Name: an intramedullary locking nail for standardized fixation of femur osteotomies to analyze normal and defective bone healing in mice
Uploaded: 2016-11-13T15:00:00.000+00:00
Duration: 7 min 41 s
Description: Watch this Scientific Journal Video about An Intramedullary Locking Nail for Standardized Fixation of Femur Osteotomies to Analyze Normal and Defective Bone Healing in Mice at JoVE.com

이 작품은 RISystem AG, 스위스 다 보스에 의해 지원되었다.

모든 절차 IACUC가 승인했다 제도적 지침 (Landesamt 대 Verbraucherschutz, Zentralstelle Amtstierärztlicher DIENST, 자르브뤼켄, 독일) 하였다. 통증 및 감염 예방 실험이 수행 될 수있는 국가 및 기관의 각각의 가이드 라인에 일치해야한다. 임플란트 및 수술 도구 1. 준비 <ol><li> 미세 수술 도구 상자에서 메스 블레이드 (사이즈 15), 작은 준비 가위, 미세 집게, 드레싱 포셉, 작은 집게, 24 게이지 (G) 27 G 바늘, 비 흡수성 5-0 봉합하고, 바늘 홀더를 선택 . </li><li> 골수 손톱, 연동 핀, 특별한 목표 장치는 질리가 본의 질리에 대한 템플릿을보고, 센터링 드릴 비트 (1 mm 직경), 드릴 비트 (0.3 mm 직경) 및 핸드 드릴의 압축을 풉니 다 (그림 2) 재료의 목록을 참조하십시오. 참고 : intramedullary 네일 (0.8 mm 직경 15.7 mm 길이)가 대퇴골에 역행 주입을위한 의료용 스테인리스 골수 내 고정 못하다. 손발톱은 근위 실 (4 mm 길이) 및 축 방향 및 회전 안정성을 달성하기 위해 인터 로킹 핀의 삽입 (0.3 mm 직경)에 대한 두 개의 구멍 (도 1)을 갖는다. </li><li> (130 ° C, 25 분간 증기 멸균)를 5 분간 소독 용액 (예를 들면, 96 % 알코올)로 임플란트 및 수술기구 모두를 노출하거나 소독. 소독 또는 살균 후 무균 조작 천으로 악기를 배치합니다. 작은 동물의 작업 테이블에 바로 인접하여 무균 조작 천 놓습니다. </li></ol> 2. 동물, 마취 및 진통 <ol><li> 연구 및 해결되어야 할 질문에 대한 필요에 따라 마우스의 변형, 연령, 성별 선택. 참고 : 14 주령의 수컷 CD-1 마우스에 대한 연구 12을 위해 사용되었다. 네일 impla에 대한35g - ntation 동물의 이상적인 체중은 25이다. </li><li> 15 ㎎ / ㎏ 자일 라진, 75 ㎎ / ㎏ 케타민의 복강 내 주사로 쥐를 마취. 발가락 핀치에 의한 마취를 확인합니다. 마취 동안 건조에서 동물의 눈을 보호하기 위해 눈 윤활제를 적용합니다. 마취 유도 후, 체온을 일정하게 유지하기위한 가열 램프하에 마우스를 놓는다. </li><li> 수술 후 3 일까지 수술 전 1 일에서 진통제에 대한 식수 (2.5 밀리그램 / 100 ml) 중 트라마돌 염산염을 적용합니다. </li></ol> 3. 수술 절차 및 네일 주입 <ol><li> 수술하기 전에, 전체 오른쪽 뒷다리를 면도와 제모 크림을 적용합니다. 5 분 후, 크림을 제거하고 물로 세척 레그. (130 ° C, 25 분간 증기 멸균) 소독 용액 (예를 들면, 96 % 알코올)로 임플란트 및 수술기구 모두를 노출하거나 소독. </li><li> 무균 조건에서 양해 각서 (MOU)를 배치작은 동물의 작업 테이블에 앙와위에서 자체. 무릎의 과두에 전방 접근을 허용하기 위해 오른쪽 무릎을 구부리고. 메스 블레이드를 사용하여 오른쪽 무릎에 5mm 내측 슬개골 절개를 수행합니다. </li><li> 미세 집게로 슬개 인대를 들어 올려 메스 블레이드로 조심스럽게 인대을 동원. 그리고, 대퇴골 과간 노치를 노출 메스 블레이드 횡 슬개골을 이동. </li><li> 골수 캐비티에 도달 할 때까지 시추하여 과간 노치를 엽니 다. <ol><li> 드릴 비트를 중심으로 1mm를 사용하여 대퇴골 축에 오프셋을 45 °로 드릴링 시작합니다. 천천히가 대퇴골의 골 축 평행까지 드릴링 동안 드릴 용 비트의 방향을 변경합니다. 골수 공동 도달하면 드릴 중지합니다. </li></ol></li><li> 과간 노치에서 뼈를 연 후, 대퇴골의 전체 길이에 걸쳐 골수 캐비티로 24 G 바늘을 삽입합니다. intrame를 연수동으로 24 G 바늘의 회전 운동을 통해 대퇴골의 dullary 공동. 24 G 바늘을 제거하고 골수 공동으로 얇은 27 G 바늘을 삽입합니다. 근위 큰 대 전자에서 대퇴골의 피질골을 관통 앞으로 바늘을 밀어 넣습니다. </li><li> 대퇴골에서 27 G 바늘을 제거합니다. 손발톱의 선단부 과두의 레벨에 도달 할 때까지 핸드 드릴을 사용하여, 연속적인 회전 및 축 방향 압력 하에서 과간 노치를 통해 골수 이식 손톱. 주 : 못의 선단부는 작은 표시가 식별 될 수있다. </li><li> 왼쪽 측면 위치에 마우스를 놓습니다. 수술 대퇴골 간부를 노출하기 위해 고관절에 무릎 관절의 측방 대퇴골의 골간을 따라 부 메스 블레이드를 사용하여 길이 피부 절개를 수행한다. </li><li> 작은 준비 가위를 사용하여, 밴드 분할하고, 측면에서 대퇴골 축 방향의 근육을 확산.대퇴골의 골간 부분이 노출 될 때까지 근육 확산. 좌골 신경을 보존합니다. <ol><li> 드레싱 집게로 뼈를 훼손하여 대퇴골의 전체 둘레를 준비합니다. 그런 다음, 드레싱 집게를 확산하여 근육을 수축과 대퇴골을 노출. </li></ol></li><li> 손발톱의 선단 조준 장치를 마운트. 이 손톱의 어댑터 플랜지에 부착 될 때까지 장치를 발전 대퇴골에 외측 위치에 조준 장치를 켜십시오. </li><li> 근위 및 원위 연동 핀 못을 연동. <ol><li> 근위 연동 핀을 시작합니다. </li><li> 핸드 드릴로 센터링 드릴 비트 (1 mm 직경)를 삽입합니다. 근위 연동 구멍 위치에서 뼈 카운터 싱크. 참고 : 카운터 싱크으로 작은 구멍이 뼈를 통해 드릴링없이 직면 피질골에 만들어집니다. 이 공동 개선 중심과 얇은 드릴 비트의 안내 (0.3 mm 직경)을 허용나중에 사용. </li><li> 핸드 드릴로 드릴 비트 (0.3 mm 직경)를 삽입합니다. 조준 장치를 사용하여 대면하고 피할 피질골 (bicortical) 모두를 통해 구멍을 드릴. 조준 장치를 통해 첫 번째 연동 핀을 삽입합니다. 즉시 상기 연동 토크가 달성 될 때 오프 연동 핀 구동축 위. </li><li> 말단 연동 핀에 대해이 절차를 반복합니다. </li></ol></li><li> 골간 절골술을 수행합니다. <ol><li> 두 연동 핀 사이의 측면에 조준 장치로 톱 가이드를 부착합니다. 질리가 식염수 연속 관개에서보고와 다음, 뼈를 보았다. 절골 종료 후, 골 가까이 일단 톱 절단. 연부 조직의 손상을 일으키는 원인이되지 않도록 조심스럽게 톱을 제거합니다. </li></ol></li><li> 마크 된 라인의 골수 손톱의 나머지 샤프트를 클립, 작은 집게로, 조준 장치를 제거합니다. </li><li> 위도에있는 근육 층을 닫습니다대한 일반적인의 대퇴골의 사이트와 단일 봉합으로 피부 폐쇄를 수행합니다. 무릎의 전방 사이트에서 슬개골의 위치를 ​​변경하고 하나의 봉합사로 근육 슬개골 건을 해결. 뿐만 아니라이 상처를 닫 하나의 봉합사를 사용합니다. </li><li> 그들이 마취에서 회복 될 때까지 가열 램프 아래에있는 동물을 유지합니다. 그들은 복부 드러 누움을 유지하기 위해 충분한 의식을 회복 할 때까지 무인 동물을 두지 마십시오. 동물 시설에서 하나의 새장에 동물을 돌려줍니다. </li><li> 매일 신중하게 동물을 모니터링합니다. 처음 3 일간은 수술 후 통증을 유지한다. 수술 후 4 일째에, 동물은 여전히 ​​고통의 증거를 보여, 경우 발성, 불안, 이동성의 부족, 신랑 실패, 비정상적인 자세, 주변에 일반의 관심 부족으로 나타낸 바와 같이, 진통제를 계속합니다. 동물들이 고통없는 경우 진통제를 종료합니다. </li></ol>

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Romano Matthys is an employee of RISystem AG, Davos, Switzerland, which produces the implants and implant-specific instruments. The other authors have no conflicts of interest.

수술 기법의 가장 중요한 단계는 손톱 조준 장치, 및 핀의 정확한 위치 결정된다. 손발톱은 과두의 수준에서 무릎 관절에 손발톱의 돌출부 무릎 (도 3 A)의 움직임을 제한 할 수 있기 때문에, 손발톱의 선단에서 현저한 들여 완전히 삽입되어야한다. 따라서, 대퇴의 크기는, 따라서, 동물의 체중을 고려하여야한다. 의사는 또한 손톱의 어댑터 플랜지를 부착 조준 장치의 최종 위치에 특?...

뼈 치유 생물학 및 세포 배양 타원체를 사용하여 시험 관내에서 조사 될 수 있고,뿐만 아니라 동물 실험 접근법을 사용하여 생체 내에서 필요하다. 대형 동물 실험은 여전히 임상 시험에 중요한 역할을하지만, 제품 또는 가설 초기 시험은 지난 10 년 동안 변경되어 현재에는 종종 작은 동물 모델 (1)에 수행된다. 이 스위치는 여러 가지 이유로 하였다. 생산 및 마우스 및 쥐의 유지 보수가 저렴 돼지와 양에 비교된다. 또한, 작은 동물 만성 실험 큰 시리즈의 성능을 용이하게 둘 짧은 재생 시간 및 정상 짧은 치료 기간을 갖는다. 마지막으로, 유전자 타겟팅 동물과 특이 적 항체의 가용성 뼈 치유 분자 기전의 분석을 허용한다. 앞에서 큰 동물 모델에서 골 접합 기술을 사용하면서 그러나 최소한 variat로 번역 될 수있다<html> 인간 또는 동물 임상 환자 치료, 소형 쥐와 생쥐의 개발 ​​및 골 유합술 기술의 응용 프로그램에서 사용되는 유사한 절차에서 이온 도전으로 밝혀졌다. 잘 역학적 환경이 현저하게 공정이 골 치유에 영향을 미치는 것으로 알려져있다. 인간의 골절 치료에서 알 수있는 바와 같이, micromovements 덜 견고한 고정 후 견고한 고정 및 연골 골화 후 intramembranous 골화를 포함한 치료의 독특한 모드에서 골절 안정화 결과의 차이. 전체 축 또는 회전 불안정성은 치유 과정이 지연 될 수 있습니다 또는 비 노조 3의 원인이 될 수 있습니다. 따라서, 우리는 마우스 및 래트에서 정교한 임플란트 시스템 및 고정술 기술을 개발할 필요가 있다고 느낀다. 이와 같이, 생체 역학적 조건은 치유 과정을 분석 할 때 유효한 결과를 보장 적절하게 표준화 될 수있다. e_content &quot;&gt; 고도의 뮤린 안정화 기술들의 상당수는 지난 몇 년 동안 소개되어 있지만, 가장 일반적으로 사용되는 기술은 여전히 ​​간단한 골수 핀이다.이 기술의 주요 단점은, 그러나, 회전 방향 및 축 방향의 부족 안정성 4. 회전 및 축 방향의 안정성을 향상시키기 위해, 골수 나사 마우스 5 대퇴골 골절을 안정화하기 위해 도입되었다. 그러나, 나사 고정 인해 뼈 조각 사이의 접촉과 압축의 필요성 뼈 결함 치유를 분석 할 수없는 회전 안정성을 유지하기 위해서는. 못을 잠금 골수 높은 축 방향 및 간단한 핀과 골수 나사 (4)에 비해 회전 안정성을 제공합니다. 때문에 질리에 대한 가이드의 가능한 고도의 재현 대퇴골 절골술은보고 정의 갭 크기를 만들 수있는 기능은 모두 정상 봉의 분석을 가능하게전자 치유와 뼈 결함 치유 6. 인해 연동 핀의 삽입으로, 못 잠금 골수 전체 중량을지지하면서도, 전체 치료 과정 동안 일정한 갭 크기를 보장한다. 여기, 우리는 디자인과 골수 잠금 손톱의 응용 프로그램에 대해보고뿐만 아니라 장점과 정상 및 지연 골 치유에 관한 실험 연구의 단점에있다.

<table><tbody><tr><td>MouseNail</td><td>RISystem AG</td><td>221,122</td><td></td></tr><tr><td>MouseNail aiming device</td><td>RISystem AG</td><td>221,201</td><td></td></tr><tr><td>MouseNail interlocking pin</td><td>RISystem AG</td><td>221,121</td><td></td></tr><tr><td>Centering bit</td><td>RISystem AG</td><td>592,205</td><td></td></tr><tr><td>Drill bit</td><td>RISystem AG</td><td>590,200</td><td></td></tr><tr><td>Gigli wire saw</td><td>RISystem AG</td><td>590,100</td><td></td></tr><tr><td>Suture (5-0 Prolene)</td><td>Ethicon</td><td>8614H</td><td></td></tr><tr><td>Forceps</td><td>Braun Aesculap AG &amp;amp;CoKG&amp;nbsp;</td><td>BD520R</td><td></td></tr><tr><td>Dressing forceps</td><td>Braun Aesculap AG &amp;amp;CoKG&amp;nbsp;</td><td>BJ009R</td><td></td></tr><tr><td>Scissors</td><td>Braun Aesculap AG &amp;amp;CoKG&amp;nbsp;</td><td>BC100R</td><td></td></tr><tr><td>Needle holder</td><td>Braun Aesculap AG &amp;amp;CoKG&amp;nbsp;</td><td>BM024R</td><td></td></tr><tr><td>24 G needle</td><td>BD Mircolance 3</td><td>304100</td><td></td></tr><tr><td>27 G needle</td><td>Braun Melsungen AG</td><td>9186182</td><td></td></tr><tr><td>Scalpel blade size 15</td><td>Braun Aesculap AG &amp;amp;CoKG&amp;nbsp;</td><td>16600525</td><td></td></tr><tr><td>Pincers</td><td>Knipex</td><td>7932125</td><td></td></tr><tr><td>Heat radiator</td><td>Sanitas</td><td>605.25</td><td></td></tr><tr><td>Depilatory cream</td><td>Asid bonz GmbH</td><td>NDXZ10</td><td></td></tr><tr><td>Eye lubricant</td><td>Bayer Vital GmbH</td><td>2182442</td><td></td></tr><tr><td>Xylazine</td><td>Bayer Vital GmbH</td><td>1320422</td><td></td></tr><tr><td>Ketamine</td><td>Serumwerke Bernburg</td><td>7005294</td><td></td></tr><tr><td>Tramadol</td><td>Gr&amp;uuml;nenthal GmbH</td><td>2256241</td><td></td></tr><tr><td>Disinfection solution (SoftaseptN)</td><td>Braun Melsungen AG</td><td>8505018</td><td></td></tr><tr><td>CD-1 mice</td><td>Charles River</td><td>22</td><td></td></tr></tbody></table>

an intramedullary locking nail for standardized fixation of femur osteotomies to analyze normal and defective bone healing in mice

뼈 치유 모델 임상 골절 치료를위한 새로운 치료 전략의 개발에 필수적이다. 또한, 마우스 모델은 외상 연구에 사용 된 더 일반적으로되고있다. 이들은 뼈 치유의 차별화 프로세스 뒤에 분자 기전의 해석 변이주 항체의 다수를 제공한다. 생체 역학적 환경을 제어하기 위해 표준화 된 잘 특성화 골 유합술 기술은 마우스에서 필수입니다. 여기, 우리는 생쥐에서 열린 대퇴골 절골술을 안정화하기 위해 디자인과 골수 손톱의 사용에보고한다. 의료용 스테인레스 스틸로 만든 못, 높은 축 방향 및 회전 강성을 제공합니다. 또한 임플란트 정의, 상수 절골술 갭의 창조는 0.00 mm에서 2.00 mm로 크기를 할 수 있습니다. 0.00 mm 및 0.25 mm의 갭 크기의 대퇴골 절골술의 손톱 안정화 잠금 골수은 연골 및 intramembranous ossificat을 통해 적절한 뼈 치유 결과이온. 위축성 비 노조에서 2.00 mm 결과의 간격 크기 대퇴골 절골술의 안정화. 따라서, 골수 내 고정 못 치유 비 치유 모델에서 사용될 수있다. 다른 오픈 뼈 치유 모델에 비해 손발톱의 사용의 또 다른 이점은 충분히 골 통합 과정을 연구하기 위해 뼈 대체 물질 및 지지체를 고정 할 수있는 가능성이다. 골수 손발톱의 사용의 단점은 폐 모델에 비해 모든 오픈 절차에 고유 침습적 수술이다. 다른 단점은 골수 외 안정화 방법에 비해 모든 골수 안정화 기술에 내재 된 골수 공동에 손상이 유도 될 수있다.

수술에 대한 전체 시간은 폐쇄 상처 약 30 분 피부 절개에서였다. 제공된 외과 용 임플란트를 사용하여 외과 스테레오 현미경없이 수행 될 수있다. 수술 후, 동물을 매일 관찰 하였다. 동물의 것도이 기간 후 통증 (발성, 불안, 이동성의 부족, 신랑 실패, 비정상적인 자세, 또는 주변에 정상적인 관심의 부족)의 증거를 보여 주었다 없기 때문에 수술 후 진통제 3 일 후에 종료?...

Watch this Scientific Journal Video about An Intramedullary Locking Nail for Standardized Fixation of Femur Osteotomies to Analyze Normal and Defective Bone Healing in Mice at JoVE.com

An Intramedullary Locking Nail for Standardized Fixation of Femur Osteotomies to Analyze Normal and Defective Bone Healing in Mice

This protocol describes an osteosynthesis technique using an intramedullary locking nail for standardized fixation of femur osteotomies, which can be used to analyze normal and defective bone healing in mice.

대퇴골 절골술의 표준화 된 고정을위한 골수 내 잠금 네일은 마우스에 정상 및 결함 뼈 치유를 분석하기

Bone healing models are essential to the development of new therapeutic strategies for clinical fracture treatment. Furthermore, mouse models are becoming ...

an-intramedullary-locking-nail-for-standardized-fixation-femur

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

13.3K 조회수.  Saarland University. This protocol describes an osteosynthesis technique using an intramedullary locking nail for standardized fixation of femur osteotomies, which can be used to analyze normal and defective bone healing in mice.

비디오: An Intramedullary Locking Nail for Standardized Fixation of Femur Osteotomies to Analyze Normal and Defective Bone Healing in Mice

Creating Rigidly Stabilized Fractures for Assessing Intramembranous Ossification, Distraction Osteogenesis, or Healing of Critical Sized Defects

Assessing modes of skeletal repair is essential for developing therapies to be used clinically to treat fractures. Mechanical stability plays a large role in healing of bone injuries. In the worst-case scenario mechanical instability can lead to delayed or non-union in humans. However, motion can also stimulate the healing process. In fractures that have motion cartilage forms to stabilize the fracture bone ends, and this cartilage is gradually replaced by bone through recapitulation of the developmental process of endochondral ossification. In contrast, if a bone fracture is rigidly stabilized bone forms directly via intramembranous ossification. Clinically, both endochondral and intramembranous ossification occur simultaneously. To effectively replicate this process investigators insert a pin into the medullary canal of the fractured bone as described by Bonnarens4. This experimental method provides excellent lateral stability while allowing rotational instability to persist. However, our understanding of the mechanisms that regulate these two distinct processes can also be enhanced by experimentally isolating each of these processes. We have developed a stabilization protocol that provides rotational and lateral stabilization. In this model, intramembranous ossification is the only mode of healing that is observed, and healing parameters can be compared among different strains of genetically modified mice 5-7, after application of bioactive molecules 8,9, after altering physiological parameters of healing 10, after modifying the amount or time of stabilization 11, after distraction osteogenesis 12, after creation of a non-union 13, or after creation of a critical sized defect. Here, we illustrate how to apply the modified Ilizarov fixators for studying tibial fracture healing and distraction osteogenesis in mice.

This article describes a method for stabilizing long bone fractures that is based on the application of modified Ilizarov external fixators 1-3. After application of the fixators and creation of the bone injury, healing can be assessed, distraction osteogenesis can be performed, or non-union or critical sized defect can be created and used to study therapeutic interventions.

산만 Osteogenesis, 또는 임계 크기의 결함의 치유는 Intramembranous 골화을 평가하기위한 Rigidly 안정 골절 만들기

Assessing modes of skeletal repair is essential for developing therapies to be used clinically to treat fractures. Mechanical stability plays a large role ...

연구

의학

A Mini-Invasive Internal Fixation Technique for Studying Immobilization-Induced Knee Flexion Contracture in Rats

Joint contracture, resulting from a prolonged joint immobilization, is a common complication in orthopedics. Currently, utilizing an internal fixation to restrict knee joint mobility is a widely accepted model to generate experimental contracture. However, implanting application will inevitably cause surgical trauma to the animals. Aiming to develop a less invasive approach, we combined a muscle-gap separation modus with a previously reported mini-incision skill during the surgical procedure: Two mini skin incisions were made on the lateral thigh and leg, followed by performing muscle-gap separation to expose the bone surface. The rat knee joint was gradually immobilized by a preconstructed internal fixation at approximately 135&#176; knee flexion without interfering essential nerves or blood vessels. As expected, this simple technique permits rapid postoperative rehabilitation in animals. The correct position of the internal fixation was confirmed by an x-ray or micro-CT scanning analysis. The range of motion was significantly restricted in the immobilized knee joint than that observed in the contralateral knee joint demonstrating the effectiveness of this model. Besides, histological analysis revealed the development of fibrous deposition and adhesion in the posterior-superior knee joint capsule over time. Thus, this mini-invasive model may be suitable for mimicking the development of immobilized knee joint contracture.

Here, we present a protocol to describe a minimally invasive technique for knee joint immobilization in a rat model. This reproducible protocol, basing on muscle-gap separation modus and the mini-incision skill, is suitable for studying the underlying molecular mechanism of acquired joint contracture.

쥐에서 고정 유도 무릎 굴곡 계약을 연구하기위한 미니 침략적 인 내부 고정 기술

Joint contracture, resulting from a prolonged joint immobilization, is a common complication in orthopedics. Currently, utilizing an internal fixation to ...

발생학

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.

마우스의 간단한 임계 크기의 대퇴 결함 모델

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.

마우스의 분절 대퇴 임계 크기 결함 모델의 설립은 플레이트 고정술에 의해 안정화

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 ...

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

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The amount of the different tissues in the callus provides important information on the fracture healing progress. Available in vivo techniques to longitudinally monitor the callus tissue development in preclinical fracture-healing studies using small animals include digital radiography and &#181;CT imaging. However, both techniques are only able to distinguish between mineralized and non-mineralized tissue. Consequently, it is impossible to discriminate cartilage from fibrous tissue. In contrast, magnetic resonance imaging (MRI) visualizes anatomical structures based on their water content and might therefore be able to noninvasively identify soft tissue and cartilage in the fracture callus. Here, we report the use of an MRI-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice. The experiments demonstrated that the fixator and a custom-made mounting device allow repetitive MRI scans, thus enabling longitudinal analysis of fracture-callus tissue development.

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

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.

Vivo에서 골절 뼈 쥐 마우스 대 퇴 골에 대 한 MRI-호환 Osteosynthesis 장치를 사용 하 여 치료 기간 동안 굳은 살 개발의 평가

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The ...

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.

정형 외과 수술 후 인지 기능 장애 및 조직 재생 연구의 마우스 모델

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.

Endochondral 골절 표준화 된 Biomechanical 조건 하에서 쥐에서 치유를 분석 하는 최소 침 습 모델

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.

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

Fracture Apparatus Design and Protocol Optimization for Closed-stabilized Fractures in Rodents

The reliable generation of consistent stabilized fractures in animal models is essential for understanding the biology of bone regeneration and developing therapeutics and devices. However, available injury models are plagued by inconsistency resulting in wasted animals and resources and imperfect data. To address this problem of fracture heterogeneity, the purpose of the method described herein is to optimize fracture generation parameters specific to each animal and yield a consistent fracture location and pattern. This protocol accounts for variations in bone size and morphology that may exist between mouse strains and can be adapted to generate consistent fractures in other species, such as rat. Additionally, a cost-effective, adjustable fracture apparatus is described. Compared to current stabilized fracture techniques, the optimization protocol and new fracture apparatus demonstrate increased consistency in stabilized fracture patterns and locations. Using optimized parameters specific to the sample type, the described protocol increases the precision of induced traumas, minimizing the fracture heterogeneity typically observed in closed-fracture generation procedures.

The goal of the protocol is to optimize the fracture generation parameters to yield consistent fractures. This protocol accounts for the variations in bone size and morphology that may exist between animals. Additionally, a cost-effective, adjustable fracture apparatus is described.

골절 기구 디자인 및 설치류에서 폐쇄 안정 골절에 대 한 프로토콜 최적화

The reliable generation of consistent stabilized fractures in animal models is essential for understanding the biology of bone regeneration and developing ...

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.

안정화 핀을 가진 마우스 대퇴골의 횡단 골절

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 ...