Name: A Surgical Procedure for Resecting the Mouse Rib: A Model for Large-Scale Long Bone Repair (Video) | JoVE Journal
Uploaded: 2015-01-21T15:00:00.000+00:00
Duration: 8 min 42 s
Description: Learn a surgical rib resection technique in mice for studying large-scale skeletal repair. This method enables observation of cellular and molecular events during bone healing in vivo.

ai_agent_IP

ai_agent

maintenance

instantTrialDurationMinutes

isBiopharma

enableNewMyJoveSectionDesign

educationFeatures

seoMetaApi

ai_copilot_institution_blacklist

ai_chat_institution_blacklist

freeAccessWindow

ssoRoleCapture

scheduledMaintenanceBanner

joveLabsFeatures

disable_autoplay_for_bots

extended_trial_duration

5 minutes

We thank members of the Mariani lab for critical reading of the manuscript. Our funding sources were: the Baxter Medical Scholar Research Fellowship (to M.K.S.), USC undergraduate fellowships and the Provost, Dean Joan M. Schaeffer, and Rose Hills Fellowships, (to M.K.S.). We also acknowledge a CIRM BRIDGES fellowship through Pasadena City College (to T.T.T). and the James H. Zumberge Research and Innovation Fund, the USC Regenerative Medicine Initiative, and the NIAMS NIH under Award Number R21AR064462 (to F.V.M).

NOTE: All procedures are in accordance with an animal protocol approved by the Institutional Animal Care and Use Committee (IACUC) at the University of Southern California.
1. Preparation for Surgery
<ol>
	<li>Lay out the necessary surgical tools and supplies (sterilized, items 1 - 18 in Materials list) and arrange a dissecting microscope and a fiber optics trans-illuminator in a hood or on a downdraft table.</li>
	<li>Place a sexually mature mouse (20 - 30 g, 1 - 3 months old) in an induction chamber with 4% isoflurane to induce anesthesia.</li>
	<li>Confirm anesthesia with a paw and/or tail pinch test.</li>
	<li>Apply ointment to the mouse&#8217;s eyes to avoid eye dryness during the surgery.</li>
	<li>Move the mouse to the microscope stage. Place the mouse into the nose cone of the maintenance hose and position laterally. Provide a warming pouch adjacent to the animal to help maintain body temperature. 
	NOTE: Position of the mouse is dependent on personal preference and/or handedness of the surgeon., either left or right side is fine.</li>
	<li>Adjust isoflurane to 2 - 3% for maintenance, and watch breathing rate.</li>
	<li>Inject buprenorphine (0.05 mg/kg) subcutaneously (lateral flank of the leg) for post-surgery pain with a 25 G needle. The leg may twitch which is a normal response.</li>
</ol>
2. Opening Incision
<ol>
	<li>Detect the desired rib by palpation, and clear the area of hair with a shaver. 
	NOTE: Ribs 8-10 are recommended since ribs 1-7 are closer to the lungs and 11-13 are considerably shorter. Counting the ribs should begin with the most rostral rib as number 1.</li>
	<li>Prepare the area with alternating povidone-iodine and isopropanol (70%) swabbing (3x is often recommended by animal use protocols).</li>
	<li>Cut a 2 cm incision through the skin, directly above and parallel to the desired rib with medium sized microsurgery scissors. Incise through the underlying muscle and fat layers.</li>
	<li>Place all 3 layers (skin, muscle, fat) into a retractor to expose the surgical area, while minimizing the size of the incision.</li>
</ol>
3. Excising the Rib
<ol>
	<li>Cut through the intercostal muscles overlying the desired section of rib with a 5.0 mm scalpel. Position this incision around 5 mm proximal to the chondrocostal joint as this is where the rib is not as highly curved. Carefully separate the muscle from the bone with fine tip forceps.</li>
	<li>To create a resection while retaining the periosteum in the animal, cut through the periosteum, along the length of rib with a 5.0 mm scalpel. Carefully separate the periosteum from the underlying bone laterally with fine tip forceps. Proceed with caution, as the periosteum is very delicate and has a gelatinous consistency.</li>
	<li>Next make a cross-section through the bone at one end with fine micro-scissors. If needed, measure the resection with a reticle in the microscope or a ruler type gauge. Then carefully lift the bone out of the periosteum and cut the other end. 
	NOTE: Proceed with caution. This is the most delicate step, as pulling up without excising the rib below will tear the pleural membrane and result in a pneumothorax. If the pleural membrane is torn this will be readily evident as the interior of the thoracic cavity will become visible.</li>
	<li>If bleeding occurs as a result of cutting the bone, apply pressure at the cut end with a cotton-tipped swab for 4 - 5 sec to stop the bleeding. 
	NOTE: On the rare occasion that bleeding continues, it may be necessary to stop and euthanize the animal as significant blood loss will compromise recovery (without fluid replacement, the maximum blood volume which can be safely be lost is 10% of the total blood volume or 7.7 - 8 &#181;l/g. For a 25 g mouse, this is equivalent to around 180 - 200 &#181;l9).</li>
	<li>Immediately place the removed rib into 4% PFA for future analysis. 
	NOTE: If the resection of both the bone and the periosteum is to be carried out, omit the periosteum incision and separation (step 3.2). Since the periosteum is very tightly attached to the pleural membrane proceed cautiously, teasing the rib away from the pleural membrane with forceps to avoid tearing.</li>
</ol>
4. Closing the Incision
<ol>
	<li>Suture the intercostal muscles over the top of the remaining periosteal sleeve with 9-0 sutures (2 sutures are usually sufficient). Place the sutures directly above the cut ends of the rib, to act as indicators for surgery location.</li>
	<li>Remove the retractor. Suture the overlying muscle and fat with 9-0 sutures (3 - 4 sutures are usually sufficient). In mice with particularly thick layers of muscle and/or fat, suture each layer separately (e.g., 1 layer of sutures for the muscular layer and 1 layer of sutures for the fatty layer).</li>
	<li>Close the skin with 7-0 sutures (4 - 5 sutures are usually sufficient).</li>
	<li>Secure the incision with suture glue, pinching the edges together with large forceps.</li>
	<li>Slowly wean the mouse off isoflurane by first adjusting to 1% for few minutes and then turn off.</li>
	<li>Place the mouse under a heat lamp and leave until consciousness is regained, 5 - 10 min. After consciousness is regained and throughout the healing period, the mouse should move and ambulate normally and show no signs of distress.</li>
</ol>
5. Recovery and Analysis
<ol>
	<li>Provide post-operative pain management including the administration of buprenorphine in an oral gelatin form at 0.5 mg/kg every 12 hr for 48 hr. Oral administration does not require restraining the animal which might cause pain to the thoracic region.</li>
	<li>Maintain the mouse in a cage with free access to food and water during the healing period. After the incision is dry, female animals can be co-housed while males must remain isolated to prevent fighting.</li>
	<li>After the healing period, follow the approved procedure for euthanasia. Remove the rib cage for fixation and analysis.</li>
</ol>

<ol><li>Philip, S. J., Kumar, R. J., Menon, K. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Morphological+study+of+rib+regeneration+following+costectomy+in+adolescent+idiopathic+scoliosis%5BTitle%5D)+AND+%22Eur+Spine+J%22%5BJournal%5D)">Morphological study of rib regeneration following costectomy in adolescent idiopathic scoliosis.</a> Eur Spine J. 14 (8), 772-776 (2005).</li>
<li>Munro, I. R., Guyuron, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Split-rib+cranioplasty%5BTitle%5D)+AND+%22Ann+Plast+Surg%22%5BJournal%5D)">Split-rib cranioplasty.</a> Ann Plast Surg. 7 (5), 341-346 (1981).</li>
<li>Taggard, D. A., Menezes, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Successful+use+of+rib+grafts+for+cranioplasty+in+children.+Pediatric+neurosurgery%5BTitle%5D)+AND+%22%22%5BJournal%5D)">Successful use of rib grafts for cranioplasty in children. Pediatric neurosurgery.</a> 34 (3), 149-155 (2001).</li>
<li>Head, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Prevention+of+Regeneration+fo+the+Ribs%3A+A+problem+in+thoracic+surgery%5BTitle%5D)+AND+%22Archives+of+Surgery%22%5BJournal%5D)">Prevention of Regeneration fo the Ribs: A problem in thoracic surgery.</a> Archives of Surgery. 14 (6), 1215-1221 (1927).</li>
<li>Kawanabe, Y., Nagata, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((A+new+method+of+costal+cartilage+harvest+for+total+auricular+reconstruction%3A+part+I.+Avoidance+and+prevention+of+intraoperative+and+postoperative+complications+and+problems%5BTitle%5D)+AND+%22Plastic+and+reconstructive+surgery%22%5BJournal%5D)">A new method of costal cartilage harvest for total auricular reconstruction: part I. Avoidance and prevention of intraoperative and postoperative complications and problems.</a> Plastic and reconstructive surgery. 117 (6), 2011-2018 (2006).</li>
<li>Cheung, K. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((An+externally+fixed+femoral+fracture+model+for+mice%5BTitle%5D)+AND+%22Journal+of+orthopaedic+research+%3A+official+publication+of+the+Orthopaedic+Research+Society%22%5BJournal%5D)">An externally fixed femoral fracture model for mice.</a> Journal of orthopaedic research : official publication of the Orthopaedic Research Society. 21 (4), 685-690 (2003).</li>
<li>Matthys, R., Perren, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Internal+fixator+for+use+in+the+mouse%5BTitle%5D)+AND+%22Injury%22%5BJournal%5D)">Internal fixator for use in the mouse.</a> Injury. 40, Suppl 4. S103-S109 (2009).</li>
<li>Cooper, G. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Testing+the+critical+size+in+calvarial+bone+defects%3A+revisiting+the+concept+of+a+critical-size+defect%5BTitle%5D)+AND+%22Plastic+and+reconstructive+surgery%22%5BJournal%5D)">Testing the critical size in calvarial bone defects: revisiting the concept of a critical-size defect.</a> Plastic and reconstructive surgery. 125 (6), 1685-1692 (2010).</li>
<li><a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Ask+the+Vet%5BTitle%5D)+AND+%22JAX+NOTES%22%5BJournal%5D)">Ask the Vet.</a> JAX NOTES. 499, Available from:
 
 <a target="_blank" href="http://jaxmice.jax.org/jaxnotes/archive/499c.html">http://jaxmice.jax.org/jaxnotes/archive/499c.html</a> (2005).</li>
<li>Flecknell, P. A., Roughan, J. V., Stewart, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Use+of+oral+buprenorphine+%28%27buprenorphine+jello%27%29+for+postoperative+analgesia+in+rats--a+clinical+trial%5BTitle%5D)+AND+%22Laboratory+animals%22%5BJournal%5D)">Use of oral buprenorphine ('buprenorphine jello') for postoperative analgesia in rats--a clinical trial.</a> Laboratory animals. 33 (2), 169-174 (1999).</li>
<li>Rigueur, D., Lyons, K. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Whole-mount+skeletal+staining%5BTitle%5D)+AND+%22Methods+in+molecular+biology%22%5BJournal%5D)">Whole-mount skeletal staining.</a> Methods in molecular biology. 1130, 113-121 (2014).</li>
<li>Evans, D. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Contribution+of+somitic+cells+to+the+avian+ribs%5BTitle%5D)+AND+%22Developmental+biology%22%5BJournal%5D)">Contribution of somitic cells to the avian ribs.</a> Developmental biology. 256 (1), 114-126 (2003).</li>
<li>Colnot, C., Thompson, Z., Miclau, T., Werb, Z., Helms, J. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Altered+fracture+repair+in+the+absence+of+MMP9%5BTitle%5D)+AND+%22Development%22%5BJournal%5D)">Altered fracture repair in the absence of MMP9.</a> Development. 130 (17), 4123-4133 (2003).</li>
<li>Lu, C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Cellular+basis+for+age-related+changes+in+fracture+repair%5BTitle%5D)+AND+%22Journal+of+orthopaedic+research+%3A+official+publication+of+the+Orthopaedic+Research+Society%22%5BJournal%5D)">Cellular basis for age-related changes in fracture repair.</a> Journal of orthopaedic research : official publication of the Orthopaedic Research Society. 23 (6), 1300-1307 (2005).</li>
<li>Zilberman, Y., Gafni, Y., Pelled, G., Gazit, Z., Gazit, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Bioluminescent+imaging+in+bone%5BTitle%5D)+AND+%22Methods+in+molecular+biology%22%5BJournal%5D)">Bioluminescent imaging in bone.</a> Methods in molecular biology. 455, 261-272 (2008).</li>
<li>Pelled, G., Gazit, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Imaging+using+osteocalcin-luciferase%5BTitle%5D)+AND+%22Journal+of+musculoskeletal%22%5BJournal%5D)">Imaging using osteocalcin-luciferase.</a> Journal of musculoskeletal. 4 (4), 362-363 (2004).</li>
<li>Elefteriou, F., Yang, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Genetic+mouse+models+for+bone+studies--strengths+and+limitations%5BTitle%5D)+AND+%22Bone%22%5BJournal%5D)">Genetic mouse models for bone studies--strengths and limitations.</a> Bone. 49 (6), 1242-1254 (2011).</li>
<li>Srour, M. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Natural+large-scale+regeneration+of+rib+cartilage+in+a+mouse%5BTitle%5D)+AND+%22J.+Bone+Miner%22%5BJournal%5D)">Natural large-scale regeneration of rib cartilage in a mouse.</a> J. Bone Miner. , (2014).</li>
</ol>

The authors have nothing to disclose

When first learning this protocol, determining where to locate the initial incision can be challenging. However, practice on euthanized mice helps the surgeon learn where to place the initial incision and expose the desired rib to be resected. Working on cadavers also improves the fine-motor skills required to remove the rib portion with or without the periosteum. In addition, someone new to this procedure might find manipulating the fine tools and thin sutures to be difficult. While tying off, excess pulling on the sutu...

Debilitating skeletal injury, chronic osteoarthritis, and the severe problems associated with reconstructive surgery impact economic productivity, family well-being, and quality of life. While small breaks and lesions can heal fairly well, humans are not capable of repairing large defects and therefore must rely on reconstructive procedures&#160;to restore structure and function. Reconstruction may involve allogeneic or heterogeneic grafts, morcellized bone, implanted scaffolds, or distraction osteogenesis. Unfortunately, not only are there persistant morbidity factors associated with these treatments but the original strength of the repaired bone is rarely attained. Thus, new clinical approaches are needed.
One-way to develop innovative methods to treat segmental defects is to study situations in which large-scale repair occurs naturally. Amphibians famously can regenerate skeletal elements, while mammals are considered limited in this ability. However, since the early part of the 20th century, a few reports of regeneration in the human rib have been published suggesting that humans may not be so limited1-4 . Currently this phenomenon is best known by plastic surgeons who use rib material for jaw, face and ear reconstruction, but it is not more broadly appreciated5. In order to study this repair in more detail, we have developed a surgical model using the mouse. Using this protocol, researchers can identify the innate factors involved and use this information to facilitate skeletal healing in other locations.
There are many advantages to using the ribs as a model for studying skeletal repair. First, the surrounding ribs provide a natural fixator (as compared to resection of the femur6,7). This decreases the morbidity risk of internal and external fixators and simplifies the surgical procedure. Secondly, the thin muscular layers of the chest wall provide for easy access and excellent visibility which make the assay comparable to the convenience of calvarial resections8. Thirdly, in contrast to the calvariae which form by intramembranous ossification, the ribs form by endochondral ossification and grow in length via extension at growth plates located at either end of a central diaphysis. Therefore, repair of the ribs may be more comparable to repair of the long bones of the appendicular skeleton. Furthermore, we have found that compared to the femur, the periosteum of the rib is thicker and can be more easily manipulated. Thus, investigators who wish to assay bone repair for the purpose of studying the periosteum or testing cell therapies, pharmacological agents, and/or tissue scaffolds may find this surgical model useful. In summary, this rib resection model provides a context within which to study natural large-scale bone repair in mammals as no such model in general use currently exists.

<table><tbody><tr><td>Medium sized micro-dissection scissors (Vannas-T&amp;uuml;bingen Spring Scissors 5 mm)</td><td>Fine Science Tools</td><td>15003-08</td><td></td></tr><tr><td>Fine micro-dissection scissors (Vannas Spring Scissors - 2mm Cutting Edge)</td><td>Fine Science Tools</td><td>15000-04</td><td>curved tip is beneficial</td></tr><tr><td>Micro-scalpel 5.0 mm</td><td>Fine Science Tools</td><td>10315-12</td><td>other fine scalpels can be substituted</td></tr><tr><td>Dumont 55 forceps</td><td>Fine Science Tools</td><td>11295-51</td><td></td></tr><tr><td>Retractor&amp;nbsp;</td><td>Fine Science Tools</td><td>17004-05</td><td>adjustability is convenient</td></tr><tr><td>Micro-needle holders</td><td>Fine Science Tools</td><td>12060-01</td><td></td></tr><tr><td>9.0 nylon sutures (Ethilon), taper point best</td><td>Ethicon</td><td>2819G or similar</td><td>taper point best but reverse cutting is also good</td></tr><tr><td>7.0 prolene sutures (Prolene)</td><td>Ethicon</td><td>8700H or similar</td><td>6-0 can be used too, needle point can vary</td></tr><tr><td>Large forceps (Adson Forceps)</td><td>Fine Science Tools</td><td>11006-12</td><td>other brands are fine</td></tr><tr><td>Lubricant Eye Ointment (Akwa Tears)</td><td>Akorn</td><td>17478-062-35</td><td></td></tr><tr><td>Suture glue (GLUture Topical Tissue Adhesive)</td><td>Abbot</td><td>32046-01</td><td>has excellent working time</td></tr><tr><td>Shaver</td><td>Wahl</td><td>9918-6171 or similar</td><td></td></tr><tr><td>Clamp lamp</td><td>Zoo Med</td><td>LF-5</td><td></td></tr><tr><td>Infrared Bulb, 75W</td><td>Zoo Med</td><td>RS-75</td><td></td></tr><tr><td>RC&lt;sup&gt;2&lt;/sup&gt; Rodent Anesthesia System</td><td>VetEquip&amp;nbsp;</td><td>922100</td><td></td></tr><tr><td>IsoFlo (Isoflurane)</td><td>Abbot</td><td>05260-05</td><td></td></tr><tr><td>Buprenorphine (Buprenex)</td><td>Reckitt Benckiser</td><td>12496-0757-1</td><td></td></tr><tr><td>Betadine</td><td>Purdue Frederick</td><td>67618015017</td><td></td></tr><tr><td>Flavored Gelatin, raspberry</td><td>Jell-O</td><td>B000E1FYL0</td><td>made up firm, to the consistency of &#039;jigglers&#039;</td></tr></tbody></table>

a surgical procedure for resecting the mouse rib: a model for large-scale long bone repair

A Surgical Procedure for Resecting the Mouse Rib: A Model for Large-Scale Long Bone Repair

This protocol introduces researchers to a new model for large-scale bone repair utilizing the mouse rib. The procedure details the following: preparation of the animal for surgery, opening the thoracic body wall, exposing the desired rib from the surrounding intercostal muscles, excising the desired section of rib without inducing a pneumothorax, and closing the incisions. Compared to the bones of the appendicular skeleton, the ribs are highly accessible. In addition, no internal or external fixator is necessary since the adjacent ribs provide a natural fixation. The surgery uses commercially available supplies, is straightforward to learn, and well-tolerated by the animal. The procedure can be carried out with or without removing the surrounding periosteum, and therefore the contribution of the periosteum to repair can be assessed. Results indicate that if the periosteum is retained, robust repair occurs in 1 - 2 months. We expect that use of this protocol will stimulate research into rib repair and that the findings will facilitate the development of new ways to stimulate bone repair in other locations around the body.

Animals typically recover quickly from this procedure, the incision heals well, and the animals exhibit normal behavior. The procedure uses commercially available supplies and after practice, can be completed in under 1 hr. Before using this procedure to generate experimental data, it is important to analyze the resection at the day 0 time point to determine if any large bone fragments might remain within the resection zone. One way to assess proper procedure is to assess the animals by X-ray or microCT imaging postopera...

The overall goal of this procedure is to successfully resect a portion of bone from the rib of a mouse. The procedure was developed as a model to study large-scale long bone repair.

This protocol introduces researchers to a new model for large-scale bone repair utilizing the mouse rib. The procedure details the following: preparation ...

The goal of this procedure is to create a large scale resection in the bony portion of a rib as a model for skeletal repair. This is accomplished by first creating and opening an incision through the skin on the lateral side of the animal. The second step is to expose the rib bone that is to be removed.Next, a portion of the rib bone is removed either along with the surrounding connective tissue sheath or the periosteum, or leaving the periosteum behind. The final step is to close the incisions and allow the animal to recover. Ultimately, this model for bone repair can be used to show the cellular and molecular events that occur during large scale skeletal repair.In a mammal Visual demonstration of this method is critical as manipulating the surgical tools requires precision and successful surgery involves knowing where the different tissues are located. Demonstrating the procedure will be Marissa Roar and Nikita Tripper and any both graduate students in my laboratory. After preparing the surgical area and the necessary sterile tools and supplies, induce anesthesia by placing a ture mouse into an induction chamber with 4%ice of fluorine.Once the mouse is anesthetized, apply ointment to the eyes of the mouse and confirm the depth of anesthesia. By performing a toe pinch, there should be no reaction to the toe pinch. Place the mouse on or adjacent to a heat pad on the stage of a dissecting microscope with the nose cone of the anesthesia maintenance hose placed over the nose of the mouse.Ensure that the mouse is in a lateral position. Adjust the isof fluorine to two to 3%for maintenance and watch the breathing rate. Then inject buprenorphine subcutaneously via a 25 gauge needle.For preemptive post-surgery pain, the leg may twitch, which is a normal response. First, select the desired rib by palpating from the most rostral rib, which is counted as number one. In this video, a portion of rib number 10 will be removed.Shave the area over the rib and then clean with povidone iodine, followed by isopropanol three times After creating a two centimeter incision through the skin, muscle and fat layers above the rib to be excised, place all three layers into a retractor to expose the surgical area while minimizing the size of the incision. Then use a five millimeter scalpel to cut through the intercostal muscles above the rib at a location, five millimeter proximal to the chocos joint where the rib is not as highly curved. Then carefully separate the muscle from the bone with fine tip forceps.Next to create a resection while retaining the periosteum in the animal, cut through the periosteum along the length of the rib. Then use fine tip forceps to very carefully separate the delicate periosteum laterally from the underlying bone. If needed, measure the resection with a ridicule in the microscope or a ruler type gauge.Then use fine micro scissors to make a cross-section through the bone. At one end, carefully lift the bone out of the periosteum and cut the other end Immediately. Place the removed rib into 4%paraldehyde for future analysis.If bleeding occurs as a result of cutting the bone, apply pressure at the cut end with a cotton tipped swab for four to five seconds to stop the bleeding. To close the incision, suture the intercostal muscles over the top of the remaining periosteal sleeve with three or 4 9 0 discontinuous sutures. Place the sutures directly above the cut ends of the rib to act as indicators for surgery location.Remove the retractor and then suture the overlying muscle and fat with nine zero discontinuous sutures. Suturing each layer separately in mice with particularly thick layers of muscle or fat. Close the skin with four to 5 7 0 sutures, and then secure the incision with tissue adhesive.Pinching the edges together with large forceps. Slowly wean the mouse off isof fluorine by adjusting the concentration to 1%for a few minutes, and then turn off the isof fluorine. Place the mouse under a heat lamp and leave until consciousness is regained.After consciousness is regained and throughout the healing period, the mouse should move and ambulate normally and show no signs of distress. Once the incision is dry, house the mouse in a clean cage with free access to food and water. Females can be co housed, but males must be housed alone to prevent fighting, provide postoperative pain management, including the administration of buprenorphine in an oral genin form at 0.5 milligrams per kilogram every 12 hours for 48 hours.Oral administration does not require restraining the animal, which might cause pain to the thoracic region. This image shows a 4.5 millimeter rib resection collected immediately after surgery. No bone fragments are left behind.The sample was stained with a zarin red and cleared in 80%glycerol. The arrowhead indicate the location of the cut ends. This is the removed rib portion from the rib cage in the previous image without the periosteum attached.The histological section stained with hematin and eoin shown here illustrates that there is minimal periosteum attached to the bone as indicated by the yellow arrowhead on the left and no periosteum to the right. The yellow asterisk indicates the bone marrow cavity. This image shows a removed rib portion with the periosteum still attached, and this histological section shows the intact periosteum overlying the bone as indicated by yellow bracket.The yellow asterisk again indicates the bone marrow cavity. This image shows a skeletal preparation after two months of healing following resection with the intact periosteum inside the mouse, complete repair of the region can be observed. This histological section of the skeletal preparation just shown demonstrates a restoration of the lamella bone walls and the bone marrow cavity.This image shows a skeletal preparation following three months of healing. After removing both a rib portion and the surrounding periosteum, there was only minimal repair at the cut end shown at the left of the panel. The yellow arrowhead points to the chocos joint.This histological section of the previously shown skeletal preparation shows granulation, tissue adipose and muscle cells filling in the site of resection. Once mastered, this technique can be performed in under one hour if done properly following this procedure. Other methods like creating a resection in the cartilage portion of the rib can be performed in order to answer additional questions like, how does large scale repair of cartilage occur?After watching this video, you should have a good understanding of how to create a resection in the bone portion of the rib in order to study skeletal repair.I.

a-surgical-procedure-for-resecting-mouse-rib-model-for-large-scale

Research

JoVE Journal

Medicine

One constraint of preclinical research in the field of bone repair is the lack of experimental control over the local mechanical environment within a healing bone lesion. We report the design and use of an external fixator for bone repair with the ability to change fixator stiffness in vivo.

Adjustable Stiffness, External Fixator for the Rat Femur Osteotomy and Segmental Bone Defect Models

Animal models are frequently employed to mimic serious bone injury in biomedical research. Due to their small size, establishment of stabilized bone lesions in mice are beyond the capabilities of most research groups. Herein, we describe a simple method for establishing and analyzing experimental femoral defects in mice.

A Simple Critical-sized Femoral Defect Model in Mice

Although mouse models are invaluable tools for bone tissue engineering, models of long bone defects are sparse. This need motivated development of the present protocol which uses a locking plate with four screws and a dedicated jig to perform and stabilize a reproducible, femoral, critical-size defect with low morbidity.

Establishment of a Segmental Femoral Critical-size Defect Model in Mice Stabilized by Plate Osteosynthesis

The evaluation of tissue development in the fracture callus during endochondral bone healing is essential to monitor the healing process. Here, we report the use of a magnetic resonance imaging (MRI)-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice.

In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur

This protocol describes a mouse model of orthopedic surgery that has been used to study mechanisms of postoperative neuroinflammation and behavioral changes, and when combined with parabiosis, to study tissue regeneration during aging.

A Mouse Model of Orthopedic Surgery to Study Postoperative Cognitive Dysfunction and Tissue Regeneration

This protocol describes a minimally invasive osteosynthesis technique using an intramedullary screw for standardized stabilization of femur fractures, which can be used to analyze endochondral bone healing in mice.

A Minimally Invasive Model to Analyze Endochondral Fracture Healing in Mice Under Standardized Biomechanical Conditions

We present a mouse tibial distraction osteogenesis model developed using a custom-made distractor. The use of a mouse as an analysis target is advantageous for advancing research.

A Mouse Distraction Osteogenesis Model

The murine closed femoral fracture model is a powerful platform to study fracture healing and novel therapeutic strategies to accelerate bone regeneration. The goal of this surgical protocol is to generate unilateral closed femoral fractures in mice using an intramedullary steel rod to stabilize the femur.

The Generation of Closed Femoral Fractures in Mice: A Model to Study Bone Healing

This protocol describes a method to perform fractures on adult mice and monitor the healing process.

Transverse Fracture of the Mouse Femur with Stabilizing Pin

This protocol describes a surgical procedure for the establishment of a diaphyseal fracture in the femur of mice, which is stabilized with an intramedullary wire, for fracture healing studies.

Establishing a Diaphyseal Femur Fracture Model in Mice

This protocol presents a comprehensive and efficient method for analyzing mandibular bone repair. We describe a reproducible technique for non-stabilized mandibular fracture in a mouse model, allowing analysis of the process of endochondral bone repair with minimal tissue damage and bone loss.

Less-Invasive Technique for Non-stabilized Mandibular Fracture in Mouse Models

Learn a surgical rib resection technique in mice for studying large-scale skeletal repair. This method enables observation of cellular and molecular events during bone healing in vivo.

A Surgical Procedure for Resecting the Mouse Rib: A Model for Large-Scale Long Bone Repair

Explore More Videos