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

Denna studie stöddes av R01DE023348 (RHK) från NIDCR / NIH och fakulteten Research Grant (RHK) från rådet för forskning av Academic senaten i Los Angeles uppdelningen av University of California.

Möss köptes från Jackson Laboratory och förvaras i en patogen-fri vivarium i UCLA Avdelningen för försöksdjursmedicin (DLAM). Experimenten utfördes enligt de godkända institutionella riktlinjer från djurkommittén förbundskanslerns forskning (ARC # 2016-037). 1. Mus Anesthetization <ol><li> Använd åtta veckor gamla kvinnliga C57 / BL6-möss (n = 3). </li><li> Söva mössen med användning av ketamin (80 till 120 mg / kg av mus vikt) / xylazin (5 mg / kg mus vikt) lösningar och administrera intrap...

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För närvarande finns det flera olika experimentella modeller för att validera effekter in vivo av dentala material, ställningar, eller tillväxtfaktorer på odontogena differentiering av dentala massa stamceller (DPSC) 13. Dessa modeller inkluderar ektopisk autolog transplantation av DPSC in i ett organ, såsom njurkapseln, eller subkutan transplantation av DPSC in i immunförsvagade möss med byggnadsställningar 14,15. Emellertid är dessa metoder begränsade i det att deras odontoge...

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 height="21" style="height:21px;width:164px;">BM-LED stereo microscope</td>
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			<td style="width:173px;">Microscope&#160;</td>
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			<td height="21" style="height:21px;">Optima MCX-LED&#160;</td>
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			<td>1700588-001</td>
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			<td height="21" style="height:21px;">OptiBond SoloPlus</td>
			<td>Kerr</td>
			<td>29669</td>
			<td>Adhesives</td>
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			<td height="21" style="height:21px;">Coltolux LED</td>
			<td>Coltene/whaledent Inc.</td>
			<td>C7970100115</td>
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			<td height="21" style="height:21px;">Characterization tint</td>
			<td>Bisco</td>
			<td>T-14012</td>
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			<td>Breuker</td>
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			<td height="21" style="height:21px;">Microm</td>
			<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>
			<td>7221</td>
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			<td>Thermo Scientific</td>
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			<td height="21" style="height:21px;">Cytoseal 60</td>
			<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.

Här visade vi steg-för-steg-anvisningar för att utföra pulpaöverkappning på möss tänder. En av de viktigaste aspekterna av pulpaöverkappning i möss är att ha lämplig apparat. I detta avseende har den mikroskop med en 10X power förstoring är väsentligt (Figur 1A). För att skapa en klass-I-liknande preparat i tanden, använde vi en ¼-runda burr i en elektrisk höghastighetshandstycke vid 200.000 rpm (Figur 1B). Alternativt kan varje annan ...

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 steg-för-steg-metod för att utföra direkt pulpaöverkappning på möss tänder för utvärdering av pulpal sårläkning och reparativa dentin formation in vivo.

Utveckling av en direkt Pulp-tak modell för utvärdering av pulpa sårläkning och Reparativ Dentin bildning hos möss

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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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

Hornhinnans epitel nötning med okulär Burr som modell för hornhinnan sårläkning

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 slutna lårbensfrakturer hos möss: en modell för att studera benläkning

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 sårläkning korneal orgel kultur modell för 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

Inriktning grå Rami Communicantes i selektiv kemiska ländryggen Sympathectomy

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