Ultrastructural and histochemical examination of alveolar bone at the pressure areas of rat molars submitted to continuous orthodontic force

It is usually believed that repair in alveolar bone during orthodontic movement occurs after decreasing of force. However, we have recently observed signs of repair in previously resorbed cementum from human teeth exposed to continuous forces. In order to test the hypothesis that bone resorption and deposition occur concomitantly at the pressure areas, a continuous 15 cN force was applied in a buccal direction to upper first molars from eight 2.5-month-old male Wistar rats for 3 d (n=4) and 7 d (n=4). As a control, two additional rats did not have their molars moved. Maxillae were fixed in 2% glutaraldehyde + 2.5% formaldehyde, under microwave irradiation, decalcified in ethylenediaminetetraacetic acid, and processed for transmission electron microscopy. Specimens from one rat from each group were processed for tartrate-resistant acid phosphatase (TRAP) histochemistry. At both the times studied, the alveolar bone surface at the pressure areas showed numerous TRAP-positive osteoclasts, which were apposed to resorption lacunae. In addition, osteoblasts with numerous synthesis organelles were present in the neighboring areas overlying an organic matrix. Thus, this study provides evidence that the application of continuous forces produces concomitant bone resorption and formation at the pressure areas in rat molars.

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

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

Differential expression of osteoblast and osteoclast chemmoatractants in compression and tension sides during orthodontic movement

Orthodontic tooth movement is achieved by the remodeling of alveolar bone in response to mechanical loading, and is supposed to be mediated by several host mediators, such as chemokines. In this study we investigated the pattern of mRNAs expression encoding for osteoblast and osteoclast related chemokines, and further correlated them with the profile of bone remodeling markers in palatal and buccal sides of tooth under orthodontic force, where tensile (T) and compressive (C) forces, respectively, predominate. Real-time PCR was performed with periodontal ligament mRNA from samples of T and C sides of human teeth submitted to rapid maxillary expansion, while periodontal ligament of normal teeth were used as controls. Results showed that both T and C sides exhibited significant higher expression of all targets when compared to controls. Comparing C and T sides, C side exhibited higher expression of MCP-1/CCL2, MIP-1 alpha/CCL3 and RANKL, while T side presented higher expression of OCN. The expression of RANTES/CCL5 and SDF-1/CXCL12 was similar in C and T sides. Our data demonstrate a differential expression of chemokines in compressed and stretched PDL during orthodontic tooth movement, suggesting that chemokines pattern may contribute to the differential bone remodeling in response to orthodontic force through the establishment of distinct microenvironments in compression and tension sides. (C) 2008 Elsevier Ltd. All rights reserved.

Growth hormone receptor and IGF-1 receptor immunoreactivity during orthodontic tooth movement in the prednisolone-treated rat

Bone remodeling during tooth movement is regulated by local and systemic factors. Two regulators of bone metabolism are growth hormone (GH) and insulin-like growth factor-I (IGF-1). Their effects are mediated via binding to GH receptor (GHR) and IGF-I receptor (IGF-IR) in target tissues. Corticosteroids may affect the activity of these growth factors. This study examined the effect of prednisolone on GHR and IGF-IR expression in dental tissues following orthodontic tooth movement. The corti ticosteroid-treated group (N = 6) was administered prednisolone ( 1 mg/kg,) daily and the control group (N = 6) received equivalent volumes of saline. An orthodontic force (30 g) was applied to the maxillary first molar. Animals were sacrificed 12 days postappliance insertion. Sagittal sections of the first molar were stained for GHR and IGF-IR immunoreactivity. GHR and IGF-IR cell counts were elevated following appliance-treatment. Orthodontic tooth movement appeared to up-regulate GHR and IGF-IR immunoreactivity, but this up-regulation was reduced following prednisolone treatment. The suppression of GHR and IGF-I immunoreactivity in steroid-treated animals infers the mechanism whereby bone resorption and deposition, necessary for orthodontic tooth movement, may be inhibited by prednisolone. However, at 12 days postappliance insertion. no difference in orthodontic tooth movement was observed following low-dose prednisolone treatment.

Myeloperoxidase activity is increased in gingival crevicular fluid and whole saliva after fixed orthodontic appliance activation

Introduction: Orthodontic tooth movement uses mechanical forces that result in inflammation in the first days. Myeloperoxidase (MPO) is an enzyme found in polymorphonuclear neutrophil (PMN) granules, and it is used to estimate the number of PMN granules in tissues. So far, MPO has not been used to study the inflammatory alterations after the application of orthodontic tooth movement forces. The aim of this study was to determine MPO activity in the gingival crevicular fluid (GCF) and saliva (whole stimulated saliva) of orthodontic patients at different time points after fixed appliance activation. Methods: MPO was determined in the GCF and collected by means of periopaper from the saliva of 14 patients with orthodontic fixed appliances. GCF and saliva samples were collected at baseline, 2 hours, and 7 and 14 days after application of the orthodontic force. Results: Mean MPO activity was increased in both the GCF and saliva of orthodontic patients at 2 hours after appliance activation (P<0.02 for all comparisons). At 2 hours, PMN infiltration into the periodontal ligament from the orthodontic force probably results in the increased MPO level observed at this time point. Conclusions: MPO might be a good marker to assess inflammation in orthodontic movement; it deserves further studies in orthodontic therapy. (Am J Orthod Dentofacial Orthop 2010;138:613-6)

Effects of Externally Applied, Cyclical, Low Magnitude Forces on Orthodontic Tooth Movement in Rats

Orthodontic tooth movement requires external orthodontic forces to be converted to cellular signals that result in the coordinated removal of bone on one side of the tooth (compression side) by osteoclasts, and the formation of new bone by osteoblasts on the other side (tension side). The length of orthodontic treatment can take several years, leading to problems of caries, periodontal disease, root resorption, and patient dissatisfaction. It appears that the velocity of tooth movement is largely dependent on the rate of alveolar bone remodeling. Pharmacological approaches to increase the rate of tooth movement are limited due to patient discomfort, severe root resorption, and drug-induced side effects. Recently, externally applied, cyclical, low magnitude forces (CLMF) have been shown to cause an increase in the bone mineral density of long bones, and in the growth of craniofacial structures in a variety of animal models. In addition, CLMF is well tolerated by the patient and produces no known adverse effects. However, its application in orthodontic tooth movement has not been specifically determined. Since factors that increase alveolar bone remodeling enhance the rate of orthodontic tooth movement, we hypothesized that externally applied, cyclical, low magnitude forces (CLMF) will increase the rate of orthodontic tooth movement. In order to test this hypothesis we used an in vivo rat orthodontic tooth movement model. Our specific aims were: Specific Aim 1: To develop an in vivo rat model for tooth movement. We developed a tooth movement model based upon two established rodent models (Ren and Yoshimatsu et al, See Figure 1.). The amount of variation of tooth movement in rats exposed to 25-60 g of mesial force activated viii from the first molar to the incisor for 4 weeks was calculated. Specific Aim 2: To determine the frequency dose response of externally applied, cyclical, low magnitude forces (CLMF) for maximal tooth movement and osteoclast numbers. Our working hypothesis for this aim was that the amount of tooth movement would be dose dependent on the frequency of application of the CLMF. In order to test this working hypothesis, we varied the frequency of the CLMF from 30, 60, 100, and 200 Hz, 0.4N, two times per week, for 10 minutes for 4 weeks, and measured the amount of tooth movement. We also looked at the number of osteoclasts for the different frequencies; we hypothesized an increase in osteoclasts for the dose respnse of different frequencies. Specific Aim 3: To determine the effects of externally applied, cyclical, low magnitude forces (CLMF) on PDL proliferation. Our working hypothesis for this aim was that PDL proliferation would increase with CLMF. In order to test this hypothesis we compared CLMF (30 Hz, 0.4N, two times per week, for 10 minutes for 4 weeks) performed on the left side (experimental side), to the non-CLMF side, on the right (control side). This was an experimental study with 24 rats in total. The experimental group contained fifteen (15) rats in total, and they all received a spring plus a different frequency of CLMF. Three (3) received a spring and CLMF at 30 Hz, 0.4N for 10 minutes. Six (6) received a spring and CLMF at 60 Hz, 0.4N for 10 minutes. Three (3) received a spring and CLMF at 100 Hz, 0.4N for 10 minutes. Three (3) received a spring and CLMF at 200 Hz, 0.4N for 10 minutes. The control group contained six (6) rats, and received only a spring. An additional ix three (3) rats received CLMF (30 Hz, 0.4N, two times per week, for 10 minutes for 4 weeks) only, with no spring, and were used only for histological purposes. Rats were subjected to the application of orthodontic force from their maxillary left first molar to their left central incisor. In addition some of the rats received externally applied, cyclical, low magnitude force (CLMF) on their maxillary left first molar. micro-CT was used to measure the amount of orthodontic tooth movement. The distance between the maxillary first and second molars, at the most mesial point of the second molar and the most distal point of the first molar (1M-2M distance) were used to evaluate the distance of tooth movement. Immunohistochemistry was performed with TRAP staining and BrdU quantification. Externally applied, cyclical, low magnitude forces (CLMF) do appear to have an effect on the rate, while not significant, of orthodontic tooth movement in rats. It appears that lower CLMF decreases the rate of tooth movement, while higher CLMF increases the rate of tooth movement. Future studies with larger sample sizes are needed to clarify this issue. CLMF does not appear to affect the proliferation in PDL cells, and has no effect on the number of osteoclasts.

Rapid maxillary expansion causes neuronal activation in brain structures of rats

A correlation between pain sensation and neuronal c-fos expression has been analyzed following experimental rapid maxillar expansion (RME). Adult male Wistar rats were anaesthetized and divided into three groups: animals that received an orthodontic apparatus, which was immediately removed after the insertion (control), animals that received an inactivated orthodontic apparatus (without force), and animals that received an orthodontic apparatus previously activated (140 g force). After 6, 24, 48, or 72 h, the animals were re-anaesthetized, and perfused with 4% paraformaldehyde. The brains were removed, fixed, and sections containing brain structures related to nociception were processed for Fos protein immunohistochemistry (IHC). The insertion of the orthodontic apparatus with 140 g was able to cause RME that could be seen by radiography. The IHC results showed that the number of activated neurons in the different nuclei changed according to the duration of appliance insertion and followed a temporal pattern similar to that of sensations described in clinics. The animals that received the orthodontic apparatus without force did not show RME but a smaller c-fos expression in the same brain structures. In conclusion, we demonstrate that orthodontic force used for palate disjunction activates brain structures that are related to nociception, and that this activation is related to the pain sensation described during orthodontic treatment. (c) 2008 Elsevier Inc. All rights reserved.

Biomecánica da movimentação ortodôntica--resposta inicial dos tecidos periodontais.

There occurs a biological response of the tissues of dental support, in answer to the external physiological forces and those realized during clinical treatments with orthodontical purposes. These forces differ from the first ones because they are continuous and time dependent. A great dental mobility is related to the degree of tissue organization of the periodontium system and the orthodontical movement must utilize this exceptional capacity of renewal and adaptation of the periodontium structures adequately. Therefore, through histological means a search was made to evaluate the succession of alterations of the periodontium system after the application of an orthodontical force on the molars of young rats and to interpret the standards of horizontal mobility and their consequences on the periodontium structures, biologically. An orthodontic force was applied on young rats utilizing steel wire placed in a ring form on a contact point between the first and second lower molars. The animals were sacrificed after 30 minutes, 1, 2, 6, 12, 24, 48, 72, 96 and 168 hours after the placement of the metal ring. After technical preparation, the microscopic slides were examined and the results were compared. In all the sections there was evidence of an intense metabolic activity. A gradual evolution of modification on the phenomenons had occurred.

Expanding the spectrum of PTH1R mutations in patients with primary failure of tooth eruption

Objectives: Primary failure of tooth eruption (PFE) is a rare autosomal-dominant disease characterized by severe lateral open bite as a consequence of incomplete eruption of posterior teeth. Heterozygous mutations in the parathyroid hormone 1 receptor (PTH1R) gene have been shown to cause PFE likely due to protein haploinsufficiency. To further expand on the mutational spectrum of PFE-associated mutations, we report here on the sequencing results of the PTH1R gene in 70 index PFE cases. Materials and methods: Sanger sequencing of the PTH1R coding exons and their immediate flanking intronic sequences was performed with DNA samples from 70 index PFE cases. Results: We identified a total of 30 unique variants, of which 12 were classified as pathogenic based on their deleterious consequences on PTH1R protein while 16 changes were characterized as unclassified variants with as yet unknown effects on disease pathology. The remaining two variants represent common polymorphisms. Conclusions: Our data significantly increase the number of presently known unique PFE-causing PTH1R mutations and provide a series of variants with unclear pathogenicity which will require further in vitro assaying to determine their effects on protein structure and function. Clinical relevance: Management of PTH1R-associated PFE is problematic, in particular when teeth are exposed to orthodontic force. Therefore, upon clinical suspicion of PFE, molecular DNA testing is indicated to support decision making for further treatment options. © 2013 Springer-Verlag Berlin Heidelberg.

Experimental model of tooth movement in mice: A standardized protocol for studying bone remodeling under compression and tensile strains

During orthodontic tooth movement (OTM), alveolar bone is resorbed by osteoclasts in compression sites (CS) and is deposited by osteoblasts in tension sites (TS). The aim of this study was to develop a standardized OTM protocol in mice and to investigate the expression of bone resorption and deposition markers in CS and TS. An orthodontic appliance was placed in C57BL6/J mice. To define the ideal orthodontic force, the molars of the mice were subjected to forces of 0.1 N, 0.25 N, 0.35 N and 0.5 N. The expression of mediators that are involved in bone remodeling at CS and TS was analyzed using a Real-Time PCR. The data revealed that a force of 0.35 N promoted optimal OTM and osteoclast recruitment without root resorption. The levels of TNF-alpha, RANKL, MMP13 and OPG were all altered in CS and TS. Whereas TNF-a and Cathepsin K exhibited elevated levels in CS. RUNX2 and OCN levels were higher in TS. Our results suggest that 0.35 N is the ideal force for OTM in mice and has no side effects. Moreover, the expression of bone remodeling markers differed between the compression and the tension areas, potentially explaining the distinct cellular migration and differentiation patterns in each of these sites. (C) 2012 Elsevier Ltd. All rights reserved.

COMPARAÇÃO ENTRE A APLICAÇÃO ÚNICA E FRACIONADA DO LASER DE BAIXA INTENSIDADE NA MOVIMENTAÇÃO DENTÁRIA EM RATOS

O laser de baixa intensidade (LBI) tem demonstrado efeitos bioestimulatórios na movimentação ortodôntica, acelerando a resposta celular e, reduzindo o tempo de tratamento. Entretanto, o uso do LBI seria mais efetivo com maior potência, menor tempo (única irradiação), ou a junção da menor potência e tempo em aplicação fracionada? Este estudo visou quantificar a movimentação de molares de 64 ratos e suas alterações histológicas, submetidos a uma força ortodôntica, liberada por meio de uma mola fechada de níquel-titânio (Niti/25gf), calibrada em 20gf. Os animais foram divididos em quatro grupos (cada grupo com 16 ratos), de acordo com os seguintes protocolos de irradiação do laser GaAlAs(780nm): grupo controle , submetidos à movimentação ortodôntica mas sem aplicação do LBI ; grupo total 1 (P=60mW/DE=15J/cm²/Et=6J/t=100seg), com a movimentação ortodôntica associada à única aplicação do LBI laser (P=60mW/DE=15J/cm²/Et=6J/t=100seg) no dia zero. Grupo total 2 em que o LBI foi aplicado no dia 0, segundo o protocolo P=20mW/DE=15J/cm²/E=6J/t=300seg;,; e Grupo fracionado em que o LBI foi aplicado nos dias 0, 3 e 7, empregando-se o protocolo (P=20mW/DE=5J/cm²/E=2J/t=100seg). Os sacrifício dos ratos foi realizado em quatro momentos (dias 1,4,8 e 15), sendo que 4 ratos de cada grupo foi sacrificado em cada um dos dias. A quantidade de movimentação ortodôntica foi mensurada in loco por meio de paquímetro digital, antes da instalação das molas e, imediatamente após o sacrifício dos animais. Para o exame histológico, as maxilas foram removidas, preparadas e coradas pelo sistema H/E. Os resultados foram avaliados pela Análise de Variância, seguida do teste Tukey (p<0,05). O grupo fracionado apresentou maior movimentação dentária frente aos demais nos dias 4 e 8, sendo que os grupos total 1 e 2 demonstraram maior movimentação dentária que o controle somente no dia 8. Não houve diferença estatisticamente significante entre os grupos total 1 e 2 em todo o período avaliado. Este estudo concluiu que, o laser de baixa intensidade acelerou a movimentação dentária, salientando-se o efeito da aplicação fracionada em relação às aplicações únicas.

A UTILIZAÇÃO DO LASER DE BAIXA INTENSIDADE NA MOVIMENTAÇÃO DENTÁRIA E NA SENSIBILIDADE DOLOROSA APÓS APLICAÇÃO DE FORÇA ORTODÔNTICA

O presente estudo objetivou avaliar a eficácia do Laser de Baixa Intensidade (LBI) na aceleração da movimentação dentária e na diminuição da dor frente à aplicação de força ortodôntica. A amostra foi composta por 19 pacientes, sendo doze do sexo feminino e sete do sexo masculino, com idade inicial média de 14,69 anos, todos com indicação para extrações de primeiros pré-molares. Destes, 66 caninos foram submetidos à retração inicial, sendo que 33 receberam aplicação de laser e 33 foram considerados controle. Utilizou-se o Laser de Baixa Intensidade de arseneto de gálio e alumínio, com comprimento de onda de 780nm, na dosimetria de 40mW;10J/cm2;10s/ponto, aplicado apenas uma vez ao mês em dez pontos, sendo cinco por vestibular e cinco por lingual/palatino. Modelos de gesso foram confeccionados durante todos os meses de retração dos caninos, que teve duração de quatro meses, sendo, posteriormente, digitalizados para se mensurar a quantidade de movimentação de um lado em relação ao outro, utilizando-se como referência as papilas incisivas. Para a avaliação da dor experimentada pelos pacientes, os mesmos foram orientados a preencher uma escala analógica visual (VAS) que variava de 0 a 10 , em que zero significava nenhuma dor e dez significava dor insuportável, nos intervalos de 12, 24, 48 e 72 horas após a aplicação da força ortodôntica. Foi mensurado o apinhamento de todas as hemiarcadas dos pacientes na fase inicial, medindo-se a distância entre os pontos de contato de cada dente. Para a verificação do padrão de normalidade, empregou-se o teste de Kolmogorov-Smirnov, sendo que para comparar o lado irradiado com o lado não irradiado foi utilizado o teste t pareado, exceto para a variável razão caninos/molares , analisada pelo teste não paramétrico de Wilcoxon (p<0,05). Os resultados mostraram que em relação ao apinhamento dentário, os lados irradiado e não irradiado apresentaram-se compatíveis. Além disso, não houve diferença estatisticamente significante entre a quantidade de retração dos caninos irradiados comparados aos não irradiados, o mesmo acontecendo com a sensibilidade dolorosa experimentada pelos pacientes. Concluiu-se assim que o LBI na dosimetria e forma como foi utilizado não foi eficiente na aceleração da movimentação dentária nem na redução da dor experimentada pelos pacientes frente às forças ortodônticas.

Assessment of force decay in orthodontic elastomeric chains: An in vitro study

Introduction: Elastomeric materials are considered important sources of orthodontic forces. Objective: To assess force degradation over time of four commercially available orthodontic elastomeric chains (Morelli, Ormco, TP and Unitek). Methods: The synthetic elastics were submerged in 37 oC synthetic saliva and stretched by a force of 150 g (15 mm - Morelli and TP; 16mm - Unitek and Ormco). With a dynamometer, the delivered force was evaluated at different intervals: 30 minutes, 7 days, 14 days and 21 days. The results were subjected to ANOVA and Tukey's test. Results: There was a force decay between 19% to 26.67% after 30 minutes, and 36.67% to 57% after 21 days of activation. Conclusions: TP elastomeric chains exhibited the smallest percentage of force decay, with greater stability at all time intervals tested. Meanwhile, the Unitek chains displayed the highest percentage of force degradation, and no statically significant difference was found in force decay between Ormco and Morelli elastomeric chains during the study period.