Este trabalho avaliou o comportamento eletroquímico de três ligas de níquel-cromo e duas de cobalto-cromo in vitro e in vivo. Nos estudos in vitro foram fabricados eletrodos de trabalho a partir destas ligas. Estes eletrodos ficaram imersos em meio de NaCl 0,15 mol.L-1 a 36,5°C e foram submetidos às técnicas do potencial de circuito aberto, polarização potenciostática anódica e cronoamperometria. Para os estudos in vivo foram selecionados quinze pacientes com implantes já osseointegrados e aguardando a instalação das próteses. Esses pacientes foram distribuídos aleatoriamente em cinco grupos, correspondentes às cinco ligas diferentes. Deste modo, as estruturas protéticas dos implantes foram fundidas utilizando as cinco ligas. Foi utilizada a microscopia eletrônica de varredura (MEV) e a espectroscopia por dispersão de energia (EDS) como métodos de análise das superfícies linguais metálicas das próteses, antes da instalação em ambiente oral e após seis meses de permanência. Vistas por MEV, as ligas duas de cobalto-cromo Starloy C e Remanium 2001 e duas de níquel-cromo Wiron 99 e Remanium CSe apresentaram superfícies heterogêneas caracterizadas por uma fase semelhante à matriz da liga e a outra fase, composta por elementos de maior densidade atômica, correspondente à fase interdendrítica. Após seis meses, não houve sinais de corrosão e alterações significativas na composição destas ligas. As ligas de Co-Cr avaliadas, sob o ponto de vista eletroquímico, indicam comportamento catódico quando unidas ao implante sendo um fator indicativo de proteção à corrosão. Dentro dos limites dos estudos in vitro, as ligas de Ni-Cr apresentaram um filme passivo com ampla faixa de potencial em que se mantêm passiva e ausência de corrosão localizada. A liga de níquel-cromo Dan Ceramalloy apresentou a superfície heterogênea com quatro áreas distintas (área 1 rica em titânio; área 2 rica em níquel; área 3 correspondente à matriz; área 4 rica em molibdênio). Após seis meses, as três próteses metalo-cerâmicas apresentaram as regiões correspondentes à fase 2, rica em níquel, com depressões em baixo relevo, sugerindo a oxidação com a liberação de íons níquel. Nos estudos in vitro a liga Dan Ceramalloy apresenta potencial de corrosão mais negativo, uma faixa passiva de potencial mais estreita e um filme menos protetor, quando comparada com as demais ligas estudadas. Apesar de não ter sido observado clinicamente o processo de corrosão das ligas, os estudos in vitro e in vivo deste material, permitiram verificar o seu comportamento eletroquímico e sugerir a descontinuidade do seu emprego em estruturas protéticas de implantes orais. Os estudos in vitro e in vivo apresentaram concordância para todos os materiais estudados. / In this study, the electro-chemical behavior of three nickel-chrome and two cobalt chrome alloys was studied in vitro and in vivo. In the in vitro studies, working electrodes made of these alloys were fabricated. These electrodes were immersed in a medium of NaCl 0.15 mol.L-1 at 36.5°C and were submitted to the open circuit potential, anodic potentiostatic polarization and chronoamperometry techniques. For the in vivo studies, 15 patients whose implants had osseointegrated, and who were waiting for dental implant placement, were selected. These patients were randomly distributed into five groups, corresponding to the five different alloys. Thus the prosthetic implant superstructures were cast, using the five alloys. Scanning electron microscopy (SEM) and energy dispersive spectroscopy (EDS) were used as methods to analyze the metal lingual surfaces of the prostheses, before they were placed in the oral environment, and after six months in place. Visualized by SEM, the two cobalt-chrome alloys Starloy C and Remanium 2001, and two types of nickel-chrome - Wiron 99 and Remanium Cse - presented heterogeneous surfaces characterized by a phase similar to that of the alloy matrix, and the other phase composed of elements of greater atomic density, corresponding to the interdendritic phase. After six months, there were no signs of corrosion and significant alterations in the composition of these alloys. The Co-Cr alloys evaluated, from an electrochemical point of view, indicated a cathodic behavior when united to the implant, this being a factor indicative of protection against corrosion. Within the limits of the in vitro studies, the Ni-Cr alloys presented a passive film with an ample range of potential in which they remained passive, with absence of corrosion due to pitting. The nickel-chrome alloy Dan Ceramalloy presented a heterogeneous surface with four distinct areas (Area 1 rich in titanium; Area 2 rich in nickel; Area 3 corresponding to the matrix; Area 4 rich in molybdenum). After six months, the three metal-ceramic dental prostheses presented the regions corresponding to Phase 2, rich in nickel, with depressions in low relief, suggesting oxidation with nickel ion release. In the in vitro studies, the alloy Dan Ceramalloy presented a more negative corrosion potential, a narrower range of passive potential and a less protective film, when compared with the other alloys studied. In despite of the process of corrosion of the alloys not having been clinically observed, the in vitro and in vivo studies of this material allowed verification of its electrochemical behavior, and suggest that its use in prosthetic superstructures for oral implants should be discontinued. The in vitro and in vivo studies were shown to be in agreement for all the materials studied.

Abstract Osteoporosis is a disease characterized by abnormalities in the amount and architectural arrangement of bone tissue, which leads to impaired skeletal strength and increased susceptibility to fractures. Type 5 tartrate-resistant acid phosphatase (TRACP, AcP5) has been suggested to participate directly in bone resorption. In this study, baculovirus expression vector system in insect cells was used to gain large amounts of recombinant type 5 acid phosphatase for structure determination, structure-based functional studies and production of monoclonal antibodies. Active and inactive forms of the enzyme were separated from each other by cation-exchange chromatography, and characterized. The enzyme was crystallized and the three-dimensional structure was determined. Based on the three-dimensional structure of the active site five different enzyme variants were constructed, produced in insect cells, and purified. The wild type enzyme and the mutated forms were characterized, and their kinetic parameters were determined. The importance of amino acids that were expected to be essential for the acid phosphatase activity was confirmed. The acid phosphatase activity and reactive oxygen species generating activity of this dual enzyme proved to exploit different amino acids in their reaction mechanisms. Further studies are needed to clarify the physiological substrates of TRACP in vivo. The findings of this study could form a base for construction of inhibitors for TRACP that could be useful therapeutic agents for osteoporosis and related bone disorders.

Please read the abstract on page 3 in the dissertation. / Dissertation (MSc)--University of Pretoria, 2011. / Chemical Pathology / unrestricted

A erupção dentária depende de uma coordenada interação entre o germe dentário e o tecido ósseo da cripta que o envolve. Para que seja formada a via eruptiva, a reabsorção da porção oclusal da cripta óssea por osteoclastos é indispensável. Os bisfosfonatos são drogas com reconhecida capacidade de inibir a atividade clástica e foram empregados no presente estudo a fim de interferir no tecido ósseo da cripta alveolar durante a formação e erupção de molares de ratos. Doses diárias dos bisfosfonatos alendronato ou etidronato de 2,5 e 8 mg/kg, respectivamente, foram administradas a ratos recém nascidos. Os controles foram injetados com solução salina. Nos períodos de 4, 8, 14, 21 e 28 dias, as maxilas foram fixadas em 2,5% de formaldeído + 2% de glutaraldeído, 4% de formaldeído + 0,1% de glutaraldeído ou fixador Zamboni, descalcificadas em EDTA a 4,13% e processadas para análise em microscopias de luz, eletrônica de transmissão e confocal, histoquímica TRAP, imunocitoquímica para OPN, BSP, RANK, RANKL e OPG. Alguns espécimes não foram descalcificados para análise em microscopia eletrônica de varredura, ou congelados em nitrogênio líquido para extração e análise e da expressão de proteínas por Western Blotting. O etidronato ocasionou alterações no metabolismo ósseo da cripta que resultaram no atraso da erupção e da formação radicular em relação ao controle. O alendronato aumentou o número de osteoclastos no osso alveolar, porém a maioria apresentou estado latente, o que diminuiu a reabsorção óssea da cripta ao redor do germe dentário e impediu a erupção dos molares e a formação radicular. A expressão de RANKL, molécula ativadora dos osteoclastos, durante o início do processo eruptivo, diminuiu em comparação ao controle. Com a diminuição da remodelação óssea, o tecido apresentou distribuição de OPN e BSP típica de osso primário. Os resultados demonstram que a reabsorção óssea é importante em todos os pontos da cripta e não apenas em sua porção oclusal durante a formação da via eruptiva, para que a formação e erupção dentária ocorram adequadamente. / Tooth eruption depends on coordinated interactions between the tooth germ and the surrounding bony crypt. The formation of the eruptive pathway requires the resorption of the occlusal portion of the bony crypt by osteoclasts. The bisphosphonates are drugs with known capability to inhibit clastic activity, and were employed in the present study with the aim of interfering in the alveolar bony crypt during the formation and eruption of rat molars. Daily alendronate or etidronate 2.5 and 8 mg/kg doses, respectively, were administered to newborn rats. The controls were injected with saline solution. At 4, 8, 14, 21 and 28 days, the maxillae were fixed in 2.5% formaldehyde + 2% glutaraldehyde, 4% formaldehyde + 0,1% glutaraldehyde or Zambonis fixative, decalcified in 4.13% EDTA and processed for light, confocal and transmission electron microscopy analyzes, TRAP histochemistry, and immunocytochemistry for OPN, BSP, RANK, RANKL and OPG. Some specimens were left undecalcified for scanning electron microscopy analyzes, or frozen in liquid nitrogen for protein extraction and Western Blotting protein expression analyzes. Etidronate occasioned alterations in the alveolar bony crypt metabolism that resulted in the delay of tooth eruption. Alendronate increased osteoclast number in the alveolar bone; however, most of them were latent, which decreased the resorption of the bony crypt surrounding the tooth germ and impeded the eruption and root formation of the molars. The expression of RANKL, an osteoclast-activating molecule, was decreased. The inhibition of the bone remodeling resulted in typical primary bone OPN and BSP labeling pattern. These results demonstrate that bone resorption at the entire surface of the crypt, and not only during the eruptive pathway formation, is crucial for adequate tooth eruption and formation.

Osteopetrosis is a metabolic bone disease in mammals characterized by a generalized skeletal sclerosis caused by reduced bone resorption. This reduced bone resorption is manifested in afflicted animals by abnormal bone shape, reduced or absent marrow cavities, extramedullary hemopoiesis, abnormal mineral homeostasis and absent or delayed tooth eruption. The available osteopetrotic animal mutations have been a constant source of fruitful investigations concerning the systemic regulation of osteoclastogenesis and bone metabolism. Tooth eruption, on the other hand, is a localized manifestation of the timely activation of bone resorption and bone formation on opposite sides of an erupting tooth. Its rate-limiting step is the speed of bone resorption to form the eruption pathway. In this dissertation, we used two osteopetrotic rat mutations, toothless (tl) and microphthalmia blanc (mib), to investigate the abnormal development of osteoclasts and tooth eruption in mutant rats with an emphasis on the role of systemic and local factors. The significant contributions to this work are listed below. 1. In the toothless rat, a mutation lacking erupted dentition due to severely reduced bone resorption, colony-stimulating factor-1 (CSF-1) promoted tooth eruption but this was delayed compared to normal rats. Eruption was accompanied by changes in the populations of tartrate-resistant acid phosphatase-positive (TRAP+) mononuclear cells in the dental follicle and TRAP+ osteoclasts on adjacent alveolar bone surfaces. These cell populations were dramatically increased in treated mutants compared to untreated tl rats, but the timing of their appearance was delayed compared to normal littermates. This lag in the appearance of osteoclasts and their precursors corresponded to the delay in eruption of first molars in treated tl rats. 2. CSF-1 also accelerated the eruption of molars in normal rats. CSF-1 increased the number of TRAP+ mononuclear cells in the dental follicle and TRAP+ osteoclasts on adjacent alveolar bone surfaces, but had no effect on the timing of their appearance in normal rats. 3. Our data revealed a differential effect on tooth eruption of the growth factors CSF-1 and epidermal growth factor (EGF). CSF-1 accelerated eruption of molars in normal rats, but had no effect on incisor eruption. On the other hand, EGF accelerated incisor eruption; but did not affect molar eruption in normal rats. 4. We have described the mechanism for the transient, mild form of osteopetrosis inherited by mib rats. Mutant animals possess a typical sclerosis at birth, which diminished--but was not resolved--during the first postnatal month. These characteristics are caused by early reductions in osteoclast number and function which improve to normal levels by 4 weeks. Osteoclast numbers were severely reduced in mib rats between birth and 2 weeks, but improved to near normal levels by 4 weeks. Neonatal abnormalities in osteoclast function included reduced staining for the functional enzymes TRAP and TrATPase, decreased levels of mRNA for both TrATPase and CAll, and inability to form a well-developed ruffled border. None of these defects were apparent after the first postnatal month. 5. Finally, we have shown that the dental abnormalities caused by the mild, transient form of osteopetrosis in mib rats are limited to incisor defects and delayed eruption of all teeth. Histologic and radiographic examination of mutant incisors revealed that, contrary to the situation in normal rats, the apex of the incisors of mib rats failed to extend past the first molar region to the third molar. The incisor apex of newborn mib rats was misshaped due to ankylosis of incisor matrices with alveolar bone. This ankylosis was temporary, being resolved by the third postnatal day. The delayed eruption of incisors in mib rats and abnormal shape and occlusion of these teeth in older animals is a consequence of the temporary ankylosis in newborn rats.

No description available.

Les eicosanoïdes sont des médiateurs importants qui encadrent et régulent les fonctions osseuses. Leur production est sous la tutelle des phospholipases A[indice inférieur 2] qui permettent la relâche d’acide arachidonique et de lysophospholipides puis de leur métabolisme subséquent des membranes cellulaires. Les PLA[indice inférieur 2] sécrétées ont également comme particularité de pouvoir exercer leurs effets directement via leurs récepteurs membranaires comme ligand. Malgré l’implication connue des prostaglandines sur les fonctions ostéoclastiques et dans plusieurs processus pathologiques résultants en érosion osseuse, les phospholipases A[indice inférieur 2] ostéoclastiques restent inconnues et leurs rôles, spéculatifs. Les études présentées démontrent la présence de la cPLA[indice inférieur 2]-α et de la sPLA[indice inférieur 2] IIA chez les ostéoclastes humains. Par contre, leur expression semble différer selon l’état de l’os. En effet, la cPLA[indice inférieur 2]-α semble exprimée ubiquitairement, mais la sPLA[indice inférieur 2] IIA n’est détectable que dans l’os fœtal ou atteint de la maladie de Paget et n’est pas exprimée dans l’os sain, ostéoporotique ou arthrosique. La sPLA[indice inférieur 2] IIA semble donc exprimée en condition de fort remodelage osseux. Dans notre modèle, la cPLA[indice inférieur 2]-α revêt un rôle anti-ostéoclastogénique. En effet, la cPLA[indice inférieur 2]- α produit des écosanoïdes qui inhibent l’ostéoclastogenèse. Ces derniers sensibilisent les ostéoclastes à l’apoptose. En revanche, un certain degré d’activation de la cPLA[indice inférieur 2]-α est requis pour la résorption osseuse, car son inhibition bloque la résorption osseuse en désorganisant les anneaux d’actine requis pour la résorption, et ce, de façon dépendante de la production d’acide arachidonique. En ce qui a trait à la sPLA[indice inférieur 2] IIA, elle stimule l’ostéoclastogenèse indépendamment de son activité catalytique, probablement via l’un de ses récepteurs membranaires. D’autre part, elle confère une résistance à l’apoptose autant chez les ostéoclastes matures que chez leurs précurseurs CD14+. Par contre, l’inhibition de la sPLA[indice inférieur 2] IIA bloque la résorption osseuse indépendamment du remodelage du cytosquelette d’actine, probablement via leur apoptose. Les résultats inclus dans cette thèse semblent donc démontrer la présence de deux PLA[indice inférieur 2]s chez les ostéoclastes humains ainsi que leur attribuer des rôles en physiologie et pathologie osseuse. Ces évidences pourraient faire des PLA[indice inférieur 2] de nouvelles cibles thérapeutiques pour le traitement de pathologies ostéo-articulaires, dont la maladie osseuse de Paget. // Abstract : Eicosanoïds are important mediators of bone biology. The first regulated enzymes in the biosynthetic pathway of eicosanoids are the PLA[subscript 2]s, which release arachidonic acid and lysophospholipids from membrane phospholipids. Further metabolism of arachidonic acid will lead, among other things, to the synthesis of prostaglandins, which deeply influence bone metabolism. Secreted PLA[subscript 2]s (sPL[subscript 2]) may also act independently of their catalytic activity, as a ligand to their membrane bound receptors. Even though PLA[subscript 2]s have been associated with joint and bone pathologies, their presence and functions were never investigated in osteoclasts (OCs), the principal cell responsible for bone destruction. Our study established the presence of cPLA[subscript 2] and sPLA[subscript 2] in human OCs, but demonstrated a contrast in their expression. cPLA[subscript 2] seems to be expressed ubiquitously, contrary to sPLA[subscript 2] whose expression is restricted to OCs from foetal bone and bone suffering from Paget disease. There is no trace of sPLA[subscript 2] in healthy bone or bone suffering from osteoarthrosis or osteoporosis, thus sPLA[subscript 2] seems tightly linked to high bone turnover. In our model, cPLA[subscript 2] exerted an anti-osteoclastogenic effect. Indeed, cPLA[subscript 2] produces eicosanoïds that inhibit osteoclastogenesis and sensitize OCs to apoptosis. Nevertheless, a minimum cPLA[subscript 2] activity is required since complete cPLA[subscript 2] inhibition blocks osteoclast mediated bone resorption. Its inhibition leads to disorganization of the OC cytoskeleton and inhibits the actin ring formation required for bone resorption in an arachidonic acid dependent fashion. On the other hand, sPLA[subscript 2] stimulates osteoclastogenesis independently of its catalytic activity; probably via interaction with one of its membrane bound receptors. sPLA[subscript 2] decreases OC and their CD14+ precursors’ sensibility to apoptosis. Moreover, sPLA[subscript 2] inhibition inhibits bone resorption independently of actin cytoskeleton remodeling, probably by inducing mature OC apoptosis. Together, these results demonstrate the presence of two PLA[subscript 2]s in human OCs and highlight their important roles in bone physiology and pathophysiology. Highlighting those functions could eventually lead to the elaboration of new strategies to control hyperosteoclastic states by targeting PL[subscript 2]s.

L’arthrose (OA) est une maladie articulaire dégénérative à l’étiologie complexe et diverse. Les travaux de ces dernières années ont démontré que l’OA est une pathologie affectant tous les tissus de l’articulation incluant le cartilage, la membrane synoviale et l’os sous-chondral. L’OA se traduit par une déstructuration et une perte de fonctionnalité de l’articulation, et est principalement caractérisée par une perte de cartilage articulaire. L’inflammation de la membrane synoviale joue un rôle déterminant dans la progression de l’OA, toutefois elle serait secondaire à la dégradation du cartilage. De plus, l’os sous-chondral est également le siège de nombreuses transformations lors de l’OA. Il est fortement suggéré que ces changements ne correspondent pas seulement à une conséquence, mais pourraient être une cause du développement de l’OA impliquant une communication entre ce tissu et le cartilage. Il est maintenant bien établi que les voies inflammatoires et cataboliques jouent un rôle crucial dans l’OA. C’est pourquoi, nous avons étudié l’implication d’une nouvelle famille de récepteurs membranaires, les PARs, et plus particulièrement le PAR-2 dans les voies physiopathologiques de l’OA. Notre hypothèse est que l’activation de PAR-2 au cours de l’OA est un phénomène majeur du développement/progression de la maladie faisant du récepteur PAR-2 un candidat privilégié pour le développement de nouvelles approches thérapeutiques ciblant non seulement le cartilage mais aussi l’os sous-chondral. Pour cette étude, nous avons travaillé in vitro avec des chondrocytes (Cr) et des ostéoblastes (Ob) OA respectivement du cartilage et de l’os sous-chondral du condyle fémoral humain. Nos résultats ont démontré que PAR-2 était plus exprimé dans les Cr et les Ob OA que dans les cellules normales. Par ailleurs, PAR-2 est régulé positivement par certains facteurs retrouvés au cours de l’OA comme l’IL-1β, le TNF-α et le TGF-β dans les Cr OA, et par l’IL-1β, le TNF-α et la PGE2 dans les Ob OA. De plus, les principaux facteurs cataboliques et inflammatoires, soit la MMP-1, la MMP-13 et la COX-2 sont produits en quantité plus élevée suite à l’activation du récepteur dans le cartilage OA. De même, l’activation de PAR-2 dans les Ob OA conduit à une production accrue de facteurs pro-résorptifs tels que RANKL, l’IL-6, la MMP-1 et la MMP-9, et à l’augmentation de l’activité pro-résorptive de ces cellules. En outre, dans les deux types tissulaires étudiés, l’activation de PAR-2 augmente l’activité de certaines protéines de la famille des MAPKinases comme Erk1/2, p38 et JNK. Finalement, nous avons conclu notre étude en employant un modèle in vivo d’OA induite chez la souris sauvage et déficiente pour le gène PAR-2. Nos résultats ont démontré que l’absence d’expression et de production de PAR-2 influençait le processus inflammatoire et les changements structuraux affectant à la fois le cartilage et l’os sous-chondral, conduisant à un ralentissement du développement de l’OA. Nos travaux de recherche ont donc permis de montrer que le récepteur PAR-2 est un élément majeur du processus OA en agissant sur les voies cataboliques et inflammatoires du cartilage, et sur le remodelage tissulaire de l’os sous-chondral. Mots-clés : Arthrose, chondrocyte, cartilage, ostéoblaste, os sous-chondral, PAR-2, MMPs, COX, ILs, RANKL, résorption osseuse, MAPKinase, catabolisme, inflammation / Osteoarthritis (OA) is a complex degenerative articular disease. Recent studies have shown that OA is a pathology affecting all the tissues of the joint including the cartilage, the synovial membrane and the subchondral bone. OA is regarded as destruction and a loss of functionality of joint, and is mainly characterized by a loss of articular cartilage. The synovial inflammation also plays a key role in the progression of OA; however, it is believed to be secondary to cartilage degradation. In addition, there are also in the subchondral bone numerous changes which occur during the course of the disease. It is strongly suggested that these changes in subchondral bone are not just a consequence but could be a cause of the development of OA, thus involving a cross-talk between this tissue and the cartilage. It is now well established that inflammatory and catabolic pathways play a crucial role during OA. This is why we have engaged the study of the involvement of a new family of membranous receptors, the PARs, and more particularly PAR-2 during the pathophysiological process of OA. Our hypothesis is that PAR-2 activation during OA course is a major phenomenon for the development/progression of the disease, and that PAR-2 seems a suitable candidate for the development of new therapeutic approaches targeting not only the cartilage but also the subchondral bone. Thus, we have performed in vitro studies on OA chondrocyte (Cr) and osteoblasts (Ob) cells issued respectively from human femoral condyle cartilage and subchondral bone. Our results showed that PAR-2 was expressed at a higher level in the OA Cr and Ob compared to normal cells. Moreover, PAR-2 is found to be positively regulated by some factors present during the course of OA, such as IL-1β, TNF-α and TGF-β in the OA Cr, and IL-1β, TNF-α and PGE2 in the OA Ob. In addition, the main catabolic and inflammatory factors including MMP-1, MMP-13 and COX-2 are enhanced following PAR-2 receptor activation in the OA cartilage. Similarly, the activation of PAR-2 in OA Ob lead to an increased production of pro-resorptive factors such as RANKL, IL-6, MMP-1 and MMP-9, and an increased pro-resorptive activity of these cells. In addition, in both tissue types, PAR-2 activation increases the activity of certain protein MAPKinases family such as Erk1/2, p38 and JNK. Finally, we have performed an in vivo study using an OA induced model in wild-type and PAR-2 gene knock-out mice. Our results demonstrated that the absence of PAR-2 expression and production influenced the inflammatory process and structural changes affecting the cartilage and the subchondral bone, leading to a slowing down of OA development. Our research investigation have bring to light that PAR-2 receptor is a key element during OA process by acting on the cartilage catabolic and inflammatory pathways as well as the tissue remodelling of the subchondral bone. Keywords : Osteoarthritis, chondrocyte, cartilage, osteoblast, subchondral bone, PAR-2, MMPs, COX, ILs, RANKL, bone resorption, MAPKinase , catabolism, inflammation

Tensão elevada no osso pode provocar reabsorção óssea. O objetivo deste estudo é avaliar por meio do método dos elementos finitos tridimensionais as áreas de concentração de tensões no osso basal após realização dos orifícios para inserção dos implantes e com implantes osseointegrados, em mandíbulas com três níveis de reabsorção e prótese mucosa-suportada. Foram confeccionados 7 modelos por meio do software de CAD: Rhinoceros 4.0 SR9; 1 (controle)- mandíbula com 15mm de altura e sem orifício; 2- mandíbula com 15mm de altura e 2 orifícios de 3,75mm de diâmetro; 3- mandíbula com 15mm e 2 implantes osseointegrados; 4- mandíbula com 20mm e 2 orifícios; 5- mandíbula com 20mm e 2 implantes osseointegrados 6- mandíbula com 26mm e 2 orifícios e 7- mandíbula com 26mm e 2 implantes osseointegrados. A força aplicada foi de 60N no sentido axial, nos dentes artificiais da prótese removível total reembasada com 2mm de material macio. Após a discretização dos modelos foi realizado o ensaio pelo método dos elementos finitos por meio do Software NEiNastran for Windows Version 10.0.3.997. Os resultados evidenciaram aumento da tensão no osso cortical em relação ao medular. Mandíbulas sem perfuração apresentaram picos de tensão mais elevados que as com implantes osseointegrados. Nas mandíbulas com orifícos os picos de tensão se localizaram no sentido vestíbulo-lingual enquanto nas com implantes esses picos de tensão foram no sentido mésio-distal. Quanto mais reabsorvida a mandíbula maior a tensão transmitida pela base da prótese removível total ao orifício. Conclui-se que as mandíbulas com 15 e 20 mm de altura com orifícios apresentaram maiores picos de tensão que as com implantes osseointegrados, enquanto nas com 26mm de altura não houve diferença entre ambas condições / Increased pressure in the bone can cause resorption. The purpose of this study is to evaluate areas of stress concentration in the basal mandibular bone after perforation for inserting the implants and dental implants in mandibles with three levels of resorption and muco-supported dental prosthesis using the three-dimensional finite element method. 7 models were produced through CAD software: Rhinoceros 4.0 SR9; 1 (control) -15mm jaw height and without perforation; 2- 15mm jaw height and two perforations of 3.75 mm in diameter; 3- 15mm jaw height with 2 osseointegrated dental implants; 4- 20mm jaw height and 2 perforations; 5- 20mm jaw heigth with two osseointegrated dental implants; 6- 26mm jaw height and two perforation; and 7- 26mm jaw height with two osseointegrated dental implants. The force of 60N was applied in the axial direction to the artificial teeth denture provisional recover with 2mm soft material for edentolous tissue conditioner. After discretization of the test models it was performed a trial using the finite element method through NEiNastran Software for Windows Version 10.0.3.997. The results showed increased tension in cortical bone in relation to the medular bone. Mandibles without perforation showed stress peaks higher than those with dental implants. In the jaws with drilling the stress peaks were located in the buccal-lingual direction while in those with implant they were in the mesial-distal direction. The more resorbed jaw greater the tension transmitted by the base of full removable denture to the drilling. The study concluded that the jaws 15 and 20 mm height perforated had higher stress peaks than the ones with dental implants while in the 26mm height perforated no difference between the two conditions was observed

A acidose metabólica causa perda de mineral ósseo e a estimulação mecânica causa remodelamento ósseo adaptativo. A reabsorção óssea que caracteriza essas mudanças ósseas depende da acidificação extracelular pela secreção vetorial de H+ pelos osteoclastos. A H+-ATPase vacuolar em paralelo com o trocador Cl-/H+ (CLC7) são os mecanismos conhecidos envolvidos na reabsorção óssea, entretanto, os osteoclastos também expressam canais para H+ dependentes a voltagem. Este trabalho foi realizado para avaliar a contribuição dos canais para H+ na função celular visando à compreensão de seu relacionamento com a H+-ATPase vacuolar e o CLC7 (1); analisar se o fluxo de fluido extracelular modifica a secreção de H+ (2) e avaliar a diferenciação dos osteoclastos in vitro sob acidose metabólica devido à redução do HCO3- (3). Osteoclastos de ratos Wistar foram obtidos diretamente dos animais ou foram diferenciados in vitro (com M-CSF e RANKL) e semeados sobre vidro, plástico ou substratos mineralizados em <font face=\"Symbol\">&#945;-MEM + 10% SFB, em pH 7,4 ou 6,9, e então mantidos em incubadora com 5% CO2, a 37<font face=\"Symbol\">&#176;C. A diferenciação celular foi avaliada pela contagem de células TRAP-positivas ou de núcleos marcados por DAPI. A secreção de H+ foi avaliada por epifluorescência, utilizando-se BCECF-AM, sensível a pH. Os registros do pH intracelular foram feitos na vigência de soluções tamponadas por HEPES, na ausência de CO2/HCO3- (pH 7,4, 300 mOsm/L H2O, a 37<font face=\"Symbol\">&#176;C), na presença ou ausência de perfusão contínua de fluido extracelular a uma velocidade de 5 ml/min. Na ausência de perfusão, os osteoclastos exibiram variações cíclicas do pHi (acidificação e alcalinização espontâneas), com período de 12 a 45 minutos (n = 35) e amplitude de 0,12 a 1,43 unidades de pHi. As oscilações não foram abolidas por concanamicina (100 mM) (n = 3), por NPPB (100 <font face=\"Symbol\">mM) (n = 3), na ausência de Na+ extracelular (n = 5) ou na ausência de Cl- extracelular (n = 3). O fluxo de fluido aboliu as oscilações e a ausência de Cl- extracelular modificou significativamente seu padrão. Na ausência de perfusão, a secreção de H+ após acidificação intracelular induzida foi abolida por Zn2+ (100 <font face=\"Symbol\">mM) (n = 5). Além disso, na presença de perfusão, a secreção de H+ após acidificação intracelular induzida foi abolida por NPPB (n = 4) e não foi abolida por bafilomicina (200 nm) (n = 3). A acidose metabólica não modifica o número de osteoclastos diferenciados in vitro, entretanto, o tratamento das culturas com Zn2+ causou redução do numero de células mononucleares e aumento relativo do número de osteoclastos multinucleados em relação ao controle tanto em pH 7,4 quanto em pH 6,9. / Metabolic acidosis can cause a loss of bone mineral and the mechanic stimulation can cause adaptative bone remodeling. The bone resorption characteristic of these bone changes aforementioned depends on the extracellular acidification by osteoclastmediated proton secretion. The H+ secretion by vacuolar H+-ATPase together with Clsecretion through a Cl-/H+ exchanger (CLC7) are the known mechanisms involved in the bone resorption; however, osteoclasts also express voltage-gated proton channels. The proposed aims of these work were to evaluate the contribution of proton channels in the osteoclast function for better understanding its relation with vacuolar H+-ATPase and CLC7 (1); to analyze whether the flow of extracellular fluid modifies the H+ secretion or not (2); and to analyse the osteoclast differentiation in vitro under metabolic acidosis due to HCO3- reduction (3). Osteoclasts were freshly isolated or generated from bone marrow precursor cells (using M-CSF and RANK- L) from of Wistar rats. The cells were placed on glass coverslips, plastic coverslips, or on mineralized substrate in <font face=\"Symbol\">&#945;-MEM + 10% FBS, pH 7.4 or 6.9, and then maintained in a 5% CO2 incubator at 37<font face=\"Symbol\">&#176;C. The differentiation was analyzed by counting of TRAP-stained cells or DAPIstained nuclei. The H+ secretion was analysed by epifluorescence, using the pHsensitive dye BCECF-AM. The intracellular pH record was done using a standard HEPES-buffered solution free of CO2/HCO3- (pH 7.4, 300 mOsm/L H2O, at 37<font face=\"Symbol\">&#176;C), with or without continuous perfusion of extracellular fluid at a rate of 5 ml/min. In the absence of perfusion, the osteoclasts exhibit cyclic pHi variations (spontaneous acidification and alkalinization), with a period of 12 to 45 minutes (n = 35) and amplitude difference between maximal and minimal pHi of 0.12 to 1.43 units pHi. These oscillations were not abolished in the presence of oncanamycin (100 mM) (n = 3), NPPB (100 <font face=\"Symbol\">mM) (n = 3), in the absence of Na+ (n = 5) or in the absence of Cl- (n = 3) in the extracellular solution. The fluid flow itself abolished the pH oscillations and the absence of extracellular Cl- modifies significantly these patterns. In the absence of perfusion, the H+ secretion after induced intracellular acidification was abolished by Zn2+ (100 <font face=\"Symbol\">mM) (n = 5). In addition, in the presence of perfusion, the H+ secretion after induced intracellular acidification was abolished by NPPB (n = 4) and was not abolished by bafilomycin (200 nm) (n = 3). Metabolic acidosis does not modify the number of osteoclasts differentiated in vitro, however, when the cell culture was treated with Zn2+, there was a significant reduction in the number of mononuclear cells and a relative increase in the number of multinucleated osteoclasts compared to control, both in pH 7.4 and pH 6.9 medium.