Assessing peri-implant bone microarchitecture: conventional vs. osseodensification drilling - ex vivo analysis

Abstract: The aim was to evaluate primary implant stability and bone microarchitecture in two drilling situations, by comparing the conventional technique (CT) and osseodensification (OD) (Versah Burs - Jackson - Mississippi - USA). The implant insertion torque (IT), implant stability quotient (ISQ), and the peri-implant trabecular microstructure were assessed on bone fragments obtained from pig's tibia (n=12), divided between CT (n=6) and OD (n=6). After the drilling procedure, the implants were installed (3.5x8.5 mm, Epikut - SIN - São Paulo - Brazil). The IT and ISQ were measured using a digital torque wrench and resonance frequency analysis. Then, the bone fragments containing the implants were removed with a trephine and analyzed by Microtomography (µCT, 8.0 µm). The comparison between groups was performed using the unpaired t-test (α=0.05). The results revealed that OD promotes a higher insertion torque (CT: 7.67±2.44 Ncm; OD: 19.78±5.26 Ncm) (p=0.0005), although ISQ was not different (CT: 61.33±4.66; OD: 63.25±4.58) (p=0.48). There was a significant increase in peri-implant bone volume (CT: 23.17±3.39 mm3; OD: 32.01±5.75 mm3) (p=0.008), and trabecular parameters: separation (CT: 0.4357±0.03 mm; OD: 0.3865±0.04 mm) (p=0.0449), number (CT: 1.626±0.18 1/mm; OD: 1.946±0.13 1/mm) (p=0.007), and thickness (CT: 0.1130±0.009 mm; OD: 0.1328±0.015 mm) (p=0.02). Structure model index (SMI) data demonstrate no significant differences between groups (CT: 1.7±0.2; OD: 1.4±0.4) (p=0.12). In conclusion, OD increases the insertion torque values and promotes beneficial changes regarding bone microarchitecture compared with CT, revealing more peri-implant bone volume with consequent higher primary stability.

Resumo O objetivo deste estudo foi avaliar a estabilidade de implante e a microarquitetura óssea em duas técnicas de fresagem, comparando a técnica convencional (CT) e a osseodensificação (OD) (Versah Burs - Jackson - Mississippi - EUA). O torque de inserção do implante (IT), quociente de estabilidade primária (ISQ) e a estrutura trabecular peri-implantar foram avaliados em fragmentos ósseos obtidos de tíbia de porco (n=12), divididos entre CT (n=6) e OD (n=6). Após o procedimento de fresagem, foram instalados implantes (3,5x8,5 mm, Epikut - SIN - São Paulo - Brasil). O IT e o ISQ foram aferidos por meio de um torquimetro digital e análise de frequência de ressonância. Em seguida, os fragmentos ósseos contendo os implantes foram removidos com trefina e analisados ​​por microtomografia computadorizada (µCT, 8,0 µm). A comparação entre os grupos foi realizada por meio do teste-t não-pareado (α=0.05). Os resultados revelaram que a OD promove maior torque de inserção (CT: 7,67 ± 2,44º Ncm; OD: 19,78 ± 5,26 Ncm) (p=0,0005), embora a estabilidade primária não tenha sido diferente (CT: 61.33 ± 4.66; OD:63.25 ± 4.58) (p=0,48). Houve um aumento significativo no volume ósseo peri-implantar (CT: 23,17±3,39 mm3; OD: 32,01±5,75 mm3) (p=0,0089) e parâmetros trabeculares: separação (CT: 0,4357 ± 0,03 mm; OD: 0,3865 ± 0,04 mm) (p=0,0449), número (CT: 1,626 ± 0,18 1/mm; OD: 1,946 ± 0,13 1/mm) (p=0,007) e espessura (CT: 0,1130 ± 0,009 mm; OD: 0,1328 ± 0,015 mm) (p=0,02) O índice de modelo estrutural (SMI) não demostrou diferença estatisticamente significativa (p=0.1228). Concluindo, OD apresenta maiores valores de torque de inserção e promove mudanças benéficas na microarquitetura óssea em comparação com a TC, revelando maior volume ósseo peri-implantar.

Cortical microarchitecture and remodeling-associated gene expression related to oral cancer prognosis

Abstract The objective of this study was to assess the remodeling-associated gene expression in the mandible of patients diagnosed with oral squamous cell carcinoma (OSCC), investigating the cortical microarchitecture, and their influence on disease-free survival (DFS) and overall survival (OS) rates. A total of twenty-four patients who underwent mandibulectomy for OSCC treatment had two bone fragments harvested from the mandible for gene expression (RANK, RANKL, OPG, and SOST), and microarchitecture analysis, including bone volume, surface, mineral density, degree of anisotropy, and fractal dimension. The prognosis of the patients was assessed. The results revealed that RANK, RANKL, and SOST were predominantly downregulated, while OPG was completely downregulated. Tumors located adjacent to the posterior region of the mandible (p = 0.02), with a bone mineral density below 1.03 g/cm3 HA (p = 0.001), and a bone volume less than 86.47% (p = 0.03) were associated with poor outcomes. In conclusion, bone-remodeling-associated genes exhibited downregulation in the cortex of the mandible in OSCC patients. Additionally, the tumor's location within the mandible, bone volume, and cortical bone mineral density were identified as factors impacting DFS.

Effect of micro-CT acquisition parameters and individual analysis on the assessment of bone repair

Abstract This study aimed to investigate whether two acquisition parameters, voxel size and filter thickness, used in a micro-computed tomography (micro-CT) scan, together with the examiner's experience, influence the outcome of bone repair analysis in an experimental model. Bone defects were created in rat tibiae and scanned using two voxel sizes of 6- or 12-µm and two aluminum filter thickness of 0.5- or 1-mm. Then, bone volume fraction (BV/TV) and trabecular thickness (Tb.Th) were analyzed twice by two groups of operators: experienced and inexperienced examiners. For BV/TV, no significant differences were found between scanning voxel sizes of 6 and 12 µm for the experienced examiners; however, for the inexperienced examiners, the analysis performed using a 12-µm voxel size resulted in higher BV/TV values (32.4 and 32.9) than those acquired using a 6-µm voxel size (25.4 and 24.8) (p < 0.05). For Tb.Th, no significant differences between the analyses performed by experienced and inexperienced groups were observed when using the 6-µm voxel size. However, inexperienced examiners' analysis revealed higher Tb.Th values when using the 12-µm voxel size compared with 6 µm (0.05 vs. 0.03, p < 0.05). Filter thickness had no influence on the results of any group. In conclusion, voxel size and operator experience affected the measured Tb.Th and BV/TV of a region with new bone formation. Operator experience in micro-CT analysis is more critical for BV/TV than for Tb.Th, whereas voxel size significantly affects Tb.Th evaluation. Operators in the initial phases of research training should be calibrated for bone assessments.

Implant site preparation-related bone microdamage: An alternative ex vivo analysis of drilling speed protocols / Microdanos ósseos gerados na preparação do sítio de instalação de implantes: uma alternativa para análise ex vivo de diferentes protocolos de velocidade de perfuração

ABSTRACT Objective: The aim of this study was to evaluate bone microdamage in sites prepared for implant placement by using an ex vivo model with three drilling rotation speeds. Methods: Bovine bone ribs were used for the creation of 18 osteotomy sites at different rotation speeds: 1200 rpm, 800 rpm, and 400 rpm. Specimens were stained with xylenol orange and prepared for histological analysis by using fluorescence and polarized light microscopies. Bone microdamage was evaluated by number and based on total bone area, as follows: microfracture density (Fr.D = n/mm2), microcrack morphology (diffuse or linear), and density (Cr.D = n/mm2), and presence of bone chips. To complement the analysis, linear microcracks were assessed by using confocal microscopy for three-dimensional visualization. Results: Bone microdamage on the osteotomy sites included microcracks, diffuse damages, microfracture, and bone chip formation. There was an association between bone microdamage and cancellous bone (p 0.0016), as well as a positive correlation between Fr.D and Cr.D (p 0.05, r 0.54). BM occurrence was not different between the three rotation speeds. In 3D, the height of the microcrack depth was 60.81 µm. Conclusion: Bone microdamage occurs during osteotomy, and the ex vivo model used was effective for the assessment of these biomechanical parameters. In addition, microdamage was not influenced by the drilling rotation speed in this experimental condition.

RESUMO Objetivo: O objetivo desse estudo foi avaliar os microdanos ósseos em locais preparados para a instalação de implantes utilizando um modelo ex vivo acessando três velocidades de rotação de perfuração. Métodos: Fragmentos ósseos de costela bovina foram utilizados para a criação de 18 sítios de osteotomia em diferentes velocidades de rotação: 1200 rpm, 800 rpm e 400 rpm. As amostras foram coradas com Alaranjado de Xilenol e preparadas para análise microscópica em fluorescência e luz polarizada. Os microdanos ósseos foram avaliados em número e calibrados com base na área total óssea: densidade de microfraturas (Fr.D = n/mm2), morfologia (difusa ou linear) e densidade de microtrincas (Cr.D = n / mm2) e presença de espículas ósseas. Para complementar a análise, microtrincas lineares foram avaliadas por meio de microscopia confocal para visualização tridimensional. Resultados: Os microdanos ósseos incluíram microtrincas, danos difusos, microfraturas e formação de espículas. Houve uma associação entre MO e localização em osso esponjoso (p=0,0016), bem como uma correlação positiva entre Fr.D e Cr.D (p=0,05, r = 0,54). A ocorrência de microdanos ósseos não foi diferente entre as três velocidades de rotação utilizadas. Em 3D, a profundidade maior da microfissura atingiu 60,81 µm. Conclusão: Microdanos ósseos ocorrem durante a osteotomia e podem ser acessados em um modelo ex vivo na condução de experimentos em biomecânica. Sugere-se que a presença de microdanos não é influenciada pela velocidade de rotação durante a perfuração.

Cortical bone modifications after radiotherapy: cortex porosity and osteonal changes evaluated over time

Abstract Aiming to evaluate cortical bone microarchitecture and osteonal morphology after irradiation, twelve male New Zealand rabbits were used. The animals were divided: control group (no radiation-NIr); and 3 irradiated groups, sacrificed after: 7 (Ir7d); 14 (Ir14d) and 21 (Ir21d) days. A single radiation dose of 30 Gy was used. Computed microtomography analyzed the cortical microarchitecture: cortical thickness (CtTh), bone volume (BV), total porosity (Ct.Po), intracortical porosity (CtPo-cl), channel/pore number (Po.N), fractal dimension (FD) and degree of anisotropy (Ct.DA). After scan, osteonal morphology was histologically assessed by means: area and perimeter of the osteons (O.Ar; O.p) and of the Haversian canals (C.Ar; C.p). Microtomographic analysis were performed by ANOVA, followed by Tukey and Dunnet tests. Osteon morphology analyses were performed by Kruskal-Wallis, and test Dunn's. Cortical thickness was significant difference (p<0.010) between the NIr and irradiated groups, with thicker cortex at Ir7d (1.15±0.09). The intracortical porosity revealed significant difference (p<0.001) between irradiated groups and NIr, with lower value for Ir7d (0.29±0.09). Bone volume was lower in Ir14d compared to control. Area and perimeter of the osteons were statistically different (p<0.0001) between NIr and Ir7d. Haversian canals also revealed lower values (p<0.0001) in Ir7d (80.57±9.3; 31.63±6.5) compared to NIr and irradiated groups. Cortical microarchitecture was affected by radiation, and the effects appear to be time-dependent, mostly regarding the osteons morphology at the initial days. Cortex structure in Ir21d revealed similarities to control suggesting that microarchitecture resembles normal condition after a period.

Resumo Com o objetivo de avaliar a microarquitetura óssea cortical e a morfologia dos osteons após irradiação, foram utilizados doze coelhos machos da Nova Zelândia. Os animais foram divididos: grupo controle (sem radiação-NIr); e 3 grupos irradiados, sacrificados após: 7 (Ir7d); 14 (Ir14d) e 21 (Ir21d) dias. Foi utilizada uma dose única de radiação de 30 Gy. A microtomografia computadorizada analisou a microarquitetura cortical: espessura cortical (CtTh), volume ósseo (BV), porosidade total (Ct.Po), porosidade intracortical (CtPo-cl), número de canal/ poro (Po.N), dimensão fractal (DF) e grau de anisotropia (Ct.DA). Após a varredura, a morfologia dos osteosn foi avaliada histologicamente por meio de: Área e perímetro do osteon (O.Ar; O.p) e dos canais de Havers (C.Ar; C.p). A análise microtomográfica foi realizada por ANOVA, seguida pelos testes de Tukey e Dunnet. As análises morfológicas do osteon foram realizadas por Kruskal-Wallis e testadas por Dunn. A espessura cortical foi diferente (p<0,010) entre os grupos controle e irradiados, com córtex mais espesso no Ir7d (1,15±0,09). A porosidade intracortical revelou diferenças significativas (p<0,001) entre os grupos irradiados e o controle, com menor valor para Ir7d (0,29±0,09). O volume ósseo foi menor no Ir14d em relação ao controle. Área e perímetro do osteon foi diferente (p<0,0001) entre o controle e Ir7d. Os canais haversianos também revelaram valores mais baixos (p<0,0001) em Ir7d (80.57±9.3; 31.63±6.5) em relação ao controle e demais grupos irradiados. A microarquitetura cortical é afetada pela radiação e os efeitos parecem ser dependentes do tempo, principalmente em relação à morfologia dos osteons nos dias iniciais. A estrutura cortical em Ir21d revelou semelhanças com o controle, sugerindo que a microarquitetura se assemelha à condição normal após um período.

Use of micro-computed tomography for bone evaluation in dentistry

Abstract Micro computed tomography (µCT) follows the same principle of computed-tomography used for patients, however providing higher-resolution. Using a non-destructive approach, samples can be scanned, and each section obtained is used to build a volume using tridimensional reconstruction. For bone analysis, it is possible to obtain information about the tissue's microarchitecture and composition. According to the characteristics of the bone sample (e.g. human or animal origin, long or irregular shape, epiphysis or diaphysis region) the pre-scanning parameters must be defined. The resolution (i.e. voxel size) should be chosen taking into account the features that will be evaluated, and the necessity to identify inner structures (e.g. bone channels and osteocyte lacunae). The region of interest should be delimited, and the threshold that defines the bone tissue set in order to proceed with binarization to separate the voxels representing bone from the other structures (channels, resorption areas, and medullary space). Cancellous bone is evaluated by means of the trabeculae characteristics and their connectivity. The cortex is evaluated in relation to the thickness and porosity. Bone mineral density can also be measured, by the amount of hydroxyapatite. Other parameters such as structure-model-index, anisotropy, and fractal dimension can be assessed. In conclusion, intrinsic and extrinsic determinants of bone quality can be assessed by µCT. In dentistry, this method can be used for evaluating bone loss, alterations in bone metabolism, or the effects of using drugs that impair bone remodeling, and also to assess the success rate of bone repair or surgical procedures.

Resumo A microtomografia computadorizada segue o mesmo princípio da tomografia computadorizada utilizada para avaliação dos pacientes, mas neste caso, é empregada para pequenas amostras com alta resolução. De forma não destrutiva, as amostras podem ser escaneadas, e cada fatia obtida é organizada de forma seriada para formar um volume tridimensional (3D). Para análise óssea, é possível obter informações de microarquitetura e composição mineral, permitindo avaliação distinta entre diferentes sítios. De acordo com as características de cada amostra óssea, como amostras de humanos, animais, ossos longos ou achatados, epífise ou diáfise, etc, devem ser definidos os parâmetros pré-escaneamento com a resolução desejada, levando em consideração quais informações serão extraídas da avaliação. Depois do escaneamento e da reconstrução, deve-se proceder com a seleção da região de interesse (ROI), e depois seguir com o processo de binarização, que se caracteriza pela escolha de um limiar que define os voxels que compõem a região de osso e àqueles que compõem a região dos buracos (canais, áreas de reabsorção e espaço medular). No osso trabecular e no reparo os parâmetros avaliados se baseiam nas características das trabéculas e sua conectividade. No osso cortical os parâmetros estão relacionados com a espessura e porosidade. Além dos parâmetros de microarquitetura, também é possível avaliar a densidade mineral óssea, calculada por volume de hidroxiapatita. Outros parâmetros também podem ser mensurados, utilizando técnicas computacionais como a análise de textura. Parâmetros intrínsecos e extrínsecos da qualidade óssea podem ser avaliados pela microtomografia computadorizada. Na odontologia, este método pode ser empregado em estudos que objetivem avaliar doenças, alterações metabólicas e medicamentos com repercussão no metabolismo ósseo, e na avaliação do processo de reparo e de técnicas cirúrgicas.

Efeito do diabetes mellitus tipo I na organização espacial das trabéculas ósseas: análise por meio do processo de esqueletonização / Effects of type I diabetes mellitus on the spatial organization of bone trabeculae: analysis by skeletonization

Introdução: Diabetes mellitus é uma doença metabólica que afeta vários órgãos-alvo, incluindo os ossos. Objetivo: Avaliar pelo método de esqueletonização o efeito do Diabetes mellitus tipo I (DM1) na microarquitetura de osso esponjoso. Material e métodos: Quatorze ratos Wistar foram divididos em: Saudável (S, n=7) e Diabético (D, n=7). O DM1 foi induzido por meio de injeção endovenosa de estreptozotocina no grupo D, sendo a confirmação da condição realizada por checagem do nível glicêmico. Os animais foram sacrificados após 35 dias da indução no grupo D, juntamente com os do grupo S. As epífises femorais foram seccionadas, removidas, desmineralizadas e incluídas em parafina. Dois cortes (5 µm) foram obtidos, corados em Hematoxilina e Eosina, e analisados ao Microscópio de Luz. Foi realizada a delimitação interativa das trabéculas ósseas, seguido pelo processo de binarização utilizando threshold global, feita por dois operadores distintos. Depois, foi realizado o processo de esqueletonização para acesso às características das trabéculas e da rede de interconexão entre elas. Os parâmetros avaliados foram: Área óssea em micrômetros quadrados (B.Ar, seguido pela proporção em porcentagem BV/TV), Índice de Modelo estrutural (SMI), Dimensão Fractal (FD), Número de trabéculas (Tb.N), Número de ramos (B.N), Número total de junções (Junc.N), Média de pontos terminais (End.p), Média de extensão de cada ramo (R.Le) e Número de junções triplas (Triple.points.N). Resultados: Houve diferença significante apenas no parâmetro SMI para os diferentes operadores (p<0,0001), sendo o mesmo retirado da análise entre diabetes vs saudável. Houve diferença significante na quantidade óssea, sendo maior no grupo S (0,46±0,09) comparado ao grupo D (0,41±0,07) (p=0,0082). Os demais parâmetros não mostraram diferença significante. Conclusão: Conclui-se que a área óssea no grupo saudável é maior em comparação ao DM1. Dentro das limitações deste estudo, parece que a distribuição espacial das trabéculas e suas características de interconexão não são alteradas no diabetes.

Introduction: Diabetes is a metabolic disease that affects several target-organs, including bone. Objective: Analyze the effects of Diabetes Mellitus Type 1 (DM1) on the trabecular bone microarchitecture by using the skeletonization process. Material and methods: Fourteen Wistar rats were divided in two groups: Health (S, n=7) and Diabetic (D, n=7). DM1 was induced with streptozotocin in D group, and glycemic levels were tested on peripheral blood samples. After 35 days, the animals were euthanized and had their femurs removed. The epiphysis were decalcified and embedded in paraffin. Five microns sections were stained in Hematoxylin and Eosin, and analyzed at the light microscope. Bone trabeculae were manually delimited, and then the binarization process with a global threshold was performed for each image. The whole process were conducted by two operators separately. Skeletonization was applied to binary images in order to evaluate the trabeculae characteristics and their network. Bone area (B.Ar), Bone proportion (BV/TV) Strucutre Model Index (SMI), Fractal Dimension (FD), Trabeculae number (Tb.N), Mean branches (B.N), Mean junction points (Junc.N), Mean End-points (End.p), Mean branches length (B.Le), and Mean triple points (Triple.points.N) were evaluated. Results: There was a significant difference only for SMI between different operators (p<0.0001), being this parameter excluded for the evaluation between health and diabetic groups. There was a significant difference between S and D for bone area, with S (0.46±0.09) higher than D (0.41±0.07) (p=0.0082). The other parameters analyzed were not significantly different. Conclusion: Bone trabecular area was higher in health compared with diabetes. Within the limitations of this study, one could suggest that there are no alterations of the spatial distribution of the trabeculae with their network and their inner structural characteristics.

Changes in cortical bone channels network and osteocyte organization after the use of zoledronic acid

Objective The aim of this study was to evaluate the effects of zoledronic acid (ZA) on the cortical bone channels network (CBCN) and osteocyte organization in relation to the bone channels. Materials and methods Eighteen male Wistar rats were divided into control (CG) and test groups (TG). Twelve animals from TG received 3 ZA doses (7.5 µg/kg), and 6 animals from CG did not receive any medication. TG animals were euthanized at 14 (n = 6) and 75 (n = 6) dadys after drug injection. CBCN was analyzed in mandibles and tibias using computational routines. The osteocyte organization was qualitatively evaluated in tibias using a three-dimensional reconstruction of images from serial histological sections. Results Significant differences in CBCN of tibia were found between the treated and untreated rats, with a wider range of sizes and shapes of the channels after the use of ZA (channels area p = 0.0063, channels area SD p = 0.0276) and less bone matrix (bone volume p = 0.0388). The alterations in the channels’ morphology were more evident at 75 days after the drug injection (channels perimeter p = 0.0286). No differences were found in mandibles CBCN. The osteocyte distribution revealed more variable patterns of cell distribution in ZA groups, with non-homogeneous distribution of cells in relation to the bone channels. Conclusion Zoledronic acid induces structural changes in CBCN and modifies the osteocyte arrangement in cortical bone in the tibia; also, the variability in the morphology of bone channels became more evident after a certain time of the use of the drug.

Efeitos do ácido zoledrônico na rede de canais, no sistema lacunocanalicular e nas propriedades biomecânicas do osso cortical / Zoledronic acid effects in the cortical bone channels network, lacunocanalicular system and biomechanical properties

O objetivo do presente trabalho foi avaliar a microarquitetura, caracterizando a rede de canais e o sistema lacunocanalicular, e também, as propriedades biomecânicas, do osso cortical de ratos Wistar tratados com Ácido Zoledrônico. Vinte e um ratos machos foram divididos da seguinte forma: 12 animais alocados no grupo de animais submetidos ao experimento no quarto mês de idade e 9 alocados no grupo de animais submetidos ao experimento no nono mês de idade. Dentro destes grupos, os mesmos foram divididos em grupos controle e teste, sendo que no grupo teste os animais receberam três doses, no esquema de 7,5 g/kg, semanalmente. Após o uso da medicação, três animais foram eutanasiados nos respectivos períodos, após a última dose: 14 dias, 42 dias e 75 dias. Todos os animais tiveram suas mandíbulas, tíbias e fêmures dissecados. As tíbias e mandíbulas foram descalcificadas e processadas para inclusão em parafina, enquanto que, os fêmures foram armazenados para os testes biomecânicos. Imagens histológicas digitalizadas, provenientes das mandíbulas e tíbias, foram analisadas utilizando o processo de segmentação interativa, acessando as características da microarquitetura da rede de canais. O processo de reconstrução tridimensional, para caracterização do sistema lacunocanalicular, foi realizado nos cortes seriados proveniente das tíbias.

Os fêmures foram analisados com os ensaios de microdureza, compressão e flexão em três pontos. Os resultados revelaram que a microarquitetura da rede de canais não foi alterada nas mandíbulas, no entanto, nas tíbias, houve diferença significante apenas nos animais de 4 meses de idade, nos quesitos área dos canais (p 0.0031), desvio-padrão da área (p 0.0338), perímetro dos canais (p 0.0068), desvio padrão do perímetro (p 0.0297) e porcentagem de matriz (p 0.0235). Os testes biomecânicos revelaram resultados que indicavam uma melhora nas características analisadas, que foi correlacionada apenas com o aumento da idade e não com o uso do ácido zoledrônico. A visualização tridimensional possibilitou analisar a distribuição dos osteócitos e dos canais, caracterizando o sistema lacunocanalicular, e constatou-se que nos animais de 4 meses o uso da droga causou uma descontinuidade na estabilidade da distribuição dos osteócitos. Conclui-se que o ácido zoledrônico não interferiu na microarquitetura da rede de canais do osso cortical das mandíbulas, enquanto que nas tíbias, nos animais jovens, houve uma modificação na microarquitetura e no sistema lacunocanalicular, revelando uma topologia mais heterogênea quando comparadas ao grupo controle. As propriedades biomecânicas não sofreram alterações quanto ao uso da droga, sendo evidenciada melhora nos parâmetros analisados relacionada apenas ao aumento de idade nos animais.

The aim of this study was to evaluate the microstructure, characterizing the bone channels network and the lacunocanalicular system, also, the biomechanical properties of the cortical bone of Wistar rats treated with Zoledronic Acid. Twenty-one male rats were separated as follows: 4-month-old (12 animals) and 9 month-old (9 animals). For each age they were divided in control and test group, and the test group received a 7.5 g/kg dose of the drug, once a week, for 3 weeks. In the 4 month-old test group they were euthanized at 14, 42 and 75 days after the drug injection and in the 9-month-old test group they were sacrificed at 14 and 75 days after medication. All animals had their mandible, tibia and femur dissected, and then, the mandible and tibia were decalcified and embedded in paraffin, and, the femur were stored for mechanical tests. Histological images were analyzed through developed algorithms using the SCILAB mathematical environment, getting percentage of bone matrix, ROI areas, ROI perimeters, their standard deviations and Lacunarity, as well, the images were accessed and processed to tridimensional visualization, to characterize the lacunocanalicular system. The femurs were analyzed by microhardness, three point bending and compression testing.

The results revealed that the bone channels network in the mandible was not affected by the drug, whereas the tibia in the 4-month-old animals revealed a statistical difference between control and test groups, with the test group revealing more heterogeneity in the channels topography, as seen in results of bone matrix (p=0.0235), ROI area (p=0.0031), ROI area standard-deviation (p=0.0338), ROI perimeter (p=0.0068) and ROI perimeter standard-deviation (p=0.0297). The biomechanical tests showed an increased in strength, and that this event was age-related and not depending on the use of the drug. The tridimensional visualization allowed to analyze the osteocyte distribution in relation to the channels position, characterizing the lacunocanalicular system, and, it was found that the 4-month-old animals the drug caused a discontinuity in the stability of osteocyte distribution. In conclusion, the zoledronic acid not interfere in the bone channels network in the cortical bone of the mandible, however, in the tibia of the young animals, theres been an alteration in bone channels network and in the lacunocanalicular system, revealing more heterogeneity in the topography in these animals. The biomechanical properties were not altered after the use of the drugs, and the increased in strength being age-related.

Detection of single and mixed colonization of Candida species in patients with denture stomatitis

Aim: To evaluate the profile of the colonization by Candida spp. using presumptive identification to classify the patients with denture stomatitis as having single or mixed colonization, correlating with oral and systemic status. Methods: The CHROMagar Candida™ Medium (CC) for yeast culture and exfoliative cytology was used to identify colonization by Candida spp. and distinguish the different species of the Candida genus from patients with denture stomatitis (DS) and denture wearers without DS (control group). In addition, colonization was correlated with specific habits, such as tobacco and alcohol use, as well as with the use of systemic drugs. Results: Direct swabbing of whole unstimulated saliva (WUS) and palatal mucosa revealed colonization in 97.3% of the patients with DS. In the control group, 55.0% patients presented colonization. The presumptive identification found C. albicans as the most prevalent between both groups, respectively in 89.4% of the DS group and 40.0% from the control group. Regarding the nonalbicans species in the DS group, the most frequent were C. krusei (31.5%), C. glabrata (21.0%) C. tropicalis (15.7%) and Candida spp (2.6%). Smokers presented 90% of mixed isolates, and no C.albicans single colonization in the DS group, with statistically significant difference between smokers and non-smokers (p=0.0051). In the control group, the non-albicans species were C.glabrata (23.0%) and C.tropicalis (23.0%). The results of cytology from the DS group showed positive results in 22.2% of the cases. Conclusions: The use of CC was effective as a complementary method for the diagnosis of colonization by Candida spp. and DS, with the additional advantage of enabling a rapid presumptive identification of the specie. Smoking seemed to play a role in the colonization of oral mucosa by mixed albicans and non-albicans species. Mixed colonization seems to be more prevalent between patients with DS.

Tumor odontogênico queratocístico envolvendo o seio maxilar: relato de caso / Non-syndromic keratocystic odontogenic tumor involving the maxillary sinus: case report

Introdução: O tumor odontogênico queratocístico é raramente encontrado em maxilla. Relato do Caso: Este artigo descreve o relato de um caso de uma paciente de 60 anos de idade, leucoderma, sexo feminino, com um crescimento assintomático na região de maxila posterior diagnosticada como tumor odontogênico queratocístico. Também é discutido o diagnóstico e a propedêutica para este tipo de lesão.

Introduction: The odontogenic keratocyst tumor is rarely found in maxilla. Case Report: This article describes a report of a case of a 60-year-old Caucasian female with an asymptomatic growth in the region of the posterior maxilla tumor diagnosed as odontogenic keratocyst. We discuss the diagnosis and workup for this type of injury.