Os autores gostaria de agradecer Dr. David Rowe para gentilmente fornecer os ratos transgénicos e Li Chen pela assistência histológica.
A pesquisa reportada nesta publicação foi apoiada pelo Instituto Nacional de Dental & pesquisa Craniofacial do institutos nacionais da saúde sob prêmio número K08DE025914 e pela Associação Americana de fundação ortodôntico para Sumit Yadav.

<ol>
	<li>LeResche, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiology+of+Temporomandibular+Disorders:+Implications+for+the+Investigation+of+Etiologic+Factors.">Epidemiology of Temporomandibular Disorders: Implications for the Investigation of Etiologic Factors.</a> Crit Rev Oral Biol Med. 8 (3), 291-305 (1997).</li><li>Chen, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Altered+functional+loading+causes+differential+effects+in+the+subchondral+bone+and+condylar+cartilage+in+the+temporomandibular+joint+from+young+mice.">Altered functional loading causes differential effects in the subchondral bone and condylar cartilage in the temporomandibular joint from young mice.</a> Osteoarthr Cartil. 17 (3), 354-361 (2009).</li><li>Pirttiniemi, P., Kantomaa, T., Sorsa, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+decreased+loading+on+the+metabolic+activity+of+the+mandibular+condylar+cartilage+in+the+rat.">Effect of decreased loading on the metabolic activity of the mandibular condylar cartilage in the rat.</a> Eur J Orthod. 26 (1), 1-5 (2004).</li><li>Chavan, S. J., Bhad, W. A., Doshi, U. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparison+of+temporomandibular+joint+changes+in+Twin+Block+and+Bionator+appliance+therapy:+a+magnetic+resonance+imaging+study.">Comparison of temporomandibular joint changes in Twin Block and Bionator appliance therapy: a magnetic resonance imaging study.</a> Prog Orthod. 15 (57), (2014).</li><li>Dutra, E. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+and+Matrix+Response+of+the+Mandibular+Condylar+Cartilage+to+Botulinum+Toxin.">Cellular and Matrix Response of the Mandibular Condylar Cartilage to Botulinum Toxin.</a> PLoS ONE. 11 (10), 0164599 (2016).</li><li>Benjamin, M., Ralphs, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biology+of+fibrocartilage+cells.">Biology of fibrocartilage cells.</a> Int Rev Cytol. 233, 1-45 (2004).</li><li>Shen, G., Darendeliler, M. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Adaptive+Remodeling+of+Condylar+Cartilage-+A+Transition+from+Chondrogenesis+to+Osteogenesis.">The Adaptive Remodeling of Condylar Cartilage- A Transition from Chondrogenesis to Osteogenesis.</a> J Dent Res. 84 (8), 691-699 (2005).</li><li>Utreja, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+and+matrix+response+of+temporomandibular+cartilage+to+mechanical+loading.">Cell and matrix response of temporomandibular cartilage to mechanical loading.</a> Osteoarthr Cartil. 24 (2), 335-344 (2016).</li><li>Kaul, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Effect+of+Altered+Loading+on+Mandibular+Condylar+Cartilage.">The Effect of Altered Loading on Mandibular Condylar Cartilage.</a> PLoS ONE. 11 (7), 0160121 (2016).</li><li>Dyment, N. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-Throughput,+Multi-Image+Cryohistology+of+Mineralized+Tissues.">High-Throughput, Multi-Image Cryohistology of Mineralized Tissues.</a> J Vis Exp. , e54468 (2016).</li><li>Kawamoto, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+a+new+adhesive+film+for+the+preparation+of+multi-purpose+fresh-frozen+sections+from+hard+tissues,+whole-animals,+insects+and+plants.">Use of a new adhesive film for the preparation of multi-purpose fresh-frozen sections from hard tissues, whole-animals, insects and plants.</a> Arch Histol Cytol. 66 (2), 123-143 (2003).</li><li>Hayman, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tartrate-resistant+acid+phosphatase+(TRAP)+and+the+osteoclast/immune+cell+dichotomy.">Tartrate-resistant acid phosphatase (TRAP) and the osteoclast/immune cell dichotomy.</a> Autoimmunity. 41 (3), 218-223 (2008).</li></ol>

Os autores têm sem interesses financeiros concorrentes.

Este manuscrito descreve métodos para a medição de morfometria e análise celular dos côndilos mandibulares murino e mandíbulas. As medições radiográficas morfométricas também podem ser usadas para analisar outros ossos de pequenos animais experimentais. Além disso, a análise celular (célula quantificação e mapeamento de distância de cartilagem) não estão limitada a do côndilo mandibular roedor, mas pode ser usado para quantificar a cortes histológicos de tecidos diversos.
...

A TMJ é uma articulação de carga exclusiva localizada na região craniofacial e é formada de fibrocartilagem. O MCC da TMJ é essencial para a função da articulação, incluindo o movimento da mandíbula sem impedimentos ao falar e mastigar, mas geralmente é afetado por doenças degenerativas, incluindo osteoartrite1. A TMJ tem a capacidade de adaptação a estímulos externos e alterações de carga, levando a alterações estruturais e celulares para os componentes do MCC2,3,4,5. As propriedades de carga do MCC podem ser explicadas pelas interações entre seus componentes, incluindo a água, a rede de colágeno e densamente proteoglicanos. O MCC tem quatro zonas distintas de celulares que expressam diferentes tipos de proteínas de colágeno e não-colágeno: 1) a zona superficial ou articular; 2) a zona proliferativa, composta de células mesenquimais indiferenciadas e que responde ao carregar as demandas; 3) a zona de prehypertrophic, composto de condrócitos maduros expressando colágeno tipo 2; e 4) a hipertrófica de zona, a região onde os condrócitos hipertróficas expressando colágeno tipo 10 morrer e sofrer calcificação. A região não-mineralizado é rica em proteoglicanos que proporcionam resistência a forças compressivas6.
Há contínua mineralização na zona hipertrófica do MCC, onde ocorre a transição da condrogênese a osteogênese, garantindo a robusta estrutura mineral do osso subcondral do côndilo mandibular7. As mudanças celulares em regiões não mineralizadas e mineralizadas, finalmente, levam a alterações morfológicas e estruturais no côndilo mandibular e maxilar inferior. Manutenção da homeostase de todas as regiões celulares do MCC e a mineralização da parte subcondral são essenciais para a saúde, capacidade load-bearing e integridade da TMJ.
O modelo de rato transgénico colágeno múltipla (como descrito por Utreja et al) 8 é uma ótima ferramenta para usar para entender mudanças na expressão do colágeno, porque todos os transgenes são expressas em MCC. Para uma avaliação aprofundada e histológica, manchas histológicas são utilizadas para estudar a deposição de matriz, mineralização, proliferação celular, apoptose, e expressão de proteínas para as camadas de células diferentes do MCC.
Neste manuscrito, histológico e análises morfométricas são usadas para avaliar as alterações estruturais e celulares no MCC e subcondral osso do côndilo mandibular de ratos. Além disso, um método de quantificação de célula, para a análise de imagens histológicas fluorescentes e para mapear as corrediças do microscópio de luz, é descrito. A TMJ estática à compressão carregando o método, que provoca alterações celulares e morfológicas na MCC e subcondral osso9, também é ilustrada para validar nossos métodos.
Os métodos descritos aqui podem ser usados para determinar morfométricos e as alterações histológicas no côndilo mandibular e maxilar inferior de roedores ou para analisar outras regiões de ossificação endocondral e a morfologia dos tecidos mineralizados adicionais.

<table><tbody><tr><td>MX20 Radiography System</td><td>Faxitron X-Ray LLC&amp;nbsp;</td><td></td><td></td></tr><tr><td>Digimizer Image software&amp;nbsp;</td><td>MedCalc Software</td><td></td><td></td></tr><tr><td>Shandon Cryomatrix embedding resin</td><td>Thermo Scientific</td><td>6769006</td><td></td></tr><tr><td>Manual microscope Axio Imager Z1</td><td>Carl Zeiss</td><td>208562</td><td></td></tr><tr><td>yellow fluorescent protein filter&amp;nbsp; - EYFP</td><td>Chroma Technology Corp</td><td>49003</td><td></td></tr><tr><td>cyan fluorescent protein filter - ECFP</td><td>Chroma Technology Corp</td><td>49001</td><td></td></tr><tr><td>red fluoresecent protein filter - Cy5</td><td>Chroma Technology Corp</td><td>49009</td><td></td></tr><tr><td>sodium acetate anhydrous</td><td>Sigma-Aldrich</td><td>S2889</td><td></td></tr><tr><td>sodium L-tartrate dibasic dihydrate&amp;nbsp;</td><td>Sigma-Aldrich</td><td>228729</td><td></td></tr><tr><td>sodium nitrite&amp;nbsp;</td><td>Sigma-Aldrich</td><td>237213</td><td></td></tr><tr><td>ELF97 substrate</td><td>Thermo Fisher Scientific</td><td>E6600</td><td></td></tr><tr><td>ClickiT EdU Alexa Fluor 594 HCS kit</td><td>Life Technologies</td><td>C10339&amp;nbsp;</td><td>includes EdU (5-ethynyl-2&#039;-deoxyuridine)&amp;nbsp;</td></tr><tr><td>DAPI (4&#039;,6-Diamidino-2-Phenylindole, Dihydrochloride)</td><td>Thermo Scientific</td><td>D1306</td><td></td></tr><tr><td>Sodium phosphate dibasic&amp;nbsp;</td><td>Sigma-Aldrich</td><td>S3264</td><td></td></tr><tr><td>Sodium phosphate monobasic&amp;nbsp;</td><td>Sigma-Aldrich</td><td>71505</td><td></td></tr><tr><td>Toluidine Blue O&amp;nbsp;</td><td>Sigma-Aldrich</td><td>T3260</td><td></td></tr><tr><td>Adobe Photoshop&amp;nbsp;</td><td>Adobe Systems Incorporated</td><td></td><td></td></tr><tr><td>Phosphate buffered saline tablets (PBS)</td><td>Research Products International</td><td>P32080-100T</td><td></td></tr><tr><td>CNA Beta III Nickel-Free Archwire</td><td>Ortho Organizers, Inc.</td><td></td><td></td></tr><tr><td>GraphPad Prism&amp;nbsp;</td><td>GraphPad Software, Inc.</td><td></td><td></td></tr></tbody></table>

a morphometric and cellular analysis method for the murine mandibular condyle

Articulação temporomandibular (ATM) tem a capacidade de adaptação a estímulos externos, e carregar as alterações pode afetar a posição dos côndilos, bem como os componentes estruturais e celulares da cartilagem condilar mandibular (MCC). Este manuscrito descreve métodos para analisar estas mudanças e um método para alterar o carregamento da TMJ em camundongos (i.e., à compressão estática TMJ carregando). A avaliação estrutural ilustrada aqui é uma abordagem simples morfométricos que usa o software Digimizer e é executada em radiografias de ossos pequenos. Além disso, a análise de celulares muda levando a alterações na expressão do colágeno, a remodelação óssea, a divisão celular, e proteoglycan distribuição no MCC é descrita. A quantificação dessas mudanças em cortes histológicos - contando os pixels fluorescentes positivos usando imagem de software e o mapeamento de distância de medição e manchado área com Digimizer - também é demonstrado. Os métodos mostrados aqui não são limitados a TMJ murino, mas podem ser usados em ossos adicionais de pequenos animais experimentais e em outras regiões de ossificação endocondral.

Estatísticas descritivas foram realizadas para examinar a distribuição de medidas morfométricas (comprimento mandibular, condilar comprimento, largura condilar) e análise histológica. Os resultados foram comparados entre o grupo carregado (ou seja, ratos submetidos a carga compressiva com a mola de titânio beta) e grupo controle (ou seja, ratos de controle que não recebeu qualquer procedimento de correspondência). Diferenças estatisticamente significativas entr...

Watch this Scientific Journal Video about A Morphometric and Cellular Analysis Method for the Murine Mandibular Condyle at JoVE.com

A Morphometric and Cellular Analysis Method for the Murine Mandibular Condyle

Este manuscrito apresenta métodos para a Análise morfométrica e as mudanças celulares dentro do côndilo mandibular de roedores.

Um método de análise celular para o côndilo Mandibular murino e morfométricas

The temporomandibular joint (TMJ) has the capacity to adapt to external stimuli, and loading changes can affect the position of condyles, as well as the ...

a-morphometric-cellular-analysis-method-for-murine-mandibular

Research

JoVE Journal

Biology

8.2K visualizações.  University of Connecticut Health Center. Este manuscrito apresenta métodos para a Análise morfométrica e as mudanças celulares dentro do côndilo mandibular de roedores.

Vídeo: A Morphometric and Cellular Analysis Method for the Murine Mandibular Condyle

Laser Capture Microdissection of Mouse Embryonic Cartilage and Bone for Gene Expression Analysis

Laser capture microdissection (LCM) is a powerful tool to isolate specific cell types or regions of interest from heterogeneous tissues. The cellular and molecular complexity of skeletal elements increases with development. Tissue heterogeneity, such as at the interface of cartilaginous and osseous elements with each other or with surrounding tissues, is one obstacle to the study of developing cartilage and bone. Our protocol provides a rapid method of tissue processing and isolation of cartilage and bone that yields high quality RNA for gene expression analysis. Fresh frozen tissues of mouse embryos are sectioned and brief cresyl violet staining is used to visualize cartilage and bone with colors distinct from surrounding tissues. Slides are then rapidly dehydrated, and cartilage and bone are isolated subsequently by LCM. The minimization of exposure to aqueous solutions during this process maintains RNA integrity. Mouse Meckel&rsquo;s cartilage and mandibular bone at E16.5 were successfully collected and gene expression analysis showed differential expression of marker genes for osteoblasts, osteocytes, osteoclasts, and chondrocytes. High quality RNA was also isolated from a range of tissues and embryonic ages. This protocol details sample preparation for LCM including cryoembedding, sectioning, staining and dehydrating fresh frozen tissues, and precise isolation of cartilage and bone by LCM resulting in high quality RNA for transcriptomic analysis.

This protocol describes laser capture microdissection for the isolation of cartilage and bone from fresh frozen sections of the mouse embryo. Cartilage and bone can be rapidly visualized by cresyl violet staining and collected precisely to yield high quality RNA for transcriptomic analysis.

Microdissecção de captura a laser da cartilagem embrionária do rato e osso para análise de expressão gênica

Laser capture microdissection (LCM) is a powerful tool to isolate specific cell types or regions of interest from heterogeneous tissues. The cellular and ...

Biologia do Desenvolvimento

Dissection, MicroCT Scanning and Morphometric Analyses of the Baculum

Modern morphometrics provides powerful methods to quantify size and shape variation. A basic requirement is a list of coordinates that define landmarks; however such coordinates must represent homologous structures across specimens. While many biological objects consist of easily identified landmarks to satisfy the assumption of homology, many lack such structures. One potential solution is to mathematically place semi-landmarks on an object that represent the same morphological region across specimens. Here, we illustrate a recently developed pipeline to mathematically define semi-landmarks from the mouse baculum (penis bone). Our methods should be applicable to a wide range of objects.

Many biological structures lack easily definable landmarks, making it difficult to apply modern morphometric methods. Here we illustrate methods to study the mouse baculum (a bone in the penis), including dissection and microCT scanning, followed by computational methods to define semi-landmarks that are used to quantify size and shape variation.

Dissecação, digitalização MicroCT e morfométricas Análises do Baculum

Modern morphometrics provides powerful methods to quantify size and shape variation. A basic requirement is a list of coordinates that define landmarks; ...

Bioengenharia

A Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

Um Defeito modelo simples Critical porte Femoral em Ratos

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

Medicina

Analysis of Craniomaxillofacial Malformations in Mice Using Three-dimensional Microcomputed Tomography

To model craniofacial malformations caused by vitamin A deficiency (VAD), we expressed a dominant-negative retinoid receptor mutation in osteoblasts to specifically inhibit RAR transcriptional activity in mice. This approach allowed us to investigate the effects of VAD on cranial hypomineralization, mandibular deformity, and clavicular hypoplasia in clinical cases. In this study, microcomputed tomography (microCT) scanning of the craniomaxillofacial region of mice represented a valuable tool for studying the growth and development of this animal model. The manual estimation of images is both time-consuming and inaccurate. Hence, here, we present a straightforward, efficient, and accurate approach for segmenting and quantifying the microCT images of each craniomaxillofacial bone. MicroCT software was used to slice the mandible, frontal bone, parietal bone, nasal bone, premaxilla, maxilla, interparietal bone, and occipital bone of mice and measure their corresponding lengths and widths. This segmentation method can be applied to study growth and development in developmental biology, biomedicine, and other related sciences and allows researchers to analyze the effects of genetic mutations on individual craniofacial bones.

Here, we describe in depth a method, which is based on microcomputed tomography, to segment and measure 3D models of craniomaxillofacial bones in mice, for better assessment of craniomaxillofacial bone development in mice than what is possible with current methods.

Análise de malformações craniomaxilofaciais em camundongos por meio de tomografia computadorizada tridimensional

To model craniofacial malformations caused by vitamin A deficiency (VAD), we expressed a dominant-negative retinoid receptor mutation in osteoblasts to ...

Cantilever Bending of Murine Femoral Necks

Fractures in the femoral neck are a common occurrence in individuals with osteoporosis. Many mouse models have been developed to assess disease states and therapies, with biomechanical testing as a primary outcome measure. However, traditional biomechanical testing focuses on torsion or bending tests applied to the midshaft of the long bones. This is not typically the site of high-risk fractures in osteoporotic individuals. Therefore, a biomechanical testing protocol was developed that tests the femoral necks of murine femurs in cantilever bending loading to replicate better the types of fractures experienced by osteoporosis patients. Since the biomechanical outcomes are highly dependent on the flexural loading direction relative to the femoral neck, 3D printed guides were created to maintain a femoral shaft at an angle of 20&#176; relative to the loading direction. The new protocol streamlined the testing by reducing variability in alignment (21.6&#176; &plusmn; 1.5&#176;, COV = 7.1%, n = 20) and improved reproducibility in the measured biomechanical outcomes (average COV = 26.7%). The new approach using the 3D printed guides for reliable specimen alignment improves rigor and reproducibility by reducing the measurement errors due to specimen misalignment, which should minimize sample sizes in mouse studies of osteoporosis.

The present protocol describes the development of a reproducible testing platform for murine femoral necks in a cantilever bending set-up. Custom 3D printed guides were used to consistently and rigidly fix the femurs in optimal alignment.

Curva cantilever de pescoços femorais murinos

Fractures in the femoral neck are a common occurrence in individuals with osteoporosis. Many mouse models have been developed to assess disease states and ...

Standardized Histomorphometric Evaluation of Osteoarthritis in a Surgical Mouse Model

One of the most prevalent joint disorders in the United States, osteoarthritis (OA) is characterized by progressive degeneration of articular cartilage, primarily in the hip and knee joints, which results in significant impacts on patient mobility and quality of life. To date, there are no existing curative therapies for OA able to slow down or inhibit cartilage degeneration. Presently, there is an extensive body of ongoing research to understand OA pathology and discover novel therapeutic approaches or agents that can efficiently slow down, stop, or even reverse OA. Thus, it is crucial to have a quantitative and reproducible approach to accurately evaluate OA-associated pathological changes in the joint cartilage, synovium, and subchondral bone. Currently, OA severity and progression are primarily assessed using the Osteoarthritis Research Society International (OARSI) or Mankin scoring systems. In spite of the importance of these scoring systems, they are semiquantitative and can be influenced by user subjectivity. More importantly, they fail to accurately evaluate subtle, yet important, changes in the cartilage during the early disease states or early treatment phases. The protocol we describe here uses a computerized and semiautomated histomorphometric software system to establish a standardized, rigorous, and reproducible quantitative methodology for the evaluation of joint changes in OA. This protocol presents a powerful addition to the existing systems and allows for more efficient detection of pathological changes in the joint.

The current protocol establishes a rigorous and reproducible method for quantification of morphological joint changes that accompany osteoarthritis. Application of this protocol can be valuable in monitoring disease progression and evaluating therapeutic interventions in osteoarthritis.

Avaliação histomorfométrica padronizada da osteoartrite em um modelo de rato cirúrgico

One of the most prevalent joint disorders in the United States, osteoarthritis (OA) is characterized by progressive degeneration of articular cartilage, ...

3D Imaging of PDL Collagen Fibers during Orthodontic Tooth Movement in Mandibular Murine Model

Orthodontic tooth movement is a complex biological process of altered soft and hard tissue remodeling as a result of external forces. In order to understand these complex remodeling processes, it is critical to study the tooth and periodontal tissues within their 3D context and therefore minimize any sectioning and tissue artefacts. Mouse models are often utilized in developmental and structural biology, as well as in biomechanics due to their small size, high metabolic rate, genetics and ease of handling. In principle this also makes them excellent models for dental related studies. However, a major impediment is their small tooth size, the molars in particular. This paper is aimed at providing a step by step protocol for generating orthodontic tooth movement and two methods for 3D imaging of the periodontal ligament fibrous component of a mouse mandibular molar. The first method presented is based on a micro-CT setup enabling phase enhancement&#160;imaging of fresh collagen tissues. The second method is a bone clearing method using ethyl cinnamate that enables imaging through the bone without sectioning and preserves endogenous fluorescence. Combining this clearing method with reporter mice like Flk1-Cre;TdTomato provided a first of its kind opportunity to image the 3D vasculature in the PDL and alveolar bone.

We present a protocol for generating orthodontic tooth movement in mice and methods for 3D visualization of the collagen fibers and blood vessels of periodontal ligament without sectioning.

Imagem 3D de fibras de colágeno PDL durante movimento dentário ortodôntico no modelo de murina mandibular

Orthodontic tooth movement is a complex biological process of altered soft and hard tissue remodeling as a result of external forces. In order to ...

Destabilization of the Medial Meniscus and Cartilage Scratch Murine Model of Accelerated Osteoarthritis

Osteoarthritis is the most prevalent musculoskeletal disease in people over 45, leading to an increasing economic and societal cost. Animal models are used to mimic many aspects of the disease. The present protocol describes the destabilization and cartilage scratch model (DCS) of post-traumatic osteoarthritis. Based on the widely used destabilization of the medial meniscus (DMM) model, DCS introduces three scratches on the cartilage surface. The current article highlights the steps to destabilize the knee by transecting the medial meniscotibial ligament followed by three intentional superficial scratches on the articular cartilage. The possible analysis methods by dynamic weight-bearing, microcomputed tomography, and histology are also demonstrated. While the DCS model is not recommended for studies that focus on the effect of osteoarthritis on the cartilage, it enables the study of osteoarthritis development in a shorter time window, with special focus on (1) osteophyte formation, (2) osteoarthritic and injury pain, and (3) the effect of cartilage damage in the whole joint.

The present protocol describes the controlled microblade scratches on the surface of the articular cartilage after destabilizing the mouse knee by cutting the medial miniscotibial ligament. This animal model presents an accelerated form of osteoarthritis (OA) suitable for studying osteophyte formation, osteosclerosis, and early-stage pain.

Desestabilização do Menisco Medial e Cartilagem Scratch Murine Modelo de Osteoartrite Acelerada

Osteoarthritis is the most prevalent musculoskeletal disease in people over 45, leading to an increasing economic and societal cost. Animal models are ...

Um guia de modelo ortodôntico maxilar murino para análise de remodelação óssea

The Establishment of a Murine Maxillary Orthodontic Model

Due to the lack of reproducible protocols for establishing a murine maxillary orthodontic model, we present a reliable and reproducible protocol to provide researchers with a feasible tool to analyze mechanical loading-associated bone remodeling. This study presents a detailed flowchart in addition to different types of schematic diagrams, operation photos, and videos. We performed this protocol on 11 adult wide-type C57/B6J mice and harvested samples on postoperative days 3, 8, and 14. The micro-CT and histopathological data have proven the success of tooth movements coupled with bone remodeling using this protocol. Furthermore, according to the micro-CT results on days 3, 8, and 14, we have divided bone modeling into three stages: preparation stage, bone resorption stage, and bone formation stage. These stages are expected to help researchers concerned with different stages to set sample collection time reasonably. This protocol can equip researchers with a tool to carry out regenerative analysis of bone remodeling.

Autor em destaque: Insights sobre um protocolo eficiente de modelo ortodôntico maxilar murino

Here we demonstrate step-by-step a manageable, orthodontic tooth movement protocol operated on a murine maxillary model. With the explicit explanation of each step and visual demonstration, researchers can master this model and apply it to their experimental needs with a few modifications.

O estabelecimento de um modelo ortodôntico murino maxilar

Due to the lack of reproducible protocols for establishing a murine maxillary orthodontic model, we present a reliable and reproducible protocol to ...

Um método de análise celular para o côndilo Mandibular murino e morfométricas

Resumo

Explore mais vídeos

Capítulos neste vídeo

Microdissecção de captura a laser da cartilagem embrionária do rato e osso para análise de expressão gênica

Dissecação, digitalização MicroCT e morfométricas Análises do Baculum

Um Defeito modelo simples Critical porte Femoral em Ratos

Análise de malformações craniomaxilofaciais em camundongos por meio de tomografia computadorizada tridimensional

Curva cantilever de pescoços femorais murinos

Avaliação histomorfométrica padronizada da osteoartrite em um modelo de rato cirúrgico

Imagem 3D de fibras de colágeno PDL durante movimento dentário ortodôntico no modelo de murina mandibular

Desestabilização do Menisco Medial e Cartilagem Scratch Murine Modelo de Osteoartrite Acelerada

O estabelecimento de um modelo ortodôntico murino maxilar

O estabelecimento de um modelo ortodôntico murino maxilar