Indiana University-Purdue University Indianapolis (IUPUI); Graduate Endodontics, Masters degree thesis / We evaluated the antibacterial stability (shelf life) of a new radiopaque double antibiotic paste (RoDAP) loaded in a methylcellulose system with 30% w/v barium sulfate against biofilm collected from an immature tooth with necrotic pulp. Uniform radicular dentin specimens were infected with bacterial biofilm obtained from an immature tooth with a necrotic pulp and incubated anaerobically for three weeks. These samples were randomly divided into 6 experimental groups (n=7) and treated for 1 week at three time points of aged radiopaque DAP: 0 months, 3 months, and 6 months. Group 1: 1mg/mL RoDAP treatment. Group 2: 10 mg/mL RoDAP treatment. Group 3: Calcium hydroxide (Ca(OH)2) treatment. Group 4: Methylcellulose with barium sulfate. Group 5: No treatment. Group 6: No bacteria or treatment. The samples were rinsed with sterile saline to detach biofilms and then spiral plated using a biofilm disruption assay. Statistical analyses were performed using Wilcoxon rank-sum tests and Wilcoxon signed rank tests with fixed effects for treatment, time, and the treatment-by-time interaction. Treatment of infected dentin with 1 mg/mL RoDAP, 10 mg/mL RoDAP, and Ca(OH)2 demonstrated significant and substantial antibiofilm effects in comparison to untreated control groups or groups treated with placebo paste after 0, 3, and 6 months of aging. Calcium hydroxide, however, showed slightly less antibiofilm activity after 6 months of aging when compared to 0 months and 3 months of aging. This difference was statistically significant (p > 0.05). In conclusion, both concentrations of RoDAP maintained full antibacterial efficacy after 6 months of aging, while calcium hydroxide lost some antibacterial activity after a shelf life of 6 months.

Indiana University-Purdue University Indianapolis (IUPUI) / EFFECTS OF DYNAMATRIX® ON ANGIOGENIC CYTOKINE EXPRESSION FROM HUMAN DENTAL PULP FIBROBLASTS: AN IN VITRO STUDY by Joseph Benjamin Adams Indiana University School of Dentistry Indianapolis, IN Introduction: An exogenous scaffold may lead to more predictable pulp tissue regeneration and continued root formation in a regenerative endodontic procedure. DynaMatrix® is a natural membrane scaffold made of porcine small intestine, currently used in periodontal regenerative surgeries. Objective: The purpose of this study was to investigate if human dental pulp fibroblasts (HDPFs) seeded on DynaMatrix® membrane would result in an increase in the expression of angiogenic cytokines. Materials and Methods: HDPFs (75,000 per well) were seeded in 6-well plates. Three groups were tested: Group 1 (C): HDPFs in 70 media only; Group 2 (M): DynaMatrix® (Cook Biotech, Indianapolis, IN) alone in media; and Group 3 (C+M): HDPFs seeded on DynaMatrix® membranes. After 72 hours of incubation in serum positive, the conditioned media were collected and analyzed for the expression of 20 angiogenic cytokines utilizing RayBiotech Inc., arrays per the manufacturer’s instruction. The data were analyzed by ANOVA. Results: Group M was significantly higher than C for bFGF (p = 0.0023). C+M was significantly higher than M for ANG (p = 0.0104); GRO (p = 0.0003); IFN-γ (p = 0.0023); IL-6 (p = 0.0003); IL-8 (p = 0.0003); Leptin (p = 0.0003); MCP-1 (p = 0.0104); TIMP-1 (p = 0.0190); TIMP-2 (0.0123). C was significantly higher than C+M for ANG (p = 0.0104); MCP-1 (p = 0.0104); and THPO (p = 0.0308). Cytokines such as b-FGF, ANG, and leptin promote angiogenesis, and stimulate migration and proliferation of cells. Conclusion: The cytokine expression profile from the cells seeded on DynaMatrix® suggests that it might be a suitable scaffold for regenerative endodontic procedures. It could improve vascularization by increasing angiogenic cytokines in the microenvironment of the treated root canal and supporting tissue regeneration.

published_or_final_version / Dentistry / Doctoral / Doctor of Philosophy

Cone beam computed tomography (CBCT) is the recommended imaging modality of choice for evaluating previous endodontic treatment (1). The aim of this study was to compare treatment planning decisions made when evaluating previous endodontic treatment to determine if treatment planned and projected prognosis differs when digital radiography is used alone or in combination with CBCT. A retrospective chart review was conducted. Patients for whom a CBCT was taken were included in the study. Twenty-eight patients qualified. Patient’s periapical digital radiographs (Dexis©) were evaluated by 2 calibrated endodontists, a treatment plan was identified and a prognosis was projected. Later the same radiographs were viewed with CBCT scan. The CBCT provided significant information 75% of the time. CBCT provided the only information for an accurate diagnosis 17% of the time. Prognosis changed 38% of the time when CBCT was added. An unfavorable or questionable prognosis changed to favorable 30% of the time.

Orientador: Paulo Sergio Cerri / Co-orientador: Juliane Maria Guerreiro Tanomaru / Banca: Manuel de Jesus Simões / Banca: Mario Tanomaru Filho / Resumo: BiodentineTM é um biomaterial à base de silicato de cálcio produzido, segundo o fabricante, com o intuito de apresentar propriedades físico-químicas e biológicas superiores ao MTA. Nosso objetivo foi avaliar a resposta tecidual promovida por BiodentineTM (BDT) e MTA Angelus Branco em subcutâneo de ratos. Foram utilizados ratos adultos distribuídos em 3 grupos (n=20/grupo), segundo o material preenchendo os tubos de polietileno implantados no subcutâneo: BDT, MTA e GC (controle, tubos vazios). Após 7, 15, 30 e 60 dias, os implantes e os tecidos adjacentes foram processados para parafina. Cortes longitudinais das cápsulas adjacentes aos implantes foram corados com H&E, tricrômico de Masson, Picrosirius e submetidos ao Alcian Blue (AB) e reações imuno-histoquímicas para IL-6 e FGF-1. Obteve-se a densidade numérica de células inflamatórias (CI), de células imunomarcadas para IL-6 e FGF-1 e de mastócitos AB-positivos, além da porcentagem de colágeno birrefringente. Os dados foram submetidos à ANOVA e teste Tukey (p≤0,05). O número de CI e de células imunomarcadas para IL-6 e FGF-1 foi significantemente maior aos 7 dias em todos os grupos. Diferenças significantes no número de células IL-6-positivas não foram observadas entre BDT e MTA aos 30 e 60 dias; aos 60 dias, diferenças significantes no número de CI também não foram detectadas. O número de células FGF-1-positivas foi significantemente maior no grupo BDT em comparação ao grupo MTA, em todos os períodos. A densidade numérica de mastócitos e a porcentagem de colágeno aumentaram significantemente ao longo do tempo. Aos 60 dias, o número de mastócitos e o conteúdo de colágeno foram significantemente maiores nos grupos BDT e MTA, respectivamente. A redução significante do processo inflamatório, concomitante à redução na imunoexpressão para IL-6, indica que ambos os materiais são biocompatíveis. A acentuada imunoexpressão de FGF-1 aos 7 dias... / Abstract: BiodentineTM is a new calcium silicate-based biomaterial which presents improved physicochemical and biological properties compared to MTA. The aim of this study was to evaluate the tissue reaction promoted by BiodentineTM (BDT) and MTA Angelus White in rat subcutaneous. Adult rats were distributed into 3 groups (n=20/group) according to the implanted materials: BDT, MTA or CG (Control group; empty tubes). After 7, 15, 30 and 60 days, the implants and adjacent tissues were fixed and embedded in paraffin. Longitudinal sections of the capsule adjacent to implants were stained with H&E, Masson's trichrome, Picrosirius and submitted to Alcian Blue (AB). Immunohistochemical reactions for IL-6 and FGF-1 were also performed. The number of inflammatory cells (IC), IL-6 and FGF-1 immunolabeled cells, AB-positive mast cells as well as birefringent collagen percentage were obtained. Data were statistically analyzed by ANOVA and Tukey test (p≤0.05). The number of IC and IL-6 and FGF-1 immunolabeled cells were significantly high at 7 days in all groups. At 30 and 60 days, significant differences in the number of IL-6-positive cells were not detected between BDT and MTA. At 60 days, statistical difference in the IC number was not observed between BDT and MTA groups. In all periods, the number of FGF-1-positive cells was significant higher in BDT group in comparison to MTA. The numerical density of mast cells and collagen percentage increased over time. At 60 days, mast cells number and collagen content were significantly high in BDT and MTA groups, respectively. A significant reduction of inflammatory process and IL-6 immunoexpression indicates that both materials are biocompatible. Intense FGF-1 immunoexpression at 7 days suggests that this factor may be responsible, at least in part, for fibroblast proliferation and subsequent collagen formation in the capsules. Since a strong correlation between mast cells number and collagen density was detected... / Mestre

Philosophiae Doctor - PhD / Persistent root canal pathogens are one of the main causes of endodontic treatment failure. These pathogens are usually isolated in areas within the root canals that are inaccessible to mechanical instrumentation, chemical irrigants and medicaments resulting in incomplete sterilization of the root canal system. Furthermore, the development of resistant microbial species renders it difficult to disinfect the root canal system using commonly available root canal irrigants and intra-canal medicaments. Intra-canal medicaments are antimicrobial agents that are placed inside the root canal system in order to eliminate the remaining microorganisms that persist after mechanical instrumentation and irrigation. However, their antimicrobial efficacy is effective only against some of the root canal pathogens. Furthermore, the presence of tissue inhibitory factors such as dentine powder and serum albumine within the root canal system inhibits their antimicrobial activity. The use of nanoparticles as antimicrobial agents has recently attracted considerable attention especially in the medical field as a result of their unique antibacterial properties. These properties include their ability to use multiple mechanisms to eradicate microbial cells and their low potentiality to produce microbial resistance. Polymeric nanoparticles such as chitosan nanoparticles (Ch-Np) gained significant interest as a result of their biocompatible and antimicrobial properties. In medicine, several vehicles were designed to carry these antibacterial nanoparticles. Zeolites (Ze) are microporous crystalline hydrated sodium aluminosilicate material that is utilized in the chemical sciences as a carrier for various nanoparticles.

O objetivo deste estudo foi comparar o preparo de canais radiculares de dentes decíduos artificiais com o uso dos sistemas manual, rotatório e de rotação recíproca, no que diz respeito ao desgaste promovido após a instrumentação dos canais radiculares e o tempo necessário para a realização da instrumentação. Os fatores em estudo foram técnica de instrumentação em três níveis e área de avaliação no canal radicular em três níveis. Foram utilizados apenas os dentes multiradiculares (primeiro molar superior, segundo molar superior, primeiro molar inferior e segundo molar inferior), compondo 24 dentes os quais foram aleatoriamente divididos entre as três técnicas de instrumentação em estudo: manual, Sistema rotatório Mtwo e Sistema de rotação recíproca Reciproc. A unidade amostral foi conduto radicular (n=20). Os condutos radiculares foram escaneados no tomógrafo de TCFC (Accuitomo). Após a instrumentação dos dentes novas imagens foram realizadas, avaliações quantitativas foram feitas medindo-se nas imagens axiais dos canais radiculares, a espessura das paredes em três diferentes regiões: terços apical, médio e cervical. As variáveis de resposta analisadas foram cálculo do transporte do canal, cálculo da descentralização e tempo de instrumentação. Tanto para a avalição da descentralização, quanto para a avaliação do transporte, os dados foram transformados e analisados por ANOVA a 2 critérios e Teste de Tukey, para os dados da comparação dos tempos foi utilizado o teste de Kruskal-Wallis e Teste de Tukey (p<0,05). Para a avaliação do transporte do canal e da descentralização não foi encontrada diferença estatística significativa entre os tipos de instrumentação e terços radiculares (p>0,05). Na avaliação do tempo de preparo, o grupo do sistema reciprocante apresentou menor tempo de preparo em comparação com o sistema manual e rotatório (p<0,05). Desta forma conclui-se que os tipos de instrumentação avaliados promoveram um preparo similar dos canais, sem diferença entre os terços cervical, médio e apical, sendo que o sistema de rotação recíproca resultou em um menor tempo de instrumentação. / The objective of this study was to compare the preparation of root canals of artificial deciduous teeth with the use of the manual systems, rotational and reciprocal rotation, with regard the detrition promoted after instrumentation of root canals and the time necessary for the realization of instrumentation. The factors in study were the techniques of instrumentation in tree levels and the area of root canal in tree levels. Have been used only teeth multirooted (first molar, second molar, lower first molar and second molar), composing 24 teeth which were randomly divided among the three instrumentation techniques under study: manual, rotational system Mtwo and reciprocal rotation system Reciproc. The sample unit was root canal (n = 20). The root canals were scanned in tomographer CBCT (Accuitomo). After instrumentation of new images teeth were conducted quantitative evaluations were made by measuring the axial images of the root canals, the wall thickness at three different regions: apex, middle and cervical thirds. Response variables analyzed were calculating the channel transport, decentralization and calculation time instrumentation. Both for the evaluation of decentralization, as for the evaluation of transportation, data were processed and analyzed by ANOVA and Tukey 2 criteria test to the data comparing the times we used the Kruskal-Wallis test and Tukey test (p <0.05). For the evaluation of the channel transport and decentralization was no statistically significant difference between the types of instrumentation and root thirds (p> 0.05). In assessing the preparation time, the reciprocating system group showed less preparation time compared to the manual system and rotary (p <0.05). Thus it is concluded that the types of instrument reviews promoted a similar preparation of channels, no difference between the cervical, middle and apical thirds, and the reciprocal rotation resulted in a shorter time instrumentation.

Orientador: Juliana Maria Guerreiro Tanomaru / Banca: Iara Teresinha Correia Silva Sousa / Banca: Elisabeth Loshchagin Pizzolitto / Resumo: A substituição do óxido de bismuto por óxido de zircônio (ZrO2) e o acréscimo de hidroxiapatita nanoparticulada (HAn) podem favorecer propriedades do Cimento Portland (CP). Este estudo objetiva comparar propriedades físico-químicas, mecânicas e antibiofilme do CP com ZrO2, acrescido de HAn. Os materiais avaliados foram: G1 - MTA Angelus; G2 - CP/ZrO2; G3 - CP/ZrO2/HAn10%; G4 - CP/ZrO2/HAn20%. O pH foi analisado em phmetro digital, a solubilidade pela perda de massa e o tempo de presa pelo teste de Gilmore. A resistência à compressão foi avaliada por meio de ensaio mecânico. Amostras dos materiais foram radiografadas com uma escala de alumínio para análise da radiopacidade. Para a avaliação antibiofilme os materiais foram colocados em contato com biofilme de E. faecalis formado em blocos de dentina, e avaliado pela contagem de unidades formadoras de colônias (UFCmL-1). Os resultados foram submetidos aos testes ANOVA e Tukey com 5% de significância. G2 e G3 apresentaram pH mais elevado. Maior solubilidade foi observada para G3 e G4. O tempo de presa inicial foi menor para G1 e o tempo de presa final para G3. Todos os materiais apresentaram radiopacidade superior a 3 mm Al. A resistência à compressão foi maior para G2 nos períodos avaliados. Nenhum cimento foi capaz de neutralizar completamente o biofilme bacteriano, mas G3 e G4 proporcionaram maior redução bacteriana. Conclui-se que a associação CP/ZrO2 e CP/ZrO2 com 10% de HAn apresentam propriedades físico-químicas adequadas e que o acréscimo de HAn melhora a atividade antibiofilme de E faecalis. / Abstract: The bismuth oxide replacement by zirconium oxide (ZrO2) and the addition of hydroxyapatite nanoparticle may improve the Portland Cement (PC) properties. This study aimed evaluate the physicochemical, mechanical and antibiofilm of Enterococcus faecalis properties of Portland Cement (PC) association with zirconium oxide (ZrO2), added with hydroxyapatite nanoparticles (HAn). The analised materials were: G1 -MTA Angelus; G2 - PC/ZrO2; G3 - PC/ZrO2/HAn10%; G4 - PC/ZrO2/HAn20%. The pH was measure with a digital phmeter and the solubility by the weight loss. Setting time was assayed by Gilmore needle. Compressive strength was tested in universal testing machine. Materials samples were radiographed alongside an aluminum (Al) step wedge. For the antibiofilm test, the samples were place in direct contact with E. faecalis biofilm over dentine blocks, and assessed by colony forming units (CFU mL-1). The results were submitted to ANOVA and Tukey's test at 5% significance. G2 and G3 showed the higher pH in both experimental periods. Bigger solubility was noted for G3 and G4. The initial setting time was shorter for G1, and the final setting time for G3 and G4. All material presented radiopacity greater than 3 mmAl. Compressive strength was better for G2 in both experimental periods. No cement was able to completely neutralize the bacterial biofilm, however G3 and G4 exhibited higher bacteria reduction. The PC/ZrO2 and PC/ZrO2 with HAn 10% association presented the better physicochemical and mechanical properties, and the association with HAn improves the E. faecalis antibiofilm activity. / Mestre

Orientador: Paulo Sergio Cerri / Coorientador: Juliane M. Guerreiro Tanomaru / Banca: Raquel Assed Bezerra da Silva / Banca: Giulio Gavini / Banca: Gisele Faria / Banca: Mário Tanomaru Filho / Resumo: Objetivo: Avaliar as propriedades físicas, químicas e biológicas do cimento Portland (CP) associado a diferentes radiopacificadores (óxido de zircônio - OZr e óxido de nióbio - ONb) micro e nanoparticulados. Metodologia: Para isso, os grupos experimentais foram divididos em: Capítulo 1 - CP, CP+20% de óxido de bismuto (OB), CP+30% OZrmicro e CP+30% OZrnano; Capítulo 2 - CP+30% ONbmicro, CP+30% ONbnano e MTA-Angelus. Todos os materiais foram submetidos aos testes de radiopacidade, tempo de presa, pH e liberação de íons cálcio. Além disso, nos materiais do capítulo 1 foi realizado o teste de resistência à compressão após 24 horas e 21 dias. A reação tecidual provocada pelos materiais foi investigada por meio de análises morfológica, morfométrica (densidade numérica de células inflamatórias, CI) e imuno-histoquímica (número de células imunopositivas à interleucina-6, IL-6) da cápsula formada após 7, 15, 30 e 60 dias de implantação no subcutâneo dorsal. O método von Kossa, para detecção de estruturas calcificadas, também foi realizado. Resultados: Capítulo 1 - Os materiais testados apresentaram pH alcalino e liberação de íons cálcio após 1, 7, 14 e 28 dias. Os materiais CP+OB, CP+OZrmicro e CP+OZrnano apresentaram valores de radiopacidade estatisticamente superiores ao CP e similares entre si. O CP+OZr micro e o CP+OZr nano exibiram tempos de presa maiores do que CP (p≤0,05). O CP apresentou valor de resistência à compressão, após 24 horas e 21 dias, maior do que os materiais, porém, a adição de OZr promoveu médias estatisticamente superiores quando comparado ao CP+OB. Em todos os grupos, verificou-se redução significante no número de CI e de células imunopositivas à IL-6 aos 15, 30 e 60 dias em comparação à 7 dias. O número de CI e de células imunomarcadas à IL-6 nos materiais... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Objective: The aim of this study was to evaluate the effect of different micro e nanoparticulated radiopacifiers (zirconium oxide - ZrO and niobium oxide - NbO) on the physicochemical and biological properties of Portland cement (PC). Methods: The experimental groups were divided in: Chapter 1 - PC, PC+20% bismuth oxide (BO), PC+30% ZrOmicro e PC+30% ZrOnano; Chapter 2 - PC+30% NbOmicro, PC+30% NbOnano and MTA-Angelus. All materials were submitted to the tests of radiopacity, setting time, pH and calcium ion release. Besides, in the materials of chapter 1, the test of compressive strength after 24 hours and 21 days was performed. Moreover, tissue reaction promoted by all materials was evaluated in rat subcutaneous. In each rat, two polyethylene tubes filled with materials were implanted into dorsal subcutaneous. Empty tubes were used as control. After 7, 15, 30 and 60 days, the specimens were fixed and embedded in paraffin. In the HE-stained sections, the number of inflammatory cells (IC) in the capsule was obtained. Moreover, detection of interleukin-6 (IL-6) by immunohistochemistry and number of IL-6 immunolabeled cells were carried out. The von Kossa method was also performed. The differences among the groups were subjected to ANOVA with a significance level of 5%. Results: Chapter 1 - All materials presented an alkaline pH and released calcium ions after 1, 7, 14 and 28 days. The PC+BO, PC+ZrOmicro and PC+ZrOnano exhibited radiopacity values statistically superior than PC. The PC+ZrOmicro and PC+ZrOnano presented setting times higher than PC (p≤0.05). After 24 hours and 21 days, the PC showed compressive strength value significantly superior to the other materials. A higher compressive strength in the PC+ZrO (micro and nano) was found compared to PC+BO. In all groups, significant reduction in... (Complete abstract click electronic access below) / Doutor

Orientador: Idomeo Bonetti Filho / Banca: João Eduardo Gomes Filho / Banca: Alexandre Augusto Zaia / Banca: Gisele Faria / Banca: Mario Tanomaru Filho / Resumo: O objetivo do presente estudo foi avaliar a porcentagem de guta-percha, cimento endodôntico e falhas na obturação dos canais radiculares, usando as técnicas de cone único e condensação lateral ativa, após inserção de cimento endodôntico por meioou do próprio cone de guta-percha, ou de lima endodôntica, ou de lentulo. Sessenta canais radiculares de molares inferiores foram instrumentados com limas ProTaper Universal e divididos em 2 grupos: grupo I- Canais obturados pela técnica da condensação lateral ativa; grupo II- Canais obturados pela técnica do cone único. Cada grupo foi dividido em 3subgrupos (A, B e C), de acordo com o modo de introdução do cimento endodôntico no início da obturação: A-Por meio do próprio cone de guta-percha principal; B- Por meio de lentulo; C-Por meio de lima endodôntica. As raízes foram seccionadas a 2, 4 e 6mm do ápice radicular, fotografadas em microscópio digital e as imagens transferidas para o programa Image Tool 3.0, para mensuração das áreas de guta-percha, cimento endodôntico e falhas. A análise estatística dos resultados foi realizada por meio dos testes ANOVA, Kruskall-Wallis, Dunn e Mann-Whitney. Observamos que não houve diferença estatística significante na porcentagem de guta-percha, cimento endodôntico e falhas na técnica da condensação lateral ativa. Com a técnica do cone único, verificamos maiores porcentagens de área de cimento endodôntico e menores porcentagens de área de falhas, quando utilizado instrumento para a sua inserção. Com a técnica do cone único, a inserção de cimento endodôntico antes do cone diminui significativamente as falhas da obturação. Métodos diferentes de inserção do cimento endodôntico não influenciam na qualidade da obturação, quando utilizada a técnica da condensação lateral ativa / Abstract: The aim of this study was to evaluate percentages of gutta-percha, endodontic sealer and voids in radicular canals obturations using Single Cone and active Lateral Compaction, after placing the endodontic sealer through master gutta-percha cone, endodontic file or lentulo. Sixty root canals of inferior molars, after being instrumented with ProTaper Universal files, were distributed in 2 groups: group I- Root canals sealed using active lateral condensation; group II- Root canals sealed using single cone technique. Each of these groups was subdivided into 3 subgroups (A, B and C) according different methods of placing the endodontic sealer in the beginning of the filling: A- Through the master gutta-percha cone insertion; B- Through lentulo; C- Through endodontic file. The roots were transected at 2, 4 and 6mm from the root apex and photographed with digital microscope. The pictures were transferred to Image Tool 3.0 Software, in order to allow measurement of the areas of gutta-percha, endodontic sealer and voids. ANOVA, Kruskall-Wallis, Dunn and Mann-Whitney tests were applied for statistical analysis of the results, through which we observed there are no significant differences in the percentages of gutta-percha, endodontic cement and voids, when it is used Lateral Compaction. With single cone, we verified higher percentages of sealer area and lower percentages of voids, when an instrument is used to place the sealer. Furthermore, with this technique, placing the sealer using an instrument before the insertion of the gutta-percha cone reduces voids. Different sealer placement methods do not affect the quality of the filling, when Lateral Compaction is applied / Doutor