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 undersøgelse blev støttet af R01DE023348 (RHK) fra NIDCR / NIH og fakultetet Research Grant (RHK) fra Rådet om Forskning for Akademisk Senat i Los Angeles Division fra University of California.

Mus blev købt fra Jackson Laboratory og holdes i et patogen-fri vivarium i UCLA Division Laboratory Animal Medicine (DLAM). Forsøgene blev udført i overensstemmelse med de godkendte institutionelle retningslinier fra Udvalget for kansler Animal Research (ARC # 2016-037). 1. Mus bedøvelse <ol><li> Brug otte uger gammel kvinde C57 / BL6-mus (n = 3). </li><li> Bedøve mus ved anvendelse ketamin (80-120 mg / kg af muse vægt) / xylazin (5 mg / kg af muse vægt) opløsninger og administrere intraperitonealt (ip...

<ol>
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I øjeblikket er der adskillige forskellige eksperimentelle modeller at validere in vivo effekter af dentale materialer, stilladser, eller vækstfaktorer for odontogent differentiering over dental pulp stamceller (DPSC&#39;er) 13. Disse modeller inkluderer ektopisk autolog transplantation af DPSC&#39;er ind i et organ, såsom nyrekapslen eller subkutan transplantation af DPSC&#39;er til immunkompromitterede mus med stilladser 14,15. Imidlertid er disse metoder begrænset i at deres odontog...

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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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 trin-for-trin procedurer til at udføre pulp capping på mus tænder. Et af de vigtigste aspekter af pulp capping i mus er at have det rette apparat. I denne henseende har den mikroskop med en 10X forstørrelse er afgørende (figur 1A). For at oprette en klasse-I-lignende præparat i tanden, brugte vi en ¼-rund grat i en elektrisk høj hastighed håndstykke ved 200.000 rpm (figur 1B). Alternativt andre motorer, herunder dem, der bruger tryk...

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 trin-for-trin fremgangsmåde til udførelse af direkte pulp capping på mus tænder for evaluering af pulpa sårheling og reparerende dentin dannelse in vivo.

Udvikling af en Direct Pulp-capping Model for Vurdering af pulpa Sårheling og Reparative Dentin Formation 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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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

Hornhindens epitel slid med okulær Burr som en Model for hornhinden sårheling

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

Generation af lukkede femur frakturer i mus: en Model til at studere knogleheling

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 sårheling hornhinde orgel kultur Model for undersøgelser

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

Målretning grå Rami Communicantes i selektiv kemiske lumbal Sympathectomy

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