ABSTRACT
Diabetes is associated with increased glucose levels and accumulation of glycated products. It is also associated with impairment in the immune response, such as increased susceptibility to infections. In this study, we assessed the possible interactions between TLR4 and RAGE signalling on apoptosis and on the expression of inflammatory cytokines in PBMC from individuals with and without diabetes. PBMCs were isolated from seven diabetic patients and six individuals without diabetes and stimulated in vitro with bacterial LPS (1 µg/ml) associated or not with BSA-AGE (200 µg/ml). This stimulation was performed for 6 h, both in the presence and in the absence of inhibitors of TLR4 (R. sphaeroides LPS, 20 µg/ml) and RAGE (blocking monoclonal antibody). Apoptosis at early and late stages was assessed by the annexin-V/PI staining using flow cytometry. Regulation of TNF-α and IL-10 gene expression was determined by RT-qPCR. PBMCs from diabetes patients tended to be more resistant apoptosis. There were no synergistic or antagonistic effects with the simultaneous activation of TLR4 and RAGE in PBMCs from either diabetes or non-diabetes group. Activation of TLR4 is more potent for the induction of TNF-α and IL-10; RAGE signalling had a negative regulatory effect on TNF-α expression induced by LPS. TLR and RAGE do not have relevant roles in apoptosis of PBMCs. The activation of TLR has greater role than RAGE in regulating the gene expression of IL-10 and TNF-α.
Subject(s)
Apoptosis/immunology , Diabetes Mellitus/immunology , Gene Expression Regulation/immunology , Leukocytes, Mononuclear/immunology , Receptor for Advanced Glycation End Products/immunology , Toll-Like Receptor 4/immunology , Adult , Antibodies, Blocking/immunology , Antibodies, Monoclonal/immunology , Female , Glycation End Products, Advanced/metabolism , Glycation End Products, Advanced/pharmacology , Humans , Inflammation/immunology , Interleukin-10/biosynthesis , Lipopolysaccharides , Male , Middle Aged , Receptor for Advanced Glycation End Products/antagonists & inhibitors , Serum Albumin, Bovine/pharmacology , Signal Transduction/immunology , Toll-Like Receptor 4/antagonists & inhibitors , Tumor Necrosis Factor-alpha/biosynthesisABSTRACT
BACKGROUND AND OBJECTIVE: Although chronic periodontitis (CP) is a multifactorial condition, few studies have investigated the potential association of gene variants with the outcome of periodontal therapy. In a previous study, we reported that variants in the interleukin-8 (IL8) gene were associated with CP in a Brazilian population. The aim of this nonrandomized study was to investigate whether genetic susceptibility to CP, conferred by the presence of the IL8 ATC/TTC haplotype, influences the clinical outcomes of nonsurgical periodontal therapy and the IL-8 protein levels in the gingival crevicular fluid. MATERIAL AND METHODS: Forty-one individuals were grouped according to the presence (susceptible to CP; n = 21) or absence (not susceptible to CP; n = 20) of the IL8 ATC/TTC haplotype. These individuals received nonsurgical periodontal therapy from one periodontist, who was blinded to the genetic status of each patient, and follow up continued for 45 d. The clinical parameters and gingival crevicular fluid samples were collected at baseline and on day 45. The IL-8 levels were determined by an ELISA. The data were subjected to the Mann-Whitney U-test, Wilcoxon and Spearman tests and to multiple logistic-regression analysis. RESULTS: No significant differences between patients with or without the IL8 ATC/TTC haplotype were found for the outcome of nonsurgical periodontal therapy and IL-8 levels. The multiple logistic-regression analysis did not show a statistically significant association between the IL8 haplotype and the variables studied. CONCLUSION: In this longitudinal clinical study, we observed that neither the outcome of nonsurgical periodontal therapy nor the IL-8 levels were influenced by the IL8 ATC/TTC CP-susceptibility haplotype. Additional studies of CP patients from other ethnic populations are necessary to confirm these results.
Subject(s)
Adenine , Chronic Periodontitis/therapy , Cytosine , Haplotypes/genetics , Interleukin-8/genetics , Thymine , Adult , Chronic Periodontitis/genetics , Dental Plaque Index , Female , Follow-Up Studies , Genetic Predisposition to Disease/genetics , Gingival Crevicular Fluid/chemistry , Gingival Crevicular Fluid/immunology , Humans , Longitudinal Studies , Male , Middle Aged , Periodontal Attachment Loss/therapy , Periodontal Index , Periodontal Pocket/therapy , Single-Blind Method , Treatment OutcomeABSTRACT
Periodontitis is an inflammatory disease that results from an interaction between dental biofilm agents and the host immune-inflammatory response. Periodontopathogenic organisms, such as Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola, as well as the host's susceptibility, represented by the host's genetic makeup, are the key factors that influence this complex disease. Recently, we identified haplotypes in the IL4 gene that were associated with chronic periodontitis (CP). This study aimed to evaluate whether subjects with different IL4 haplotypes (TCI/CCI and TTD/CTI) would be differentially colonized by periodontopathogens and whether they would respond differently to non-surgical periodontal therapy. Thirty-nine patients carrying the IL4 haplotype of genetic susceptibility to CP (IL4+) or protection against CP (IL4-) were evaluated. Those groups were further subdivided into individuals with CP (CP IL4+ or CP IL4-) and those that were periodontally healthy (H) (H IL4+ or H IL4-). CP patients were submitted to non-surgical periodontal therapy. Clinical and microbiological analyses were performed considering the data at baseline and 45 and 90 days after periodontal therapy. Periodontopathogens levels were evaluated by absolute quantitative polymerase chain reaction (qPCR). The baseline data revealed that the total levels of periodontopathogens were higher in the CP IL4+ than in the CP IL4- groups. Clinical analyses revealed that the periodontal therapy was equally effective, independent of the subject's IL4 genetic load. The TCI/CCI IL4 haplotype, previously associated with genetic susceptibility to CP, was also associated with increased levels of periodontopathogenic bacteria, but this genetic background did not influence the response to non-surgical periodontal treatment.
Subject(s)
Chronic Periodontitis/microbiology , Interleukin-4/genetics , Adult , Bacterial Load , Bacteroides/isolation & purification , Chi-Square Distribution , Chronic Periodontitis/genetics , Chronic Periodontitis/immunology , Chronic Periodontitis/therapy , Female , Genetic Predisposition to Disease , Haplotypes , Humans , Interleukin-4/immunology , Male , Middle Aged , Porphyromonas gingivalis/isolation & purification , Treatment Outcome , Treponema denticola/isolation & purificationABSTRACT
Chronic periodontitis (CP) is considered to be a multifactorial disease influenced by microbial and genetic factors. The aim of the present study was to investigate whether the genetic susceptibility to CP in individuals with the IL8 ATC/TTC haplotype is associated with subgingival levels of periodontopathogens. Sixty-five individuals, grouped according to the presence (n = 28) or absence (n = 37) of the IL8 haplotype, were evaluated. After clinical periodontal evaluation, each group was subdivided according to the presence (CP) or absence (H) of periodontitis. Four subgingival samples were obtained from CP and two samples per subject from H patients. The levels and proportions of Porphyromonas gingivalis, Tannerella forsythia, and Treponema denticola were analyzed using quantitative real-time polymerase chain reaction (q-PCR). No differences were found in the proportion of periodontopathogenic bacteria between groups with the presence or absence of the IL8 haplotype. However, in the CP groups, the levels of periodontopathogens were significantly higher in the individuals without the IL8 haplotype than in the individuals with the IL8 haplotype. These results suggest that periodontal destruction may occur in patients who are considered to be genetically susceptible to CP with a lower microbial challenge because of the presence of the IL8 ATC/TTC haplotype than in patients without this haplotype.
Subject(s)
Bacterial Load , Bacteroidetes/isolation & purification , Chronic Periodontitis/immunology , Genetic Predisposition to Disease , Interleukin-8/genetics , Porphyromonas gingivalis/isolation & purification , Treponema denticola/isolation & purification , Adult , Bacteroidetes/immunology , Chronic Periodontitis/microbiology , Female , Haplotypes , Humans , Interleukin-8/immunology , Male , Middle Aged , Porphyromonas gingivalis/immunology , Real-Time Polymerase Chain Reaction , Treponema denticola/immunologyABSTRACT
The type of fluid used during resuscitation may have an important impact on tissue edema. We evaluated the impact of two different regimens of fluid resuscitation on hemodynamics and on lung and intestinal edema during splanchnic hypoperfusion in rabbits. The study included 16 female New Zealand rabbits (2.9 to 3.3 kg body weight, aged 8 to 12 months) with splanchnic ischemia induced by ligation of the superior mesenteric artery. The animals were randomized into two experimental groups: group I (N = 9) received 12 mL x kg-1 x h-1 lactated Ringer solution and 20 mL/kg 6% hydroxyethyl starch solution; group II (N = 7) received 36 mL x kg-1 x h-1 lactated Ringer solution and 20 mL/kg 0.9% saline. A segment from the ileum was isolated to be perfused. A tonometric catheter was placed in a second gut segment. Superior mesenteric artery (Q SMA) and aortic (Qaorta) flows were measured using ultrasonic flow probes. After 4 h of fluid resuscitation, tissue specimens were immediately removed for estimations of gut and lung edema. There were no differences in global and regional perfusion variables, lung wet-to-dry weight ratios and oxygenation indices between groups. Gut wet-to-dry weight ratio was significantly lower in the crystalloid/colloid-treated group (4.9 +/- 1.5) than in the crystalloid-treated group (7.3 +/- 2.4) (P < 0.05). In this model of intestinal ischemia, fluid resuscitation with crystalloids caused more gut edema than a combination of crystalloids and colloids.
Subject(s)
Edema/etiology , Hydroxyethyl Starch Derivatives/administration & dosage , Ischemia/therapy , Isotonic Solutions/administration & dosage , Mesenteric Vascular Occlusion/therapy , Resuscitation/methods , Animals , Disease Models, Animal , Edema/pathology , Female , Hydroxyethyl Starch Derivatives/adverse effects , Intestinal Diseases/etiology , Intestinal Diseases/pathology , Intestinal Mucosa/blood supply , Intestinal Mucosa/pathology , Ischemia/pathology , Isotonic Solutions/adverse effects , Lung Diseases/etiology , Lung Diseases/pathology , Mesenteric Vascular Occlusion/pathology , Rabbits , Random Allocation , Resuscitation/adverse effects , Ringer's Lactate , Severity of Illness Index , Splanchnic CirculationABSTRACT
The type of fluid used during resuscitation may have an important impact on tissue edema. We evaluated the impact of two different regimens of fluid resuscitation on hemodynamics and on lung and intestinal edema during splanchnic hypoperfusion in rabbits. The study included 16 female New Zealand rabbits (2.9 to 3.3 kg body weight, aged 8 to 12 months) with splanchnic ischemia induced by ligation of the superior mesenteric artery. The animals were randomized into two experimental groups: group I (N = 9) received 12 mL·kg-1·h-1 lactated Ringer solution and 20 mL/kg 6 percent hydroxyethyl starch solution; group II (N = 7) received 36 mL·kg-1·h-1 lactated Ringer solution and 20 mL/kg 0.9 percent saline. A segment from the ileum was isolated to be perfused. A tonometric catheter was placed in a second gut segment. Superior mesenteric artery (Q SMA) and aortic (Qaorta) flows were measured using ultrasonic flow probes. After 4 h of fluid resuscitation, tissue specimens were immediately removed for estimations of gut and lung edema. There were no differences in global and regional perfusion variables, lung wet-to-dry weight ratios and oxygenation indices between groups. Gut wet-to-dry weight ratio was significantly lower in the crystalloid/colloid-treated group (4.9 ± 1.5) than in the crystalloid-treated group (7.3 ± 2.4) (P < 0.05). In this model of intestinal ischemia, fluid resuscitation with crystalloids caused more gut edema than a combination of crystalloids and colloids.
Subject(s)
Animals , Female , Rabbits , Edema/etiology , Hydroxyethyl Starch Derivatives/administration & dosage , Ischemia/therapy , Isotonic Solutions/administration & dosage , Mesenteric Vascular Occlusion/therapy , Resuscitation/methods , Disease Models, Animal , Edema/pathology , Hydroxyethyl Starch Derivatives/adverse effects , Intestinal Diseases/etiology , Intestinal Diseases/pathology , Intestinal Mucosa/blood supply , Intestinal Mucosa/pathology , Ischemia/pathology , Isotonic Solutions/adverse effects , Lung Diseases/etiology , Lung Diseases/pathology , Mesenteric Vascular Occlusion/pathology , Random Allocation , Resuscitation/adverse effects , Severity of Illness Index , Splanchnic CirculationABSTRACT
Os autores avaliaram o efeito do tratamento ortopédico para correçäo de maloclusäo Classe II divisäo 1ª de Angle, e os resultados dessa correçäo, sobre a profundidade de sondagem do sulco gengival. Foram estudadas 26 crianças de 7 a 9 anos de idade de ambos os sexos, portadores desse tipo de maloclusäo, tratadas (13) e näo tratadas ortopedicamente (13), avaliadas quanto à profundidade de sondagem do sulco gengival na regiäo de incisivos superiores e inferiores, a intervalos de 3 meses por um período de 1 ano. Os dados obtidos submetidos à análise de variância, demostraram que o uso de ortopedia funcional influenciou na mensuraçäo da profundidade de sondagem com maiores valores para o grupo tratado, mantendo essa diferença mesmo após a correçäo da maloclusäo. Estes resultados levaram à conclusäo que a terapia com aparelhos ortopédicos contribui para maiores valores de profundidade de sondagem, provavelmente pelo efeito da movimentaçäo dentária, e que a correçäo da maloclusäo apesar de proporcionar melhor posicionamento de dentes e arcos, näo resultou em menores valores comparado ao grupo controle. Este efeito pode ser transitório, podendo ser minimizado ou desaparecer com a estabilizaçäo da posiçäo dentária e a instituiçäo de medidas efetivas de higiene e controle da placa bacteriana
