Name: Forfatter Spotlight: Forbedring af tandmasseforskning med forbedrede musemodeller
Uploaded: 2023-10-27T13:30:00
Description: Watch this Scientific Journal Video about Establishment of a Murine Pulp Exposure Model with a Novel Mouth-Gag for Pulpitis Research at JoVE.com

Denne undersøgelse blev finansieret af tilskud fra National Natural Science Foundation of China U21A20368 (L. Y.), 82101000 (H. W.) og 82100982 (F. L.) og af Sichuan Science and Technology Program 2023NSFSC1499 (H. W.). Alle de originale data og billeder er inkluderet i denne artikel.

De eksperimentelle procedurer i denne undersøgelse blev godkendt af den etiske komité ved West China School of Stomatology, Sichuan University (WCHSIRB-D-2021-125). Voksne C57BL/6-mus blev opnået fra Gempharmatech Experimental Animals Company, Chengdu, Kina. Hele kronen på den kæbelige første kindtand bryder ud 21 dage efter fødslen. Mus til operation skal være ældre end 21 dage med normal vitalitet11. Her blev 6 til 8 uger gamle mus brugt til modellering. Figur 1</st...

Oprettelse af en pulp inflammation musemodel ved hjælp af en ny mundgag 

<ol>
	<li>Kleinert, A., Kleinert, L., Ozimirska, M., Cha&#322;as, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Endodontium+-+together+or+separately.">Endodontium - together or separately.</a> Folia Morphol. 77 (3), 409-415 (2018).</li><li>Dhillon, H., Kaushik, M., Sharma, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regenerative+endodontics-Creating+new+horizons.">Regenerative endodontics-Creating new horizons.</a> J Biomed Mater Res B Appl Biomater. 104 (4), 676-685 (2016).</li><li>Prati, C., Pirani, C., Zamparini, F., Gatto, M. R., Gandolfi, M. 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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Regenerative+endodontics:+a+review+of+current+status+and+a+call+for+action.">Regenerative endodontics: a review of current status and a call for action.</a> J Endod. 33 (4), 377-390 (2007).</li><li>Arora, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Potential+application+of+immunotherapy+for+modulation+of+pulp+inflammation:+opportunities+for+vital+pulp+treatment.">Potential application of immunotherapy for modulation of pulp inflammation: opportunities for vital pulp treatment.</a> Int Endod J. 54 (8), 1263-1274 (2021).</li><li>Eramo, S., Natali, A., Pinna, R., Milia, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dental+pulp+regeneration+via+cell+homing.">Dental pulp regeneration via cell homing.</a> Int Endod J. 51 (4), 405-419 (2018).</li><li>Hasan, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Expression+of+Toll-like+receptor+2,+Dectin-1,+and+Osteopontin+in+murine+model+of+pulpitis.">Expression of Toll-like receptor 2, Dectin-1, and Osteopontin in murine model of pulpitis.</a> Clin Oral Investig. 27 (3), 1177-1192 (2023).</li><li>Wang, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=DDIT3+aggravates+pulpitis+by+modulating+M1+polarization+through+EGR1+in+macrophages.">DDIT3 aggravates pulpitis by modulating M1 polarization through EGR1 in macrophages.</a> Int Immunopharmacol. 120, 110328 (2023).</li><li>Richert, R., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+critical+analysis+of+research+methods+and+experimental+models+to+study+pulpitis.">A critical analysis of research methods and experimental models to study pulpitis.</a> Int Endod J. 55 (Suppl 1), 14-36 (2022).</li><li>Huang, X. F., Zhao, Y. B., Zhang, F. M., Han, P. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+study+of+gene+expression+during+tooth+eruption+and+orthodontic+tooth+movement+in+mice.">Comparative study of gene expression during tooth eruption and orthodontic tooth movement in mice.</a> Oral Dis. 15 (8), 573-579 (2009).</li><li>Kwon, S. J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thermal+irritation+of+teeth+during+dental+treatment+procedures.">Thermal irritation of teeth during dental treatment procedures.</a> Restor Dent Endod. 38 (3), 105-112 (2013).</li><li>He, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pulpal+tissue+inflammatory+reactions+after+experimental+pulpal+exposure+in+mice.">Pulpal tissue inflammatory reactions after experimental pulpal exposure in mice.</a> J Endod. 43 (1), 90-95 (2017).</li><li>Karrar, R. N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+biomarkers+for+objective+assessment+of+symptomatic+pulpitis:+A+systematic+review+and+meta-analysis.">Molecular biomarkers for objective assessment of symptomatic pulpitis: A systematic review and meta-analysis.</a> Int Endod J. 56 (10), 1160-1177 (2023).</li><li>Goldman, E., Reich, E., Abramovitz, I., Klutstein, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inducing+apical+periodontitis+in+mice.">Inducing apical periodontitis in mice.</a> J Vis Exp. (150), e59521 (2019).</li><li>Duncan, H. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Present+status+and+future+directions-Vital+pulp+treatment+and+pulp+preservation+strategies.">Present status and future directions-Vital pulp treatment and pulp preservation strategies.</a> Int Endod J. 55 (Suppl 3), 497-511 (2022).</li><li>Shi, X., Li, Z., He, Y., Jiang, Q., Yang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+different+dental+burs+for+experimental+induction+of+pulpitis+in+mice.">Effect of different dental burs for experimental induction of pulpitis in mice.</a> Arch Oral Biol. 83, 252-257 (2017).</li><li>Du, W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Indigenous+microbiota+protects+development+of+medication-related+osteonecrosis+induced+by+periapical+disease+in+mice.">Indigenous microbiota protects development of medication-related osteonecrosis induced by periapical disease in mice.</a> Int J Oral Sci. 14 (1), 16 (2022).</li></ol>

Som det ensomme bløde væv i tænderne udfylder tandpulpa en afgørende rolle i opretholdelsen af tandens bioaktivitet, men forbliver meget følsom. Bevarelsen af denne vitale pulpa er blevet den foretrukne indledende tilgang i nyere endodontiske behandlinger, hvilket kræver en omfattende forståelse af de inflammatoriske mekanismer i tandmasse16. Den spatiotemporale udsving i det inflammatoriske mikromiljø og interaktioner mellem residente celletyper i pulpitis komplicerer undersøgelsen genne...

Tandmassen, en integreret del af tanden, er ansvarlig for flere væsentlige funktioner såsom næringsstofforsyning, dentindannelse, sensorisk funktion og forsvarsreaktioner1. Ikke desto mindre er tandmassen, omgivet af hårdt væv, modtagelig for skader og skader fra dybe karies, pulpitis, traumer eller efterfølgende terapier 2,3. Fraværet af funktionel tandmasse øger risikoen for tandskrøbelighed4. Desuden kan tabet af pulpa-vitalitet i unge permanente tænder påvirke tandmodningen negativt, og nuværende proteseteknikker formår ikke at genoprette den neurale feedback, der tilbydes af sund pulpa4. Denne situation har fået forskere til at udforske alternative løsninger til håndtering af betændt papirmasse ud over blot fjernelse.
I 2007 indledte Murray et al. anvendelsen af vævsteknologi i regenerativ endodonti, hvilket udløste øget interesse for pulpbevarelse og regenerering5. Imidlertid udgør betændt pulpavæv en udfordring, da celler frigiver inflammatoriske faktorer såsom IL-6, som rekrutterer inflammatoriske celler og resulterer i cellenekrose, tab af pulpavitalitet og komplikationer i funktionel genopretning 6,7. Forståelse af inflammation og den tilhørende celledød er derfor afgørende for fremskridt i bevarelsen af vital pulp. Der er en række eksperimenter, der er blevet udført for at udforske molekylærbiologien af den betændte pulp in vivo eller in vitro 8,9. Selvom in vitro-eksperimenter som 2D- eller 3D-cellekulturer er blevet udviklet i årevis og er ved at blive modne og meget brugt til at teste pulpcellers reaktioner på inflammatoriske faktorer, kan disse eksperimenter ikke afspejle interaktionen mellem pulpavæv og det systemiske immunsystem10. Hvis fænomenet, der undersøges, stammer fra celler af anden vævsoprindelse som immun-, vaskulær- og nervesystem, vil ren pulpcellekultur føre til en blindgyde. Derfor er in vivo-eksperimenter meget nødvendige og refererende.
Mus er i stigende grad blevet et almindeligt valg inden for inflammationsforskning in vivo på grund af deres omkostningseffektivitet, høje fertilitet og vitalitet. Imidlertid er der i øjeblikket en omfattende protokol for musens pulpitis-model, som kan tjene som reference. Musenes lille størrelse og deres følsomhed over for stimulering udgør betydelige udfordringer under eksperimentelle procedurer. At observere de små tænder, der er skjult dybt inde i musens mund, kræver ofte brug af et udkragende mikroskop, på trods af den mere almindelige tilstedeværelse af skrivebordsmikroskoper i laboratorier. Fraværet af en mundåbner kræver hjælp fra andre. For at løse dette har gruppen udtænkt en mund-gag ved hjælp af let tilgængelige materialer, som har til formål at give en standardiseret og reproducerbar protokol til konstruktion af musens pulpitis-model. Denne artikel beskriver proceduren, der dækker præoperativ forberedelse, immobilisering, pulpaeksponeringskirurgi og prøveindsamling på C57-mus. Denne protokol anbefaler brugen af mundgag, der giver information om dens struktur, produktion og anvendelse for at lette andre forskere med at replikere proceduren.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Animal</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>C57/B6J mice</td>
			<td>Gempharmatech Experimental Animals Company</td>
			<td>C57/B6J</td>
			<td>For the establishment of pulp exposure</td>
		</tr>
		<tr>
			<td>Equipment</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>1 mL syringe</td>
			<td>Chengdu Xinjin Shifeng Medical Apparatus &#38; Instruments Co. LTD.</td>
			<td>SB1-074(IV)</td>
			<td>Apply in drug injection.</td>
		</tr>
		<tr>
			<td>8# C+ file</td>
			<td>Readysteel</td>
			<td>0010047</td>
			<td>Apply in exposing the roof of pulp chamber.</td>
		</tr>
		<tr>
			<td>Anesthesia Mix solution</td>
			<td></td>
			<td></td>
			<td>10% ketamine hydrochloride+ 5% xylazine + 85% sterile isotonic saline.&#160;</td>
		</tr>
		<tr>
			<td>DAPI Staining Solution</td>
			<td>Beyotime</td>
			<td>C1005</td>
			<td>Apply in immunofluorescence staining for counter-staining of nucleus.</td>
		</tr>
		<tr>
			<td>Dental high-speed dental handpiece</td>
			<td>Jing yuan electronic commerce technology</td>
			<td>WJ-422</td>
			<td>Apply in pulp exposure.</td>
		</tr>
		<tr>
			<td>Heavy wire cutter</td>
			<td>Jirui Medical Instrument Co., Ltd.</td>
			<td>none</td>
			<td>Apply inarc cutting.</td>
		</tr>
		<tr>
			<td>Hematoxylin and Eosin Stain kit</td>
			<td>Biosharp</td>
			<td>BL700B</td>
			<td>For the histological analysis of the slides.</td>
		</tr>
		<tr>
			<td>IL-6 antibody</td>
			<td>Novus</td>
			<td>NBP2-89149</td>
			<td>Apply in immunofluorescence staining to detect the inflammation of the dental pulp.</td>
		</tr>
		<tr>
			<td>Ketamine(Ketamine hydrochloride)</td>
			<td>Vet One, Boise, Idaho, USA</td>
			<td>C3N VT1</td>
			<td>100mg/kg, IP. Apply in nesthetization.</td>
		</tr>
		<tr>
			<td>Medical tap</td>
			<td>3M</td>
			<td>1530</td>
			<td>Apply in mice immobilization.</td>
		</tr>
		<tr>
			<td>Orthodontic arch wire&#160;</td>
			<td>Shanghai Wei Rong Medical Apparatus Co. LTD.</td>
			<td>K417</td>
			<td>Diameter of 8&#181;m</td>
		</tr>
		<tr>
			<td>Round dental burr (0.6 mm)</td>
			<td>Shofu global</td>
			<td>072208</td>
			<td>Apply in removing enamel and shallow layer of dentin.</td>
		</tr>
		<tr>
			<td>Young loop bending plier</td>
			<td>Jirui Medical Instrument Co., Ltd.</td>
			<td>none</td>
			<td>Apply in arc bending.</td>
		</tr>
	</tbody>
</table>

establishment of a murine pulp exposure model with a novel mouth-gag for pulpitis research

Pulpitis, en almindelig årsag til naturligt tandtab, fører til nekrose og tab af bioaktivitet i den betændte tandmasse. Optrævling af de mekanismer, der ligger til grund for pulpitis og dens effektive behandling, er et løbende fokus for endodontisk forskning. Derfor er det afgørende at forstå den inflammatoriske proces i tandmassen for at forbedre bevarelsen af papirmassen. Sammenlignet med andre in vitro-eksperimenter tilbyder en murine pulpitis model en mere autentisk og genetisk forskelligartet kontekst til at observere den patologiske progression af pulpitis. Men at bruge mus, på trods af deres omkostningseffektivitet og tilgængelighed, giver vanskeligheder på grund af deres lille størrelse, dårlige koordination og lave tolerance, hvilket komplicerer intraorale og tandbehandlinger. Denne protokol introducerer et nyt design og anvendelse af en mundgag til at blotlægge musepulpa, hvilket letter mere effektive intraorale procedurer. Mundgagen, der består af en tandbue, der er let tilgængelig for de fleste tandlæger og kan fremskynde kirurgisk forberedelse betydeligt, selv ved førstegangsprocedurer. Mikro-CT, hæmatoxylin-eosin (HE) farvning og immunfluorescensfarvning blev brugt til at identificere ændringer i morfologi og celleekspression. Formålet med denne artikel er at hjælpe forskere med at etablere en mere reproducerbar og mindre krævende procedure til at skabe en pulp-inflammationsmodel ved hjælp af denne nye mund-gag.

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.

Forfatter Spotlight: Forbedring af tandmasseforskning med forbedrede musemodeller

Etablering af en model for eksponering for musemasse med en ny mundgag til pulpitisforskning

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.

Proceduren beskrevet ovenfor blev udført på højre kæbekindstand på 3, 6-8 uger gamle C57BL/6 mus, mens venstre kæbekindtænder blev bevaret som kontrol. Histologi- og immunfluorescensresultater fra blankkontrol, 12 timers pulpitis og 24 timers pulpitis prøver blev brugt til demonstration.
Efter protokol for CT-analyse fra Goldman et al.15 blev pulpeksponering bekræftet gennem mikro-CT og rekonstruktionsmodellering i figur 4 ...

Watch this Scientific Journal Video about Establishment of a Murine Pulp Exposure Model with a Novel Mouth-Gag for Pulpitis Research at JoVE.com

Denne artikel præsenterer en strømlinet protokol til etablering af en pulpitis-model hos mus ved hjælp af en innovativ mund-gag, efterfulgt af efterfølgende histologisk analyse.

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

Individuelle eksperimenter som 2D- eller 3D-cellekulturer er blevet brugt i vid udstrækning til at teste reaktionerne af tandmasseceller. Disse eksperimenter kan ikke afspejle interaktionen mellem pulpavæv og det systemiske immunsystem. Derfor bliver in vivo-eksperimenter mere og mere populære inden for endodontisk forskning.C57-mus er i stigende grad blevet et almindeligt valg i inflammationsforskning på tandmasse på grund af deres omkostningseffektivitet, høje fertilitet og vitalitet. Men musenes lille størrelse og manglende koordination udgjorde betydelige udfordringer under eksperimentelle procedurer. Fikseringsprocessen er en vigtig faktor, der hæver den tekniske tærskel.Vi udtænkte en ny mundgag ved hjælp af let tilgængelige materialer og enkle samlinger for at hjælpe forskere med pulpaeksponeringskirurgi. Ifølge vores forskning kan denne mundgag betydeligt forkorte den tid, det tager for begyndere at immobilisere munden og udsætte den maksillære første kindtand hos mus.

establishment-murine-pulp-exposure-model-with-novel-mouth-gag-for

Research

JoVE Journal

Medicine

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 underlying pulpitis and its efficient treatment is an ongoing focus of endodontic research. Therefore, understanding the inflammatory process within the dental pulp is vital for improving pulp preservation. Compared to other in vitro experiments, a murine pulpitis model offers a more authentic and genetically diverse context to observe the pathological progression of pulpitis. However, using mice, despite their cost-effectiveness and accessibility, poses difficulties due to their small size, poor coordination, and low tolerance, complicating intraoral and dental procedures. This protocol introduces a novel design and application of a mouth-gag to expose mouse pulp, facilitating more efficient intraoral procedures. The mouth-gag, comprised of a dental arch readily available to most dentists and can significantly expedite surgical preparation, even for first-time procedures. Micro-CT, hematoxylin-eosin (HE) staining, and immunofluorescence staining were used to identify changes in morphology and cell expression. The aim of this article is to help researchers establish a more reproducible and less demanding procedure for creating a pulp inflammation model using this novel mouth-gag.

This article presents a streamlined protocol for establishing a pulpitis model in mice using an innovative mouth-gag, followed by subsequent histological analysis.

Preparation of the Mouth Gag for a Mouse Pulpitis Model

To begin, straighten an 8 micron thick orthodontic arch wire with your left hand while keeping the right hand slightly bent against the wire arc. Use a Young loop bending plier to bend the top edge of the trapezoid about eight millimeters at the midpoint of the bow wire. Ensure that point A is on the edge of the plier beak.Now hold the plier with the left hand. Clamp the free end of the bow wire with the right thumb and forefinger, then bend the bow wire from point A to make a 120-degree angle. Place the arch wire on a horizontal table to check if it is on a single plane.Leave about nine millimeters on each side of the wire and use pliers to bend the free end to a 75-degree angle. Now locate point C which is about five millimeters from point B.Bend the wire at point C to make a 105-degree angle. Similarly, bend the wire at points D and E.Clamp the IK and KJ parts simultaneously.Duplicate the A, B, C bending at points L, J, and K, then bend the free end at point J to make it vertical to the IKJ plane. Bend an extra tongue depressor for the mandibular part. Clamp point I five millimeters from point J and bend the free end to make it parallel to both the IKJ plane and the CD part.Next, leave a five millimeter gap from point I and at point H, bend the arch vertically to the IH and JIH planes. Locate point G five millimeters from H and clamp the JIHG lane. Then bend the arch at point G.Now bend the free end symmetrical to the KJ plane.The free end after point F should be symmetrical to the free end at point E.Finally, place rubber caps on the free ends.

Forberedelse af mundgag til en mus pulpitis model

Using A Mouth Gag for Pulpitis Assessment in Mice

To begin, fix an anesthetized mouse in a supine position on a fixation plate. Secure the limbs with skin tape. Compress the free ends of a prepared mouth gag with the thumb and the index finger.Fix the mouse's front incisors in the trapezoidal groove between the two arms of the mouth gag. Ensure the arm with the tongue depressor is placed on the mandibula. Check the maxillary first molar to ensure it is free of dental caries, trauma, and odontogenesis.Assess the surrounding gingiva to ensure absence of redness, swelling, or fistula. Then, check the health of the opposite teeth. Next, with a dental burr, drill the occlusal side of the maxillary first molar at 20, 000 RPM.Remove the enamel. With a syringe, drop normal saline every three minutes over the tooth to prevent overheating. Put a number 8 C+file on the lowest position of the drilled pit and pierce the last dentine to expose the pulp chamber.Clean the fragments around the tooth, then remove the mouth gag. The time taken to fix the mouth with the mouth gag and to find the maxillary first molar was significantly shortened relative to the traditional method. The pulp exposure was confirmed through micro CT and reconstruction modeling.At 24 hours post-surgery, most of the pulp tissue retained its morphology.