preparation of the mouth gag for a mouse pulpitis model

Creazione di un modello murino di infiammazione della polpa utilizzando un nuovo bavaglio orale 

The dental pulp, an integral part of the tooth, is responsible for multiple essential functions such as nutrient supply, dentin formation, sensory function, and defense reactions1. Nevertheless, the dental pulp, surrounded by hard tissue, is susceptible to injuries and damages from deep caries, pulpitis, trauma, or subsequent therapies2,3. The absence of functional dental pulp increases the risk of tooth fragility4. Moreover, the loss of pulp vitality in young permanent teeth can adversely affect tooth maturation, and current denture techniques fail to restore the neural feedback offered by healthy pulp4. This situation has led researchers to explore alternative solutions for managing inflamed pulp beyond mere removal.
In 2007, Murray et al. initiated the application of tissue engineering in regenerative endodontics, thereby sparking increased interest in pulp preservation and regeneration5. However, inflamed pulp tissue poses a challenge as cells release inflammatory factors such as IL-6, which recruit inflammatory cells and results in cell necrosis, loss of pulp vitality, and complications in functional recovery6,7. Understanding inflammation and the associated cell death is, therefore, crucial for advancements in the preservation of vital pulp. There are a number of experiments that have been conducted to explore the molecular biology of the inflame pulp in vivo or in vitro8,9. Though in vitro&#160;experiments like 2D or 3D cell cultures have been developed for years and are becoming mature and widely used to test reactions of pulp cells to inflammatory factors, these experiments cannot reflect the interaction between pulp tissue and the systemic immune system10. If the phenomenon being studied is derived from cells of other tissue origin like immune, vascular and nervous system, then pure pulp cell culture will lead to a dead end. Therefore, in vivo experiments are very necessary and referential.
Mice have increasingly become a common choice in inflammation research in vivo&#160;due to their cost-effectiveness, high fertility, and vitality. However, a comprehensive protocol for mice pulpitis model is currently absent, which can serve as a reference. The small size of mice and their sensitivity to stimulation pose significant challenges during experimental procedures. Observing the minuscule teeth concealed deep within the mouse mouth often necessitates the use of a cantilever microscope, notwithstanding the more common presence of desktop microscopes in laboratories. The absence of a mouth opener requires assistance from others. To address this, the group has devised a mouth-gag using readily available materials which aims to provide a standardized and reproducible protocol for constructing the mice pulpitis model. This article details the procedure, covering preoperative preparation, immobilization, pulp exposure surgery, and sample collection on C57 mice. This protocol recommends the use of the mouth-gag, providing information on its structure, production, and application to facilitate other researchers in replicating the procedure.

Autore in primo piano: Migliorare la ricerca sulla polpa dentale con modelli murini migliorati

Istituzione di un modello di esposizione alla polpa murina con un nuovo bavaglio orale per la ricerca sulla pulpite

Individual experiments like 2D or 3D cell cultures have been widely used to test the reactions of dental pulp cells. These experiments cannot reflect the interaction between pulp tissue and the systemic immune system. Therefore, in vivo experiments are becoming increasingly popular in endodontic research.C57 mice have increasingly become a common choice in inflammation research on dental pulp due to their cost effectiveness, high fertility and vitality. However, the small size of mice and lack of coordination posed significant challenges during experimental procedures. The fixation process is an important factor that higher the technical threshold.We devised a novel mouth gag using readily available materials and simple assemblies to assist researchers in pulp exposure surgery. According to our research, this mouth gag can significantly shorten the time required for beginners to immobilize the mouth and expose the maxillary first molar of mice.

Preparazione del bavaglio orale per un modello di pulpite murina

Per iniziare, raddrizza un filo dell'arco ortodontico di 8 micron di spessore con la mano sinistra mantenendo la mano destra leggermente piegata contro l'arco del filo. Usa una pinza piegatrice ad anello Young per piegare il bordo superiore del trapezio di circa otto millimetri nel punto medio del filo dell'arco. Assicurarsi che il punto A si trovi sul bordo del becco della pinza.Ora tieni la pinza con la mano sinistra. Bloccare l'estremità libera del cavo dell'arco con il pollice e l'indice destri, quindi piegare il filo dell'arco dal punto A per creare un angolo di 120 gradi. Posiziona il filo dell'arco su un tavolo orizzontale per verificare se si trova su un unico piano.Lascia circa nove millimetri su ciascun lato del filo e usa le pinze per piegare l'estremità libera a un angolo di 75 gradi. Ora individua il punto C che si trova a circa cinque millimetri dal punto B.Piega il filo nel punto C per creare un angolo di 105 gradi. Allo stesso modo, piegare il filo nei punti D ed E.Clamp le parti IK e KJ contemporaneamente.Duplicate la piegatura A, B, C nei punti L, J e K, quindi piegate l'estremità libera nel punto J per renderla verticale rispetto al piano IKJ. Piega un abbassalingua in più per la parte mandibolare. Bloccare il punto I a cinque millimetri dal punto J e piegare l'estremità libera per renderla parallela sia al piano IKJ che alla parte CD.Quindi, lasciare uno spazio di cinque millimetri dal punto I e nel punto H, piegare l'arco verticalmente ai piani IH e JICH. Individuare il punto G a cinque millimetri da H e bloccare la corsia JIHG. Quindi piegare l'arco nel punto G. Ora piegare l'estremità libera simmetricamente al piano KJ.L'estremità libera dopo il punto F dovrebbe essere simmetrica all'estremità libera nel punto E.Infine, posizionare i cappucci di gomma sulle estremità libere.

Research

JoVE Journal

Medicine

Preparation of the Mouth Gag for a Mouse Pulpitis Model

Establishment of a Murine Pulp Exposure Model with a Novel Mouth-Gag for Pulpitis Research

Pulpitis, a common cause of natural tooth loss, leads to necrosis and loss of bioactivity in the inflamed dental pulp. Unraveling the mechanisms ...

Ricerca

Medicina

Using A Mouth Gag for Pulpitis Assessment in Mice

Utilizzo di un bavaglio orale per la valutazione della pulpite nei topi