Method Article

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

DOI:

10.3791/65908

⸱

April 30th, 2026

 ,  ,  ,  ,  ,  ,  ,  , 
1Division of Comprehensive Care, Department of Periodontology, University of North Carolina at Chapel Hill, 2Division of Pediatric and Public Health, Adams School of Dentistry, University of North Carolina at Chapel Hill, 3Department of Epidemiology, Gillings School of Global Public Health, University of North Carolina at Chapel Hill, 4Department of Applied Physical Sciences, University of North Carolina at Chapel Hill, 5Department of Biostatistics and Health Data Science, School of Medicine, Indiana University

In This Article

Summary

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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 μCT analysis.

Abstract

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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  leading 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 "mouse dental bed" and "ligature holder"), and alveolar bone quantification using μCT 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' implementation of the simplified ligature-induced periodontitis model to test innovative periodontal therapeutics in the preclinical setting.

Introduction

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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 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 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 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 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.

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Protocol

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This protocol adhered to the Animal Research: Reporting of In Vivo Experiments (ARRIVE) guidelines22,23. All experimental procedures described in this protocol were approved by the IACUC of the University of North Carolina at Chapel Hill.

1. Experimental design

  1. Sample-size calculation
    NOTE: Because of the translational nature of the research program, the power calculation presented here is based on alveolar bone measurements of 8-12-week-old wild-type C57Bl6 mice.
    1. Analyze bone based on the linear measurements between the cementum-enamel junction and the alveolar bone crest (CEJ-ABC).
      1. Healthy mice have a mean attachment level (or CEJ-ABC measurement) of 0.2 mm, while mice with periodontitis at the 9-day post ligation timepoint have double that amount (CEJ-ABC distance of 0.4 mm)11. The alveolar bone loss does not increase significantly from 9 to 18 days post ligature placement11.
      2. Perform a power calculation analysis based on this effect size to identify the number of mice required to reveal these differences between health and disease (here, a total of 8-10 mice [Figure 1A]).
        NOTE: Power was calculated to achieve a power of between 0.8 and 0.95, α = 0.05, using a two-sample means power. Group means and standard deviations are presented in Figure 1B.
    2. To determine a therapeutic effect on bone loss, perform a different power calculation. Rather than comparing healthy mice with diseased mice, as both experimental and control groups of mice develop periodontitis in this protocol, set aside between 19 and 31 mice/group to ensure result reproducibility when testing bone loss modulation using the simplified ligature-induced periodontitis mouse model (see representative results and Figure 1C).
      NOTE: The power was calculated using the same test described above. Group means and standard deviations are presented in Figure 1D.
  2. Procedure standardization and personnel calibration
    1. Train and calibrate all personnel involved in disease induction and quantification before study initiation (intercalibration and intracalibration; r2 ≥ 0.90).
      NOTE: It is estimated that the training for ligature placement and bone loss quantification requires a period of 2-3 months (timeline presented in Table 1). Other required steps are presented as a timeline in Table 1.
    2. For the ligature placement procedure, have two trained investigators working together in the procedure room. The main researcher (intra- and inter-calibrated) remains as the operator who places the ligatures.
    3. Have the second investigator work as a procedural assistant and perform the steps listed below:
      1. Record mouse weight.
      2. Place the mouse in the anesthetic box.
      3. Place eye ointment on the mouse before handing the mouse to the operator for its placement in the "mouse dental bed".
      4. Ensure that ≥3 "ligature holders" are appropriately loaded with the ligatures and ready to use.
        NOTE: To improve efficiency, it is ideal to have ≥3 "ligature holders" appropriately loaded for usage by the operator. Expediency is recommended (i.e., a procedure time of 5-10 min/mouse), increasing the number of experiments conducted in a day and decreasing potential batch effects.
      5. Perform mouse monitoring (before and after the procedure).
        NOTE: The ligature-induced periodontitis is a non-invasive, non-surgical procedure that is similar to flossing a human tooth. Ligature placement is expected to cause very mild discomfort to the mice (mainly due to accumulation of bacteria between two teeth) but should not require any usage of analgesics. No complications are expected following ligature placement conducted by a trained individual. All animals are monitored every 2-3 days after the ligature placement. Humane endpoints (e.g. failure to groom or feed, inability to rise or ambulate, labored respiration, necrotic or infected wounds) should be listed in the approved animal protocol, but if any complications occur, veterinary staff should be consulted about appropriate actions.

2. Creation of printed tools to induce ligature periodontitis in mice

NOTE: To optimize the ligature placement, 3D printed tools named Mouse Dental Bed and Ligature Holder were developed, composed of several individual 3D printed pieces that are separately printed and further assembled manually. The blueprint specifications for these devices were previously published11 and can be used by experienced investigators to create individual STL files. The new devices in the present study were created in conjunction with the University of North Carolina at Chapel Hill BeAM Design Center. Materials can be obtained through the respective core facility or 3D printed using the STL files in Supplemental File 1.

  1. Mouse dental bed
    1. Print the Mouse Dental Bed (Figure 2A) and Ligature Holder (Figure 2B) using standard print quality on a 3D printer using 3.0 mm polylactic acid (PLA) filament. Print the dental bed in the designed orientation and the other parts with the uniform flat surfaces oriented downward against the print bed. Activate the support material option by enabling the Support Placement setting. Make sure that the support material touches the building plate first by selecting the Touching Buildplate option.
      NOTE: Use the five STL files in Supplemental File 1 to print all the required elements, including the Detached Tube, Ligature (with notch), Mouse Dental Bed (with notches), Lid, and Ligature Holder Lock. See the Table of Materials for the dimensions of each item (blueprint specifications previously presented10).
    2. After printing is complete, trim any printed supporting material parts from models using wire cutters, needle-nose pliers, or another instrument.
      NOTE: Wear safety goggles during this procedure.
  2. Mouse dental bed assembly
    1. To prepare the mouse dental bed, lay out all parts as shown (Figure 3A) and prepare the glue to be used (Figure 3B).
    2. Place a few drops of glue (Figure 3C) and attach the screws as shown (Figure 3D). Again, prepare to transfer glue (Figure 3E) to the areas in which it will be applied. Slide the slot of the printed tube (Figure 3F) in place with the barbed end facing away from the Mouse Dental Bed (this is where the hose to deliver isoflurane will attach).
    3. Secure head-stabilizing pieces in positions with glue (Figure 3F).
    4. Wipe away any excess glue and keep slight pressure on all inserted pieces for firm attachment (Figure 3F).
  3. Ligature holder adjustments
    1. To reduce discomfort when locking/unlocking the mechanism, ensure that all square/sharp edges on the exterior portion of the Ligature Holder Lock (Figure 4A) are slightly beveled at a 45° angle using a 15C scalpel blade handle, no. 3 with 15C blade.
    2. To ensure a tight fit of the locking mechanism to the Ligature Holder, square off any internal rounded corners on the Ligature Holder Lock (Figure 4B-F) using a 15C scalpel blade.
      NOTE: This will allow the lock to fully seat against the Ligature Holder and only require minor adjustments.
      1. Use a 3D print trimming tool or a 15C scalpel blade to remove "flash" remaining on the top and bottom edges even after removing most of the support, and slightly bevel the edges at 45° (similar to the bevel performed on the Ligature Holder Lock in step 1) to ensure a tightly fitting lock.
        NOTE: Do not remove too much material, but just enough for the lock to slide on without any resistance.
    3. If too thin, trim the length of each tip until a thicker cross-section is reached [at least 0.5 mm thick], ensuring that enough clearance remains between the tips to prevent impinging on the palate when placing ligatures. Ensure that the tips are at least 3 mm long after cutting (Figure 5A, shown by "CUT").
    4. On the tips of the Ligature Holder, remove any flash from the "support" that the tool was printed on to make sure that the side of the tips are flat (Figure 5B, arrow).
    5. Take out a "slice" of the side of each tip at a 45° angle to make it easier to load the ligature onto it as well as soften the edges to prevent traumatic injury to the soft tissues (Figure 5C, indicated by arrows).
    6. Ensure that each tip on the Ligature Holder includes a "V"-shaped notch in which the suture silk ligatures can rest. If this was removed in step 2.3.6 because the initial printed tip was too thin, refine it on the print or reproduce it.
      1. If this "V"-shaped notch is visible, lay the Ligature Holder on a flat surface with both tips parallel to the tabletop, place a 15C scalpel blade gently into the base of the notch, and gently rock the blade side to side such that the blade edge stays at the base of the "V" and the side of the blade touches the top walls of the "V". With the blade at the base of the notch, very carefully angle the blade towards the outer surface of the tip and repeat this motion to refine the base of the "V"-shaped notch on the outer edge, which will allow the suture thread to guide into place more easily.
        NOTE: This rocking action creates just enough movement in the filament to move it out of the way and refine the notch without removing an excessive amount of material.
      2. If this "V"-shaped notch is not visible, lay the Ligature Holder down on a flat surface with both tips perpendicular to the tabletop. While looking straight down along both tips of the Ligature Holder, place a 15C scalpel blade in the middle of the tips, and in one deliberate motion, make a shallow cut to create the start of what will end up as the "V"-shaped notch.
        NOTE: It is often easier to perform this action on each tip separately rather than trying to position the blade in the correct spot on both tips at the same time.
      3. Repeat step 2.3.7.2 if necessary, making sure that the edge of the scalpel blade is always tracking back into the same spot in the middle of the tip.
        NOTE: If the edge is the blade does not track back into the same spot, multiple cuts will be made, which will cause the filament layers to begin separating in those areas and potentially reduce the performance of the tool.
        NOTE: The depth of the "V"-shape notch must be tested periodically using a tied suture-silk ligature to determine whether the ligature is holding or if it keeps slipping off (may indicate that further refinement is necessary). The holder must be printed with the notch facing down on the buildplate and the Ligature Holder Lock oriented with the wider opening down touching the buildplate. Again, support material can be used with Support Placement set to Touching Buildplate only.
  4. Ligature holder assembled
    NOTE: After the modifications are made, assemble the Ligature Holder, and check the following:
    1. When a ligature is locked in place, ensure that the lock pinches the thread to the Ligature Holder (not seen in Figure 6A as pointed by the arrow, but seen in Figure 6B) at either the top or bottom of the lock (both if possible but this may be difficult to achieve).
      NOTE: This friction will ensure that the ligature does not slip out of place.
    2. When threading the ligature into the grooves on the Ligature Holder, if it keeps slipping from the tips, either deepen the grooves or extend them further onto the outside of the tip to create a "track" in which the thread will sit.
    3. Check if the two knots are centered (Figure 6C) and level (straight line in Figure 6D) when viewed from the front of the Ligature Holder; if not, then one groove is deeper than the other.
    4. Check that the ligature is sitting just within the notches and that it is not sitting too deep; otherwise, there is an elevated risk of the tip of the Ligature Holder contacting the soft tissues and causing trauma during ligature placement.
    5. Using a dental explorer (Figure 6E), confirm that the ligature bounces back into place without slipping when gentle force is applied between the knots (this simulates the force it will experience during ligature placement).

3. Ligature placement

NOTE: It is possible for the ligatures to be placed between murine teeth with or without the addition of human periodontal pathogens11. A time-dependent analysis of the microbial composition of the ligature (without supplementation with human bacterial species) demonstrated a shift in the microbiome community (dysbiosis) during periodontitis development12. For this reason, ligature replacement is not recommended as it may alter result reproducibility. The inclusion of bacterial species in combination with the ligature is dependent on the question being addressed in a specific study. Because of the expected 10-15% ligature loss11, ligatures are placed bilaterally in a single mouse. This approach minimizes the loss of mice during the experiment.

  1. After the mouse has been anesthetized using inhaled isoflurane, place the anesthetized mouse in the Mouse Dental Bed with its back against the bed and the nose positioned in proximity to the nose cone.
    NOTE: Mice should be anesthetized using 4% (vol/vol) Isoflurane. Isoflurane vaporizer and oxygen are filtered by a 0.2-µm air filter. Once the mouse is fully anesthetized and placed in the Mouse Dental Bed, the mouse should be maintained with a 1.5-2.5% flow through the nose cone. Mice are monitored by involuntary body movements and breath rate (~55-65 breaths per minute). No analgesic is required to be provided to mice induced with ligature periodontitis, as this is a non-invasive, non-surgical procedure that is similar to flossing a human tooth. Ligature placement is expected to cause very mild discomfort to the mice (mainly due to the accumulation of bacteria between two teeth), but should not require any usage of analgesics. No complications are expected following ligature placement conducted by a trained individual.
  2. Use the rubber bands to stabilize the mouse's head and open its mouth.
    NOTE: The band around the mandibular incisors also retracts the tongue, and the tension is passive to open the jaw without pulling on it. Visualization of the molars is made easier when the light source is directed at an angle to prevent shadows from obscuring the field of view. The mouse dental bed was designed specifically for the simplified ligature-induced periodontitis mouse model. The distance between the nose of the mouse and the cone is required to allow the mouse to have small body movements during the experimental procedure. This movement of the mouse allows appropriate monitoring of the operator’s hand maneuver while using the ligature holder and controls the amount of force used during the ligature placement. Ligature placement should take 5-10 min per mouse. Procedures should be performed in a well-ventilated hood for performing animal experiments
  3. To place a ligature on the right side, hold the Ligature Holder in the right hand and the explorer in the left.
    NOTE: If the knots on the ligature are not centered between the tips but are rather closer to one tip, then the ligature may be better suited for placement on a specific side. For example, if the knots are placed closer to the right tip of the Ligature Holder, then this ligature may be easier to place on the right side.
  4. Gently place the tip of the explorer below the interproximal contact of the first and second molars on the palatal side.
    1. Make the explorer tip first touch the teeth and 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.
    2. Make note of which end of the explorer is used for placement on the right side and which is used on the left. Ensure that the tip of the explorer hooks into the proximal contact area in an upward direction while allowing the bend of the shank to lay across the contralateral side to stabilize the head against reactionary forces during placement.
  5. Using the Ligature Holder in a horizontal angulation, retract the cheek by first positioning the Ligature Holder above the interproximal contact between the second and third molars, pushing the edge of the Ligature Holder against the cheek to move it, and then bringing the Ligature Holder into position at the interproximal contact of the first and second molars.
    NOTE: This step helps ensure that no buccal mucosa gets trapped between the ligature and the teeth, which will interfere with placement.
  6. Once the ligature is positioned over the interproximal contact between the first and second molars, rotate the handle of the Ligature Holder up to 45°, and with a combination of gentle rocking and sliding, push the ligature through the contact.
  7. Cutting the excess thread:
    1. Use a fine needle forceps to clasp the excess thread attached to the knot on the right side, pull the thread across the teeth towards the left, and using scissors that are just slightly open, slide the scissors down the thread until positioned ~1.5 mm from the knot before cutting.
      NOTE: This helps reduce the likelihood of inadvertent mucosal trauma when cutting the excess.
    2. Repeat the above with directions reversed for the other knot.
  8. Mirror the above steps from 3.3. to 3.7.2. when placing a ligature on the opposite side of the mouth.
    NOTE: Ligature collection at sacrifice is recommended, as the dysbiotic shift may be altered by different experimental conditions being tested12. To facilitate the collection, mice are placed in the Murine Dental Bed and one of the knots of the ligature is cut out followed by gentle interproximal removal with forceps. The usage of the Mouse Dental Bed during the ligature collection allows for appropriate stability of the mouse and ease of access to the ligature site.

4. Bone destruction quantification

  1. μCT analysis
    1. Open the software and click on File | Import Image…
    2. Click on Browse in the popup menu, go to the folder containing Dicom files for one scan, and select the first file.
    3. Click on Examine to find and select any other files within the folder that are part of the same scanned sample. Click OK.
    4. After the image begins to load, click on Visualize and Isosurface.
    5. Change the image threshold to 5,000 and Surface Quality Factor (%) to 55.
    6. Check boxes marked Enable Image Smoothing and Clip Surface with Current ROI and click on Update.
    7. Adjust Level Properties by right-clicking on Window/Level at the bottom (Figure 7A) and click on Edit Window/Level Properties. Select a bone density value that is standard for all the samples. To follow this protocol, use 2,000 but change it based on the bone density of the samples and keep it constant for all measurements.
    8. To manipulate planes:
      1. Use keyboard shortcuts I/J/K hide/show for the three different planes.
      2. Right-click and move the cursor to zoom in and out.
      3. Press Shift and then left-click to pan.
      4. Click Slice to move and rotate the planes.
      5. Click on the center of the plane to move the plane forward/backward or 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.
      6. Click on the edge of the plane to rotate around a line parallel to this edge in the center of the plane.
      7. Click on the corner of a plane to rotate around the plane.
    9. Orient the z-axis (blue pane at the top of the screen-the z-slice) so that the surfaces/shapes of all three teeth become visible at the same time (or as close as possible) (Figure 7B).
    10. Scroll through this same plane (blue pane at the top of the screen-z-slice) so that the roots of the 1st and 2nd molars are visible (Figure 7C). Try to pick a point that is far enough down the roots so that all surrounding anatomy can be seen, but also where the canals appear well-defined and in a straight line.
      NOTE: The plane may need to be rotated to vertically orient the canals.
    11. Orient the x-axis (vertical green line in the z-slice panel) to align with the buccal roots of the 1st and 2nd molar (try to align through the central part of each root/canal if possible; otherwise, align through the buccal root of the 1st molar and the distobuccal root of the 2nd molar) (Figure 7D).
    12. Use the orientation of the x-slice as a reference for the alignment of the y-slice (vertical blue line in the x-slice panel).
      1. Align the y-slice so that it is vertically parallel through the root of the first molar in the x-slice panel (​Figure 7E). Rotate the plane to orient the root vertically. Observe that the blue line crosses through the middle of the root.
        NOTE: In areas of bone loss (as is expected with this model), the alveolar ridge may have a defect; therefore, the midline of the root must be estimated because the surrounding bone may not be present to ideally define the middle of the socket.
    13. In the y-slice panel, measure the distance from the cementoenamel junction (CEJ) to the alveolar bone crest (ABC) by placing the cursor in the deepest point of the groove at the CEJ and pressing 1, and then by placing the cursor at the closest point of the ABC and pressing 2 as shown (Figure 7F). The 1st point is denoted by a blue arrow and the second point by a green arrow.
      1. If the arrows do not show up until both points are selected, select them by hovering the arrow over the desired areas and press the number keys on the keyboard as just clicking the mouse will not work.
      2. If the CEJ is difficult to determine, use the up and down arrow keys to scroll through adjacent slices to confirm its location.
      3. Measure multiple points of the ABC if the "closest point" is difficult to determine to confirm that there is little to no variation in measurement.
        NOTE: To confirm that the correct measurement is taken, one must see the x and z slices intersecting over the desired root and can visually confirm the measurement in the 3D view by hiding the planes (Figure 7F).
  2. Gingival expression quantification of Rankl/Opg and Rankl/Rank
    1. Collect gingival tissue as previously described9.
      NOTE: Best RNA extraction results are obtained using fresh tissue samples and RNA purification must be processed as soon as possible.
    2. Extract gingival tissue RNA by bead beating using the referenced kit and ceramic beads.Follow the instructions on the kit and store the sample at -80 °C until analysis by qRT-PCR.

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Results

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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...

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Discussion

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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's experience with this specific ligature-induced periodontitis mouse model11,12,13,

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Disclosures

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The STL files used to create the 3D printed devices in previous publication9 are under a copyright agreement with the University of North Carolina at Chapel Hill ("UNC-Mouse Surgical Bed and UNC-Ligature Holder" UNC Ref. No. 18-0024). The authors declare no competing financial interests.

Acknowledgements

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The research reported in this publication was supported by the National Institute of Dental & 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).

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Materials

List of materials used in this article
NameCompanyCatalog NumberComments
BladeIntegra4-315C15C blade
Ceramic beadsVWR10158-6080.5 mL x 1.4 mm Ceramic beads
Glue/resin materialThe Gorilla Glue Company8401519
Ligature Holder (With Notch)UNC at Chapel Hill BeAM Design Center facility corenot applicable118.01 mm x 13 mm x 13.74 mm
Ligature Holder LockUNC at Chapel Hill BeAM Design Center facility corenot applicable17.5 mm x 14.57 mm x 17.8 mm
Lulzbot mini v1.0 3D printer Lulzbot (Fargo Additive Manufacturing Equipment 3D, LLC)not applicableAleph Objects, Lulzbot Mini v1.0 model
MicroView SoftwareParallax Innovations2.5.0-3139Software used for bone measurements
Mouse Dental Bed (With Notches)UNC at Chapel Hill BeAM Design Center facility corenot applicable103.86 mm x 196.85 mm x 50.99 mm
Mouse Dental Bed TopUNC at Chapel Hill BeAM Design Center facility corenot applicable99.8 mm x 77 mm x 21.5 mm
Polylactic acid (PLA) filamentVerbatim552593.0 mm PLA
Rapidpure RNA Tissue KitMP Biomedicals112721050
Scalpel blade handleBard-Parker37103015C scalpel blade handle
Suture-silkRoboz Surgical InstrumentSUT-15-1
Taqman qPCR primersThermo FisherRank (Mm00437135_ m1), Rankl (Mm00441906_ m1), Opg (Mm00435452_ m1), Gapdh (Mm99999915_g 1)
VX-765 InvivoGen, San Diego, USA#inh-vx765-1Caspase-1 inhibitor used for host modulation

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Alveolar Bone Destruction3D Printed ToolsMouse Dental BedLigature HolderMicro CT AnalysisGingival Gene ExpressionOsteoclastogenic SignalingInflammatory Bone Destruction

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