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

Denne studien ble støttet av R01DE023348 (RHK) fra NIDCR / NIH og Fakultet stipend (RHK) fra Rådet for forskning av Det akademiske kollegium av Los Angeles Division ved University of California.

Musene ble kjøpt fra Jackson Laboratory og holdt i et patogen-fri vivarium i UCLA Division of Laboratory Animal Medicine (DLAM). Forsøkene ble utført i henhold til godkjente institusjonelle retningslinjer fra kanslerens Forsøksdyrutvalget (ARC # 2016-037). 1. Mouse anesthetization <ol><li> Bruk åtte uker gammel kvinne C57 / BL6 mus (n = 3). </li><li> Bedøve musene ved hjelp av ketamin (80-120 mg / kg musevekt) / xylazin (5 mg / kg musevekt) løsninger og administrere intraperitonealt (ip) med en dose på...

<ol>
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For tiden er det flere forskjellige eksperimentelle modeller som er tilgjengelige for å validere de in vivo virkningene av dentale materialer, stillas, eller vekstfaktorer på odontogenic differensiering av tann massen stamceller (DPSCs) 13. Disse modellene har ektopisk autolog transplantasjon av DPSCs til et organ, slik som nyrekapselen, eller subkutan transplantasjon av DPSCs inn immunkompromitterte mus med stillaser 14,15. Imidlertid er disse fremgangsmåter er begrenset ved at deres o...

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>Thermo</td>
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			<td>Thermo Scientific</td>
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			<td>Thermo Scientific</td>
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			<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.

Her viste vi den trinnvise prosedyrer for å utføre masse capping på mus tenner. En av de viktigste aspektene av papirmasse capping i mus er å ha riktig apparat. I denne forbindelse har den mikroskop med en forstørrelse 10X kraft er vesentlig (figur 1A). For å opprette en klasse-I-lignende preparat i tannen, brukte vi en ¼-runde graden i en elektrisk høyhastighetshåndstykke på 200 000 rpm (figur 1B). Alternativt kan andre motorer, inkludert de s...

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

Vi beskriver en trinn-for-trinn metode for å utføre direkte papirmasse capping på mus tenner for evaluering av pulpal sårtilheling og reparerende dentin dannelse in vivo.

Utvikling av en direkte Pulp-capping modell for evaluering av pulpal Wound Healing og reparerende Dentin Dannelse i Mus

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

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

Hornhinnen Epithelial slitasje med okulær Burr som modell for hornhinnen sårtilheling

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

Generering av lukkede femur brudd i mus: en modell for å studere bein Healing

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 sårtilheling hornhinnen Organ kultur modell for studier

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

Målretting grå Rami Communicantes i selektiv kjemiske lumbale Sympathectomy

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