Name: a simple critical-sized femoral defect model in mice
Uploaded: 2015-03-15T14:00:00.000+00:00
Duration: 9 min 41 s
Description: Watch this Scientific Journal Video about A Simple Critical-sized Femoral Defect Model in Mice at JoVE.com

私たちは、この技術の開発中に彼らの貴重な助言と支援のために、比較医学、寺院、テキサス州のスコット＆ホワイト病院部門のスタッフや獣医師に感謝します。この作品は、再生医療プログラムの資金のための研究所、スコット＆ホワイトRGP助成＃90172、NIH 2P40RR017447-07及びNIH R01AR066033-01（NIAMS）によって部分的に資金を供給した。私たちは、原稿を校正するための博士スザンヌ·ゼイトゥーニーに感謝。

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The authors have no competing financial interests.

ここで、我々は、標準的な実験および獣医装置を使用して、マウスの大腿骨の臨界サイズのピンで安定化された欠陥を生成する簡単な方法を説明する。ピンのアセンブリと、外科的処置自体は練習が必要ですが、それはよくよく訓練された生物医学研究者や獣医師の能力の範囲内である。 ピンは、技術的により実現可能な外部固定またはインターロックねじを採用し、?...

これは、治療骨折を有する患者のために外科的に50万手続きは骨移植片2の使用を含む。米国の人口の半分は65 1歳までに骨折を経験していると推定されており、この数は、ますます高齢化、人口3に上昇すると予想されている。骨が完全に瘢痕なしに治癒する能力を有するいくつかの臓器の一つであるが、プロセスは3,4失敗する場合がある。状況や処理の品質に応じて、長骨骨折の2〜30％は、非組合3,5で得られた、失敗。定義、偽関節、大きさのクリティカルまたは非組合骨損傷上のいくつかの議論が残っているものの、一般的に被検体6の自然な寿命にわたって治癒しない怪我を指します。実験目的のために、この期間は、平均的なサイズの骨損傷の完全な治癒に必要な平均時間に短縮される。非組合骨病変はNUMのために起こるerousの理由が、主な要因は、原因疾患や年齢7に批判的にサイズのギャップ、感染、貧しい血管新生、タバコの使用、または阻害osteoregenerative容量になり、極端な外傷が含まれる。非組合が正常に処理されたとしても、それは損傷の種類と8を採用アプローチに応じて、手順6万ドルを超える費用がかかる。 中程度の場合には、自家骨髄移植が用いられる。この戦略は、損傷部位でのドナー部位および移植から骨の回復を伴う。このアプローチは非常に有効であるが、利用可能なドナー由来の骨の量は限られており、手順は、多くの患者9,10において持続性疼痛をもたらす追加の手術を伴う。また、自家骨移植片の有効性は、患者の健康状態に依存する。合成材料または処理死体の骨から作られた代用骨は11-13豊富に利用可能ですが、彼 ​​らはヘクタール乏しい宿主細胞の接着性、骨伝導性減少、および免疫拒絶14の可能性を含め、重大な制限VEの。安全で、効果的であり、広く利用可能な骨再生技術が緊急に必要とされている。 骨再生の戦略を改善する当社の能力は、試験動物に深刻な骨の外傷を模倣する能力に決定的に依存しているが、大きな骨病変の発生と安定化は技術的に困難である。ほとんどの場合、深刻な長骨の外傷が自然治癒しない欠陥を確立することによって実験的に模倣される。それは種15によって変化し得るが、これは骨の断面16の直径の1.5倍よりも大きい骨セグメントを完全に除去することによって達成される。骨は、その後骨折エッジの適切な配向を維持し、移動性を可能にするために、金属インプラントで安定化される。その小さなサイズとの脆弱性に起因する彼らの長い骨は、マウスではそのような病変の確立が最も研究グループの能力を超えている。このように、長骨欠損モデルは、ラットおよび大型動物に限定される。それにもかかわらず、マウスは、それらが遺伝的に修飾されたヒト細胞および組織を拒絶しない、免疫不全株として育種することができるという点で、重要な研究の利点をもたらす。 彼らは生理的によく特徴付けているため、ヒト細胞ベースのアプリケーションでは、免疫不全マウスはで動作するように魅力的で、家に簡単に、効果的なコスト、簡単に放射線学的および組織学的に分析した。最も重要な免疫不全マウスは、ヒトを含む異なる種からの細胞を拒絶しないことである。その小さなサイズはまた、細胞または整形外科用途で実験的な足場の体積の非常に少数のテストを可能にします。いくつかのマウスモデルが整形外科用骨安定17,18の様々な程度を与えることが報告されている。これらのsyste軟骨内の治癒が19に報告されているが、外部固定器とロッキングプレートとしての安定性の非常に高いレベルをもたらすmsが主に膜内骨化によって治癒する。対照的に、未修正または部分的に固定された髄ピンを用いたものなど、いくつかのミクロおよび/ ​​またはマクロ運動を可能にするものは、一般的に軟骨内骨化20,21が優勢で治癒する。長骨の遅延組合や非組合欠陥が必要な安定化の余分なレベルに起因したマウスで達成することが特に困難である。しかし、多くのアプローチが、プレートと外部固定器22をロックする、インターロック爪と髄ピンを含む、報告されている。これらのシステムは、一般的にはうまく動作しますが、その複雑な設計与えられた彼らは、インストールすることが技術的に挑戦することができます。例えば、ガルシアら 23は、マウスにおける使用のための優雅なインターロックピン方式を考案したが、手順は、2つの別々の部位における切開を伴いピンを収容するための大腿骨の広範な修正。これらの手順は、解剖顕微鏡下で行った。 ここで、我々は、3ミリメートルの骨欠損の閉鎖を防止し、また、欠陥のオリジナルのエッジを描くように設計され、中央襟付きのシンプルな大腿骨髄ピンについて説明します。ピンは、骨自体に固定されなかったが、十分な干渉髄腔結果のピン径、およびリーマの正確なサイジングがねじれ運動（ 図1）を最小化する。近交系、年齢、性別及び歪みを一致させたマウスを慎重に選択すると、結果は簡単に放射線学的に評価することが可能で再現性の高い肥大非uniondefect 22です。また、関心領域は、再現デノボ骨形成および組織形態学的パラメータを測定するためのマイクロコンピュータ断層撮影（μCT）の後に定義することができる。ピンは、容易に入手可能なツールを使用して、我々の研究室で試作された。<img alt="図1" src="/files/ftp_upload/52368/52368fig1highres.jpg" /> 図1：実験原理分節欠損モデルの図的要約。 9〜10ミリメートルマウス大腿骨の中央の3ミリメートルのセグメントは、（ 左 ）外科的に切除されている。長さ3mm、19ゲージの外科用鋼管は、正確な中心（ 右 ）での長い9ミリメートル、22 Gステンレス管の上を通過し、接着剤で固定されている。得られたピンは、骨の3mmのセグメントを置き換える19 Gのカラー（ 以下、センター ）と大腿骨の残りの近位および遠位部分の髄運河に嵌め込まれている。

<table><tbody><tr><td colspan="4">&lt;strong&gt;Pin assembly&lt;/strong&gt;</td></tr><tr><td>Dremel rotary tool</td><td>Dremel</td><td>8220</td><td>or equivalent</td></tr><tr><td>Heavy duty cut off wheel</td><td>Dremel</td><td>420</td><td></td></tr><tr><td>Surgical tubing 19&amp;nbsp;G</td><td>Small Parts (Amazon)</td><td>B000FMZ8LY</td><td>OD 1.07&amp;nbsp;mm, ID 0.889&amp;nbsp;mm</td></tr><tr><td>Surgical tubing 21&amp;nbsp;G</td><td>Small Parts (Amazon)</td><td>B000FMZ8YQ</td><td>OD 0.82&amp;nbsp;mm, ID 0.635&amp;nbsp;mm</td></tr><tr><td>Surgical tubing 22&amp;nbsp;G</td><td>Small Parts (Amazon)</td><td>B000FMYLZS</td><td>OD 0.719&amp;nbsp;mm, ID 0.502&amp;nbsp;mm</td></tr><tr><td>Surgical tubing 23&amp;nbsp;G</td><td>Small Parts (Amazon)</td><td>B000FN0SY0</td><td>OD 0.643&amp;nbsp;mm, ID 0.444&amp;nbsp;mm</td></tr><tr><td>Cyanoacrylate adhesive</td><td>Loctite</td><td>1365882</td><td></td></tr><tr><td>Emery disc</td><td>Dremel</td><td>413</td><td></td></tr><tr><td>Rubber polishing point</td><td>Dremel</td><td>462</td><td></td></tr><tr><td>Felt polishing disc</td><td>Dremel</td><td>414</td><td></td></tr><tr><td>Gelatin sponge</td><td>Surgifoam/Ethicon</td><td>1974</td><td></td></tr><tr><td>Punch biopsy cutter</td><td>Miltex</td><td>33-34</td><td></td></tr><tr><td colspan="4">&lt;strong&gt;Surgery/post-operative&lt;/strong&gt;</td></tr><tr><td>Warm pad and circulator pump</td><td>Stryker/Thermocare</td><td>TP700, TP700C, TPP722</td><td></td></tr><tr><td>Coverage quaternary spray</td><td>Steris</td><td>1429-77</td><td></td></tr><tr><td>Bead sterilizer</td><td>Germinator/CellPoint Scentific</td><td>Germinator 500</td><td></td></tr><tr><td>Anesthesia system</td><td>VetEquip Inc</td><td>901806 or 901807/901809</td><td></td></tr><tr><td>Isofluorane anesthetic</td><td>VETone/MWI</td><td>501017, 502017</td><td></td></tr><tr><td>Surgical disinfectant</td><td>Chloraprep/CareFusion</td><td>260449</td><td></td></tr><tr><td>Surgical tools</td><td>Fine Science Tools</td><td>various</td><td>recommend German made</td></tr><tr><td>Face protection</td><td>Splash Shield</td><td>4505</td><td></td></tr><tr><td>Rechargable high speed drill</td><td>Fine Science Tools</td><td>18000-17</td><td></td></tr><tr><td>Diamond cutting wheel</td><td>Strauss Diaiond</td><td>361.514.080HP</td><td></td></tr><tr><td>Absorbable sutures&amp;nbsp;</td><td>Covidien</td><td>UM-213</td><td></td></tr><tr><td>Outer sutures</td><td>Ethicon</td><td>668G</td><td>or equivalent</td></tr><tr><td>Vetbond</td><td>3M</td><td>1469SB</td><td>or equivalent</td></tr><tr><td>Hydration gel</td><td>Clear H2O</td><td>70-01-1082</td><td></td></tr><tr><td>Diet gel</td><td>Clear H2O</td><td>72-01-1062</td><td></td></tr><tr><td>Buprenorphine</td><td>Reckitt and Benckser</td><td>12496-0757-01</td><td>controlled substance</td></tr><tr><td>Mouse igloos</td><td>Bio Serv</td><td>K3328, 3570,3327</td><td></td></tr><tr><td>Euthanasia cocktail</td><td>Euthasol/Virbac</td><td>710101</td><td>controlled substance</td></tr><tr><td colspan="4">&lt;strong&gt;Analysis&lt;/strong&gt;</td></tr><tr><td>Live animal imager&amp;nbsp;</td><td>Orthoscan</td><td>FD Pulse</td><td>or equivalent</td></tr><tr><td>Micro-CT unit and software</td><td>Bruker</td><td>Skyscan1174</td><td>or equivalent</td></tr><tr><td>Sealing film/Parafilm M</td><td>VWR or Fisher</td><td>100501-338, S37441</td><td></td></tr><tr><td colspan="4">*Generic sources are suitable for all other items such as gause, drapes, protective clothing, animal care equipment.</td></tr></tbody></table>

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.

マウスは、一般的に意識を回復し、後肢の移動に麻酔の中止後5〜10分。最初の5日間、それは個別にマウスを収容し、四肢の過剰使用を防止するための環境エンリッチメントを導入することをお勧めします。この目的のために、イグルー型の巣は、巣の建物の必要性を低減し、休息をお勧めします。また、ケージの床に食品及びヒドロゲルの提供は、ピンの変位の確率を減少させることを観察...

Watch this Scientific Journal Video about A Simple Critical-sized Femoral Defect Model in Mice at JoVE.com

A Simple Critical-sized Femoral Defect Model in Mice

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 ...

a-simple-critical-sized-femoral-defect-model-in-mice

Research

JoVE Journal

Medicine

15.0K ビュー.  Texas A&M Health Science Center. 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. 

ビデオ: A Simple Critical-sized Femoral Defect Model in Mice

Use of Human Perivascular Stem Cells for Bone Regeneration

Human perivascular stem cells (PSCs) can be isolated in sufficient numbers from multiple tissues for purposes of skeletal tissue engineering1-3. PSCs are a FACS-sorted population of 'pericytes' (CD146+CD34-CD45-) and 'adventitial cells' (CD146-CD34+CD45-), each of which we have previously reported to have properties of mesenchymal stem cells. PSCs, like MSCs, are able to undergo osteogenic differentiation, as well as secrete pro-osteogenic cytokines1,2. In the present protocol, we demonstrate the osteogenicity of PSCs in several animal models including a muscle pouch implantation in SCID (severe combined immunodeficient) mice, a SCID mouse calvarial defect and a femoral segmental defect (FSD) in athymic rats. The thigh muscle pouch model is used to assess ectopic bone formation. Calvarial defects are centered on the parietal bone and are standardly 4 mm in diameter (critically sized)8. FSDs are bicortical and are stabilized with a polyethylene bar and K-wires4. The FSD described is also a critical size defect, which does not significantly heal on its own4. In contrast, if stem cells or growth factors are added to the defect site, significant bone regeneration can be appreciated. The overall goal of PSC xenografting is to demonstrate the osteogenic capability of this cell type in both ectopic and orthotopic bone regeneration models.

Human perivascular stem cells (PSCs) are a novel stem cell class for skeletal tissue regeneration similar to mesenchymal stem cells (MSCs). PSCs can be isolated by FACS (fluorescence activated cell sorting) from adipose tissue procured during standard liposuction procedures, then combined with an osteoinductive scaffold to achieve bone formation in vivo.

骨再生のための人間の血管周囲の幹細胞の使用

Human perivascular stem cells (PSCs) can be isolated in sufficient numbers from multiple tissues for purposes of skeletal tissue engineering1-3. PSCs are ...

生物工学

A Simple Guide Screw Method for Intracranial Xenograft Studies in Mice

The grafting of human tumor cells into the brain of immunosuppressed mice is an established method for the study of brain cancers including glioblastoma (glioma) and medulloblastoma. The widely used stereotactic approach only allows for the injection of a single animal at a time, is labor intensive and requires highly specialized equipment. The guide screw method, initially developed by Lal et al.,1 was developed to eliminate cumbersome stereotactic procedures. We now describe a modified guide screw approach that is rapid and exceptionally safe; both of which are critical ethical considerations. Notably, our procedure now incorporates an infusion pump that allows up to 10 animals to be simultaneously injected with tumor cells.
To demonstrate the utility of this procedure, we established human U87MG glioma cells as intracranial xenografts in mice, which were then treated with AMG102; a fully human antibody directed to HGF/scatter factor currently undergoing clinical evaluation2-5. Systemic injection of AMG102 significantly prolonged the survival of all mice with intracranial U87MG xenografts and resulted in a number of complete cures. 
This study demonstrates that the guide screw method is an inexpensive, highly reproducible approach for establishing intracranial xenografts. Furthermore, it provides a relevant physiological model for validating novel therapeutic strategies for the treatment of brain cancers.

In order to evaluate novel therapeutic paradigms for the treatment of glioma, physiological relevant models are essential. We utilize an implantable guide screw procedure for establishment of intracranial xenograft models that is more rapid and safer than stereotactic approaches.

マウスにおける頭蓋内異種移植研究のための簡単​​なガイドスクリュー方式

The grafting of human tumor cells into the brain of immunosuppressed mice is an established method for the study of brain cancers including glioblastoma ...

医学

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 Reliable and Reproducible Critical-Sized Segmental Femoral Defect Model in Rats Stabilized with a Custom External Fixator

Orthopedic research relies heavily on animal models to study mechanisms of bone healing in vivo as well as investigate the new treatment techniques. Critical-sized segmental defects are challenging to treat clinically, and research efforts could benefit from a reliable, ambulatory small animal model of a segmental femoral defect. In this study, we present an optimized surgical protocol for the consistent and reproducible creation of a 5 mm critical diaphyseal defect in a rat femur stabilized with an external fixator. The diaphyseal ostectomy was performed using a custom jig to place 4 Kirschner wires bicortically, which were stabilized with an adapted external fixator device. An oscillating bone saw was used to create the defect. Either a collagen sponge alone or a collagen sponge soaked in rhBMP-2 was implanted into the defect, and the bone healing was monitored over 12 weeks using radiographs. After 12 weeks, rats were sacrificed, and histological analysis was performed on the excised control and treated femurs. Bone defects containing only collagen sponge resulted in non-union, while rhBMP-2 treatment yielded the formation of a periosteal callous and new bone remodeling. Animals recovered well after implantation, and external fixation proved successful in stabilizing the femoral defects over 12 weeks. This streamlined surgical model could be readily applied to study bone healing and test new orthopedic biomaterials and regenerative therapies in vivo.

In vivo mammalian models of critical-sized bone defects are essential for researchers studying healing mechanisms and orthopedic therapies. Here, we introduce a protocol for the creation of reproducible, segmental, femoral defects in rats stabilized using external fixation.

ラットのカスタム創外固定器で安定した信頼性と再現性のある重要なサイズ分節大腿骨欠損モデル

Orthopedic research relies heavily on animal models to study mechanisms of bone healing in vivo as well as investigate the new treatment techniques. ...

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 ...

ラット大腿骨の樹立と評価:空洞欠損モデル

Creating a Box-Cavity Defect Model in the Cortical Bone of Rat Femora

Severe bone defects or complex fractures can result in serious complications such as nonunion or insufficient bone healing. Tissue engineering, which involves the application of cells, scaffolds, and cytokines, is considered a promising solution for bone regeneration. Consequently, various animal models that simulate bone defects play a crucial role in exploring the therapeutic potential of tissue engineering for bone healing. In this study, we established a box-shaped cortical bone defect model in the mid-femur of rats, which could serve as an ideal model for assessing the function of biomaterials in promoting bone healing. This box-shaped cortical bone defect was drilled using an oral low-speed handpiece and shaped by a lathe needle. Post-operative micro-CT analysis was immediately conducted to confirm the successful establishment of the box-cavity cortical bone defect. The femurs on the operated side of the rats were then harvested at multiple time points post-surgery (0 days, 2 weeks, 4 weeks, and 6 weeks). The healing process of each sample&#39;s defect area was evaluated using micro-CT, hematoxylin and eosin (H&#38;E) staining, and Masson trichrome staining. These results demonstrated a healing pattern consistent with intramembranous ossification, with healing essentially complete by 6 weeks. The categorization of this animal model&#39;s healing process provides an effective in vivo method for investigating novel biomaterials and drugs that target intramembranous ossification during bone tissue defect healing.

著者スポットライト:生体材料と骨再生の評価を強化するための箱腔皮質アプローチ

Here, we present a protocol for creating a box-cavity defect in rat femoral diaphysis tissue. This model can assess biomaterial performance under biomechanical stress and explore mechanisms of bone regeneration related to intramembranous osteogenesis.

ラット大腿骨の皮質骨における箱腔欠損モデルの作成

Severe bone defects or complex fractures can result in serious complications such as nonunion or insufficient bone healing. Tissue engineering, which ...

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 ...

マウスの簡単な重要なサイズの大腿欠陥モデル

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骨再生のための人間の血管周囲の幹細胞の使用

マウスにおける頭蓋内異種移植研究のための簡単なガイドスクリュー方式

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

ラットのカスタム創外固定器で安定した信頼性と再現性のある重要なサイズ分節大腿骨欠損モデル

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

ラット大腿骨の皮質骨における箱腔欠損モデルの作成

ラット大腿骨の皮質骨における箱腔欠損モデルの作成

ラット大腿骨の皮質骨における箱腔欠損モデルの作成

安定化ピン付きマウス大腿骨の横骨折

マウスの簡単な重要なサイズの大腿欠陥モデル

要約

さらに動画を探す

この動画の章

骨再生のための人間の血管周囲の幹細胞の使用

マウスにおける頭蓋内異種移植研究のための簡単​​なガイドスクリュー方式

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

ラットのカスタム創外固定器で安定した信頼性と再現性のある重要なサイズ分節大腿骨欠損モデル

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

ラット大腿骨の皮質骨における箱腔欠損モデルの作成

ラット大腿骨の皮質骨における箱腔欠損モデルの作成

ラット大腿骨の皮質骨における箱腔欠損モデルの作成

安定化ピン付きマウス大腿骨の横骨折

マウスにおける頭蓋内異種移植研究のための簡単なガイドスクリュー方式