL), and aortic levels of nitrotyrosine expression, HEL expression and 8-hydroxydeoxyguanosine (8-OHdG) compared to the control group. Vitamin C intake significantly increased plasma vitamin C level and GSH:GSSG ratio (178% and 123%, respectively), and decreased level of serum HEL and aortic levels of nitrotyrosine, HEL and 8-OHdG (23%, 87%, 84%, and 38%, respectively). CONCLUSIONS These results suggest that vitamin C intake attenuates the degree of experimental atherosclerosis induced by periodontitis in the rat by decreasing oxidative stress.

Periodontitis‐induced lipid peroxidation in rat descending aorta is involved in the initiation of atherosclerosis

BACKGROUND AND OBJECTIVE Periodontitis is a risk factor for the development of atherosclerosis. Recent studies indicate that oxidative mechanisms, including lipid peroxidation, are involved not only in periodontitis but also in atherosclerosis. Lipid peroxidation may play an important role in the pathogenesis of atherosclerosis, particularly during its earliest stages. The purpose of this study was to investigate the relationship between lipid peroxidation induced by periodontitis and the initiation of atherosclerosis. MATERIAL AND METHODS Sixteen rats were randomly divided into two groups of eight rats each. Periodontitis was ligature-induced for 4 wk in the experimental group, whereas the control group was left untreated. After the experimental period, the mandibular first molar regions were resected and then subjected to histological analysis and measurement of hexanoyl-lysine expression as an indicator of lipid peroxidation. Descending aorta was used for measuring the levels of hexanoyl-lysine, reactive oxygen species and lipid deposits, and for real-time polymerase chain reaction microarray analysis. The level of hexanoyl-lysine was also measured in serum. RESULTS In the experimental group, the levels of hexanoyl-lysine in periodontal tissue and serum increased. Only aorta samples in the experimental group showed lipid accumulation, with increased expression of hexanoyl-lysine, reactive oxygen species and oxidative stress-related genes (including nitric oxide synthases 2 and 3), whereas the superoxide dismutase 1 gene level was down-regulated. CONCLUSION In a ligature-induced periodontitis rat model, increased lipid peroxidation was found in serum and aorta as well as in periodontal tissue. Atherosclerosis-related gene expression and histological changes were also stimulated. Periodontitis-induced lipid peroxidation in the aorta may be involved in the early stage of atherosclerosis.

Oxidative damage of rat liver induced by ligature-induced periodontitis and chronic ethanol consumption

OBJECTIVE Oxidative stress plays a central role in the initiation and progression of liver disease. Chronic ethanol consumption induces oxidative damage of the liver. Using a rat model, we previously showed that chronic administration of lipopolysaccharide and proteases to gingival sulcus induced both periodontal inflammation and liver injury. Periodontitis and ethanol consumption may have an additional effect on hepatic oxidative damage. The present study investigated the effects of periodontitis on ethanol-induced oxidative damage of the liver using a rat model. DESIGN Male Wistar rats were divided into four groups (six rats/group). During the experimental period of eight weeks, two groups were fed an ethanol-containing liquid diet, and two groups were on a pair-fed control diet. Four weeks prior to the end of the experimental period, one group from each dietary treatment was ligated to induce periodontitis, while the other group was left unligated. In order to evaluate hepatic oxidative damage, the level of hexanoyl-lysine and the ratio of reduced form glutathione/oxidized form glutathione (GSH/GSSG) was determined. The concentration of blood hexanoyl-lysine was also measured as an index of circulating lipid peroxide. RESULTS Ligature-induced periodontitis increased plasma levels of hexanoyl-lysine. In the liver, periodontitis decreased the GSH/GSSG ratio, and the combination of periodontitis and ethanol consumption induced a significant increase in hexanoyl-lysine level compared to ethanol consumption alone. CONCLUSION In the rat model, ligature-induced periodontitis increased plasma lipid peroxide, decreased the hepatic GSH/GSSG ratio and augmented ethanol-induced lipid peroxidation in the liver.

Relationship between periodontitis and hepatic abnormalities in young adults.

Abstract Objective. Obesity has been implicated as a risk factor for periodontitis and non-alcoholic fatty liver disease (NAFLD). In NAFLD, elevated alanine aminotransferase (ALT) is associated with obesity. Although a possible interrelationship between liver function and periodontitis has been reported among the middle-aged population, the correlation in young adults is little known. This study was designed to investigate the relationship between ALT and the presence of periodontitis in university students in Japan. Material and methods. Medical and oral health data were collected in a cross-sectional examination conducted by the Health Service Center of Okayama University. Systemically healthy, non-smoking students aged 18 and 19 years old (n = 2225) were included. The protocol of the United States National Health and Nutrition Examination Survey was applied. Subjects with probing pocket depth ≥ 4 mm were defined as having periodontitis. Logistic regression analysis was used to estimate the association between ALT, body mass index and periodontitis. Results. The number of subjects with periodontitis was 104 (4.7%). In males, having periodontitis was significantly associated with an increased level of ALT (≥ 41 IU/l) in logistic regression analysis (adjusted odds ratio 2.3; 95% confidence interval 1.0–5.2; p < 0.05). However, there was no significant association between periodontitis and ALT in female students. Conclusions. Elevated ALT could be a potential risk indicator for periodontitis among young males. Monitoring hepatic abnormalities to prevent periodontitis must be better understood, even in the young adult population.

Effects of Obesity on Gingival Oxidative Stress in a Rat Model

BACKGROUND Studies indicate a correlation between obesity and periodontitis. Oxidative stress is involved in the progression of periodontitis. The purpose of this study was to investigate the effects of obesity on gingival oxidative stress in a rat periodontitis model. METHODS The obese Zucker rats (n = 14) and their lean littermates (n = 14) were each divided into two groups of seven rats. In one of each group, periodontitis was induced by ligature for 4 weeks, whereas the other group was left unligated. The level of 8-hydroxydeoxyguanosine and the ratio of reduced/oxidized glutathione were determined to examine gingival oxidative stress. The serum level of reactive oxygen metabolites and the gingival gene-expression pattern related to oxidative/metabolic stress, inflammation, and cell behavior were also evaluated. RESULTS The obese rats weighed more than the lean rats at 4 weeks. Compared to lean rats, obese rats had enhanced gingival 8-hydroxydeoxyguanosine levels and a decreased ratio of reduced/oxidized glutathione in the gingival tissue, with increasing serum reactive oxygen metabolites. However, there were no significant differences in the degree of alveolar bone loss between lean and obese rats, except for teeth with and without ligatures in both rats. In addition, the periodontal lesion in obese rats showed higher 8-hydroxydeoxyguanosine levels and polymorphonuclear leukocyte infiltration than the inflamed ones in lean rats, with downregulation of multiple cytochrome P450 gene expression. CONCLUSIONS Obesity induced gingival oxidative stress with increasing serum reactive oxygen metabolites in rats. In the periodontal lesion, gene expressions related to a capacity for xenobiotic detoxification were downregulated in the obese model.

Vitamin C intake inhibits serum lipid peroxidation and osteoclast differentiation on alveolar bone in rats fed on a high-cholesterol diet

OBJECTIVE A high-cholesterol diet stimulates osteoclast differentiation, which may be induced by increased serum lipid peroxidation. The inhibition of serum lipid peroxidation by vitamin C may offer beneficial effects on osteoclast differentiation including increased expression of receptor activator of nuclear factor (NF)-kappaB ligand (RANKL) and NF-kappaB. This study investigated the effects of vitamin C intake on RANKL and NF-kappaB expression in periodontal tissue of rats fed a high-cholesterol diet. DESIGN Twenty-four rats (8 weeks old) were divided into four groups: a control group (fed a regular diet) and three experimental groups (fed a high-cholesterol diet supplemented with 0, 1 and 2 g/l vitamin C/day) in this 12-week study. Vitamin C was provided by its addition to drinking water. As an index of serum lipid peroxidation, hexanoyl-lysine (HEL) level was determined by a competitive enzyme-linked immunosorbent assay method. Immunohistological analysis was performed to evaluate RANKL and NF-kappaB expression on the alveolar bone surface. The number of tartrate-resistant acid phosphatase (TRAP)-positive osteoclasts was also counted. RESULTS Feeding a high-cholesterol diet increased not only the serum HEL level but also the number of TRAP-positive osteoclasts on the alveolar bone surface, with an increase in RANKL and NF-kappaB expression on alveolar bone surface. Intake of vitamin C reduced the serum HEL level and osteoclast differentiation, with decreasing RANKL and NF-kappaB expression. CONCLUSIONS Vitamin C intake could suppress osteoclast differentiation, including RANKL and NF-kappaB expression on the alveolar bone surface, by decreasing serum lipid peroxidation in rats fed a high-cholesterol diet.