Tratamento de lesões de bifurcação grau III pela técnica de regeneração tecidual guiada associada ou não a enxerto de matriz mineral ossea

Orientador: Antonio Fernando Martorelli de LimaDissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: Este estudo avaliou o resultado do tratamento por regeneração tecidual guiada (RTG) associada ou não a enxerto de matriz mineral óssea bovina de lesões de bifurcação grau III em molares inferiores, e foi conduzido de acordo com o comitê de ética em pesquisa da FOP-UNICAMP. Foram selecionados 20 sítios em 18 pacientes de ambos os sexos, com idade variando entre 35 e 75 anos. Os pacientes foram submetidos a instrução de higiene oral e tratamento periodontal inicial dois meses antes do procedimento cirúrgico experimental. No baseline e na reentrada cirúrgica seis meses após, foram apurados os parâmetros biométricos definidos pelo nível clínico de inserção vertical (NCI) e horizontal (NHI), nível da margem gengival (NMG), profundidade de sondagem (PS), nível do defeito ósseo vertical (NDO) e horizontal (NHD) e nível da crista óssea alveolar (NCOA), utilizando sonda computadorizada. Foram realizados retalhos mucoperiosteais para instrumentação periodontal e os defeitos, separados de forma aleatória e tratados como sítios teste (RTG + enxerto de matriz mineral óssea bovina) e controle (RTG). As imagens radiográficas digitalizadas, obtidas no momento do tratamento e na cirurgia de reentrada, foram subtraídas para verificação do ganho e da perda na Área de Preenchimento e na Densidade Óptica entre os sítios teste e controle. A hipótese de não diferença entre os valores médios para todos os parâmetros foi testada pela análise de variância. Os resultados mostraram diferença estatística significativa entre o baseline e a reentrada para PS, NCOA (P < 0,01) e NMG (P < 0,05) nos sítios controle e para NDO (P < 0,01) nos teste. Houve diferença estatística significativa (P < 0,05) para o parâmetro NCOA entre o sítios teste e controle. A hipótese nula foi rejeitada para o parâmetro NCOA mas continua válida para os demais parâmetros. Não há evidências de que a associação RTG + matriz mineral óssea seja um tratamento previsível para lesões de bifurcação grau IIIAbstract: The aim of this clinical trial was to evaluate GTR therapy with and without bovine mineralized bone matrix in mandibular class III furcation. This investigation was performed in accordance with the Ethical Committee in Research FOP-UNICAMP. Twenty defects were selected in 18 non-systemically compromised patients, 35-75 years old, non smokers. Patients were enrolled in an oral hygienic program and basic therapy was accomplished two months prior the experimental procedure, when plaque and gingival index should be lower than 20%. Horizontal (CAL-H) and Vertical Attachment Level (CAL V), Probing Depth (PD), Gingival Margin Level (GML), Horizontal (BDL-H) and Vertical Bone Defect Level (BDL-V) and Alveolar Crest Level (ACL) measurements were performed at baseline and reemtry procedures six months after using electronic probe. Mucoperiosteal flaps were raised and sites were randomized in test treated by GTR + bovine inorganic bone matrix or control treated by GTR. Digital radiographs were taken at baseline and six months after and were analyzed by subtraction to access gain or loss in Optical Density (OD) and Area (A). ANOVA was employed to test the hypothesis of no difference between mean values of the parameters for test and control groups. The results showed statistically significant difference (P < 0.01) between baseline and reentry 0.01) in test group. There was statistically significant gain in ACL (P < 0.05) for the test group while there was reduction in ACL for the control group. There was no statistically significant difference between test and contrai groups for OD and A. The null hypothesis was rejected for ACL and accepted for ali the others parameters. There is no evidence to prove that the association GTR + bovine inorganic bane matrix is a predictable treatment for class III furcationMestradoPeriodontiaMestre em Clínica Odontológic

Influencia da matriz proteica do esmalte associada ao fator de crescimento insulina-simile sobre fibroblastos do ligamento periodontal humano

Orientador : Antonio Fernando Martorelli de LimaTese (doutorado) - Universidade Estadual de Campinas, Faculdade de Odontologia de PiracicabaResumo: O conhecimento das diferenças morfológicas e proliferativas de fibroblastos do ligamento periodontal (FLP) e fibroblastos gengivais (FG) são fundamentais para o melhor entendimento do papel destas células nos eventos de homeostasia, doença e regeneração periodontal. O primeiro objetivo desse estudo foi comparar as características morfológicas, o potencial proliferativo e a síntese protéica de FLP e de FG. Os fibroblastos foram cultivados pela técnica do explante a partir de fragmentos gengival e do ligamento periodontal de um mesmo paciente. As células foram isoladas e plaqueadas para análise em microscopia de contraste de fase e microscopia óptica. O índice de proliferação celular foi determinado por contagem automática de células nos dias 1, 4, 7, 15 e 21 e pelo ensaio de incorporação de bromodioxiuridina (BrdU). A síntese de proteína total foi verificada por eletroforese em gel de poliacrilamida e zimografia. Os resultados mostraram que FLP são maiores e mais alongados que os FG em condições de subconfluência e confluência celular. Os FLP proliferam mais rapidamente que os FG nos períodos de 1, 4 e 7 dias (P<0,05). Nos períodos subsequentes de 15 e 21 dias, não houve diferença estatística significativa entre o número de células do dois tipos celulares. O índice de incorporação de BrdU demonstrou potencial proliferativo de 65,1 .:t 11,0 % para FLP e 41,2.:t 17,1% para FG (P<0,05). A síntese de proteína total verificada em nosso experimento no período de 24 horas mostrou resultados similares para FLP e FG. Nossos resultados demonstraram que FLP e FG são diferentes na morfologia e na capacidade proliferativa, porém são semelhantes na síntese protéica. O segundo objetivo deste estudo foi avaliar os efeitos da matriz protéica do esmalte (EMD), do fator de crescimento insulina-símile (IGF-I) e da associação dos dois fatores sobre o potencial proliferativo e a capacidade de adesão e migração celular de fibroblastos do ligamento periodontal humano. Esse estudo foi conduzido utilizando a análise de proliferação por contagem automática de células (Coulter Counter_Abstract: The knowledge of morphological and proliferative differences between Gingival Fibroblasts (GF) and Periodontal Ligament Fibroblast (PDL) is essential to understand the role of each cell on the homeostasis, disease process and regeneration Df the periodontium. The first objective of this study was to compare the PDL and GF morphology, proliferation rate and protein synthesis. PDL and GF were explanted from tissues Df the same patients. To characterize and compare the cells morphology, PDL and GF were plated for optical and phase contrast microscopy. The proliferation rates were determined by automated count at days 1, 4, 7, 15 and 21, and by Bromodeoxyuridine Labelling Index (BrdU). The total protein content was analyzed by means of electrophoresis in 10% polyacrylamide gel and zimography containing gelatin as substrate. PDL were greater and more elongatOO than GF in subconfluence and confluence conditions. The proliferative rate Df PDL was higher than GF at 1, 4, and 7 days (P < 0.05). At 15 and 21 days there was no statistically significant difference between cells number. The BrdU Index demonstrated a proliferative potential Df 65.1 ! 11.0 % for PDL and 41.2 :t 17.1 % for GF (P < 0.05).The synthesis of protein in a period of 24 hours was similar for both PDL and GF. Our results demonstrated that PDL and GF are different in morphology and in proliferative capacity, however, they do not differ in protein synthesis. The second objective of the present study was to evaluate the effects of the enamel matrix protein (EMD) the effects of Insulin growth factor-I (IGF-I) and the association of the these two factors on periodontal ligament fibroblasts. The study was based on the proliferation rate, and cellular attachment and migration. The proliferation rate was determined by counting the cells afier 1, 3, 7 and 10 days with the help of a automated counter (Coulter Counter). The cellular attachment was analyzed by means of colorimetric assays. Migration assays was performed in Boyden chambers for a period of 4h and analyzed by means of colorimetric assays. EMD has significantly raised the proliferation rate from a minimum concentration of 50 uglml up to maximum concentration of 200 uglml in a 24h period. The proliferation rate continuing to be raised in a period of 3,7 and 10 days (P < 0.05). IGF-I has not altered the proliferation rate at any concentration. The association of EMD + IGF-I raised the proliferation rate independently of the concentration. However, the association has not resulted on further significant effect when compared to the use of EMD alone. The cellular attachment and migration was not altered by EMD, IGF-I or EMD + IGF-I. Our results showed that EMD increased the proliferation rate of PDL but did not improve cellular attachment and migration. IGF-I did not raise neither the proliferation rate no r the cellular attachment and migration and did not have any additional effect on the EMD aloneDoutoradoPeriodontiaDoutor em Clínica Odontológic

EFEITO MUTAGÊNICO DE BAIXA DOSE DE ULTRAVIOLETA NO SISTEMA methG1 EM Aspergillus nidulans

O cncer atualmente um dos problemas mais comuns e graves. Esse grupo de doenas caracteriza-se pela formao de tumores malignos, em decorrncia de mutaes em genes responsveis pelo controle da proliferao e diferenciao celular. Assim, os agentes mutagnicos so potencialmente carcinognicos. Entre os agentes cancergenos mais conhecidos est a radiao ultravioleta, por ser um conhecido fator de risco para cncer de pele. Neste sentido, o objetivo do presente trabalho foi analisar o potencial mutagnico de baixas doses de ultravioleta no sistema methG1. Para isto, foram tratados esporos da linhagem biA1methG1 de Aspergillus nidulans com um segundo de exposio luz ultravioleta. Os resultados mostraram que, mesmo em dose mnima, este agente capaz de causar danos ao genoma, diminuindo a viabilidade e aumentando significativamente a frequncia de mutao, quando comparadas aos valores do controle (esporos no expostos ao tratamento). Estes resultados corroboram com a importncia dos cuidados preventivos e de restrio exposio demasiada a radiao ultravioleta da luz solar

Evaluation of in vitro human gingival fibroblast seeding on acellular dermal matrix

The acellular dermal matrix (ADM) was introduced in periodontology as a substitute for the autogenous grafts, which became restricted because of the limited source of donor's tissue. The aim of this study was to investigate, in vitro, the distribution, proliferation and viability of human gingival fibroblasts seeded onto ADM. ADM was seeded with human gingival fibroblasts for up to 21 days. The following parameters were evaluated: cell distribution, proliferation and viability. Results revealed that, at day 7, fibroblasts were adherent and spread on ADM surface, and were unevenly distributed, forming a discontinuous single cell layer; at day 14, a confluent fibroblastic monolayer lining ADM surface was noticed. At day 21, the cell monolayer exhibited a reduction in cell density. At 7 days, about to 90% of adherent cells on ADM surface were cycling while at 14 and 21 days this proportion was significantly reduced. A high proportion of viable cell was detected on AMD surface both on 14 and 21 days. The results suggest that fibroblast seeding onto ADM for 14 days can allow good conditions for cell adhesion and spreading on the matrix; however, migration inside the matrix was limited.A matriz dérmica acelular (MDA) foi introduzida na Periodontia como um substituto para enxertos autógenos, os quais se tornaram restritos devido à quantidade limitada de tecido doador. O objetivo deste estudo foi verificar, in vitro, a distribuição, proliferação e viabilidade de fibroblastos gengivais humanos cultivados em MDA. Fibroblastos gengivais foram cultivados sobre MDA por até 21 dias. Os seguintes parâmetros foram avaliados: distribuição, proliferação e viabilidade celular. Os resultados revelaram que, aos 7 dias, os fibroblastos estavam aderidos e espraiados na superfície da MDA, e estavam distribuídos de forma desigual, formando uma camada celular descontínua; aos 14 dias, uma monocamada confluente de fibroblastos revestindo a superfície da MDA foi observada. Aos 21 dias, a monocamada celular exibiu uma redução na densidade celular. Aos 7 dias, cerca de 90% das células aderidas na superfície da MDA estavam no ciclo celular, enquanto que aos 14 e 21 dias esse número reduziu significativamente. Uma maior proporção de células viáveis foi detectada na superfície da MDA tanto aos 14 quanto aos 21 dias. Os resultados sugerem que fibroblastos cultivados sobre a MDA por 14 dias permitem boas condições de adesão e espraiamento das células sobre a matriz, porém, a migração de células para o interior da matriz foi limitada.Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES

Mesenchymal stem cells surpass the capacity of bone marrow aspirate concentrate for periodontal regeneration

Regenerative approaches using mesenchymal stem cells (MSCs) have been evaluated to promote the complete formation of all missing periodontal tissues, e.g., new cementum, bone, and functional periodontal ligaments. MSCs derived from bone marrow have been applied to bone and periodontal defects in several forms, including bone marrow aspirate concentrate (BMAC) and cultured and isolated bone marrow mesenchymal stem cells (BM-MSCs). This study aimed to evaluate the periodontal regeneration capacity of BMAC and cultured BM-MSCs in the wound healing of fenestration defects in rats. Methodology: BM-MSCs were obtained after bone marrow aspiration of the isogenic iliac crests of rats, followed by cultivation and isolation. Autogenous BMAC was collected and centrifuged immediately before surgery. In 36 rats, fenestration defects were created and treated with suspended BM-MSCs, BMAC or left to spontaneously heal (control) (N=6). Their regenerative potential was assessed by microcomputed tomography (µCT) and histomorphometry, as well as their cell phenotype and functionality by the Luminex assay at 15 and 30 postoperative days. Results: BMAC achieved higher bone volume in 30 days than spontaneous healing (p&lt;0.0001) by enhancing osteoblastic lineage commitment maturation, with higher levels of osteopontin (p=0.0013). Defects filled with cultured BM-MSCs achieved higher mature bone formation in early stages than spontaneous healing and BMAC (p=0.0241 and p=0.0143, respectively). Moreover, significantly more cementum-like tissue formation (p&lt;0.0001) was observed with new insertion of fibers in specimens treated with BM-MSCs within 30 days. Conclusion: Both forms of cell transport, BMAC and BM-MSCs, promoted bone formation. However, early bone formation and maturation were achieved when cultured BM-MSCs were used. Likewise, only cultured BM-MSCs were capable of achieving complete periodontal regeneration with inserted fibers in the new cementum-like tissue

Oral health after breast cancer treatment in postmenopausal women

OBJECTIVE: Oral health can affect a patient’s general health and quality of life. Given the increase in breast cancer survival rates, investigations of factors influencing the quality of life of survivors have gained importance. Therefore, the objective of our study was to characterize oral health in postmenopausal breast cancer survivors. METHODS: We conducted a matched case-control study. Forty-eight women who survived breast cancer (age 62.1±9.1 years) and 48 healthy controls (age 61.8±8.6 years) were included. For each case and control, a complete oral evaluation chart was completed. RESULTS: The prevalence of chronic periodontal disease was 98% in breast cancer survivors and 87% in controls. The breast cancer survivors had a median of 16 remaining teeth, whereas controls had a median of 22 remaining teeth (p = 0.03). The percentage of sites with gingival bleeding was 16.05% (0-100%) in breast cancer survivors and 0% (0-72%) in controls (p = 0.04). CONCLUSION: Chronic periodontal disease and tooth loss were highly prevalent in postmenopausal breast cancer survivors. To improve survivors’ quality of life, a preventive oral health evaluation should be available prior to cancer treatment

Effect of biofunctionalized implant surface on osseointegration: a histomorphometric study in dogs

Among the different properties that influence bone apposition around implants, the chemical or biochemical composition of implant surface may interfere on its acceptance by the surrounding bone. The aim of this study was to investigate if a biofunctionalization of implant surface influences the bone apposition in a dog model and to compare it with other surfaces, such as a microstructured created by the grit-blasting/acid-etching process. Eight young adult male mongrel dogs had the bilateral mandibular premolars extracted and each one received 6 implants after 12 weeks, totaling 48 implants in the experiment. Four groups of implants were formed with the same microrough topography with or without some kind of biofunctionalization treatment. After histomorphometric analysis, it was observed that the modified microstructured surface with a "low concentration of the bioactive peptide" provided a higher adjacent bone density (54.6%) when compared to the other groups (microstructured + HA coating = 46.0%, microstructured only = 45.3% and microstructured + "high concentration of the bioactive peptide" = 40.7%), but this difference was not statistically significant. In conclusion, biofunctionalization of the implant surface might interfere in the bone apposition around implants, especially in terms of bone density. Different concentrations of bioactive peptide lead to different results.Entre as diferentes propriedades de uma superfície capazes de influenciar a deposição óssea ao redor de implantes, a composição química e bioquímica pode atuar no reconhecimento do tecido ósseo circundante. O presente trabalho investigou a influência da biofuncionalização de superfícies de implante na deposição óssea ao redor dos mesmos em um modelo animal, comparando-as com outras superfícies, como a microtexturizada obtida pelo processo de jateamento e ataque ácido. Metodologicamente, os pré-molares mandibulares bilaterais de 8 cães foram extraídos e após 12 semanas foram instalados 6 implantes em cada cão, constituindo uma amostra de 48 implantes. Dos 4 grupos experimentais de diferentes superfícies, todos continham a mesma microtopografia rugosa, porém possuindo ou não alguma biofuncionalização. A análise histomorfométrica revelou que a superfície microtexturizada modificada pela adição de baixa concentração peptídica obteve uma maior densidade óssea adjacente (54,6%) quando comparada aos outros grupos (microtexturizada + HA = 46%, somente microtexturizada = 45,3% e microtexturizada com adição de alta concentração peptídica = 40,7%), no entanto estas diferenças numéricas não foram estatisticamente significantes. Dentro deste contexto, conclui-se que a biofuncionalização da superfície de implantes pode interferir na aposição óssea, em particular na densidade óssea, e que diferentes concentrações peptídicas podem conduzir a diferentes resultados.FAPES

Clinical attachment loss and molecular profile of inflamed sites before treatment

Objective: To monitor early periodontal disease progression and to investigate clinical and molecular profile of inflamed sites by means of crevicular fluid and gingival biopsy analysis. Methodology: Eighty-one samples of twenty-seven periodontitis subjects and periodontally healthy individuals were collected for the study. Measurements of clinical parameters were recorded at day −15, baseline and 2 months after basic periodontal treatment aiming at monitoring early variations ofthe clinical attachment level. Saliva, crevicular fluid and gingival biopsies were harvested from clinically inflamed and non-inflamed sites from periodontal patients and from control sites of healthy patients for the assessment of IL-10, MMP-8, VEGF, RANKL, OPG and TGF-β1 protein and gene expression levels. Results:Baseline IL-10 protein levels from inflamed sites were higher in comparison to both non-inflamed and control sites (p&lt;0.05). Higher expression of mRNA for IL-10, RANK-L, OPG, e TGF-β1 were also observed in inflamed sites at day −15 prior treatment (p&lt;0.05). After the periodontal treatment and the resolution of inflammation, seventeen percent of evaluated sites still showed clinically detectable attachment loss without significant differences in the molecular profile. Conclusions: Clinical attachment loss is a negative event that may occur even after successful basic periodontal therapy, but it is small and limited to a small percentage of sites. Elevated inflammation markers of inflamed sites from disease patients reduced to the mean levels of those observed in healthy subjects after successful basic periodontal therapy. Significantly elevated both gene and protein levels of IL-10 in inflamed sites prior treatment confirms its modulatory role in the disease status

New tendencies in non-surgical periodontal therapy

The aim of this review was to update the evidence of new approaches to non-surgical therapy (NSPT) in the treatment of periodontitis. Preclinical and clinical studies addressing the benefits of adjunctive antimicrobial photodynamic therapy, probiotics, prebiotics/synbiotics, statins, pro-resolving mediators, omega-6 and -3, ozone, and epigenetic therapy were scrutinized and discussed. Currently, the outcomes of these nine new approaches, when compared with subgingival debridement alone, did not demonstrate a significant added clinical benefit. However, some of these new alternative interventions may have the potential to improve the outcomes of NSPT alone. Future evidence based on randomized controlled clinical trials would help clinicians and patients in the selection of different adjunctive therapies

Short-Term effect of COX-2 selective inhibitor as an adjunct for the treatment of periodontal disease: a clinical double-blind study in humans

Adjunctive therapeutic strategies that modulate the inflammatory mediators can play a significant role in periodontal therapy. In this double-blind, placebo-controlled study, 60 subjects diagnosed as periodontitis patients were evaluated for 28 days after periodontal treatment combined with selective cyclooxygenase-2 (COX-2) inhibitor. The experimental group received scaling and root planning (SRP) combined with the Loxoprofen antiinflammatory drug (SRP+Loxoprofen). The control group received SRP combined with placebo (SRP+placebo). Plaque index (PI), probing pocket depth (PD) and bleeding on probing (BOP) were monitored with an electronic probe at baseline and after 14 and 28 days. Both groups displayed clinical improvement in PD, PI and BOP. They also showed statistically similar values (p>0.05) of PD reduction on day 14 (0.4 mm) and on day 28 (0.6 mm). At the baseline, few deeper sites (>7 mm) from SRP+Loxoprofen group were responsible and most PD reduction was observed after 14 days (p<0.05). The percentage of remaining deep pockets (>7 mm) after 14 days in the SRP+Loxoprofen group was significantly lower (p<0.05) than in the SRP+placebo group. Loxoprofen presents potential effect as an adjunct of periodontal disease treatment, but long-term clinical trials are necessary to confirm its efficacy.Estratégias terapêuticas adjuvantes que modulam os mediadores inflamatórios podem ter função significante na terapia periodontal. Neste estudo duplo-cego controlado com placebo, 60 indivíduos diagnosticados com periodontite foram avaliados por 28 dias após tratamento periodontal combinado com inibidor seletivo de COX-2. O grupo experimental foi tratado com raspagem e alisamento radicular combinado com medicação anti-inflamatória Loxoprofeno (RAR+Loxoprofen). O grupo controle foi tratado com raspagem e alisamento radicular combinado com medicação placebo (Raspagem e alisamento radicular - RAR+placebo). Presença de placa (PI), profundidade de sondagem (PS) e sangramento à sondagem (SS) foram monitoradas com auxílio de uma sonda computadorizada no início do estudo e após 14 e 28 dias. Os dois grupos demonstraram melhora clínica em relação a PS, PI e SS. Também foi observado valores semelhantes (p>0,05) de redução da PS nos períodos de 14 dias (0,4 mm) e 28 dias (0,6 mm). No início do estudo, alguns sítios profundos (>7 mm) do grupo RAR+Loxoprofen foram os responsáveis pela maior redução da PS depois de 14 dias (p<0,05). A porcentagem de bolsas periodontais profundas >7 mm após 14 dias no grupo RAR+Loxoprofen foi significativamente inferior do que o grupo RAR+placebo (p<0.05). A medicação Loxoprofen apresenta potencial efeito adjuvante à terapia periodontal, mas estudos de longo prazo são necessários para confirmar sua eficácia.Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)FAPESPSankyo Pharma Ltda