Name: development of a direct pulp-capping model for the evaluation of pulpal wound healing and reparative dentin formation in mice
Uploaded: 2017-01-12T12:30:00
Description: Watch this Scientific Journal Video about Development of a Direct Pulp-capping Model for the Evaluation of Pulpal Wound Healing and Reparative Dentin Formation in Mice at JoVE.com

Deze studie werd ondersteund door R01DE023348 (RHK) van NIDCR / NIH en de Faculteit Research Grant (RHK) van de Raad voor onderzoek van de Academische Senaat van de afdeling Los Angeles van de Universiteit van Californië.

Muizen werden gekocht bij Jackson Laboratory en bewaard in een pathogeen-vrij vivarium in het UCLA afdeling Laboratory Animal Medicine (DLAM). De experimenten werden uitgevoerd volgens de goedgekeurde institutionele richtlijnen van het Comité van de kanselier&#39;s Animal Research (ARC # 2016-037). 1. Mouse verdoving <ol><li> Met acht weken oude vrouwelijke C57 / BL6 muizen (n = 3). </li><li> Verdoven de muizen middels ketamine (80-120 mg / kg muis gewicht) / xylazine (5 mg / kg muis gewicht) oplossingen en b...

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A pending public health crisis.</a> Am J Dent. 22 (1), 3-8 (2009).</li><li>Koliniotou-Koumpia, E., Tziafas, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pulpal+responses+following+direct+pulp+capping+of+healthy+dog+teeth+with+dentine+adhesive+systems.">Pulpal responses following direct pulp capping of healthy dog teeth with dentine adhesive systems.</a> J Dent. 33 (8), 639-647 (2005).</li><li>Tarim, B., Hafez, A. A., Cox, C. 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Momenteel zijn er verschillende experimentele modellen voor de in vivo effecten van tandheelkundige materialen, scaffolds of groeifactoren op odontogene differentiatie van dentale pulp stamcellen (DPSC&#39;s) 13 te valideren. Deze modellen omvatten ectopische autologe transplantatie van DPSC&#39;s in een orgaan, zoals de renale capsule of subcutane transplantatie van DPSC in immuungecompromitteerde muizen met steigers 14,15. Echter, deze methoden beperkt hun odontogene effect op DPSC&#39;s...

Dental caries are one of the most prevalent oral diseases and the leading cause of surgical interventions to dentitions in almost all individuals1,2. The prognosis of surgical interventions and restorations of a tooth largely depends upon proper pulpal response and successful wound healing. Indeed, dental caries that penetrate deeply through the enamel and dentin frequently lead to the exposure of the underlying pulp tissue that is often "capped" with dental materials, such as calcium hydroxide (Ca(OH)2) or hydraulic calcium-silicate cements (HCSCs), including mineral trioxide aggregates (MTA). The ultimate goal of such a pulp-capping procedure is to expedite pulpal wound healing by regenerating reparative dentin, a physical barrier that functions as a "biological seal" to protect the underlying pulp tissue and to increase the life expectancy of the tooth and the overall oral health. However, the underlying mechanism of pulpal wound healing and reparative dentin formation is not fully understood.
To better understand the mechanisms of pulpal wound healing and reparative dentin formation in vivo, several animals were previously used, including monkeys, dogs, and pigs3-5. Among them, rats are frequently used because they are relatively smaller in sizes compared to the other animals, but their teeth are large enough to perform direct pulp capping without any technical difficulties6-10. These animal models are ideal alternatives to human studies for examining pulpal responses and reparative dentin formation. However, their utilization is limited to observational studies at the cellular level, and they scarcely provide mechanistic insights during reparative dentin formation at the molecular level.
Recent technical advances in genetic engineering provided invaluable and indispensable research tools-mice that harbor a gene that is either overexpressed or deleted-that are instrumental to studying molecular mechanisms of human diseases in vivo. The numbers of different strains of transgenic or knockout mice that are strategically inducible in a cell-specific manner are continually growing in the scientific community. Therefore, examining pulpal wound healing and reparative dentin regeneration in these mice would greatly help to expedite our understanding of these processes at the molecular level. However, the use of mice is significantly dampened, as performing a pulp-capping procedure on a mouse tooth is technically challenging due to its miniature size. Here, we present our reproducible method of performing direct pulp capping in mice for the evaluation of pulpal wound healing and reparative dentin formation in vivo.

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			<td style="width:173px;">Microscope&#160;</td>
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			<td>Coltene/whaledent Inc.</td>
			<td>C7970100115</td>
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			<td>T-14012</td>
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			<td>Thermo</td>
			<td>HM355S</td>
			<td>Microtome</td>
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			<td height="21" style="height:21px;">Hematoxyline-1</td>
			<td>Thermo Scientific</td>
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			<td>Thermo Scientific</td>
			<td>8310-16</td>
			<td>Mounting solution</td>
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development of a direct pulp-capping model for the evaluation of pulpal wound healing and reparative dentin formation in mice

Dental pulp is a vital organ of a tooth fully protected by enamel and dentin. When the pulp is exposed due to cariogenic or iatrogenic injuries, it is often capped with biocompatible materials in order to expedite pulpal wound healing. The ultimate goal is to regenerate reparative dentin, a physical barrier that functions as a "biological seal" and protects the underlying pulp tissue. Although this direct pulp-capping procedure has long been used in dentistry, the underlying molecular mechanism of pulpal wound healing and reparative dentin formation is still poorly understood. To induce reparative dentin, pulp capping has been performed experimentally in large animals, but less so in mice, presumably due to their small sizes and the ensuing technical difficulties. Here, we present a detailed, step-by-step method of performing a pulp-capping procedure in mice, including the preparation of a Class-I-like cavity, the placement of pulp-capping materials, and the restoration procedure using dental composite. Our pulp-capping mouse model will be instrumental in investigating the fundamental molecular mechanisms of pulpal wound healing in the context of reparative dentin in vivo by enabling the use of transgenic or knockout mice that are widely available in the research community.

Hier toonden we de procedures stap-voor-stap om pulp uit te voeren aftopping op muizen tanden. Een van de belangrijkste aspecten van de pulp aftopping in muizen is om de juiste apparatuur hebben. In dit verband heeft de microscoop met 10x vermogenvergroting essentieel (Figuur 1A). Een klasse-I-achtige preparaat in de tand maken, gebruikten we een ¼-round burr in een elektrische hoge snelheid handstuk bij 200.000 rpm (Figuur 1B). Alternatief andere motor...

Watch this Scientific Journal Video about Development of a Direct Pulp-capping Model for the Evaluation of Pulpal Wound Healing and Reparative Dentin Formation in Mice at JoVE.com

Development of a Direct Pulp-capping Model for the Evaluation of Pulpal Wound Healing and Reparative Dentin Formation in Mice

We beschrijven een stap voor stap methode om direct pulp aftopping op muizen tanden voor de evaluatie van pulpa wondgenezing en vorming herstellende dentine in vivo.

Ontwikkeling van een Direct-Pulp aftopping model voor de evaluatie van pulpal wondgenezing en Reparative Dentin Formation in Muizen

Dental pulp is a vital organ of a tooth fully protected by enamel and dentin. When the pulp is exposed due to cariogenic or iatrogenic injuries, it is ...

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Medicine

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Muizenmodel van wondgenezing

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Muizen Endoscopie voor<em&gt; In Vivo</em&gt; Multimodal Imaging van Carcinogenese en Evaluatie van intestinale wondgenezing en ontsteking

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Artificial skin has achieved considerable therapeutic results in clinical practice. However, artificial skin treatments for wounds in diabetic patients ...

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Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The ...

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Methoden voor de beoordeling van epitheliale cel migratie tijdens het In Vitro wondgenezing op basis van microscopie

Cell migration is a mandatory aspect for wound healing. Creating artificial wounds on research animal models often results in costly and complicated ...

A Minimally Invasive Model to Analyze Endochondral Fracture Healing in Mice Under Standardized Biomechanical Conditions

Een minimaal invasieve Model voor het analyseren van Endochondral fractuur genezing in muizen onder gestandaardiseerde condities van de biomechanische

Bone healing models are necessary to analyze the complex mechanisms of fracture healing to improve clinical fracture treatment. During the last decade, an ...

Corneal Epithelial Abrasion with Ocular Burr As a Model for Cornea Wound Healing

Hoornvlies epitheliale schuren met oogbeschadigingen en/of Burr als een Model voor hoornvlies wondgenezing

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to ...

The Generation of Closed Femoral Fractures in Mice: A Model to Study Bone Healing

De generatie van gesloten femorale fracturen in muizen: een Model om te studeren bot genezing

Bone fractures impose a tremendous socio-economic burden on patients, in addition to significantly affecting their quality of life. Therapeutic strategies ...

Ex Vivo Corneal Organ Culture Model for Wound Healing Studies

Ex Vivo wondgenezing hoornvlies orgel cultuur Model voor Studies

The cornea has been used extensively as a model system to study wound healing. The ability to generate and utilize primary mammalian cells in two ...

Targeting Gray Rami Communicantes in Selective Chemical Lumbar Sympathectomy

Gericht op grijze Rami Communicantes in selectieve chemische Lumbale Sympathectomy

Chemical lumbar sympathectomy (CLS) is a commonly used, minimally invasive procedure for the treatment of conditions including ischemic diseases of the ...