Expression of monocyte chemokine receptors in diabetes after non-surgical periodontal treatment: A pilot study.

The aim of this study was to evaluate the effect of non-surgical periodontal treatment in the expression of chemokine receptors, in individuals with Periodontitis, associated or not with Diabetes. Pilot study, which included patients (n = 45) with Periodontitis, associated (n = 25) or not (n = 20) with Diabetes, submitted to the non-surgical periodontal treatment for one month. The expression of chemokine receptors CCR2, CCR5, and CX3CR1 at the mRNA level was evaluated in the peripheral mononuclear cells, as well as the expression of these receptors at the protein level was verified in monocyte subtypes (classical, intermediate, and non-classical monocytes). There was higher expression of CCR2 and CCR5 receptors at the initial visit in the group with Diabetes, with no differences for CX3CR1 (p = 0.002; p = 0.018, and p = 0.896, respectively), without differences after treatment. There was higher expression of CCR2 and CCR5 proteins in the group with Diabetes at the initial visit for classical, intermediate, and nonclassical monocytes, with no differences for CX3CR1 (CCR2: p = 0.004; p = 0.026; p = 0.024; CCR5: 0.045; p = 0.045; p = 0.013; CX3CR1: p = 0.424; p = 0.944; p = 0.392, respectively), without differences after the end of treatment. Concerning each group separately, there were reductions in the expression of CCR2 as well as CCR5 in classical, intermediate, and nonclassical monocytes, and reduction of CX3CR1 in classical monocytes after treatment in the group with Diabetes (p = 0.003; p = 0.006; p = 0.039; p = 0.007; p = 0.006; p = 0.004; p = 0.019, respectively), without differences in the group without Diabetes. The expression of the chemokine receptors CCR2 and CCR5, in patients with Periodontitis associated with Diabetes, is favorably modified after the end of the non-surgical periodontal treatment.

Modulation of monocyte subtypes in diabetes after non-surgical periodontal treatment.

OBJECTIVES: The current study aims to evaluate the effect of non-surgical periodontal treatment on the modulation of monocyte phenotype, in the presence or absence of diabetes. MATERIALS AND METHODS: The identification, quantification, and phenotypic characterization of monocyte subtypes (classical, intermediate, and non-classical) were performed by flow cytometry, at baseline and 1 month after the end of non-surgical periodontal treatment, in patients with periodontitis, associated or not with diabetes. RESULTS: There was an increase in non-classical monocytes after treatment and a reduction in intermediate monocytes, without differences for the classical subtype, regardless of the diabetes status. Furthermore, there was a reduction in intermediate monocytes and an increase in non-classical and classical monocytes after treatment in the diabetes group, while no significant differences were observed for classical, intermediate, and non-classical monocytes in the group without diabetes. Comparisons between the two groups showed significant differences for classical, intermediate, and non-classical monocytes at baseline; these differences were not found one month after treatment. CONCLUSIONS: Non-surgical periodontal treatment leads to modulation of monocytes to a less inflammatory phenotype, especially in individuals with diabetes. CLINICAL RELEVANCE: A better understanding of the role of these biomarkers in the periodontitis contex may constitute a new strategic target for a better treatment of patiens with diabetes associated to periodontitis. CLINICAL TRIAL REGISTRATION: Brazilian Registry of Clinical Trials-RBR-35szwc. Jhefferson Miranda Alves and Danielle Borges Germano contributed equality to this study and should be considered first authors.

Accuracy of guided dental implant surgery using a fully digital workflow: A case series.

STATEMENT OF PROBLEM: Computer-guided implant surgery facilitated by intraoral scanning may enhance the efficiency of the digital workflow. However, it is necessary to assess technique accuracy to evaluate the accuracy of implant placement. PURPOSE: The purpose of this clinical study was to evaluate the accuracy of a virtual computer-aided design and computer-aided manufacturing (CAD-CAM) static guided surgery technique associated with intraoral scanning in partially edentulous participants by analyzing the overlap among preoperative and postoperative cone beam computed tomography (CBCT) scans, virtual planning, and the guided surgery performed. MATERIAL AND METHODS: Eleven partially edentulous participants underwent CBCT and intraoral scanning (TRIOS3). Data were integrated into a software program (ImplantViewer 3.5) for the virtual planning of implants and 3-dimensional (3D) printing of the prototype CAD-CAM surgical guide. A total of 18 implants were placed using the CAD-CAM static computer-aided implant surgery technique (Strong SW). After 15 days, postoperative CBCT scans were made and 4 variables (angular, coronal, apical, and vertical deviation) were measured to compare the virtually planned implants and the implants placed by analyzing the overlap between preoperative and postoperative of the virtual planning and guided surgery performed using the ImplantViewer 3.5 and Rhino 6 software programs. RESULTS: Deviations were found in all parameters analyzed. The mean angular deviation was 2.68 ±1.62 degrees; mean coronal deviation, 0.82 ±0.44 mm; mean apical deviation, 1.14 ±0.44 mm; and mean vertical deviation, 0.62 ±0.44 mm. CONCLUSIONS: The implants placed using the CAD-CAM static guided surgery technique associated with intraoral scanning in partially edentulous participants exhibited angular and linear deviations when compared with virtual planning implants. However, these deviations were not clinically significant.

Topical application of lectin Artin M improves wound healing in defects created in the palatal mucosa: an in vivo study in dogs.

Previous studies have shown that topical application of lectin Artin-M accelerates wound healing in the rat oral mucosa. The aim of this study was to evaluate, by means of histology and immunohistochemistry (IHC) the effects of Artin-M on wound healing in the palatal mucosa in dogs. Three full thickness wounds of 6 mm diameter were surgically created in the palatal mucosa of twenty dogs and randomly divided into three groups according to one of the treatment assigned: Group C-Control (coagulum); Group A-Artin-M gel; Group V-Vehicle (carboxymethylcellulose 3%). Each animal received all the three experimental treatments. Afterwards, four animals were killed at 2, 4, 7, 14 and 21 days post-surgery. Wounded areas were photographed and scored for macroscopic evaluation. Biopsies were harvested and used for descriptive histological analysis, proliferating cell nuclear antigen IHC and measurement of myeloperoxidase activity. The results demonstrated faster wound closure in group A in comparison to the other groups in all the periods evaluated. Histological analyses exhibited improved re-epithelialization and collagen fiber formation resulting in faster maturation of granulation tissue in group A compared to the other groups by day 14. Treatment with Artin-M gel significantly induced cell proliferation and increased volumetric density of fibroblasts at day 2 and 4 (p < 0.05). Neutrophil infiltration in group A was significantly higher than the other groups (p < 0.05) at the same time points. Collectively, our findings demonstrated that Artin-M may potentially favor wound healing on palatal mucosa lesions via recruitment of neutrophils and promotion of cell proliferation.

Role of circulating microparticles and cytokines in periodontitis associated with diabetes.

Background: Periodontitis is a chronic inflammatory condition that affects the supporting tissues of the teeth, and can lead to serious complications such as tooth loss and systemic health problems, including diabetes, which have a bidirectional relationship with periodontitis. Circulating microparticles originate from different cell types after stimuli such as activation or apoptosis. Interleukins are related to processes in the regulation of the immune response, inflammation, and cell growth. This study aimed to evaluate circulating microparticles as well as interleukins in the plasma, at baseline and 1 month after the end of the non-surgical periodontal treatment. Methods: Samples were collected from 45 patients, with moderate to severe periodontitis with diabetes (N = 25) and without diabetes (N = 20). Microparticles were evaluated in the platelet-poor plasma by flow cytometer. Cytokine levels were evaluated by the enzyme immunoabsorption assay (ELISA). Results: Higher levels of the pro-inflammatory cytokines were found in the group with diabetes compared to the non-diabetic group both at baseline and 1 month after the end of the treatment. A higher IL-6/IL-10 ratio was found in patients with diabetes compared to the group without diabetes at T0 and T1, whereas an increased IFN-γ/IL-10 ratio was only found at T1 in patients with diabetes in comparison to the group without diabetes. In the group with diabetes, it was verified positive correlations between IL-10 and IL-6 or IFN-γ and a negative correlation between IL-6 and PMP, at T0; in contrast, in the T1, negative correlations were found between TNF-α and IL-10 or PMP. Besides, at T0, it was evidenced positive correlations both between circulating TNF-α and IL-6, and IL-10 and EMP, as well as a negative correlation between IL-10 and PMP in the group with diabetes. In addition, it was observed in T1 positive correlations between levels of TNF-α and IL-6, IFN-γ, or IL-10, and between PMP and IFN-γ, and between EMP and IL-6, TNF-α and IFN-γ in this group. Conclusion: The results suggest a modulatory effect of the periodontitis associated with diabetes, as well as the periodontal treatment, in the systemic inflammatory status of the participants of the study.

SOCS3 expression correlates with severity of inflammation, expression of proinflammatory cytokines, and activation of STAT3 and p38 MAPK in LPS-induced inflammation in vivo.

SOCS3 is an inducible endogenous negative regulator of JAK/STAT pathway, which is relevant in inflammatory conditions. We used a model of LPS-induced periodontal disease in rats to correlate SOCS3 expression with the inflammatory status. In vitro we used a murine macrophage cell line to assess the physical interaction between SOCS3 and STAT3 by coimmunoprecipitation. 30 ug of LPS from Escherichia coli were injected in the gingival tissues on the palatal aspect of first molars of the animals 3x/week for up to 4 weeks. Control animals were injected with the vehicle (PBS). The rats were sacrificed at 7, 15, and 30 days. Inflammation and gene expression were assessed by stereometric analysis, immunohistochemistry, RT-qPCR, and western blot. LPS injections increased inflammation, paralleled by an upregulation of SOCS3, of the proinflammatory cytokines IL-1 ß , IL-6, and TNF- α and increased phosphorylation of STAT3 and p38 MAPK. SOCS3 expression accompanied the severity of inflammation and the expression of proinflammatory cytokines, as well as the activation status of STAT3 and p38 MAPK. LPS stimulation in a macrophage cell line in vitro induced transient STAT3 activation, which was inversely correlated with a dynamic physical interaction with SOCS3, suggesting that this may be a mechanism for SOCS3 regulatory function.

Dental implants in patients seropositive for HIV: A 12-year follow-up study.

BACKGROUND AND OVERVIEW: Patients seropositive HIV and AIDS represent a group of patients who experience longer longevity at the expense of effective therapies for infection control and related opportunistic diseases. However, the prolonged use of these drugs is often associated with adverse events, which theoretically may influence dental management and the long-term stability of dental implants. The objective of this study was to prospectively evaluate a group of HIV-positive people from a previous study who had received dental implants for 12 years after oral rehabilitation and functional loading. CASE DESCRIPTION: Nine patients with a total of 18 implants participated in this study. Viral load was undetectable in 8 patients, with 1 who had 48 copies/milliliter. The cluster of differentiation 4 T lymphocyte count ranged from 227 through 1,000 cells/cubic millimeter, mean (standard deviation [SD]) 564 (271.13) cells/mm3. Five of the 9 (55.5%) patients had visible plaque, and 5 (55.5%) had bleeding on probing with no implant mobility. Radiographs obtained at 6 months, 12 months, and 12 years of functional loading showed mean (SD) marginal bone losses of 0.32 (0.23) mm, 0.37 (0.23) mm, and 2.43 (1.48), respectively. CONCLUSION AND PRACTICAL IMPLICATIONS: These results suggest that dental implant treatment in HIV-positive patients achieved long-term survival, with a success rate comparable with that observed in healthy patients, indicating that implant rehabilitation is not a contraindication for HIV-positive patients.

The Importance of Histopathological Examination to the Final Diagnosis of Peripheral Odontogenic Tumors: A Case Report of a Peripheral Odontoma.

A 30-year-old Caucasian man presented with an 18-month history of an asymptomatic calcified mass, located on the buccal side of the alveolar ridge. Medical records did not present any underlying conditions. On intraoral examination, the lesion was located on the right side of the maxilla, showing mucosal fenestration with mineralized tissue measuring approximately 1 cm in diameter. Radiographic examination showed multiple radiopaque masses. Incisional biopsy was performed, and histological analysis revealed a presence of enamel matrix, dentin, and cementum, resembling tooth-like structures. Surgical removal was offered after the diagnostic confirmation of peripheral odontoma, but the patient refused because of the asymptomatic nature of the lesion.

Bone autografts & allografts placed simultaneously with dental implants in rabbits.

OBJECTIVE: This study compared stability, removal torque, bone implant contact (BIC) and area (BA) of implants installed simultaneously with onlay autografts or allografts in rabbits' tibias. MATERIAL AND METHODS: Total of 18 rabbits were used in this study. Fresh frozen allografts were obtained from six animals at T(-6). Two implants with autogenous grafts (Group 1) or allografts (Group 2) were simultaneously inserted into both sides of the tibiae in a vertical periimplant defect model at T0. The resonance frequency (ISQ) was measured in implant proximal epiphysis on the day of installation of T0 and T18 (18 weeks post-surgery). At T18 the removal torque was assessed at the distal implants, the implants' proximal epiphysis and surrounding bone were harvested to perform histomorphometric analysis. The BIC and BA within the limits of the implants threads were evaluated. RESULTS: The ISQ revealed a statistically significant difference between T0 and T18 in each group (p = 0.024, p = 0.003). The removal torque indicates that there was no significant difference between the two groups (p = 0.47). No significant differences were observed between the groups regarding both BIC (p = 0.3713) and the BA (p = 0.3883). CONCLUSION: Both grafts and implants demonstrated the same stability, torque removal and the BIC and BA.

Vertical Bone Augmentation Using Deproteinized Bovine Bone Mineral, Absorbable Collagen Sponge, and Recombinant Human Bone Morphogenetic Protein-2: An In Vivo Study in Rabbits.

PURPOSE: The objective of this investigation was to assess vertical bone augmentation using deproteinized bovine bone mineral (DBBM) infused or not with recombinant human bone morphogenetic protein (rhBMP-2) in rabbit tibiae. MATERIALS AND METHODS: A total of 18 female rabbits (New Zealand) received two blocks of DBBM in each tibia. The DBBM blocks were randomly assigned into four experimental groups: DBBM (only the bone graft); DBBM associated with absorbable collagen sponge (ACS); DBBM plus rhBMP-2 (1.5 mg/mL); and DBBM infused with rhBMP-2 (1.5 mg/mL) in an ACS carrier. Animals were sacrificed after 12 weeks, and the tibiae containing the DBBM blocks were dissected and analyzed radiographically (microcomputed tomography [micro-CT]), histologically, and immunohistochemically. RESULTS: Micro-CT analysis showed a considerable increase in bone volume (BV) and BV/tissue volume in the rhBMP-2/ACS group compared with all the others. Trabeculae thickness also increased in the rhBMP-2/ACS group compared with the DBBM/ACS group. Trabecular number, separation, and bone mineral density were not different among groups. Histomorphometric evaluation showed increased newly formed bone in the rhBMP-2/ACS group compared with the DBBM and DBBM/ACS groups. The amount of residual bone graft was statistically higher in the rhBMP-2 groups compared with the DBBM/ACS group. Immunohistochemical analysis showed that the vascular endothelial growth factor (VEGF) expression was more intense in the rhBMP-2/ACS group compared with the DBBM/ACS group. The immunopositivity for type 1 collagen tended to be higher in the two groups with rhBMP-2. CONCLUSION: Collectively, the results of this study suggest that the addition of rhBMP-2 in an ACS carrier placed on top of the DBBM graft enhanced bone formation in this animal model.

Evaluation of periodontal condition and risk in patients with chronic kidney disease on hemodialysis.

OBJECTIVE: To establish a profile of periodontal conditions in chronic kidney disease patients on hemodialysis and their periodontal risk. METHODS: We included 115 patients on hemodialysis. Clinical periodontal parameters assessed were: plaque index, gingival index, probing depth and clinical attachment level. Patients were classified according to presence/absence and severity of periodontal disease and periodontal risk. RESULTS: In 107 dentate patients (93%) the plaque index was 1.53±0.78, the gingival index was 0.95±0.85, the probing depth was 2.2±0.6mm and the clinical attachment level was 3.18±1.75mm. We observed that 1 patient (0.94%) did not have periodontal disease, 55 patients (51.40%) had slight, 28 (26.17%) moderate and 23 (21.49%) severe periodontal disease. Among 107 patients, 37 (34.58%) had low risk, 35 (32.71%) moderate risk and 35 (32.71%) high risk. Patients with severe periodontal disease showed 104.5 more chance of high risk compared with low risk individuals (odds ratio: 104.5; 95%CI: 10.7-1017.2; p<0.0001). CONCLUSION: Most of patients with chronic renal disease presented periodontal disease, indicating the presence of chronic inflammatory and infection process that may influence in systemic conditions. A prevention and interventionist approach in this population is needed, especially to emphasize the importance of oral health. The periodontal risk assessment is a useful tool to create individualized periodontal therapies and to improve general health condition. OBJETIVO: Traçar um perfil das condições periodontais de pacientes com doença renal crônica em hemodiálise e seu risco periodontal. MÉTODOS: Foram incluídos no estudo 115 pacientes em hemodiálise. Os parâmetros clínicos periodontais avaliados foram: índice de placa, índice gengival, profundidade de sondagem e perda de nível de inserção clínico. Os pacientes foram classificados de acordo com a presença e a gravidade da doença periodontal, bem como de acordo com o risco periodontal. RESULTADOS: Quanto aos parâmetros clínicos, 107 pacientes dentados (93%) apresentaram média de índice de placa de 1,53±0,78 e gengival de 0,95±0,85, profundidade de sondagem de 2,2±0,6mm e nível de inserção clínica de 3,18±1,75mm. Em relação à doença periodontal, 1 (0,94%) era saudável, 55 (51,40%) apresentavam periodontite leve, 28 (26,17%) moderada e 23 (21,49%) avançada. Com relação ao risco, dos 107 pacientes, 37 apresentavam risco baixo, 35 moderado e 35 alto. Os pacientes com doença periodontal avançada apresentaram 104,5 vezes mais chance de ter alto risco comparado ao baixo (odds ratio: 104,5; IC95%: 10,7-1017,2; p<0,0001). CONCLUSÃO: A maioria dos pacientes com doença renal crônica apresentou doença periodontal, indicando um processo infeccioso e inflamatório crônico, que pode influenciar na condição sistêmica. Evidencia-se a necessidade de uma abordagem preventiva e intervencionista nesta população, enfatizando a importância da saúde bucal. A avaliação do risco periodontal seria uma ferramenta na elaboração de terapias periodontais individualizadas para uma melhor condição de saúde geral.

Improvement of an anterior infrabone defect using combined periodontal and orthodontic therapy: A 6-year follow-up case report.

Extensive intraosseous lesions represent a clinical challenge for the periodontist. Sites with bone defects have been shown to be at higher risk of periodontitis progression in patients who had not received periodontal therapy. Thus, the aim of this case report was to describe a novel approach for the treatment of 1-walled intraosseous defect by combining nonsurgical periodontal therapy and orthodontic movement toward the bone defect, avoiding regenerative and surgical procedures. A 47-year-old woman underwent the proposed procedures for the treatment of her left central incisor with 9 mm probing depth and 1-walled intraosseous defect in its mesial aspect. Initially, basic periodontal therapy with scaling and root planning was accomplished. Two months later, an orthodontic treatment was planned to eliminate the intraosseous lesion and to improve the interproximal papillary area. Orthodontic root movement toward the osseous defect was performed for 13 months with light forces. After 6 years postoperative it was concluded that combined basic periodontal therapy and orthodontic movement was capable of eliminating the intraosseous defect and improve the esthetics in the interproximal papillary area between the central incisors.

Orthodontic force increases interleukin-1ß and tumor necrosis factor-α expression and alveolar bone loss in periodontitis.

BACKGROUND: The present study aims to evaluate the effects of orthodontic movement (OM) on the periodontal tissues of rats with ligature-induced periodontal disease. METHODS: Eighty-eight rats were divided into four groups: 1) negative control (sham operated); 2) periodontal disease; 3) OM; and 4) periodontal disease followed by OM (OMP). Rats were sacrificed 3 hours or 1, 3, or 7 days after OM commencement. Bone volume fraction (BVF) and bone mineral density (BMD) were assessed in hemimaxillae by microcomputed tomography analysis. Expression of the proinflammatory cytokines interleukin (IL)-1ß and tumor necrosis factor (TNF)-α were evaluated in gingival samples by quantitative polymerase chain reaction and enzyme-linked immunosorbent assay, and in the furcation region by immunohistochemistry analysis (IHC). RESULTS: The OMP group had lower BVF and BMD levels compared to the other groups at day 7 (P <0.05). Maximum messenger ribonucleic acid expression of both cytokines was observed in the OMP group at day 1 (P <0.05). In the same period, all proteins were expressed in high levels for all test groups compared to the control group. The number of cells positive for IL-1ß and TNF-α by IHC was highest in the OMP group at day 1, with progressive reduction thereafter. CONCLUSION: The results suggest that OM acts synergistically with periodontal disease in periodontal breakdown through upregulation of proinflammatory cytokines.

Removal of dental surgical bur from maxillary sinus: a case report

The most commonly performed surgical procedure in Oral and Maxillofacial Surgery practices are the removal of impacted third molars. Extensive training, skill and experience allow this procedure to be performed in an atraumatic approach. The aim of this study was to drawing attention to the importance of the correct management of the complications cases of foreign body inside maxillary sinus after surgical removal of maxillary third molars. This is an unusual clinical case of a dental surgical bur accidentally displacement into the maxillary sinus during an upper third molar extraction surgery. After removal, the clinical case showed a satisfactory repair emphasizing the importance of a meticulous clinical examination to achieve a correct diagnosis and an appropriate treatment plan, which is essential for a favorable prognosis

Evaluation of periodontal condition and risk in patients with chronic kidney disease on hemodialysis / Avaliação da condição e risco periodontal em pacientes com doença renal crônica em hemodiálise

ABSTRACT Objective To establish a profile of periodontal conditions in chronic kidney disease patients on hemodialysis and their periodontal risk. Methods We included 115 patients on hemodialysis. Clinical periodontal parameters assessed were: plaque index, gingival index, probing depth and clinical attachment level. Patients were classified according to presence/absence and severity of periodontal disease and periodontal risk. Results In 107 dentate patients (93%) the plaque index was 1.53±0.78, the gingival index was 0.95±0.85, the probing depth was 2.2±0.6mm and the clinical attachment level was 3.18±1.75mm. We observed that 1 patient (0.94%) did not have periodontal disease, 55 patients (51.40%) had slight, 28 (26.17%) moderate and 23 (21.49%) severe periodontal disease. Among 107 patients, 37 (34.58%) had low risk, 35 (32.71%) moderate risk and 35 (32.71%) high risk. Patients with severe periodontal disease showed 104.5 more chance of high risk compared with low risk individuals (odds ratio: 104.5; 95%CI: 10.7-1017.2; p<0.0001). Conclusion Most of patients with chronic renal disease presented periodontal disease, indicating the presence of chronic inflammatory and infection process that may influence in systemic conditions. A prevention and interventionist approach in this population is needed, especially to emphasize the importance of oral health. The periodontal risk assessment is a useful tool to create individualized periodontal therapies and to improve general health condition.

RESUMO Objetivo Traçar um perfil das condições periodontais de pacientes com doença renal crônica em hemodiálise e seu risco periodontal. Métodos Foram incluídos no estudo 115 pacientes em hemodiálise. Os parâmetros clínicos periodontais avaliados foram: índice de placa, índice gengival, profundidade de sondagem e perda de nível de inserção clínico. Os pacientes foram classificados de acordo com a presença e a gravidade da doença periodontal, bem como de acordo com o risco periodontal. Resultados Quanto aos parâmetros clínicos, 107 pacientes dentados (93%) apresentaram média de índice de placa de 1,53±0,78 e gengival de 0,95±0,85, profundidade de sondagem de 2,2±0,6mm e nível de inserção clínica de 3,18±1,75mm. Em relação à doença periodontal, 1 (0,94%) era saudável, 55 (51,40%) apresentavam periodontite leve, 28 (26,17%) moderada e 23 (21,49%) avançada. Com relação ao risco, dos 107 pacientes, 37 apresentavam risco baixo, 35 moderado e 35 alto. Os pacientes com doença periodontal avançada apresentaram 104,5 vezes mais chance de ter alto risco comparado ao baixo (odds ratio: 104,5; IC95%: 10,7-1017,2; p<0,0001). Conclusão A maioria dos pacientes com doença renal crônica apresentou doença periodontal, indicando um processo infeccioso e inflamatório crônico, que pode influenciar na condição sistêmica. Evidencia-se a necessidade de uma abordagem preventiva e intervencionista nesta população, enfatizando a importância da saúde bucal. A avaliação do risco periodontal seria uma ferramenta na elaboração de terapias periodontais individualizadas para uma melhor condição de saúde geral.

Association of haplotypes in the IL8 gene with susceptibility to chronic periodontitis in a Brazilian population.

BACKGROUND: Interleukin 8 (IL-8) is a chemokine related to the initiation and amplification of acute and chronic inflammatory processes. Polymorphisms in the IL8 gene have been associated with inflammatory diseases. We investigated whether the -845(T/C) and -738(T/A) single nucleotide polymorphisms (SNPs) in the IL8 gene, as well as the haplotypes they form together with the previously investigated -353(A/T), are associated with susceptibility to chronic periodontitis. METHODS: DNA was extracted from buccal epithelial cells of 400 Brazilian individuals (control n=182, periodontitis n=218). SNPs were genotyped by the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method. Disease associations were analyzed by the chi(2) test, Exact Fisher test and Clump program. Haplotypes were reconstructed using the expectation-maximization algorithm and differences in haplotype distribution between the groups were analyzed to estimate genetic susceptibility for chronic periodontitis development. RESULTS: When analyzed individually, no SNPs showed different distributions between the control and chronic periodontitis groups. Although, nonsmokers carrying the TTA/CAT (OR=2.35, 95% CI=1.03-5.36) and TAT/CTA (OR=6.05, 95% CI=1.32-27.7) haplotypes were genetically susceptible to chronic periodontitis. The TTT/TAA haplotype was associated with protection against the development of periodontitis (for nonsmokers OR=0.22, 95% CI=0.10-0.46). CONCLUSION: Although none of the investigated SNPs in the IL8 gene was individually associated with periodontitis, some haplotypes showed significant association with susceptibility to, or protection against, chronic periodontitis in a Brazilian population.

Avaliação do efeito da Artin M no processo de reparação em mucosa mastigatória / Evaluation of Artin M on wound healing in palatal mucosa

Artin M é uma lectina purificada de sementes de Artocarpus heterophyllus que, recentemente, mostrou-se capaz depromover aceleração da cicatrização de lesões por queimadura de pele ou por abrasão da córnea em ratos e coelhos. O objetivo desta tese foi avaliar os efeitos da Artin M no processo de reparação da mucosa mastigatória bucal, por meio de modelos de estudo in vivo. Primeiro estudo: Três feridas cirúrgicas circulares de 6 mm de diâmetro foram criadas na mucosa palatina de 20 cães, e divididas aleatoriamente em 3 grupos de acordo com os tratamentos: C ­ controle (não tratados), A - Artin M, V - veículo. Quatro animais de cada grupo foram sacrificados após 2, 4, 7, 14 e 21 dias pós tratamento e suas maxilas analisadas clinicamente quanto ao padrão de cicatrização, seguido de análise histológica, imuno-histoquímica para antígeno nuclear de proliferação celular (PCNA) e atividade de mieloperoxidase. Clinicamente, o grupo A demonstrou melhor cicatrização em todos os períodos quando comparada aos outros grupos (p<0,05). A análise histológica mostrou ter havido no grupo A maior estimulação na produção de fibras colágenas, maturação do tecido de granulação e organização do epitélio. A imunolocalização de PCNA mostrou uma maior tendência na proliferação celular em lesões do grupo A principalmente nos dias iniciais (p<0,05). O influxo de neutrófilos mostrou-se estatisticamente aumentado no grupo A quando comparado aos outros grupos nos dias 2 e 4 (p<0,05). Conclui-se que a Artin M promoveu aceleração na reparação das feridas na mucosa mastigatória em cães, via recrutamento de neutrófilos e indução da proliferação celular. É bem conhecido o envolvimento de mediadores biológicos como citocinas e fatores de crescimento liberados pelas diferentes células no processo de reparação tecidual. Portanto, tendo em vista os resultados obtidos no primeiro modelo visamos avaliar a resposta ao tratamento tópico do gel de Artin M na mucosa mastigatória de rato. Segundo estudo: Feridas cirúrgicas circulares de 4 mm de diâmetro foram criados na mucosa palatina de 72 ratos Wistar, divididos aleatoriamente de acordo com o tratamento realizado nas lesões em 3 grupos: C ­ controle (não tratados), A -Artin M, V - veículo. Após 3, 5 e 7 dias pós-tratamento, 8 animais de cada grupo foram sacrificados e suas hemimaxilas foram divididas para realização da análise histológica; imuno-histoquímica para PCNA, bFGF, VEGF e TGF-ß; expressão protéica das citocinas pró-inflamatórias IL-1, IL-6 e TNFα, e dos fatores de crescimento VEGF e TGF-ß por ELISA. A análise histológica mostrou que no grupo A houve estimulação na regeneração e organização do epitélio, na produção de fibras colágenas e maturação do tecido de granulação e diminuição estatisticamente significante no infiltrado inflamatório no dia 7 quando comparado aos outros grupos (p<0,05). A análise imuno-histoquímica mostrou maior expressão de TGF-ß e VEGF no dia 3 (p<0,05) e maior proliferação celular no dia 5 (p<0,05) no grupo A. Além disso, maior expressão protéica de TGF-ß e VEGF nas lesões tratadas pelo Artin M foi confirmada pela ELISA no dia 3 (p<0,05). Diante dos resultados obtidos, concluiu-se que a aplicação tópica de gel de Artin M promoveu a reparação da mucosa mastigatória de ratos através estimulação na produção de TGF-ß e VEGF, proliferação celular, deposição de colágeno e reepitelização

Artin M, lectin from Artocarpus heterophyllus seeds, has been demonstrated to stimulate recruitment and activation of neutrophil and mast cells. Furthermore, it has been shown to accelerate the process of wound healing on burn injuries and corneal epithelial lesions in rats and rabbits, respectively. The aim of this study was to evaluate the effects of Artin M on wound healing in palatal mucosa in two animal models. Study 1: Three wounds of 6 mm full thickness were surgically created in the hard palate mucosa of twenty dogs. The wounds of each animal were randomly divided into three groups according to one of the treatment assigned: C­ Control (nontreated), A- Artin M gel, and V­ Vehicle (carboxymethylcellulose 3% gel). Four animals per group were sacrificed at 2, 4, 7, 14 and 21 days post-surgery. Before sacrifice, wounded areas were photographed and scored for macroscopic healing evaluation. Afterwards, maxillary tissue were harvested and divided to perform histological analysis, immunohistochemical staining against Proliferating Cell Nuclear Antigen (PCNA) and measurement of myeloperoxidase activity. Clinical analysis showed that wound closure was accelerated in group A in comparison to other groups in all periods. Histological features showed enhanced reepithelization and collagen fiber formation resulting in faster maturation of granular tissue in group A compared to the other groups, by day 14. Artin M treatment significantly induced cells proliferation at day 2 and 4 (p<0.05). Neutrophils infiltration in the group A was significantly higher than in the other groups at days 2 and 4 (p<0.05). The results suggest that Artin M gel may potentially facilitate wound healing on palatal mucosa via the recruitment of neutrophils and promotion of cells proliferation. It is well known that cytokines and growth factors released from different cells coordinate the healing process. Therefore, based on previous results we evaluated the effects of topical application of Artin M on wound healing in palatal mucosa of rats. Study 2: Seventy-two rats were divided into three groups according to one of the treatment assigned: C­ Control (non-treated), A- Artin M gel, and V­ Vehicle (carboxymethylcellulose 3% gel). A 4 mm full thickness wound was surgically created on the palatal mucosa of each animal. Eight animals per group were sacrificed at 3, 5, and 7 days post-surgery. Maxillary biopsies were harvested and divided to perform histological analysis, imunohistochemical analysis for bFGF, PCNA TGF-ß and VEGF and ELISA assay to evaluate levels of IL-1, IL-6, TNFα, TGF-ß and VEGF. Histological features showed faster reepithelization in group A, significant decrease in inflammatory cells infiltration by day 7 (p<0.05) and increased collagen fiber formation resulting in faster maturation of granular tissue compared to the other groups. Also, Artin M significantly induced cells proliferation at day 5 (p<0.05). Significantly higher expression of TGF-ß and VEGF were observed in group A compared to C at day 3 (p<0.05). In conclusion, the single application of Artin M gel has shown to be effective in the healing of oral mucosa wound in rats by enhancing TGF-ß and VEGF release, cell proliferation, reepithelization, collagen deposition and arrangement of collagen fibers. These findings may partially explain potential pharmacological actions of Artin M on promoting wound healing process

Investigação de polimorfismos no promotor do gene interleucina 8 em indivíduos com periodontite / Investigation of polymorphisms in the promoter region of interleukin 8 in Brazilian individuals with periodontitis

O estudo investigou se há associação individual dos polimorfismos –353(A/T), -738(T/A) e -845(T/C) na região promotora do gene interleucina 8 (IL8), bem como de seus haplótipos, com suscetibilidade à periodontite em indivíduos Brasileiros. Foram selecionados 500 indivíduos de ambos os gêneros (Grupo Controle n=224 e Grupo Periodontite n=276) que procuraram atendimento na Faculdade de Odontologia de Araraquara. A partir do DNA obtido de células da mucosa oral, os polimorfismos -738(T/A) e -845(T/C) foram analisados por PCR-RFLP (Polymerase Chain Reaction – Restriction Fragment Length Polymorphism), e o -353(A/T) por SSP-PCR (Sequence Specific Primer – PCR). Os fragmentos obtidos foram submetidos à eletroforese vertical em gel de poliacrilamida a 10%. Para analisar a freqüência dos genótipos e alelos foi utilizado o teste χ2, ou o programa CLUMP para os polimorfismos que mostraram alelos raros. Os haplótipos foram estimados pelo programa ARLEQUIN e a diferença na distribuição deles entre os grupos foi investigada por meio do programa CLUMP. Os resultados da análise individual dos polimorfismos não evidenciaram associação com a periodontite. Em relação aos haplótipos, as freqüências dos mesmos como alelos e/ou genótipos entre Grupo Controle e Grupo Periodontite apresentaram diferenças estatisticamente significantes considerando a população total e também a não fumante. Os indivíduos com o haplótipo CTA apresentaram-se 2,14 vezes mais susceptíveis à doença periodontal (p=0,0012, OR=2,14; 95% CI =1,6-3,38). Concluiu-se que houve associação dos haplótipos formados pelos polimorfismos –845(T/C), –738(T/A) e –353(A/T) no gene IL8 com a suscetibilidade a periodontite na população brasileira estudada.

The purpose of this study was to investigate whether there is an individual association of the –845(T/C), –738(T/A) and –353(A/T) single nucleotide polymorphisms (SNPs) in the IL8 gene, as well as their haplotypes with susceptibility to periodontitis in Brazilian population. DNA was extracted from buccal epithelial cells from 500 Brazilian individuals (Control group n = 224, Periodontitis group n = 276). The –845 and –738 SNPs were genotyped by PCR-RFLP method (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism). The –353 SNP was investigated using the Sequence Specific Primers Polymerase Chain Reaction method (SSP-PCR). Differences of the allelic and genotypic frequencies were assessed by Chi-squared test or by using the CLUMP program for rare alleles. Haplotypes were estimated using the ARLEQUIN program. Differences in the haplotype frequencies between the studied groups were assessed by the CLUMP program. No differences were observed in the allelic and genotypic distribution of all the investigated SNPs between control and periodontitis groups when they were analysed individually. The haplotypes distribution in the periodontitis and controls groups was significantly different both considering the total casuistic or the non-smokers subgroup only. Individuals with CTA haplotype seemed to be over twice more likely to develop periodontitis than individuals with other haplotypes (OR=2.14, 95% CI =1.36-3.38). The analysis of haplotypes constructed from –845(T/C), – 738(T/A) and –353 (A/T) polymorphisms in the IL8 gene demonstrated association with susceptibility to periodontitis in Brazilian individuals.

Investigação de polimorfismos no promotor do gene Interleucina 8 em indivíduos com periodontite / Investigation of polymorphisms in the promoter region of interleukin 8 in Brazilian individuals with periodontitis

O estudo investigou se há associação individual dos polimorfismos ­353(A/T), -738(T/A) e -845(T/C) na região promotora do gene interleucina 8 (IL8), bem como de seus haplótipos, com suscetibilidade à periodontite em indivíduos Brasileiros. Foram selecionados 500 indivíduos de ambos os gêneros (Grupo Controle n=224 e Grupo Periodontite n=276) que procuraram atendimento na Faculdade de Odontologia de Araraquara. A partir do DNA obtido de células da mucosa oral, os polimorfismos -738(T/A) e -845(T/C) foram analisados por PCR-RFLP (Polymerase Chain Reaction ­ Restriction Fragment Length Polymorphism), e o -353(A/T) por SSP-PCR (Sequence Specific Primer ­ PCR). Os fragmentos obtidos foram submetidos à eletroforese vertical em gel de poliacrilamida a 10%. Para analisar a freqüência dos genótipos e alelos foi utilizado o teste 2 , ou o programa CLUMP para os polimorfismos que mostraram alelos raros. Os haplótipos foram estimados pelo programa ARLEQUIN e a diferença na distribuição deles entre os grupos foi investigada por meio do programa CLUMP. Os resultados da análise individual dos polimorfismos não evidenciaram associação com a periodontite. Em relação aos haplótipos, as freqüências dos mesmos como alelos e/ou genótipos entre Grupo Controle e Grupo Periodontite apresentaram diferenças estatisticamente significantes considerando a população total e também a não fumante. Os indivíduos com o haplótipo CTA apresentaram-se 2,14 vezes mais susceptíveis à doença periodontal (p=0,0012, OR=2,14; 95% CI =1,6-3,38). Concluiu-se que houve associação dos haplótipos formados pelos polimorfismos ­845(T/C), ­738(T/A) e ­353(A/T) no gene IL8 com a suscetibilidade a periodontite na população brasileira estudada

The purpose of this study was to investigate whether there is an individual association of the ­845(T/C), ­738(T/A) and ­353(A/T) single nucleotide polymorphisms (SNPs) in the IL8 gene, as well as their haplotypes with susceptibility to periodontitis in Brazilian population. DNA was extracted from buccal epithelial cells from 500 Brazilian individuals (Control group n = 224, Periodontitis group n = 276). The ­845 and ­738 SNPs were genotyped by PCR-RFLP method (Polymerase Chain Reaction-Restriction Fragment Length Polymorphism). The ­353 SNP was investigated using the Sequence Specific Primers Polymerase Chain Reaction method (SSP-PCR). Differences of the allelic and genotypic frequencies were assessed by Chi-squared test or by using the CLUMP program for rare alleles. Haplotypes were estimated using the ARLEQUIN program. Differences in the haplotype frequencies between the studied groups were assessed by the CLUMP program. No differences were observed in the allelic and genotypic distribution of all the investigated SNPs between control and periodontitis groups when they were analysed individually. The haplotypes distribution in the periodontitis and controls groups was significantly different both considering the total casuistic or the non-smokers subgroup only. Individuals with CTA haplotype seemed to be over twice more likely to develop periodontitis than individuals with other haplotypes (OR=2.14, 95% CI =1.36-3.38). The analysis of haplotypes constructed from ­845(T/C), ­ 738(T/A) and ­353 (A/T) polymorphisms in the IL8 gene demonstrated association with susceptibility to periodontitis in Brazilian individuals