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).

注：すべての手順は、南カリフォルニア大学の施設内動物管理使用委員会（IACUC）によって承認された動物のプロトコルに従っている。 外科1.準備<ol><li>必要な手術器具や消耗品（滅菌、アイテム1 - 材料のリストの18）をレイアウトし、フード内やダウンドラフトテーブルの上に解剖顕微鏡および光ファイバトランスイルミを手配。 </li><li>麻酔を誘導するために、4％イソフルランで誘導チャンバー内に（3ヶ月 - - 30グラム、1〜20）性的に成熟したマウスを置きます。 </li><li>足および/またはテールピンチテストで麻酔を確認してください。 </li><li>手術中に目の乾燥を避けるために、マウスの目に軟膏を適用します。 </li><li>顕微鏡ステージにマウスを移動します。メンテナンスホースのノーズコーンにマウスを置き、横方向に配置する。体温を維持するために動物に隣接した温暖化ポーチを提供します。 NOTE：マウスの​​位置は個人的な好み、および/または外科医の利き手に依存している、左または右のいずれかの側は問題ありません。 </li><li>メンテナンスのため、3％、および呼吸数を見て - 2にイソフルランを調整します。 </li><li> 25 G針を有する手術後の痛みのためにブプレノルフィン（0.05 mg / kgを）皮下（脚の横脇腹）を注入します。脚部は正常な反応であるけいれんがあります。 </li></ol> 2.オープニング切開<ol><li>触診が希望するリブを検出し、シェーバーで毛のエリアをクリア。 注：リブ1-7が肺に近いと11-13がかなり短いので、リブ8-10を推奨します。肋骨を数えるのは数1のような最も吻側リブで始める必要があります。 </li><li> （3Xは、多くの場合、動物使用プロトコルによって推奨）ポビドンヨードとイソプロパノール（70％）スワブを交互にエリアを準備します。 </li><li>真上中小マイクロサージェリーのはさみで希望の肋骨に平行、皮膚を介して2cmの切開をカット。 Incis下の筋肉と脂肪の層を介して電子。 </li><li>切開部のサイズを最小限に抑えながら、手術領域を露出させるために開創器にすべての3つの層（皮膚、筋肉、脂肪）を配置します。 </li></ol> 3.リブを切除<ol><li> 5.0ミリメートルのメスでリブの所望の部分の上にある肋間筋を切断。リブとして非常に湾曲していない場合に、このようなchondrocostal共同の5 mmの近位の周りのこの切開がある。慎重に先端の細いピンセットで骨から筋肉を分離する。 </li><li>動物に骨膜を保持しながら、切除を作成するために、5.0ミリメートルメスとリブの長さに沿って、骨膜を切断。慎重に先端の細い鉗子で横方向に下にある骨から骨膜を分離する。骨膜は非常にデリケートですし、ゼラチン状の一貫性を持っていたように、注意して進みます。 </li><li>次に細かいマイクロハサミで一方の端部に骨の断面を作る。必要に応じて、との切除を測定顕微鏡や定規タイプのゲージでレチクル。その後、慎重に骨膜から骨を持ち上げ、もう一方の端をカット。 注：慎重に進みます。これは胸膜を引き裂くと気胸になります下の肋骨を切除せずに引き上げなど、ほとんどの繊細なステップ、である。胸膜が破れた場合胸腔の内部が見えるようになるので、これは容易に明らかであろう。 </li><li>出血を止めるために5秒 - 出血が骨の切断の結果として発生した場合、4用綿棒との切断端に圧力をかける。 注：出血が続くことをまれに、それはかなりの血液損失が回復が損なわれるように動物を停止し、安楽死が必要な場合があり（流体を交換することなく、安全に失われることができる最大血液量は、全血液容量の10％であるまたは7.7 - 。）を200μl9 - 8μL/ gの25グラムのマウスの場合、これは約180に相当します。 </li><li>すぐにに削除されたリブを配置今後の分析のために4％PFA。 注：骨と骨膜の両方の切除を行うべきである場合、骨膜切開分離（ ステップ3.2）を省略する。骨膜は非常に緊密胸膜に取り付けられているのでティアリングを回避するために、ピンセットで離れて胸膜から肋骨をからかって、慎重に進んでください。 </li></ol> 4.切開を閉じる<ol><li> 9-0縫合糸（2縫合糸は通常十分です）と残りの骨膜スリーブの上に肋間筋を縫合。手術の場所のための指標として機能するように、直接、リブの切断端の上に縫合糸を配置します。 </li><li>開創器を取り外します。 （ - 4縫合通常は十分である3）9-0縫合糸で覆っている筋肉と脂肪を縫合。筋肉および/ ​​または脂肪の特に厚い層を有するマウスでは、個別の層を縫合（ 例えば、筋肉層の縫合糸の1層および脂肪層のための縫合糸の1層）。 </li><li>閉じ（ - 5縫合糸、通常は十分であり、4）は7-0縫合糸で皮膚。 </li><li>大鉗子で一緒に端をつまんで、縫合糸接着剤で切開を固定します。 </li><li>ゆっくりと最初の数分間、1％に調整することで、オフイソフルランマウスを離乳してからオフにしてください。 </li><li> 10分 - 、5加熱ランプ下にマウスを置き、意識が回復されるまで残す。意識が回復した後、治癒期間中、マウスが移動し、正常に歩き回ると苦痛の兆候を示さないはずである。 </li></ol> 5.回復と分析<ol><li> 0.5ミリグラムの経口ゼラチン状のブプレノルフィンの投与を含む術後疼痛管理を提供/ 48時間ごとに12時間をキロ。経口投与は、胸部に痛みを引き起こす可能性が動物を拘束する必要はありません。 </li><li>治癒期間中に食物と水を自由にケージにマウスを維持します。切開が乾燥した後、雌の動物は、雄ながら同時収容することができるsは戦いを防ぐために分離したままにします。 </li><li>治癒期間の後、安楽死のために承認された手順に従ってください。固定および分析のために胸郭を削除します。 </li></ol>

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The authors have nothing to disclose

最初のこのプロトコルを学習するとき、どこに挑戦することができます最初の切開を見つけるには、決定する。しかし、安楽死させたマウスでの練習は、外科医が最初の切開を置き、切​​除されるべき所望のリブを公開する場所を学​​ぶことができます。死体に作業することも骨膜の有無にかかわらずリブ部分を除去するために必要な細かい運動能力を向上させます。また、この手順に?...

骨格傷害、慢性変形性関節症を衰弱させる、と再建手術に関連した重大な問題は、経済生産性、家族の幸福、そして生活の質に影響を与える。小休憩と病変がかなりよく癒すことができますが、人間は大きな欠陥を修復することができないため、構造と機能を回復するために再建手続きに頼らなければならない。復興は、同種またはheterogeneic移植片、morcellized骨、移植された足場、または伸延骨形成を伴ってもよい。残念ながら、だけでなく、これらの治療法はなく、修復骨の本来の強さはほとんど達成されていないに関連した永続的な罹患率の要因があります。このため、新たな臨床アプローチが必要とされている。 分節欠損を治療するための革新的な方法を開発する一方向は、大規模な修復が天然に存在する状況を研究することである。哺乳類が目に限定されると考えている間に両生類は有名、骨格要素を再生成することができます能力である。しかし、20 世紀の初頭以来、人間のリブにおける再生の報告はほとんど人間が1-4そのように限定されないかもしれないことを示唆して発表されている。現在、この現象は、最高の顎、顔、耳再構成のために隔壁材料を使用する外科医に知られているが、それは、より広く5が認識されない。より詳細には、この修復を研究するために、我々は、マウスを使用して、外科的モデルを開発した。このプロトコルを使用して、研究者は、関連する生来の因子を特定し、他の場所で骨治癒を促進するためにこの情報を使用することができる。 骨修復を研究するためのモデルとしてのリブを使用することには多くの利点がある。 （大腿骨6,7の切除と比較して）まず、周囲のリブは自然な固定器を提供する。これは、内部​​と外部固定の罹患リスクを減少させ、手術手順を簡素化します。胸のWAの第二に、薄い筋肉層LL頭蓋冠切除8の利便性の検定に匹敵する簡単なアクセスと優れた視認性を提供する。第三に、膜内骨化によってその形、軟骨内骨化によってリブ形と中央骨幹のいずれかの端部に位置する成長板に拡張を介して長さに成長する頭蓋冠とは対照的である。そのため、リブの修理は、四肢骨格の長骨の修理により匹敵し得る。さらに、我々は、大腿骨と比べて、リブの骨膜が厚く、より簡単に操作することができることを見出した。このように、骨膜を研究または細胞治療、薬理学的薬剤、および/または組織足場をテストする目的で、骨修復を検定したい研究者らは、この外科的モデルが役に立つでしょう。要約すると、このリブ切除モデルは、一般的に使用されるそのようなモデルは、現在存在しないように、哺乳動物において天然の大規模骨修復を研究するためにその中にコンテキストを提供する。

<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

このプロトコルは、マウスのリブを利用した大規模な骨修復のための新しいモデルに研究者を紹介しています。 、周囲の肋間筋から目的の肋骨を露出させ、胸部体壁を開いて気胸を誘発することなく、リブの所望の部分を切除し、切開部を閉じる、手術のための動物の準備：手順は、以下の詳細を示します。四肢骨格の骨と比較すると、リブは非常にアクセス可能です。また、全く内部または外部固定器は、隣接するリブは自然な固定を提供するので必要ありません。手術は、市販品を使用して、学ぶことは簡単であり、そして動物によって十分に許容さ。手順は、または周囲の骨膜を除去することなく行うことができるので、修理の骨膜の寄与を評価することができる。 2ヶ月 - 結果は、骨膜が保持されている場合、強固な修復が1で発生したことを示している。私たちは、このプロトコルの使用が期待リブの補修に及び所見は体の周りの他の場所で骨修復を刺激するための新しい方法の開発を容易にするような研究を刺激する。

動物は、一般的に切開がうまく癒す、このプロシージャからすぐに回復し、動物は正常な挙動を示す。手順は、市販品を使用して練習した後、1時間の下で完了させることができる。実験データを生成するために、この手順を使用する前に、任意の大きい骨片を切除ゾーン内に残っているかどうかを判断するために、0日目の時点で切除を分析することが重要である。これは、典型的には、特?...

Watch this Scientific Journal Video about A Surgical Procedure for Resecting the Mouse Rib: A Model for Large-Scale Long Bone Repair at JoVE.com

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

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.

マウスリブを切除するための外科的手順：大規模ロング骨修復のためのモデル

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

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

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

13.0K ビュー.  Keck School of Medicine, University of Southern California. 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.

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

A Mouse Distraction Osteogenesis Model

Distraction osteogenesis (DO) is a surgical procedure that involves skeletal tissue regeneration without cell transplantation. A DO model consists of the following three phases: the latency phase after osteotomy and placement of the external distractor; the distraction phase, wherein the separated bone ends are gradually and continuously distracted; and the consolidation phase. This custom-made distractor used for DO is comprised of two incomplete acrylic resin rings and an expansion screw. The process was initiated by making a mold with silicone impression material and then creating the custom-made distractor. Dental resin was poured into the formwork made of silicone impression material, and it was allowed to polymerize to create the incomplete resin rings required for the custom-made distractor. These rings were fixed with an expansion screw using transparent resin. The custom-made distractor created via this approach was attached to the tibia of mice. The tibia was fixed to the device using one pair of 25 G needles proximally, one pair of 27 G needles distally, and acrylic resin. After a latency period of 5 days, distraction was initiated at a rate of 0.2 mm/12 h. The lengthening was continued for 8 days, resulting in a total gap of 3.2 mm. The mice were sacrificed 4 weeks after distraction. Bone formation in the distraction gap was confirmed using both radiography and histology.

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.

マウス気晴らし骨形成モデル

Distraction osteogenesis (DO) is a surgical procedure that involves skeletal tissue regeneration without cell transplantation. A DO model consists of the ...

発生生物学

Mouse Pneumonectomy Model of Compensatory Lung Growth

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to disability or death. Injury-based animal models of lung repair and regeneration are limited by injury-specific responses making it difficult to differentiate changes related to the injury response and injury resolution from changes related to lung repair and lung regeneration. However, use of animal models to identify these repair and regeneration signaling pathways is critical to the development of new therapies aimed at improving pulmonary function following lung injury. The mouse pneumonectomy model utilizes compensatory lung growth to isolate those repair and regeneration signals in order to more clearly define mechanisms of alveolar re-septation. Here, we describe our technique for performing mouse pneumonectomy and sham pneumonectomy. This technique may be utilized in conjunction with lineage tracing or other transgenic mouse models to define molecular and cellular mechanism of lung repair and regeneration.

Mouse pneumonectomy is a commonly employed model of compensatory lung growth. This procedure can be used in conjunction with lineage tracing or transgenic mouse models to elucidate underlying mechanisms.

代償肺の成長のマウス肺モデル

In humans, disrupted repair and remodeling of injured lung contributes to a host of acute and chronic lung disorders which may ultimately lead to ...

医学

A Simple Critical-sized Femoral Defect Model in Mice

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all limb bone fractures fail to heal completely due to the extent of the trauma, disease, or age of the patient. Our ability to improve bone regenerative strategies is critically dependent on the ability to mimic serious bone trauma in test animals, but the generation and stabilization of large bone lesions is technically challenging. In most cases, serious long bone trauma is mimicked experimentally by establishing a defect that will not naturally heal. This is achieved by complete removal of a bone segment that is larger than 1.5 times the diameter of the bone cross-section. The bone is then stabilized with a metal implant to maintain proper orientation of the fracture edges and allow for mobility. 
Due to their small size and the fragility of their long bones, establishment of such lesions in mice are beyond the capabilities of most research groups. As such, long bone defect models are confined to rats and larger animals. Nevertheless, mice afford significant research advantages in that they can be genetically modified and bred as immune-compromised strains that do not reject human cells and tissue.
Herein, we demonstrate a technique that facilitates the generation of a segmental defect in mouse femora using standard laboratory and veterinary equipment. With practice, fabrication of the fixation device and surgical implantation is feasible for the majority of trained veterinarians and animal research personnel. Using example data, we also provide methodologies for the quantitative analysis of bone healing for the model.

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.

マウスの簡単な重要なサイズの大腿欠陥モデル

While bone has a remarkable capacity for regeneration, serious bone trauma often results in damage that does not properly heal. In fact, one tenth of all ...

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

The use of tissue-engineered bone constructs is an appealing strategy to overcome drawbacks of autografts for the treatment of massive bone defects. As a model organism, the mouse has already been widely used in bone-related research. Large diaphyseal bone defect models in mice, however, are sparse and often use bone fixation which fills the bone marrow cavity and does not provide optimal mechanical stability. The objectives of the current study were to develop a critical-size, segmental, femoral defect in nude mice. A 3.5-mm mid-diaphyseal femoral ostectomy (approximately 25% of the femur length) was performed using a dedicated jig, and was stabilized with an anterior located locking plate and 4 locking screws. The bone defect was subsequently either left empty or filled with a bone substitute (syngenic bone graft or coralline scaffold). Bone healing was monitored noninvasively using radiography and in vivo micro-computed-tomography and was subsequently assessed by ex vivo micro-computed-tomography and undecalcified histology after animal sacrifice, 10 weeks postoperatively. The recovery of all mice was excellent, a full-weight-bearing was observed within one day following the surgical procedure. Furthermore, stable bone fixation and consistent fixation of the implanted materials were achieved in all animals tested throughout the study. When the bone defects were left empty, non-union was consistently obtained. In contrast, when the bone defects were filled with syngenic bone grafts, bone union was always observed. When the bone defects were filled with coralline scaffolds, newly-formed bone was observed in the interface between bone resection edges and the scaffold, as well as within a short distance within the scaffold. 
The present model describes a reproducible critical-size femoral defect stabilized by plate osteosynthesis with low morbidity in mice. The new load-bearing segmental bone defect model could be useful for studying the underlying mechanisms in bone regeneration pertinent to orthopaedic applications.

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.

プレート骨接合によって安定化マウスにおけるセグメント別大腿臨界サイズ欠陥モデルの確立

The use of tissue-engineered bone constructs is an appealing strategy to overcome drawbacks of autografts for the treatment of massive bone defects. As a ...

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

Surgery is commonly used to improve and maintain quality of life. Unfortunately, in vulnerable patients such as the elderly, complications may occur and significantly diminish the outcome. Indeed, after routine orthopedic surgery to repair a fracture, as many as 50% of elderly patients suffer from neurologic complications like delirium. Also, the capacity to heal and regenerate tissue after surgery decreases with age, and can impact the quality of fracture repair and even osseous integration of implants. Thus, a better understanding of mechanisms that drive these age-dependent changes could provide strategic targets to minimize risk for such complications and optimize outcomes. Here, we introduce a clinically relevant mouse model of tibial fracture. The postoperative changes in these mice mimic some of the cognitive impairments commonly observed after routine orthopedic surgery in humans. Briefly, an incision is performed in the right hind limb under strictly aseptic conditions. Muscles are disassociated, and a 0.38-mm stainless steel pin is inserted into the upper crest of the tibia, inside the intramedullary canal. Osteotomy is then performed, and the wound is stapled. We have used this model to investigate the effects of surgical trauma on postoperative neuroinflammation and behavioral changes. By applying this fracture model in combination with parabiosis, a surgical model in which 2 mice are anastomosed, we have studied cells and secreted factors that systemically rejuvenate organ function and tissue regeneration after injury. By following our step-by-step protocol, these models can be reproduced with high fidelity, and can be adapted to interrogate many biologic pathways that are altered by surgical trauma.

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.

整形外科術後の高次脳機能障害と組織再生を研究するマウス モデル

Surgery is commonly used to improve and maintain quality of life. Unfortunately, in vulnerable patients such as the elderly, complications may occur and ...

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

Bone healing models are necessary to analyze the complex mechanisms of fracture healing to improve clinical fracture treatment. During the last decade, an increased use of mouse models in orthopedic research was noted, most probably because mouse models offer a large number of genetically-modified strains and special antibodies for the analysis of molecular mechanisms of fracture healing. To control the biomechanical conditions, well-characterized osteosynthesis techniques are mandatory, also in mice. Here, we report on the design and use of a closed bone healing model to stabilize femur fractures in mice. The intramedullary screw, made of medical-grade stainless steel, provides through fracture compression an axial and rotational stability compared to the mostly used simple intramedullary pins, which show a complete lack of axial and rotational stability. The stability achieved by the intramedullary screw allows the analysis of endochondral healing. A large amount of callus tissue, received after stabilization with the screw, offers ideal conditions to harvest tissue for biochemical and molecular analyses. A further advantage of the use of the screw is the fact that the screw can be inserted into the femur with a minimally invasive technique without inducing damage to the soft tissue. In conclusion, the screw is a unique implant that can ideally be used in closed fracture healing models offering standardized biomechanical conditions.

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.

癒しの標準化された生体力学的条件下でマウスの軟骨破壊を分析する低侵襲モデル

Bone healing models are necessary to analyze the complex mechanisms of fracture healing to improve clinical fracture treatment. During the last decade, an ...

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

Bone fractures impose a tremendous socio-economic burden on patients, in addition to significantly affecting their quality of life. Therapeutic strategies that promote efficient bone healing are non-existent and in high demand. Effective and reproducible animal models of fractures healing are needed to understand the complex biological processes associated with bone regeneration. Many animal models of fracture healing have been generated over the years; however, murine fracture models have recently emerged as powerful tools to study bone healing. A variety of open and closed models have been developed, but the closed femoral fracture model stands out as a simple method for generating rapid and reproducible results in a physiologically relevant manner. The goal of this surgical protocol is to generate unilateral closed femoral fractures in mice and facilitate a post-fracture stabilization of the femur by inserting an intramedullary steel rod. Although devices such as a nail or a screw offer greater axial and rotational stability, the use of an intramedullary rod provides a sufficient stabilization for consistent healing outcomes without producing new defects in the bone tissue or damaging nearby soft tissue. Radiographic imaging is used to monitor the progression of callus formation, bony union, and subsequent remodeling of the bony callus. Bone healing outcomes are typically associated with the strength of the healed bone and measured with torsional testing. Still, understanding the early cellular and molecular events associated with fracture repair is critical in the study of bone tissue regeneration. The closed femoral fracture model in mice with intramedullary fixation serves as an attractive platform to study bone fracture healing and evaluate therapeutic strategies to accelerate healing.

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.

マウスで閉じた大腿骨骨折の生成: 骨の治癒を研究するためのモデル

Bone fractures impose a tremendous socio-economic burden on patients, in addition to significantly affecting their quality of life. Therapeutic strategies ...

Transverse Fracture of the Mouse Femur with Stabilizing Pin

Fracture repair is an essential function of the skeleton that cannot be reliably modeled in vitro. A mouse injury model is an efficient approach to test whether a gene, gene product or drug influences bone repair because murine bones recapitulate the stages observed during human fracture healing. When a mouse or human breaks a bone, an inflammatory response is initiated, and the periosteum, a stem cell niche surrounding the bone itself, is activated and expands. Cells residing in the periosteum then differentiate to form a vascularized soft callus. The transition from the soft callus to a hard callus occurs as the recruited skeletal progenitor cells differentiate into mineralizing cells, and the bridging of the fractured ends results in the bone union. The mineralized callus then undergoes remodeling to restore the original shape and structure of the healed bone. Fracture healing has been studied in mice using various injury models. Still, the best way to recapitulate this entire biological process is to break through the cross-section of a long bone that encompasses both cortices. This protocol describes how a stabilized, transverse femur fracture can be safely performed to assess healing in adult mice. A surgical protocol including detailed harvesting and imaging techniques to characterize the different stages of fracture healing is also provided.

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

安定化ピン付きマウス大腿骨の横骨折

Fracture repair is an essential function of the skeleton that cannot be reliably modeled in vitro. A mouse injury model is an efficient approach to test ...

生物学

Establishing a Diaphyseal Femur Fracture Model in Mice

Bones have a significant regenerative capacity. However, fracture healing is a complex process, and depending on the severity of the lesions and the age and overall health status of the patient, failures can occur, leading to delayed union or nonunion. Due to the increasing number of fractures resulting from high-energy trauma and aging, the development of innovative therapeutic strategies to improve bone repair based on the combination of skeletal/mesenchymal stem/stromal cells and biomimetic biomaterials is urgently needed. To this end, the use of reliable animal models is fundamental to better understanding the key cellular and molecular mechanisms that determine the healing outcomes. Of all the models, the mouse is the preferred research model because it offers a wide variety of transgenic strains and reagents for experimental analysis. However, the establishment of fractures in mice may be technically challenging due to their small size. Therefore, this article aims to demonstrate the procedures for the surgical establishment of a diaphyseal femur fracture in mice, which is stabilized with an intramedullary wire and resembles the most common bone repair process, through cartilaginous callus formation.

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.

マウスにおける骨幹大腿骨骨折モデルの確立

Bones have a significant regenerative capacity. However, fracture healing is a complex process, and depending on the severity of the lesions and the age ...

縫合糸拡張マウスにおける引張荷重下における骨リモデリングの3D可視化

A 3-D Visualization Technique for Bone Remodeling in a Suture Expansion Mouse Model

Craniofacial sutures play a crucial role beyond being fibrous joints connecting craniofacial bones; they also serve as the primary niche for calvarial and facial bone growth, housing mesenchymal stem cells and osteoprogenitors. As most craniofacial bones develop through intramembranous ossification, the sutures' marginal regions act as initiation points. Due to this significance, these sutures have become intriguing targets in orthopedic therapies like spring-assisted cranial vault expansion, rapid maxillary expansion, and maxillary protraction. Under orthopedic tracing force, suture stem cells are rapidly activated, becoming a dynamic source for bone remodeling during expansion. Despite their importance, the physiological changes during bone remodeling periods remain poorly understood. Traditional sectioning methods, primarily in the sagittal direction, do not capture the comprehensive changes occurring throughout the entire suture. This study established a standard mouse model for sagittal suture expansion. To fully visualize bone remodeling changes post-suture expansion, the PEGASOS tissue clearing method was combined with whole-mount EdU staining and calcium chelating double labeling. This allowed the visualization of highly proliferating cells and new bone formation across the entire calvarial bones following expansion. This protocol offers a standardized suture expansion mouse model and a 3-D visualization method, shedding light on the mechanobiological changes in sutures and bone remodeling under tensile force loading.

著者スポットライト:骨リモデリングを視覚化するためのPEGASOS組織透明化技術 

This protocol presents a standardized suture expansion mouse model and a 3-D visualization method to study the mechanobiological changes of the suture and bone remodeling under tensile force loading.

縫合糸拡張マウスモデルにおける骨リモデリングのための3次元可視化技術

Craniofacial sutures play a crucial role beyond being fibrous joints connecting craniofacial bones; they also serve as the primary niche for calvarial and ...

マウスリブを切除するための外科的手順：大規模ロング骨修復のためのモデル

要約

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マウス気晴らし骨形成モデル

代償肺の成長のマウス肺モデル

マウスの簡単な重要なサイズの大腿欠陥モデル

プレート骨接合によって安定化マウスにおけるセグメント別大腿臨界サイズ欠陥モデルの確立

整形外科術後の高次脳機能障害と組織再生を研究するマウスモデル

癒しの標準化された生体力学的条件下でマウスの軟骨破壊を分析する低侵襲モデル

マウスで閉じた大腿骨骨折の生成: 骨の治癒を研究するためのモデル

安定化ピン付きマウス大腿骨の横骨折

マウスにおける骨幹大腿骨骨折モデルの確立

縫合糸拡張マウスモデルにおける骨リモデリングのための3次元可視化技術

マウスリブを切除するための外科的手順：大規模ロング骨修復のためのモデル

要約

さらに動画を探す

この動画の章

マウス気晴らし骨形成モデル

代償肺の成長のマウス肺モデル

マウスの簡単な重要なサイズの大腿欠陥モデル

プレート骨接合によって安定化マウスにおけるセグメント別大腿臨界サイズ欠陥モデルの確立

整形外科術後の高次脳機能障害と組織再生を研究するマウス モデル

癒しの標準化された生体力学的条件下でマウスの軟骨破壊を分析する低侵襲モデル

マウスで閉じた大腿骨骨折の生成: 骨の治癒を研究するためのモデル

安定化ピン付きマウス大腿骨の横骨折

マウスにおける骨幹大腿骨骨折モデルの確立

縫合糸拡張マウスモデルにおける骨リモデリングのための3次元可視化技術

整形外科術後の高次脳機能障害と組織再生を研究するマウスモデル