Name: Optimization of the Ligature-induced Periodontitis Mouse Model for Preclinical Research (Video) | JoVE Journal
Uploaded: 2026-04-30T14:10:00.000+00:00
Duration: 9 min 26 s
Description: Learn a simplified ligature-induced periodontitis mouse model using 3D-printed tools for efficient, reproducible induction of bone loss. This protocol enables rapid preclinical testing of periodontal therapeutics with micro-CT quantification.

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The research reported in this publication was supported by the National Institute of Dental &#38; Craniofacial Research of the National Institutes of Health under Award Numbers K01DE027087 (JTM) and R01DE032830 (JTM). The work was also supported by the National Institute of Allergy and Infectious Diseases of the National Institutes of Health under award number R01-AI153265 (KVS). The content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health. Additionally, the research reported here is also supported by the Fulbright/CAPES program finance code 001 (TA).

Fabricating 3D Printed Tools in Ligature-Induced Periodontitis Studies in Mice

<ol><li>Offenbacher, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Periodontal+diseases%3A+pathogenesis%5BTitle%5D)+AND+%22Ann+Periodontol%22%5BJournal%5D)">Periodontal diseases: pathogenesis.</a> Ann Periodontol. 1 (1), 821-878 (1996).</li>
<li>Alves, T., Swanson, K. V., Girnary, M. S., Freire, M., Moss, K., Divaris, K., Beck, J., Santos, P. C. P., Brickey, W. J., Wrobel, J. A., Min, H. Y., Syed, M., Morelli, T., Seaman, W. T., Preisser, J. S., Hu, D., Preisser, H., Makhanova, N., Susin, C., Vias, N. P., Styner, M., Zhang, S., Pirih, F. Q., Chang, J., Gill, H. S., Lietzan, A. D., Webster-Cyriaque, J., Ting, J. P. Y., Marchesan, J. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Inflammasome+targeting+for+periodontitis+prevention+is+sex+dependent%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">Inflammasome targeting for periodontitis prevention is sex dependent.</a> Proc Natl Acad Sci U S A. 122 (44), e2507092122(2025).</li>
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<li>Hajishengallis, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Illuminating+the+oral+microbiome+and+its+host+interactions%3A+animal+models+of+disease%5BTitle%5D)+AND+%22FEMS+Microbiol+Rev%22%5BJournal%5D)">Illuminating the oral microbiome and its host interactions: animal models of disease.</a> FEMS Microbiol Rev. 47 (3), fuad018(2023).</li>
<li>Graves, D. T., Fine, D., Teng, Y. T., Van Dyke, T. E., Hajishengallis, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+use+of+rodent+models+to+investigate+host-bacteria+interactions+related+to+periodontal+diseases%5BTitle%5D)+AND+%22J+Clin+Periodontol%22%5BJournal%5D)">The use of rodent models to investigate host-bacteria interactions related to periodontal diseases.</a> J Clin Periodontol. 35 (2), 89-105 (2008).</li>
<li>Hasturk, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((RvE1+protects+from+local+inflammation+and+osteoclast-+mediated+bone+destruction+in+periodontitis%5BTitle%5D)+AND+%22FASEB+J%22%5BJournal%5D)">RvE1 protects from local inflammation and osteoclast- mediated bone destruction in periodontitis.</a> FASEB J. 20 (2), 401-403 (2006).</li>
<li>Hasturk, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Phase+IIa+clinical+trial+of+complement+C3+inhibitor+AMY-101+in+adults+with+periodontal+inflammation%5BTitle%5D)+AND+%22J+Clin+Invest%22%5BJournal%5D)">Phase IIa clinical trial of complement C3 inhibitor AMY-101 in adults with periodontal inflammation.</a> J Clin Invest. 131 (23), e152973(2021).</li>
<li>Maekawa, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Genetic+and+intervention+studies+implicating+complement+C3+as+a+major+target+for+the+treatment+of+periodontitis%5BTitle%5D)+AND+%22J+Immunol%22%5BJournal%5D)">Genetic and intervention studies implicating complement C3 as a major target for the treatment of periodontitis.</a> J Immunol. 192 (12), 6020-6027 (2014).</li>
<li>Maekawa, T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Antagonistic+effects+of+IL-17+and+D-resolvins+on+endothelial+Del-1+expression+through+a+GSK-3beta-C%2FEBPbeta+pathway%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">Antagonistic effects of IL-17 and D-resolvins on endothelial Del-1 expression through a GSK-3beta-C/EBPbeta pathway.</a> Nat Commun. 6, 8272(2015).</li>
<li>Hajishengallis, G., Hasturk, H., Lambris, J. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Contributing+authors.+C3-targeted+therapy+in+periodontal+disease%3A+moving+closer+to+the+clinic%5BTitle%5D)+AND+%22Trends+Immunol%22%5BJournal%5D)">Contributing authors. C3-targeted therapy in periodontal disease: moving closer to the clinic.</a> Trends Immunol. 42 (10), 856-864 (2021).</li>
<li>Marchesan, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+experimental+murine+model+to+study+periodontitis%5BTitle%5D)+AND+%22Nat+Protoc%22%5BJournal%5D)">An experimental murine model to study periodontitis.</a> Nat Protoc. 13 (10), 2247-2267 (2018).</li>
<li>Ribeiro, A. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Oral+biofilm+dysbiosis+during+experimental+periodontitis%5BTitle%5D)+AND+%22Mol+Oral+Microbiol%22%5BJournal%5D)">Oral biofilm dysbiosis during experimental periodontitis.</a> Mol Oral Microbiol. 37 (6), 256-265 (2022).</li>
<li>Jiao, Y., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Induction+of+bone+loss+by+pathobiont-mediated+Nod1+signaling+in+the+oral+cavity%5BTitle%5D)+AND+%22Cell+host+%26+microbe%22%5BJournal%5D)">Induction of bone loss by pathobiont-mediated Nod1 signaling in the oral cavity.</a> Cell host & microbe. 13 (5), 595-601 (2013).</li>
<li>Marchesan, J. T., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Role+of+inflammasomes+in+the+pathogenesis+of+periodontal+disease+and+therapeutics%5BTitle%5D)+AND+%22Periodontol+2000%22%5BJournal%5D)">Role of inflammasomes in the pathogenesis of periodontal disease and therapeutics.</a> Periodontol 2000. 82 (1), 93-114 (2000).</li>
<li>Chadwick, J. W., Glogauer, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Robust+ligature-induced+model+of+murine+periodontitis+for+the+evaluation+of+oral+neutrophils%5BTitle%5D)+AND+%22J+Vis+Exp%22%5BJournal%5D)">Robust ligature-induced model of murine periodontitis for the evaluation of oral neutrophils.</a> J Vis Exp. (155), e59667(2020).</li>
<li>Collins, F. S., Tabak, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Policy%3A+NIH+plans+to+enhance+reproducibility%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">Policy: NIH plans to enhance reproducibility.</a> Nature. 505 (7485), 612-613 (2014).</li>
<li>Landis, S. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+call+for+transparent+reporting+to+optimize+the+predictive+value+of+preclinical+research%5BTitle%5D)+AND+%22Nature%22%5BJournal%5D)">A call for transparent reporting to optimize the predictive value of preclinical research.</a> Nature. 490 (7419), 187-191 (2012).</li>
<li>Swanson, K. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Interferon+activated+gene+204+%28Ifi204%29+protects+against+bone+loss+in+experimental+periodontitis%5BTitle%5D)+AND+%22J+Periodontol%22%5BJournal%5D)">Interferon activated gene 204 (Ifi204) protects against bone loss in experimental periodontitis.</a> J Periodontol. 93 (9), 1366-1377 (2022).</li>
<li>Sun, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((IL-10+dampens+an+IL-17-mediated+periodontitis-associated+inflammatory+network%5BTitle%5D)+AND+%22J+Immunol%22%5BJournal%5D)">IL-10 dampens an IL-17-mediated periodontitis-associated inflammatory network.</a> J Immunol. 204 (8), 2177-2191 (2020).</li>
<li>Offenbacher, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((GWAS+for+Interleukin-1beta+levels+in+gingival+crevicular+fluid+identifies+IL37+variants+in+periodontal+inflammation%5BTitle%5D)+AND+%22Nat+Commun%22%5BJournal%5D)">GWAS for Interleukin-1beta levels in gingival crevicular fluid identifies IL37 variants in periodontal inflammation.</a> Nat Commun. 9 (1), 3686(2018).</li>
<li>Apaza Alccayhuaman, K. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((FasL+is+a+catabolic+factor+in+alveolar+bone+homeostasis%5BTitle%5D)+AND+%22J+Clin+Periodontol%22%5BJournal%5D)">FasL is a catabolic factor in alveolar bone homeostasis.</a> J Clin Periodontol. 50 (3), 396-405 (2023).</li>
<li>Percie du Sert, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((The+ARRIVE+guidelines+2.0%3A+updated+guidelines+for+reporting+animal+research%5BTitle%5D)+AND+%22BMJ+Open+Sci%22%5BJournal%5D)">The ARRIVE guidelines 2.0: updated guidelines for reporting animal research.</a> BMJ Open Sci. 4 (1), e100115(2020).</li>
<li>Percie du Sert, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Reporting+animal+research%3A+Explanation+and+elaboration+for+the+ARRIVE+guidelines+2.0%5BTitle%5D)+AND+%22PLoS+Biol%22%5BJournal%5D)">Reporting animal research: Explanation and elaboration for the ARRIVE guidelines 2.0.</a> PLoS Biol. 18 (7), e3000411(2020).</li>
<li>Marchesan, J. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Inflammasomes+as+contributors+to+periodontal+disease%5BTitle%5D)+AND+%22J+Periodontol%22%5BJournal%5D)">Inflammasomes as contributors to periodontal disease.</a> J Periodontol. 91 Suppl (1), S6-S11 (2020).</li>
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The STL files used to create the 3D printed devices in previous publication9&#160;are under a copyright agreement with the University of North Carolina at Chapel Hill (&#34;UNC-Mouse Surgical Bed and UNC-Ligature Holder&#34; UNC Ref. No. 18-0024). The authors declare no competing financial interests.

The present study provides insightful information and detailed, step-by-step guidance on the simplified ligature-induced periodontitis mouse model protocol previously presented10. The details included in the current paper can dramatically impact reproducibility and are supported by the group&#39;s experience with this specific ligature-induced periodontitis mouse model11,12,13,<sup class="xref"...

Periodontal disease is a complex inflammatory process that leads to the destruction of tooth-supporting structures including the gingiva, the periodontal ligament, the cementum, and the alveolar bone. For periodontal bone loss to develop, the breakage of the gingival barrier and bacterial invasion need to occur, in addition to the recruitment of inflammatory cells, and the transition from an innate-dominated to an acquired-dominated immune response1. The final imbalance between osteoblasts and osteoclasts will result in alveolar bone loss via its destruction2. The hallmark phenotypic finding that differentiates the diagnosis of periodontal disease from reversible gingival inflammation (gingivitis) is the loss of alveolar bone. Therefore, the amount of tooth-supporting alveolar bone is a key therapeutic outcome in the preclinical setting.
No single animal model can replicate all biological aspects required for the development of different therapeutics3,4,5. The ligature-induced periodontitis mouse model develops quantifiable bone destruction in a reasonably short amount of time (3-9 days)5, making this a cost-effective model to test novel therapeutic interventions for periodontitis prevention and modulation. Despite its&#160;success in supporting the development of phase II-III human clinical trials6,7,8,9,10, ligature-induced periodontitis in mice is not commonly utilized as a preclinical model. To overcome&#160;technical challenges of the classic model in which the ligature surrounds the tooth, the simplified ligature-induced periodontitis mouse model11,12 was established. The simple modification of the classic model is centered on a focused bone destruction site, between rather than around teeth. The benefits of this modification are multifold, including a shorter length of time required for 1) personnel training, 2) technical ease of the experimental procedure, and 3) reproducibility of&#160;bone quantification. Additionally, it was discovered that several microbiological and immunological events that develop and manifest under this simplified model are aligned with the current understanding of common human periodontal disease2,11,12,13. Last, the effective drug-induced bone loss prevention previously reported by us2,14 further supports the usage of the simplified ligature-induced periodontitis model as an in vivo tool to evaluate key aspects of novel periodontal therapies.
An innovative aspect of the simplified ligature-induced periodontitis model has been the development of 3D-printed tools to more easily facilitate the disease induction process11,12. While intuitive for some, investigators have reported an inability to assemble the 3D-printed tools15, which may further impede or delay animal study initiation. The present study expands upon the information presented in our&#160;previous protocol study11and provides detailed information to allow the successful implementation of the simplified ligature-induced periodontitis model in a laboratory setting. Details included in this current protocol are part of a complex array of factors that may contribute to the lack of result reproducibility of scientific findings15,16,17. The consistent model usage and reliable bone resorption identified in the multiple studies2,11,12,14,18 and by collaborative research groups13,19,20,21 over the past 10 years support the importance of disseminating detailed information to promote reproducibility in the scientific community.

<table><tbody><tr><td>Blade</td><td>Integra</td><td>4-315C</td><td>15C blade</td></tr><tr><td>Ceramic beads</td><td>VWR</td><td>10158-608</td><td>0.5 mL x 1.4 mm Ceramic beads</td></tr><tr><td>Glue/resin material</td><td>The Gorilla Glue Company</td><td>8401519</td><td></td></tr><tr><td>Ligature Holder (With Notch)</td><td>UNC at Chapel Hill BeAM Design Center facility core</td><td>not applicable</td><td>118.01 mm x 13 mm x 13.74 mm</td></tr><tr><td>Ligature Holder Lock</td><td>UNC at Chapel Hill BeAM Design Center facility core</td><td>not applicable</td><td>17.5 mm x 14.57 mm x 17.8 mm</td></tr><tr><td>Lulzbot mini v1.0 3D printer&amp;nbsp;</td><td>Lulzbot (Fargo Additive Manufacturing Equipment 3D, LLC)</td><td>not applicable</td><td>Aleph Objects, Lulzbot Mini v1.0 model</td></tr><tr><td>MicroView Software</td><td>Parallax Innovations</td><td>2.5.0-3139</td><td>Software used for bone measurements</td></tr><tr><td>Mouse Dental Bed (With Notches)</td><td>UNC at Chapel Hill BeAM Design Center facility core</td><td>not applicable</td><td>103.86 mm x 196.85 mm x 50.99 mm</td></tr><tr><td>Mouse Dental Bed Top</td><td>UNC at Chapel Hill BeAM Design Center facility core</td><td>not applicable</td><td>99.8 mm x 77 mm x 21.5 mm</td></tr><tr><td>Polylactic acid (PLA) filament</td><td>Verbatim</td><td>55259</td><td>3.0 mm PLA</td></tr><tr><td>Rapidpure RNA Tissue Kit</td><td>MP Biomedicals</td><td>112721050</td><td></td></tr><tr><td>Scalpel blade handle</td><td>Bard-Parker</td><td>371030</td><td>15C scalpel blade handle</td></tr><tr><td>Suture-silk</td><td>Roboz Surgical Instrument</td><td>SUT-15-1</td><td></td></tr><tr><td>Taqman qPCR primers</td><td>Thermo Fisher</td><td>Rank (Mm00437135_ m1), Rankl (Mm00441906_ m1), Opg (Mm00435452_ m1), Gapdh (Mm99999915_g 1)</td><td></td></tr><tr><td>VX-765</td><td>&amp;nbsp;InvivoGen, San Diego, USA</td><td>#inh-vx765-1</td><td>Caspase-1 inhibitor used for host modulation</td></tr></tbody></table>

optimization of the ligature-induced periodontitis mouse model for preclinical research

Optimization of the Ligature-induced Periodontitis Mouse Model for Preclinical Research

The experimental model of ligature-induced periodontitis in mice has played a fundamental role in understanding periodontal disease pathogenesis; yet, it has not been fully leveraged for translational and therapeutic applications. Similar to human disease, ligature-induced periodontitis in mice develops via a series of distinct microbial, inflammatory, and osteo-immunological events. The ligature-induced alveolar bone destruction progresses in a reasonably short period and is easily quantifiable, making it a cost-effective disease model for preclinical studies.
To overcome the technical challenge of the classic ligature model, the establishment of the simplified ligature-induced periodontitis mouse model was curated. Rather than inducing disease in the entire circumference of two adjacent teeth, the simplified disease model is centered on inflammatory bone destruction only in the interproximal region of two posterior teeth, which is also the region that common periodontal disease initiates in humans. Because it employs three dimensional (3D)-printed tools for ligature placement, the approach is fast and effective&#160; leading&#160;to a procedural time of 5-10 min per mouse. While osteoclastogenic signaling is detected after 3 days of ligature placement, bone destruction consistently develops at 9 days post ligation.
Here, the simplified ligature model protocol is elaborated by highlighting important elements specific to study design (i.e., power calculation, procedure standardization, calibration), development of the 3D printed tools (i.e., the &#34;mouse dental bed&#34; and &#34;ligature holder&#34;), and alveolar bone quantification using &#956;CT analysis. Gingival gene expression of Rank, Rankl, and Opg, a master regulatory triad of osteoclastogenesis, is used as indirect evidence of therapeutic effectiveness. The methods presented in this paper will guide investigators&#39; implementation of the simplified ligature-induced periodontitis model to test innovative periodontal therapeutics in the preclinical setting.

aRepresentative data analysis of linear alveolar bone measurements from WT female and male mice at baseline (health) and 9 days post ligation are presented in Figure 1A. The power calculation indicates that a total of 10 mice (4-5 mice per group) are needed to detect differences between these groups if the primary outcome considered is linear bone levels measured by μCT (Figure 1B). However, a power calculation for preclinical studies requires that both groups o...

The ligature-induced periodontitis mouse model is an effective and increasingly popular tool for studying periodontal disease. Detailed guidance is presented for implementing a simplified version of the model focused on interproximal alveolar bone destruction, including laboratory study design, 3D-printed tool production, and bone quantification using &#956;CT analysis.

The experimental model of ligature-induced periodontitis in mice has played a fundamental role in understanding periodontal disease pathogenesis; yet, it ...

Our lab investigates periodontal disease, examining how inflammation causes gingival and bone damage in different groups of individuals. We aim to hinder gingivitis and periodontitis by targeting novel players and novel pathways of disease. The simplified ligature-induced periodontitis mouse model utilizes a double-knotted single thread in between the teeth, utilizing 3D-printed tools, the mouse dental bed and the ligature holder, which happens to be less complex than traditional models.This approach uses the technique of inhalation of anesthesia, which happens to result in a quicker recovery for mice, less than 10 minutes, compared to that of more traditional models that use an injectable anesthesia, where the recovery time can be around 30 minutes. To prepare the mouse dental bed assembly, lay out all dental bed parts and the glue on the working platform. Then, secure the head-stabilizing pieces in position using the glue.Using a number three blade on a 15C scalpel blade handle, bevel the sharp edges on the exterior portion of the ligature holder lock at a 45-degree angle to reduce discomfort during locking or unlocking. Square off any internal rounded corners on the ligature holder lock. Remove the flash from the top and bottom edges of the 3D print, and bevel these edges at 45 degrees.Take out a slice of each tip of the ligature holder at a 45-degree angle to prevent traumatic injury to the soft tissues. Ensure each tip on the ligature holder includes a V-shaped notch. If this V-shaped notch is visible, place a 15C scalpel blade gently into the base of the notch.Now, carefully angle the blade towards the outer surface of the tip and refine the base of the V-shaped notch on the outer edge. If the V-shaped notch is not visible, place a 15C scalpel blade in the middle of the ligature holder tip, and in one deliberate motion, make a shallow cut to create a V-shaped notch. While threading the ligature into the grooves, ensure that two knots are centered and level when viewed from the front of the ligature holder.Using a dental explorer, confirm that the ligature bounces back into place without slipping when gentle force is applied between the knots. For ligature placement, position the anesthetized mouse in the mouse dental bed with its back against the bed and the nose positioned in proximity to the nose cone. Using rubber bands, stabilize the mouse's head and open its mouth.To place a ligature on the right side, hold the ligature holder in the right hand and the explorer in the left. Gently place the tip of the explorer below the interproximal contact of the first and second molars on the palatal side. Using the explorer tip, first touch the teeth, then, slide down and rest against the gingiva to avoid traumatizing the soft tissue.Rest the explorer handle in a horizontal angulation once properly positioned. Ensure the correct usage of the explorer tip for ligature placement. Hook the explorer into the proximal contact area, and stabilize the head against reactionary forces.Retract the cheek using the ligature holder, and position the ligature over the interproximal contact between the first and second molars. Next, rotate the handle of the ligature holder up to 45 degrees, and with a combination of gentle rocking and sliding, push the ligature through the contact. Using fine needle forceps, clasp the excess thread attached to the knot on the right side.Pull the thread across the teeth towards the left, and slide the scissors down the thread until positioned approximately 1.5 millimeters from the knot before cutting. For micro CT analysis, open the software and click on File, then Import image. In the pop-up menu, click on Browse.Go to the folder containing DICOM files for one scan and select the first file. Click on Examine to find and select any other files within the folder for the same scanned sample. Then, click OK.Once the images begin to load, click on Visualize and Isosurface.Change the Image Threshold to 5, 000 and the Surface Quality Factor percent to 55. Check boxes marked Enable Image Smoothing and Clip Surface with current ROI, and click on Update. To adjust level properties, right-click on Window/Level at the bottom, and click on Edit Window/Level Properties.Set the bone density value to 2, 000 and keep it constant for all measurements. To manipulate planes, use keyboard shortcuts IJK hide and show for the the three different planes. Right-click and move the cursor to zoom in and out.Press Shift and left-click to pan. Then, click Slice to move and rotate the planes. Next, click on the center of the plane to move the plane forward or backward.Alternatively, place the cursor in the center of the desired plane, and use the up and down arrow keys to move the plane one slice at a time. Click on the edge of the plane to rotate around a line parallel to this edge in the center of the plane. Click on the corner of a plane to rotate around the plane.Next, orient the Z slice, seen in blue at the top of the screen, so that the surfaces of all three teeth become visible at the same time or as close as possible. Then, scroll through the same Z slice plane until the roots of the first and second molars become visible, ensuring the surrounding anatomy and canals are well-defined. Align the x-axis with the buccal roots of the first and second molar through the central part of each root canal.Alternatively, align through the buccal root of the first molar and the distobuccal root of the second molar. Next, use the orientation of the X slice as a reference for the alignment of the Y slice. Align the Y slice vertically parallel through the root of the first molar in the X slice panel.Rotate the plane to orient the root vertically, and observe that the blue lines cross through the middle of the root. In the Y slice panel, to measure the distance from the cemento-enamel junction, or CEJ, to the alveolar bone crest, or ABC, place the cursor in the deepest point of the groove at the CEJ and press 1. Then, place the cursor at the closer point of the ABC and press 2.A blue arrow denotes the first point, and the second point is denoted by a green arrow. If the arrows indicating measurement points do not appear immediately, hover the arrow over the desired areas and press the number keys on the keyboard. If the CEJ is difficult to determine, use the up and down arrow keys to scroll through the adjacent slices to confirm its location.When determining the closest point of the ABC is difficult, measure multiple points to ensure consistent measurement. Despite our success with our protocols paper, new researchers have still found difficulty in the assembly and utilization of the 3D-printed tools, the mouse dental bed, the ligature holder. In order to address this, we have created a step-by-step video which walks researchers through the assembly and the utilization of these 3D-printed tools so that they can be used in a laboratory setting.Our research team has utilized clinical studies and the simplified ligature-induced periodontitis model to uncover how a group of proteins named inflammasome can be used to halt the development of inflammation and periodontitis.

optimization-ligature-induced-periodontitis-mouse-model-for

Research

JoVE Journal

Immunology and Infection

Here, we present a protocol to locally induce apical periodontitis in mice. We show how to drill a hole in the mouse's tooth and expose its pulp, in order to cause local inflammation. Analysis methods to investigate the nature of this inflammation, such as micro-CT and histology, are also demonstrated.

Inducing Apical Periodontitis in Mice

This article presents a protocol for establishing a ligature-induced model of murine periodontitis involving multiple maxillary molars, resulting in larger areas of the involved gingival tissue and bone for subsequent analysis as well as reduced animal usage. A technique to assess oral neutrophils in a manner analogous to human subjects is also described.

Robust Ligature-Induced Model of Murine Periodontitis for the Evaluation of Oral Neutrophils

In this study, a rat model of induction of periodontitis is presented via a combination of retentive ligature and repetitive injections of lipopolysaccharide derived from Porphyromonas gingivalis, over 14 days around the first maxillary molars. The ligation and LPS injection techniques were effective in inducing peridontitis, resulting in alveolar bone loss and inflammation.

Induction of Periodontitis via a Combination of Ligature and Lipopolysaccharide Injection in a Rat Model

This is a report on an experimental model of ligature-induced peri-implantitis in mice. We describe all surgical steps, from pre- and post-operative management of the animals, extractions, implant placement, and ligature-induced peri-implantitis.

Experimental Model of Ligature-Induced Peri-Implantitis in Mice

This study describes a modified 6-0 nylon ligation method for inducing periodontitis in mice, which is highly reproducible and represents an alternative for researchers to study periodontal disease from its development to its pathological consequences.

Murine Model of Advanced Periodontitis Induced by Nylon Ligature in the Second Upper Molar

Neisseria meningitidis is a human specific pathogen that infects blood vessels. In this protocol human microvessels are introduced into a mouse by grafting human skin onto immunocompromised mice. Bacteria adhere extensively to the human vessels, leading to vascular damage and development of the purpuric rash typically observed in human cases.

Humanized Mouse Model to Study Bacterial Infections Targeting the Microvasculature

The current study describes the development and applications of a genetically engineered assay system based on the transfection of rat basophilic leukemia cells with the equine FcεRIα gene. Transfected cells express a functional receptor where the release of mediators of the allergic response can be activated by IgE and antigen.

Development of an in vitro model system for studying the interaction of Equus caballus IgE with its high-affinity receptor Fc&#949;RI

Inflammation plays a central role in the pathogenesis of ischemic stroke. Increasing evidence suggests that it acts as a double-edged sword which exacerbates early brain injury, but also contributes to later repair. This protocol describes the isolation of immune cells from the ischemic brain and their subsequent flow cytometric phenotyping.

Isolation and Flow Cytometric Analysis of Immune Cells from the Ischemic Mouse Brain

This study describes an imaging-based micro-neutralization assay to analyze the antigenic relationships between viruses. The protocol employs a flatbed scanner and has four steps, including titration, titration quantitation, neutralization, and neutralization quantitation. The assay works well with current circulating influenza A(H1N1)pdm09, A(H3N2), and B viruses.

Optimization of a Quantitative Micro-neutralization Assay

The method described here allows for the visualization of IL-8 promoter-dependent inflammation activation in the lungs of mice through non-invasive bioluminescence imaging (BLI). The same animal can be subjected to BLI multiple times for up to two months from the time of delivery of the luciferase reporter construct.

An IL-8 Transiently Transgenized Mouse Model for the In Vivo Long-term Monitoring of Inflammatory Responses

Learn a simplified ligature-induced periodontitis mouse model using 3D-printed tools for efficient, reproducible induction of bone loss. This protocol enables rapid preclinical testing of periodontal therapeutics with micro-CT quantification.

Optimization of the Ligature-induced Periodontitis Mouse Model for Preclinical Research

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