Name: development of a direct pulp-capping model for the evaluation of pulpal wound healing and reparative dentin formation in mice
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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

Este estudio fue apoyado por R01DE023348 (RHK) del NIDCR / NIH y la Beca de Investigación de la Facultad (RHK) del Consejo de Investigación del Senado Académico de la División de la Universidad de California en Los Ángeles.

Los ratones fueron adquiridos de Jackson Laboratory y se mantienen en un vivero libre de patógenos en la División de UCLA de Medicina de Laboratorio Animal (DLAM). Los experimentos se llevaron a cabo de acuerdo con las directrices institucionales aprobados desde el Comité de Investigación Animal del Canciller (ARC # 2016-037). 1. Ratón anestesia <ol><li> Utilizar ratones hembra C57 / BL6 de ocho semanas de edad (n = 3). </li><li> Anestesiar los ratones utilizando ketamina (80 a 120 mg / kg de peso de rat?...

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Actualmente, hay varios modelos experimentales disponibles para validar los efectos in vivo de los materiales dentales, andamios, o factores de crecimiento sobre la diferenciación odontogénico de células madre de la pulpa dental (DPSCs) 13. Estos modelos incluyen el trasplante autólogo de ectópico DPSCs en un órgano, tales como la cápsula renal, o trasplante subcutáneo de DPSCs en ratones inmunocomprometidos con andamios 14,15. Sin embargo, estos métodos están limitados en que su...

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;">Coltolux LED</td>
			<td>Coltene/whaledent Inc.</td>
			<td>C7970100115</td>
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			<td>T-14012</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>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.

A continuación, mostramos los procedimientos paso a paso para llevar a cabo el recubrimiento pulpar de los dientes ratones. Uno de los aspectos clave de recubrimiento pulpar en ratones es tener el aparato apropiado. En este sentido, teniendo el microscopio con un aumento de potencia de 10X es esencial (Figura 1A). Para crear una preparación de Clase-I-al igual que en el diente, se utilizó una fresa de ¼ de vuelta en una pieza de mano de alta velocidad eléctrica a 20...

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

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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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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

Cicatrización de heridas de abrasión epitelial corneal con rebabas Ocular como un modelo de córnea

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

La generación de fracturas femorales cerradas en ratones: un modelo para el estudio del hueso curativo

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 modelo de cultura órgano córneo para la curación de heridas estudios

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

Targeting Communicantes grises del Rami en Sympathectomy Lumbar químico selectivo

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