Gli autori vorrei ringraziare il Dr. David Rowe per gentilmente fornire i topi transgenici e Li Chen per l'assistenza istologico.
La ricerca riportata in questa pubblicazione è stata sostenuta dal National Institute of Dental & Craniofacial Research del National Institutes of Health, sotto Premio numero K08DE025914 e dalla associazione americana di Fondazione ortodontico per 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>

Gli autori non hanno nessun concorrenti interessi finanziari.

Questo manoscritto descritto metodi per la misurazione di morfometrica e analisi cellulare del murini condili mandibolari e mandibole. Le misure morfometriche radiografica è utilizzabile anche per analizzare altre ossa da piccoli animali da esperimento. Inoltre, l'analisi cellulare (cellule quantificazione e mappatura di distanza di cartilagine) non sono limitati al condilo mandibola roditore, ma può essere utilizzato per quantificare le sezioni istologiche di numerosi tessuti.
Modelli di to...

TMJ è un unico giunto portante che si trova nella regione craniofacial ed è costituito di fibrocartilagine. Il MCC del TMJ è essenziale per la funzione articolare, compreso movimento della mascella senza ostacoli mentre parlando e masticare, ma esso è influenzato dalle malattie degeneranti, compreso l'osteoartrite1. L'ATM ha la capacità di adattarsi a stimoli esterni e le alterazioni di carico, portando a cambiamenti strutturali e cellulari ai componenti del MCC2,3,4,5. Le proprietà portanti di MCC possono essere spiegate dalle interazioni tra i suoi costituenti, compresa l'acqua, la rete di collagene e densamente proteoglicani. Il MCC ha quattro distinte zone cellulari che esprimono diversi tipi di proteine di collagene e non-collagene: 1) l'area superficiale o articolare; 2) la zona proliferativa, composta di cellule mesenchimali indifferenziate e che risponde al carico di richieste; 3) la zona di prehypertrophic, composto da condrociti maturi esprimendo il collagene di tipo 2; e 4) l'ipertrofica di zona, la regione dove i condrociti ipertrofici esprimendo collagene tipo 10 die e sottoposti a calcificazione. La regione non mineralizzato è ricca di proteoglicani che offrono resistenza alle forze di compressione6.
C'è la mineralizzazione continua alla zona ipertrofica del MCC, dove si verifica la transizione da Condrogenesi ad osteogenesi, garantendo la robusta struttura minerale dell'osso subcondrale del condilo mandibolare7. Cambiamenti cellulari nelle regioni unmineralized e mineralizzate in ultima analisi, portano a cambiamenti morfologici e strutturali nel condilo mandibolare e mandibola. Mantenimento dell'omeostasi cellulare tutte le regioni di MCC e la mineralizzazione della porzione subchondral sono essenziali per la salute, capacità di carico e l'integrità dell'ATM.
Il modello di topo transgenico collagene multipli (come descritto da Utreja et al.) 8 è un ottimo strumento da utilizzare per comprendere i cambiamenti nell'espressione di collagene poiché tutti i transgeni sono espressi in MCC. Per un'approfondita valutazione istologica, macchie istologiche sono utilizzate per studiare la deposizione di matrice, mineralizzazione, proliferazione delle cellule e apoptosi, come pure l'espressione della proteina presso gli strati differenti delle cellule del MCC.
In questo manoscritto, istologico e le analisi morfometriche vengono utilizzate per valutare i cambiamenti strutturali e cellulari nell'osso MCC e subcondrale del condilo mandibola di topi. Inoltre, un metodo di quantificazione delle cellule, per analizzare le immagini istologiche fluorescente e per il mapping di vetrini da microscopio chiaro, è descritto. Compressione statica TMJ caricamento metodo che causa cambiamenti cellulari e morfologiche al MCC e subchondral dell'osso9, è illustrata anche per convalidare i nostri metodi.
I metodi descritti qui possono essere utilizzati per determinare morfometriche e cambiamenti istologici nel condilo mandibolare e mandibola di roditori o di analizzare altre regioni di ossificazione endochondral e la morfologia dei tessuti mineralizzati aggiuntive.

<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

Il giunto temporomandibular (TMJ) ha la capacità di adattarsi agli stimoli esterni, e cambiamenti di carico possa influenzare la posizione dei condili, così come i componenti strutturali e cellulari della cartilagine del condilo mandibola (MCC). Questo manoscritto descrive metodi per l'analisi di questi cambiamenti e un metodo per alterare il caricamento dell'ATM in topi (cioè, alla compressione statica TMJ caricamento). La valutazione strutturale qui illustrata è un approccio semplice morfometriche che utilizza il software Digimizer e viene eseguito nelle radiografie di piccole ossa. Inoltre, l'analisi del cellulare cambia leader alle alterazioni nell'espressione del collagene, rimodellamento osseo, divisione cellulare, e distribuzione di proteoglycan in MCC è descritto. La quantificazione di questi cambiamenti nelle sezioni istologiche - contando i pixel fluorescenti positivi utilizzando software e il mapping di distanza di misurazione di immagine e zona macchiata con Digimizer - è dimostrato anche. I metodi qui illustrati non sono limitati a TMJ murino, ma ha potuto essere usati su altre ossa di piccoli animali da esperimento e in altre regioni di ossificazione endochondral.

Statistiche descrittive sono state effettuate per esaminare la distribuzione delle misure morfometriche (lunghezza mandibular, condylar lunghezza, larghezza condylar) e le analisi istologiche. I risultati sono stati confrontati fra il gruppo caricato (cioè, i topi sottoposti a carico di compressione con la molla in titanio beta) e il gruppo di controllo (cioè, i topi di controllo che non hanno ricevuto alcuna procedura di corrispondenza). Differenze statisticamente sig...

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

Questo manoscritto presenta metodi per l'analisi morfometrica e cambiamenti cellulari all'interno del condilo mandibolare di roditori.

Un metodo di analisi cellulare per condilo mandibolare murino e morfometrica

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 ...

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Research

JoVE Journal

Biology

8.2K Views.  University of Connecticut Health Center. Questo manoscritto presenta metodi per l'analisi morfometrica e cambiamenti cellulari all'interno del condilo mandibolare di roditori.

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

Visualization of Chondrocyte Intercalation and Directional Proliferation via Zebrabow Clonal Cell Analysis in the Embryonic Meckel&#8217;s Cartilage

Development of the vertebrate craniofacial structures requires precise coordination of cell migration, proliferation, adhesion and differentiation. Patterning of the Meckel&#39;s cartilage, a first pharyngeal arch derivative, involves the migration of cranial neural crest (CNC) cells and the progressive partitioning, proliferation and organization of differentiated chondrocytes. Several studies have described CNC migration during lower jaw&#160;morphogenesis, but the details of how the chondrocytes achieve organization in the growth and extension of Meckel&#8217;s cartilage remains unclear. The sox10 restricted and chemically induced Cre recombinase-mediated recombination generates permutations of distinct fluorescent proteins (RFP, YFP and CFP), thereby creating a multi-spectral labeling of progenitor cells and their progeny, reflecting distinct clonal populations. Using confocal time-lapse photography, it is possible to observe the chondrocytes behavior during the development of the zebrafish Meckel&#8217;s cartilage.
Multispectral cell labeling enables scientists to demonstrate extension of the Meckel&#8217;s chondrocytes.&#160;During extension phase of the Meckel&#8217;s cartilage, which prefigures the mandible, chondrocytes intercalate to effect extension as they stack in an organized single-cell layered row. Failure of this organized intercalating process to mediate cell extension provides the cellular mechanistic explanation for hypoplastic mandible that we observe in mandibular malformations.

Cell organization of craniofacial bones has long been hypothesized but never directly visualized. Multi-spectral cell labeling and in vivo live imaging allows visualization of dynamic cell behavior in zebrafish lower jaw. Here, we detail the protocol to manipulate Zebrabow transgenic fish and directly observe cell intercalation and morphological changes of chondrocytes in the Meckel&#8217;s cartilage.

Visualizzazione di condrociti intercalazione e direzionale proliferazione via Zebrabow Analisi cellulare clonale nella cartilagine del Embryonic Meckel

Development of the vertebrate craniofacial structures requires precise coordination of cell migration, proliferation, adhesion and differentiation. ...

Ricerca

Biologia dello sviluppo

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.

Microdissezione laser di cattura della cartilagine embrionale del topo e dell'osso per l'analisi dell'espressione genica

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

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.

Dissezione, microCT scansione e morfometriche analisi del Baculum

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

Bioingegneria

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.

Analisi delle malformazioni craniomaxillo-facciali nei topi mediante tomografia microcomputerizzata tridimensionale

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.

Flessione a sbalzo dei colli femorali murini

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.

Valutazione istomormetrica standardizzata dell'osteoartrite in un modello di topo chirurgico

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

Medicina

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.

Imaging 3D delle fibre di collagene PDL durante il movimento ortodontico dei denti nel modello murino mandibolare

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.

Destabilizzazione del menisco mediale e del modello murino di graffio cartilagineo di osteoartrite accelerata

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

Una guida al modello ortodontico mascellare murino per l'analisi del rimodellamento osseo

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.

Autore in primo piano: Approfondimenti su un efficiente protocollo del modello di ortodonzia mascellare murina

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.

La creazione di un modello ortodontico mascellare murino

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

Un metodo di analisi cellulare per condilo mandibolare murino e morfometrica

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Capitoli in questo video

Visualizzazione di condrociti intercalazione e direzionale proliferazione via Zebrabow Analisi cellulare clonale nella cartilagine del Embryonic Meckel

Microdissezione laser di cattura della cartilagine embrionale del topo e dell'osso per l'analisi dell'espressione genica

Dissezione, microCT scansione e morfometriche analisi del Baculum

Analisi delle malformazioni craniomaxillo-facciali nei topi mediante tomografia microcomputerizzata tridimensionale

Flessione a sbalzo dei colli femorali murini

Valutazione istomormetrica standardizzata dell'osteoartrite in un modello di topo chirurgico

Imaging 3D delle fibre di collagene PDL durante il movimento ortodontico dei denti nel modello murino mandibolare

Destabilizzazione del menisco mediale e del modello murino di graffio cartilagineo di osteoartrite accelerata

La creazione di un modello ortodontico mascellare murino

La creazione di un modello ortodontico mascellare murino