이 작품은 NIAMS에서 R01-AR053645에 의해 재정 지원되었다.

모든 절차는 UCSF 기관 동물 케어의 승인 및위원회를 사용하여 국가 지침을 준수했다. 1. 수술에 앞서 Fixators의 작성 <ol><li> 안정 파괴를 만들기 전에 한 맞춤 설계 외부 고정 장치를 조립해야합니다. 맞춤 디자인 외부 고정 프레임은 세 스테인리스 # 0 / 56 스레드 ​​막대, 8 # 56분의 2 육각 너트, 17 일치하는 볼트 (중소 부품 주식 회사, 마이애미 호수, FL 3)에 의해 안정화이 알루미늄 고리 구성되어. </li><li> 두 알루미늄 고리가 파괴 당 필요합니다. 반지는 # 56분의 2 육각 너트 (작은 부품 주식 회사, 마이애미 호수, FL)과 일치하는 볼트의 적절한 구멍에 4를 조립하려면, 이들은 느슨하게 유지됩니다. 이것은 근위 반지 될 것입니다. 말초 반지 3 볼트와 너트 조립하려면 다시 완전히 조여하지 않습니다. </li></ol> 2. 마취, 파괴 창조, 그리고 Fixator 신청 <ol><li> 쥐 (연령, 성별, 그리고 몸무게는 50mg/ml 케타민을, 0.5의 intraperitoneal 주입하여 anesthetized) 사용자가되어 결정됩니다 MG / ML Medetomidine (0.03 ML / 마우스). </li><li> 생쥐의 눈에 눈 윤활제를 적용하고 절차를 수행하는 동안 눈을 보호하기 위해 자신의 머리에 걸쳐 테이프 조각을 넣으십시오. </li><li> 동물이 절차의 나머지를위한 손난로에 배치됩니다. </li><li> 고정시키다 넷 0.25 mm 멸균 곤충 핀 (Anticorro : 파인 과학 도구, 포스터 시티, CA)를 사용하여 왼쪽 경골의 근위 및 말초 metaphyses Dremel 도구에 핀을 부착하고 뼈를 통해 그들을 드릴로. </li><li> 핀은 지향 경골의 긴​​ 축에 직각, 그리고 서로 90 °입니다. 최소한 10 근위 및 말초 핀 사이의 음. </li><li> 핀을 향해 안쪽으로 향하게 견과류와 근위 핀 위의 첫 번째 고리를 둡니다. 알루미늄 링 내에서 중앙 위치에 경골을 지키는 고리에 핀을 고정합니다. </li><li> 디 아래에있는 두 번째 반지를 배치stal 가시는 예전과 육각 너트를 사용하여 고리에 핀을 고정하십시오. 말초 fixator로 네 번째 너트 볼트 조합을 추가합니다. </li><li> 해당 장치와 함께 벤딩 세 지점에서 가로 파괴를 만듭니다. 우리는 Bonnarens 및 Einhorn 4 중력 시스템을 사용합니다. 블록에 걸쳐 경골의 posterolateral 표면을 거두길 드롭 암 따라 미리 정해진 높이로 중량을 올리고 무게를 떨어뜨려 그것이 연락처 무딘 골절을 생산하는 경골의 anterolateral 측면을 말씀.에게 높이는 무게는 경골의 독점적 단일 골절을 보장하기 위해 마우스의 각 변형에 대해 결정하는 욕구에서 제외됩니다. </li><li> 골절은 X-레이에 의​​해 확인된다. </li><li> 골절을 확인한 후, 세 스테인리스 # 0 / 56 스레드 봉 고리에 붙어있다. 말초 월말부터 시작하면 매를 삽입하고 막대를 확보 말초 링을 향해 너트에 실을 꿰다. 그 다음 막대로 다른 견과류를 넣어, 근위 반지는이 N 위에 앉을 것이다UT. 두 개의 고리가 동등한 거리에서 개최되며 핀을을하지 않아서 있는지 확인하십시오. 삼분의 일 너트로 막대에 근위 링을 고정합니다. </li><li> 진통제에 대한 buprenorphine의 피하 주사 (1.0 밀리그램 / kg)은 수술 후 즉시 부여되며, 필요에 따라 동물이 고통과 주어진 buprenorphine에 대해 감시합니다. </li><li> 마취는 Atipamezole를 관리하여 반대하고 있으며, 그들이 반응하고 ambulating 때까지 동물 모니터링하고 있습니다. </li></ol> 3. 산만 Osteogenesis (참조 : 12,13)를 수용이 절차를 수정하려면 산만 osteogenesis은 간단합니다. 고리는 스레드 봉에​​ 의해 위치에 개최되며, 장소에 막대를 들고 너트를 설정하여 고리가 분리되어 이동이 가능합니다. <ol><li> 지연 :이 모델을 사용하여 표준 산만 osteogenesis 내용 fixators가 경골에 놓여진 5 일의 잠복기 동안 혼자 남겨. </li><li> 산만 : 혼란의 osteogenesis는 안정 막대 ¼ 차례 (0.25 ㎜) 매일 칠일에 너트를 회전하여 수행할 수 있습니다. </li><li> 성숙 : 나폴레옹 브리지는 10 일까지 관찰하고 완전한 통합은 혼란 완료 후 27일에 의해 관찰된다. </li></ol> 4. 임계 크기의 결함의 생성 이전 다음 수정을 설명한대로 중요한 크기의 결함을 만들려면 외부 fixators이 적용됩니다. <ol><li> 절차를 시작하기 전에 피부가 70 % 에탄올로 세척한다. </li><li> fixators는 결함을 만드는 완전히 이전에 적용됩니다. </li><li> 마우스 그러면 중간 경골을 시각화하기 위해 해부 현미경으로 배치되고, 절개 (5~7mm)이 메스에 # 10 칼날로 이루어집니다. </li><li> 근육이 부드럽게 가로와 경골의 중간 diaphysis을 폭로 구분합니다. </li><li> 뼈 3mm 세그먼트 그런 다음 중간 diaphysis 사용하는 가위에서 제거됩니다. t를 사용그는 가위 지점, 뼈가 서서히 잘라 냈다되고 뼈 조각이 필요한 경우 집게로 제거됩니다. 케어는 큰 기계적 힘으로 경골를 생기죠 피하기 위해 가위 팁으로 연결됩니다. </li><li> 이들은 치유를 촉진있을 수 있기 때문에 더 뼈 조각은 없습니다 않도록 외과 사이트를 검사합니다. </li><li> 경골의 무딘 끝을 주위 근육을 교체하십시오. </li><li> 6-0 Polyamine의 Monofilament로부터 2-3 봉합과 피부에 절개를 닫습니다. </li><li> 마취가 반전되고 동물 감시 고통과 이전에 설명된대로 진통제를 제공. </li></ol> 5. 대표 결과 제대로 적용하면 외부 fixators 뛰어난 감소 (Figs. 1, 2)과 폐쇄 골절 tibial보다 엄격한 안정성을 제공합니다. 그러나, 경우에 불충 분한 감소 (뼈 종료 또는 여러 골절 사이에 명확한 및 대형 격차가 있습니다 (그림 3 발생 </strong&gt;), 그리고 이러한 생쥐는 분석에서 제외됩니다. 골절은 주로 intramembranous 골화 (그림 4)를 통해 치료하는이 방법을 사용하여 안정. 골절이 안정화되지 않은 경우 반대로, 대형 연골에 굳은살이 파괴 격차 (그림 5)에서 형성되며, 이것은 intramembranous 골화 과정을 통해 뼈로 대체됩니다. <img alt="그림 1" src="/files/ftp_upload/3552/3552fig1.jpg" /> 그림 1. tibial 골절을 안정화하는 데 사용되는 외부 고정 장치를 설명 Radiographs. 방사선 사진은 잘 정렬된 뼈 세그먼트 (화살촉)를 보여주는 골절 후 촬영. <img alt="그림 2" src="/files/ftp_upload/3552/3552fig2.jpg" /> 그림은 2. fixator 후 마우스의 이미지가 적용되었습니다. <img alt="그림 3" src="/files/ftp_upload/3552/3552fig3.jpg" /> 그림 3. 방사선 사진에서 F 이후 이동racture는 고르지과 조각난 뼈 세그먼트 (화살촉)를 보여줍니다. <img alt="그림 4" src="/files/ftp_upload/3552/3552fig4.jpg" /> 그림 4. 안정 골절 intramembranous 골화 통해 치유. 안정 파괴의 Trichrome 염색법은 파괴 사이트에서 몇 가지 새로운 뼈 (B)를 보여줍니다. 스케일 막대 = 500 μm의. <img alt="그림 5" src="/files/ftp_upload/3552/3552fig5.jpg" /> 그림 5. 비 안정화 골절은 endochondral 골화 통해 치유. 비 안정 파괴의 Trichrome 염색법은 (b)는 파괴 사이트에서 연골 (C)와 뼈를 보여줍니다. 스케일 막대 = 500 μm의.

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우리는 공개 할게 없다.

본즈는 기계적 안정성 (14에서 검토)에 따라 서로 다른 두 modalities에 의해 치료. 골절의 두 끝을 다리를 놓으려고 뼈로 대체됩니다 파괴 격차에 불안정 큰 연골 템플릿 양식을 떠날 때. Proximally 및 distally 휴식 시간에, 뼈가 골막과 endosteum 이내 intramembranous 골화 직접 형성하고 있습니다. 대조적으로의 안정 골절의 치유가 intramembranous 골화 3를 통해 독점적으로 발생합니다. 그러나,이 ...

<table border="1"><tbody><tr><td> 시약의 이름 </td><td> 회사 </td><td> 카탈로그 번호 </td><td> 댓글 </td></tr><tr><td> 0.25mm 곤충 핀 </td><td> 파인 과학 도구 </td><td> 26000-25 </td><td> 정전 알루 스틸, 0.25mm로드 직경 4cm 길이 </td></tr><tr><td> 스테인레스 스틸 16 진수 너트 </td><td> 소형 부품 </td><td> # 2-56 </td><td> 1 / 8 &quot;길이, 인치 당 56 스레드 </td></tr><tr><td> 스테인레스 스틸 16 진수 너트 </td><td> 소형 부품 </td><td> # 0-80 </td><td> 1 / 8 &quot;길이, 인치 당 80 스레드 </td></tr><tr><td> 스테인레스 머신 스크류 </td><td> 소형 부품 </td><td> # 0-80 </td><td> 1 / 8 &quot;길이, 인치 당 80 스레드 </td></tr><tr><td> 스테인레스 스틸 기계 스레드로드를 잘라 </td><td> 소형 부품 </td><td> # 0-80 </td><td> 6 &quot;길이, 인치 당 80 스레드 </td></tr><tr><td> 18-8 스테인레스 스틸 헤드 머신 SCREW </td><td> McMaster-카 </td><td></td><td> 2-56 스레드, 6분의 3 &quot;길이 </td></tr><tr><td> 외부 고정 장치 </td><td> 기계 공장 </td><td> 맞춤 디자인 </td><td></td></tr></tbody></table>

creating rigidly stabilized fractures for assessing intramembranous ossification, distraction osteogenesis, or healing of critical sized defects

골격 수리 모드를 평가하는 것은 골절을 치료하는 임상적으로 사용되는 치료법을 개발하기 위해 필수적입니다. 기계적 안정성이 뼈 부상 치유에 큰 역할을한다. 최악의 경우에는 기계적인 불안정성은의 지연이나 노동 조합에 속하지 않은 인간으로 이어질 수 있습니다. 그러나, 운동은 또한 치유 과정을 자극 수 있습니다. 파괴 뼈 끝이 안정화 모션 연골 형태를 가지고 있고,이 연골이 서서히 endochondral 골화의 발달 과정의 요점의 반복을 통해 뼈로 대체됩니다 골절합니다. 뼈가 골절이 rigidly 안정화되면 반대로, 뼈가 intramembranous 골화 통해 직접 형성하고 있습니다. 임상적으로 두 endochondral 및 intramembranous 골화 동시에 발생합니다. 효과적으로 처리 조사관은 Bonnarens 사에 의해 설명된대로 골절 뼈 골수의 운하에 핀을 삽입 복제합니다. rotatio을 허용하면서이 실험 방법은 우수한 측면 안정성을 제공합니다nal 불안정이 지속합니다. 그러나 이러한 두 개의 프로세스를 조절 메커니즘에 대한 우리의 이해도 실험적으로 이러한 프로세스를 각각 분리하여 향상될 수 있습니다. 우리는 회전과 수평 안정화를 제공 안정화 프로토콜을 개발했습니다. 이 모델에서는 intramembranous 골화가 관찰되고 치유 전용 모드이며, 치유의 매개 변수는 치유 10의 생리적 매개 변수를 변경 후 bioactive 분자 8,9의 신청 후 5-7 유전자 변형 생쥐의 다양한 변종,,,간에 비교할 수 산만 osteogenesis 12 이후 안정화 11 금액이나 시간을 수정 후 노동 조합에 속하지 않은 13 생성 후, 또는 임계 크기의 결함 생성 후. 여기서는 생쥐의 tibial 골절 치료와 산만 osteogenesis 공부를 위해 수정된 Ilizarov의 fixators을 적용하는 방법을 보여줍니다.

Watch this Scientific Journal Video about Creating Rigidly Stabilized Fractures for Assessing Intramembranous Ossification, Distraction Osteogenesis, or Healing of Critical Sized Defects at JoVE.com

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

이 문서는 수정된 Ilizarov 외부의 fixators의 적용을 기반으로 긴 뼈 골절을 안정화하는 방법을 설명합니다<sup&gt; 1-3</sup&gt;. fixators과 뼈가 손상 창조 신청 후, 치유가 과세 수 산만 osteogenesis가 수행될 수도 있고, 노동 조합에 속하지 않은, 주요 크기의 결함이 만들어지고 치료 중재를 공부하는 데 사용할 수 있습니다.

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 ...

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JoVE Journal

Medicine

17.3K 조회수.  University of California, San Francisco. 이 문서는 수정된 Ilizarov 외부의 fixators의 적용을 기반으로 긴 뼈 골절을 안정화하는 방법을 설명합니다 1-3. fixators과 뼈가 손상 창조 신청 후, 치유가 과세 수 산만 osteogenesis가 수행될 수도 있고, 노동 조합에 속하지 않은, 주요 크기의 결함이 만들어지고 치료 중재를 공부하는 데 사용할 수 있습니다.

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

Repair of a Critical-sized Calvarial Defect Model Using Adipose-derived Stromal Cells Harvested from Lipoaspirate

Craniofacial skeletal repair and regeneration offers the promise of de novo tissue formation through a cell-based approach utilizing stem cells. Adipose-derived stromal cells (ASCs) have proven to be an abundant source of multipotent stem cells capable of undergoing osteogenic, chondrogenic, adipogenic, and myogenic differentiation. Many studies have explored the osteogenic potential of these cells in vivo with the use of various scaffolding biomaterials for cellular delivery. It has been demonstrated that by utilizing an osteoconductive, hydroxyapatite-coated poly(lactic-co-glycolic acid) (HA-PLGA) scaffold seeded with ASCs, a critical-sized calvarial defect, a defect that is defined by its inability to undergo spontaneous healing over the lifetime of the animal, can be effectively show robust osseous regeneration. This in vivo model demonstrates the basis of translational approaches aimed to regenerate the bone tissue - the cellular component and biological matrix. This method serves as a model for the ultimate clinical application of a progenitor cell towards the repair of a specific tissue defect.

This protocol describes the isolation of adipose-derived stromal cells from lipoaspirate and the creation of a 4 mm critical-sized calvarial defect to evaluate skeletal regeneration.

Lipoaspirate에서 수확 지방 - 파생 Stromal 셀을 사용하여 임계 크기 Calvarial 결함 모델의 수리

Craniofacial skeletal repair and regeneration offers the promise of de novo tissue formation through a cell-based approach utilizing stem cells. ...

연구

의학

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

상처 치유의 생쥐 모델

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

Assessing Changes in Volatile General Anesthetic Sensitivity of Mice after Local or Systemic Pharmacological Intervention

One desirable endpoint of general anesthesia is the state of unconsciousness, also known as hypnosis. Defining the hypnotic state in animals is less straightforward than it is in human patients. A widely used behavioral surrogate for hypnosis in rodents is the loss of righting reflex (LORR), or the point at which the animal no longer responds to their innate instinct to avoid the vulnerability of dorsal recumbency. We have developed a system to assess LORR in 24 mice simultaneously while carefully controlling for potential confounds, including temperature fluctuations and varying gas flows. These chambers permit reliable assessment of anesthetic sensitivity as measured by latency to return of the righting reflex (RORR) following a fixed anesthetic exposure. Alternatively, using stepwise increases (or decreases) in anesthetic concentration, the chambers also enable determination of a population's sensitivity to induction (or emergence) as measured by EC50 and Hill slope. Finally, the controlled environmental chambers described here can be adapted for a variety of alternative uses, including inhaled delivery of other drugs, toxicology studies, and simultaneous real-time monitoring of vital signs.

Loss of the righting reflex has long served as a standard behavioral surrogate for unconsciousness, also called hypnosis, in laboratory animals. Alterations in volatile anesthetic sensitivity caused by pharmacological interventions can be detected with a carefully controlled high-throughput assessment system, which may be adapted for delivery of any inhaled therapeutic.

One desirable endpoint of general anesthesia is  the state of unconsciousness, also known as hypnosis. Defining the hypnotic state in animals is  less ...

Minimally Invasive Thumb-sized Pterional Craniotomy for Surgical Clip Ligation of Unruptured Anterior Circulation Aneurysms

Less invasive surgical approaches for intracranial aneurysm clipping may reduce length of hospital stay, surgical morbidity, treatment cost, and improve patient outcomes. We present our experience with a minimally invasive pterional approach for anterior circulation aneurysms performed in a major tertiary cerebrovascular center and compare the results with an aged matched dataset from the Nationwide Inpatient Sample (NIS). From August 2008 to December 2012, 22 elective aneurysm clippings on patients &#8804;55 years of age were performed by the same dual fellowship-trained cerebrovascular/endovascular neurosurgeon. One patient (4.5%) experienced transient post-operative complications. 18 of 22 patients returned for follow-up imaging and there were no recurrences through an average duration of 22 months. A search in the NIS database from 2008 to 2010, also for patients aged &#8804;55 years of age, yielded 1,341 hospitalizations for surgical clip ligation of unruptured cerebral aneurysms. Inpatient length of stay and hospital charges at our institution using the minimally invasive thumb-sized pterional technique were nearly half that of NIS (length of stay: 3.2 vs 5.7 days; hospital charges: $52,779 vs. $101,882). The minimally invasive thumb-sized pterional craniotomy allows good exposure of unruptured small and medium-sized supraclinoid anterior circulation aneurysms. Cerebrospinal fluid drainage from key subarachnoid cisterns and constant bimanual microsurgical techniques avoid the need for retractors which can cause contusions, localized venous infarctions, and post-operative cerebral edema at the retractor sites. Utilizing this set of techniques has afforded our patients with a shorter hospital stay at a lower cost compared to the national average.

Minimally invasive thumb-sized pterional craniotomy for aneurysm clipping has afforded our patients with a shorter hospital stay at a lower cost compared to the national average.

최소 침습 엄지 손가락 크기의 Pterional 개두술을 파열 전방 순환 동맥류의 수술 클립 결찰에 대한

Less invasive surgical approaches for intracranial aneurysm clipping may reduce length of hospital stay, surgical morbidity, treatment cost, and improve ...

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 ...

Direct Mouse Trauma/Burn Model of Heterotopic Ossification

Heterotopic ossification (HO) is the formation of bone outside of the skeleton which forms following major trauma, burn injuries, and orthopaedic surgical procedures. The majority of animal models used to study HO rely on the application of exogenous substances, such as bone morphogenetic protein (BMP), exogenous cell constructs, or genetic mutations in BMP signaling. While these models are useful they do not accurately reproduce the inflammatory states that cause the majority of cases of HO. Here we describe a burn/tenotomy model in mice that reliably produces focused HO. This protocol involves creating a 30% total body surface area partial thickness contact burn on the dorsal skin as well as division of the Achilles tendon at its midpoint. Relying solely on traumatic injury to induce HO at a predictable location allows for time-course study of endochondral heterotopic bone formation from intrinsic physiologic processes and environment only. This method could prove instrumental in understanding the inflammatory and osteogenic pathways involved in trauma-induced HO. Furthermore, because HO develops in a predictable location and time-course in this model, it allows for research to improve early imaging strategies and treatment modalities to prevent HO formation.

An Achilles tenotomy and burn injury model of heterotopic ossification allows for the reliable study of trauma induced ectopic bone formation without the application of exogenous factors.

직접 마우스 외상 / 이소성 골화의 모델을 굽기

Heterotopic ossification (HO) is the formation of bone outside of the skeleton which forms following major trauma, burn injuries, and orthopaedic surgical ...

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 ...

Scleral Cross-linking Using Riboflavin and Ultraviolet-A Radiation for Prevention of Axial Myopia in a Rabbit Model

Myopic individuals, especially those with severe myopia, are at higher-than-normal risk of cataract, glaucoma, retinal detachment and chorioretinal abnormalities. In addition, pathological myopia is a common irreversible cause of visual impairment and blindness1-3. Our study demonstrates the effect of scleral crosslinking using riboflavin and ultraviolet-A radiation on the development of axial myopia in a rabbit model. The axial length of the eyeball was measured by A-scan ultrasound in New Zealand white rabbits aged 13 days (male and female). The eye then underwent 360&#176; conjunctival peritomy with scleral crosslinking, followed by tarsorrhaphy. Axial elongation was induced in 13 day-old New Zealand rabbits by suturing their right eye eyelids (tarsorrhaphy). The eyes were divided into quadrants, and every quadrant had two scleral irradiation zones, each with an area of 0.2 cm&#178; and a radius of 4 mm. Crosslinking was performed by dropping 0.1% dextran-free riboflavin-5-phosphate onto the irradiation zones 20 sec before ultraviolet-A irradiation and every 20 sec during the 200 sec irradiation time. UVA radiation (370 nm) was applied perpendicular to the sclera at 57 mW/cm&#178; (total UVA light dose, 57 J/cm&#178;). Tarsorrhaphies were removed on day 55, followed by repeated axial length measurements. This study demonstrates that scleral crosslinking with riboflavin and ultraviolet-A radiation effectively prevents occlusion-induced axial elongation in a rabbit model.

We demonstrate the effect of scleral crosslinking with riboflavin and UVA on an axial elongation rabbit eye. Axial elongation was induced in 13 day-old New Zealand rabbits (male and female) by suturing their right eye eyelids (tarsorrhaphy).

공막 래빗 모델에 축 방향 근시의 예방 리보플라빈 및 자외선-A 방사선을 사용하여 교차 결합

Myopic individuals, especially those with severe myopia, are at higher-than-normal risk of cataract, glaucoma, retinal detachment and chorioretinal ...

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 ...

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

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