This work was supported by a Wisconsin Partnership Program New Investigator Award (ID 2832), a National Heart, Lung, Blood Institute R01 Grant (HL-068673) and a T32 training Grant (HL-110853). We thank Dr. Melina Kibbe&#8217;s group at Northwestern University for providing helpful information.

참고 : 윤리 문 : 모든 절차는 위스콘신 대학에서 기관 동물 케어 및 사용위원회에 의해 승인 된 실험 동물의 관리 및 사용을위한 설명서 (국립 아카데미 프레스, 8 판, 2011), 및 프로토콜을 준수합니다. 모든 수술 (유량 2 mL / 분, 흡입을 통해) 이소 플루 란 마취 하에서 수행하고, 모든 노력은 고통을 최소화 하였다. 동물은 점차적으로 CO 2로 채워진 챔버에서 안락사시켰다. 전신 마취 및 피부 절개 1. 유도 <ol><li> 23-28g의 12-16주의 나이와 무게 남성 C57BL / 6 마​​우스를 사용합니다. 폐회로 마취 장치를 통해 이소 플루 란으로 마취 동물. 그 발가락을 곤란하게하여 적절한 마취를 확인합니다. 건조 함을 방지하기 위해 눈에 일부 인공 눈물 연고를 적용합니다. </li><li> 약간 납치 왼쪽 뒷다리와 무릎 관절 SL와 부정사 위치에 마우스를 놓습니다ightly 근육이 수축. 왼쪽 허벅지 지역 주변의 머리를 면도하고 클로르헥시딘으로 피부를 소독, 멸균면 팁 어플리케이터를 사용하여 세 번인가. 수술 중 멸균 악기를 유지하기 위해 뜨거운 구슬 살균기를 사용합니다. </li><li> 왼쪽 다리를 통해 멸균 거즈 드레이프를 놓습니다. 사타구니 (그림 1A)에 무릎 관절에서 왼쪽 허벅지 중앙에 1.5 cm 직선, 세로 피부 절개를합니다. </li></ol> 대퇴 동맥과 나뭇 가지 2. 노출 주 : 그림 1B는 허벅지에 총 혈관 해부학을 보여줍니다. 피부 절개가 제대로 허벅지 중앙의 정중선을 따라 이루어질 때 표재성 대퇴 동맥 (SFA)를 해부 현미경으로 관찰 할 수있다. <ol><li> 미세 핀셋 다음 루프 9-0 나일론 봉합사와 SFA와 SFA를 덮는 얇은 근막을 엽니 다. 주 :이 공정에서, 대퇴 신경 용이로두고그 벽은 쉽게 관통하고 및 출혈 너무 얇은 때문에 IDE,하지만 인접 대퇴 정맥은주의가 필요합니다. 키 기술은 동맥 자체에 집중할 수있다; 한 절개가 가까운 동맥에 수행 될 때, 실수로 정맥의 손상을 피할 수있다. </li><li> SFA가 반복되면, 모기 집게와 봉합을 잡고 약간 아래쪽으로 당깁니다. 이 기동 쉽게 추가 해부한다,하지만 너무 무리하게 잡아 당기지 마십시오 동맥의 손상을 방지 할 수 있습니다. </li><li> 근위 9-0 나일론 봉합사와 대퇴 동맥 (CFA)를 노출 및 루프하는 동맥을 해부 계속합니다. 참고 : 일반적으로 상복부 동맥 (그림 1B)에 점 말단에서 CFA를 반복하는 와이어 삽입을위한 충분하다. 같은 방법으로 모기 집게로 CFA 봉합 루프를 잡습니다. </li><li> 위로 CFA 봉합 루프를 당겨 SFA 봉합 루프는 아래 SFA과 깊은 대퇴 동맥 (DFA)의 분기점을 발견 할 수 있습니다. DFA, CFA의 주요 지점,허벅지 근육 (그림 1B)를 향해 간다. 주 : 와이어 나중에 DFA에서 삽입됩니다. </li><li> 특별한주의와 루프 DFA는 인접한 정맥은 동맥 아래에 얽힌되기 때문이다. 정맥 출혈 발생시, 거즈로 30 내지 60 초 간단한 압축은 일반적으로 지혈을 제조하기에 충분하다. </li><li> DFA는 9-0 나일론 봉합사 반복되면, 세 봉합 루프 (그림 2A) 올바른 위치에 있는지 확인하십시오. </li><li> 말단 가능한 한 분기에서 지점에서 동맥을 결찰하는 DFA의 해부를 진행합니다. 여분의 봉합사를 절단하지만 양단은 DFA는 전선을 삽입하기 쉽게 안정화 모기 겸자로 유지 될 수 있도록 일정 길이를 남겨. </li></ol> 3. 와이어 부상 만들기 <ol><li> DFA를 통해 와이어를 삽입 <ol><li> 임시 지혈을 설정하는 CFA, SFA, 및 DFA 봉합 스트레인. </li><li> DF에 작은 구멍을 확인microscissors와. 0.015 인치 직경을 갖는 와이어 충분히 큰 구멍을 팽창하는 곡면 microtweezers의 선단을 삽입 (도 2b는 동맥과 와이어 간의 크기 차이를 보여준다). <ol><li> DFA에 구멍을하기 전에, CFA 봉합 루프를 풀고 부드럽게 클램프 / 무딘 팁 microtweezers와 DFA를 여러 번 짠다. 이 기동 와이어 삽입을 용이하게하기 위해 현저 동맥 직경이 확장된다. PBS로 철저하게 선을 습기를하면 부드러운 삽입을 위해 중요하다. </li><li> DFA에 구멍을 만들 때, 동맥 벽을자를 수 있지만, 작은 V 자 모양의 닙을 확인하고 조작을위한 뾰족한 microtweezers와 플랩을 끼이지 않습니다. </li></ol></li><li> DFA에 적절하게-팽창 구멍을 통해 와이어를 삽입합니다. CFA 주위 봉합 루프를 완화하면서 조심스럽게 장골 동맥으로 와이어를 진행합니다. 5-8mm가 대부분의 경우에 모든 CFA 길이를 커버하기에 충분하다 (도 2C </strong&gt;). 동맥 천공은 보통 마우스의 손실을 초래으로 강제로 전선을 삽입하지 마십시오. 길이 약 15 mm에 와이어를 단축하는 것은 처리에 도움이됩니다. </li></ol></li><li> 최소 1 내부에 와이어를 남겨 <ol><li> 최적의 길이로 와이어를 삽입 한 후, 동맥 벽을 잡아 늘 1 분 정도 둡니다. 건조를 방지하기 위해 PBS로 와이어 및 동맥을 적셔. 또 9-0 나일론 봉합사이 대기 시간 동안 와이어 삽입 지점에 인접한 DFA 주위에 배치 될 수있다. 이 봉합 와이어 제거 다음 DFA에 구멍을 닫습니다 나중에 사용됩니다. </li></ol></li><li> 와이어를 검색하고 혈액의 흐름을 다시 시작 <ol><li> 선박 삽관 집게와 와이어를 잡고 그것을 조심스럽게 당겨; 와이어가 원활하게 검색 할 수 있어야합니다. 부드러운 회전 전선을 후퇴, 일반적으로 동맥 벽에 와이어의 부착으로 인한 저항을 극복하는 것은 도움이된다. 또, 강력한 기동 동물 손실이 발생할 수 있습니다. </li><li>와이어를 검색 따고는 9-0 나일론 봉합사와 분기의 시점에서 DFA를 결찰. SFA에 혈액 유출을 위태롭게 분기, 너무 가까이 결찰하지 마십시오. </li><li> 정상적인 혈액의 흐름을 재개 SFA와 CFA 봉합 루프를 풉니 다. CFA 와이어 삽입 (그림 2D)에 의해 잘 확장합니다. </li></ol></li></ol> 4. 피부 폐쇄 및 수술 후 케어 <ol><li> 제어 출혈. 마이너 돌아 다니며 간단한 압축에 의해 중단 될 수있다 몇 가지 주요 동맥 출혈은 11-0 나일론 봉합사와 바늘이 필요합니다. 이 봉합 동맥은 결국 동맥 폐색 발생합니다. </li><li> 피부 스테이플러 (보통 3-4 스테이플)와 피부 창상을 닫습니다. 클로르헥시딘 수술 솔루션을 적용하여 멸균 상처 유지하십시오. </li><li> 수술 후 치료를 적용합니다. 부 프레 노르 핀의 피하 주사와 마우스의 고통을 제어 할 수 있습니다. 이 SUF을 회복 할 때까지 온난화 패드에 마우스를 유지흉골 드러 누움을 유지하기 위해 의식을 ficient. 다른 동물의 회사가없는 깨끗한 케이지의 집이 완전히 회복 될 때까지. </li></ol>

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The authors declare no competing financial interests.

와이어 부상 절차는, 우리가 12~16주의 연령 수컷 C57BL / 6 마우스를 사용하는만큼 그 해부학 적 구조가 유사한 된 바와 같다.이 논문 4 생쥐 균주 모두에 적용 가능하다. 우리가 우리의 예비 연구에서 배운 바와 같이 10 주 미만 쥐의 대퇴 동맥은 종종 와이어 삽입은 매우 도전 너무 작습니다. 한편, 16주보다 이전 마우스에서 와이어 삽입 기술적 쉽지만 이상의 가변 신생 내막 과형성 초래하는...

In the era of expanded application of endovascular treatments for various cardiovascular diseases, restenosis after angioplasty is one of the major problems for patients undergoing such treatments. Damage to the vascular endothelium at the time of angioplasty, in concert with the atherosclerotic background, induces excessive smooth muscle cell proliferation in the medial layer, resulting in neointimal hyperplasia.1,2 A viable animal model that recapitulates post-angioplasty neointimal hyperplasia is, thus, important not only for the investigation of disease mechanisms but also for the development of effective therapeutics to treat this pathology.
Mice represent an excellent model animal to recapitulate neointimal hyperplasia for the following reasons: the genetic backgrounds of experimental mice are well established; a wide variety of genetically-modified strains are available;2 obtaining littermates of the same background is easy; and the cost of the animals is relatively low. Arterial ligation model and wire injury model are the two most common mouse models of mechanically-induced neointimal hyperplasia. The arterial ligation model is easy to create, but physiologically dissimilar to the actual angioplasty procedure. The wire injury model closely mimics actual angioplasty procedures but is technically difficult due to the small size of mouse arteries.2,3 Sata et al. first described a wire injury method for mouse femoral arteries based on the anatomical structure of the vasculature and the use of proper-sized flexible wire. Utilizing this technique, they succeeded in reproducibly inducing neointimal hyperplasia in various strains of mice.4
Although femoral wire injury is a well-established model, some of the technical aspects of the technique are highly challenging compared to other models such as ligation.5 The purpose of this paper is to describe our mouse wire injury model procedures in detail, which is a modified version of Sata&#8217;s original method. We have made two main modifications: 1) Looping only the arteries, and 2) No lidocaine use.

<table><tbody><tr><td>Fixed Core Wire Guide</td><td>Cook</td><td>G02426</td><td>Diameter 0.015 inch, Straight</td></tr><tr><td>Dilation Forceps</td><td>Roboz</td><td>RS-4927</td><td>Curved, blunt tips</td></tr><tr><td>Dumont Tweesers #5</td><td>World Precision Instruments</td><td>14095</td><td>Straight, sharp tips</td></tr><tr><td>Dumont Vessel Cannulation Forceps</td><td>World Precision Instruments</td><td>503373</td><td></td></tr><tr><td>McPherson-Vannas Scissors</td><td>World Precision Instruments</td><td>501234</td><td></td></tr><tr><td>Mosquito Forceps</td><td>World Precision Instruments</td><td>501291</td><td></td></tr><tr><td>Ethilon Nylon Suture 9-0</td><td>Ethicon</td><td>7717G</td><td>9-0, Black Nylon Monofilament</td></tr><tr><td>Micro AROSuture, Sterile 11-0, 70 micrometers, MET Point</td><td>AROSurgical</td><td>VT4A00N07</td><td>11-0, Black Nylon Monofilament</td></tr><tr><td>3M Precise Multi-Shot DS Disposable Skin Stapler</td><td>3M</td><td>DS-25</td><td></td></tr></tbody></table>

a murine model of arterial restenosis: technical aspects of femoral wire injury

아테롬성 동맥 경화증으로 인한 심장 혈관 질환은 선진국에서 사망의 주요 원인이다. 동맥 경화 반으로 인해 개발 성립 플라크 파열로, 혈관 내강 좁아, 심근 경색이나 뇌졸중 등의 각종 합병증 선도 정상적인 혈류를 중단. 이러한 혈관 성형술과 같은 병원 혈관 절차에서 일반적으로 루멘을 다시 수행한다. 그러나, 이러한 치료법은 필연적 과도한 치유 반응과 혈관 재 협착 (재 협착)을 일으키는 루멘 내로 연장 내막 플라크의 발전을 유발, 혈관 벽뿐만 아니라, 혈관 내피 세포를 손상. 재 협착은 동맥 경화 혈관 내 치료 실패의 주요 원인이 남아있다. 따라서, 재 협착의 전임상 동물 모델은 병진 혈관 개입 접근법뿐만 아니라 병리 생리 학적 메카니즘을 조사하는 매우 중요하다. 여러 쥐 experime 중이 밀접하게 내피 세포와 혈관 벽을 모두 상해를당한 혈관 성형술 절차를 닮았다고 ntal 모델은 대퇴 동맥 와이어 부상 널리 후 혈관 성형술 재 협착의 연구에 가장 적합한으로 허용됩니다. 그러나, 많은 연구자들은 기술적 난이도 높은 수준으로이 모델을 사용하는 어려움이있다. 이 금속 선재가 눈에 띄는 신생 내막을 유도하기에 충분한 손상을 생성하기 위해, 약 와이어보다 3 배 더 얇은 대퇴 동맥에 삽입 될 필요가 있기 때문에 주로이다. 여기서 우리는 효율적으로이 모델의 주요 기술적 인 어려움을 극복하기 위해 필수적인 외과 세부 사항을 설명합니다. , 제시된 절차에 따라 수행함으로써 마우스 대퇴 동맥 와이어 부상이 쉽게된다. 친숙하면 전체 공정을 20 분 내에 완료 될 수있다.

수술을 다음과 같은 네 가지 주, 와이어 부상 대퇴 동맥을 수확하고 신생 내막 형성을 분석하는 파라핀 블록 표본을합니다. 상술 한 바와 같이 전신 마취를 유도한다. 바로 가슴을 열고 좌심실에 20 게이지 나비 바늘을 삽입합니다. 주입 병에 선을 연결합니다. 4 % 파라 포름 알데히드 (일반적으로 10 ~ 20 ml의 각) 다음 PBS의 점적 주입을 통해 관류 고정을 수행합니다. 동물은 다음 와이어 부상 C...

Watch this Scientific Journal Video about A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury at JoVE.com

A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury

재 협착 마우스 대퇴 동맥 와이어 손상 모델 기술적 도전이다. 이 프로토콜에서 우리는 성공적으로 재 협착의 연구에 대한 일관성있는 신생 내막을 유도하는 와이어 부상을 수행하기위한 주요 기술 세부 사항은 필수 보여줍니다.

동맥 재 협착의 쥐 모델 : 대퇴 와이어 부상의 기술적 측면

Cardiovascular disease caused by atherosclerosis is the leading cause of death in the developed world. Narrowing of the vessel lumen, due to ...

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Research

JoVE Journal

Medicine

20.1K 조회수.  University of Wisconsin School of Medicine and Public Health.  재 협착 마우스 대퇴 동맥 와이어 손상 모델 기술적 도전이다. 이 프로토콜에서 우리는 성공적으로 재 협착의 연구에 대한 일관성있는 신생 내막을 유도하는 와이어 부상을 수행하기위한 주요 기술 세부 사항은 필수 보여줍니다. 

비디오: A Murine Model of Arterial Restenosis: Technical Aspects of Femoral Wire Injury

Murine Spinotrapezius Model to Assess the Impact of Arteriolar Ligation on Microvascular Function and Remodeling

The murine spinotrapezius is a thin, superficial skeletal support muscle that extends from T3 to L4, and is easily accessible via dorsal skin incision. Its unique anatomy makes the spinotrapezius useful for investigation of ischemic injury and subsequent microvascular remodeling. Here, we demonstrate an arteriolar ligation model in the murine spinotrapezius muscle that was developed by our research team and previously published1-3. For certain vulnerable mouse strains, such as the Balb/c mouse, this ligation surgery reliably creates skeletal muscle ischemia and serves as a platform for investigating therapies that stimulate revascularization. Methods of assessment are also demonstrated, including the use of intravital and confocal microscopy. The spinotrapezius is well suited to such imaging studies due to its accessibility (superficial dorsal anatomy) and relative thinness (60-200 &mu;m). The spinotrapezius muscle can be mounted en face, facilitating imaging of whole-muscle microvascular networks without histological sectioning. We describe the use of intravital microscopy to acquire metrics following a functional vasodilation procedure; specifically, the increase in arterilar diameter as a result of muscle contraction. We also demonstrate the procedures for harvesting and fixing the tissues, a necessary precursor to immunostaining studies and the use of confocal microscopy.

We demonstrate a novel arterial ligation model in murine spinotrapezius muscle, including a step-by-step procedure and description of required instrumentation. We describe the surgery and relevant outcome measurements relating to vascular network remodeling and functional vasodilation using intravital and confocal microscopy.

Microvascular 기능 및 리모델링에 Arteriolar 결합의 영향을 평가하기 위해 손쥐 Spinotrapezius 모델

The murine spinotrapezius is a thin, superficial skeletal support muscle that extends from T3 to L4, and is easily accessible via dorsal skin incision. ...

연구

의학

A Murine Model of Stent Implantation in the Carotid Artery for the Study of Restenosis

Despite the considerable progress made in the stent development in the last decades, cardiovascular diseases remain the main cause of death in western countries. Beside the benefits offered by the development of different drug-eluting stents, the coronary revascularization bears also the life-threatening risks of in-stent thrombosis and restenosis. Research on new therapeutic strategies is impaired by the lack of appropriate methods to study stent implantation and restenosis processes. Here, we describe a rapid and accessible procedure of stent implantation in mouse carotid artery, which offers the possibility to study in a convenient way the molecular mechanisms of vessel remodeling and the effects of different drug coatings.

A model of stent implantation in mouse carotid artery is described. Compared to other similar methods, this procedure is very rapid, simple and accessible, offering the possibility to study in a convenient way the vascular wall reaction to different drug-eluting stents and the molecular mechanisms of restenosis.

재 협착의 연구에 대한 경동맥 스텐트 시술의 쥐과 모형

Despite  the considerable progress made in the stent development in the last decades,  cardiovascular diseases remain the main cause of death in western ...

Generation and 3-Dimensional Quantitation of Arterial Lesions in Mice Using Optical Projection Tomography

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating important information on the pathogenesis of lesion formation and the action of novel therapies. Use of atherosclerosis-prone mice, surgical methods of lesion induction, and dietary modification has dramatically improved understanding of the mechanisms that contribute to disease development and the potential of new treatments.
Classically, analysis of lesions is performed ex vivo using 2-dimensional histological techniques. This article describes application of optical projection tomography (OPT) to 3-dimensional quantitation of arterial lesions. As this technique is non-destructive, it can be used as an adjunct to standard histological and immunohistochemical analyses.
Neointimal lesions were induced by wire-insertion or ligation of the mouse femoral artery whilst atherosclerotic lesions were generated by administration of an atherogenic diet to apoE-deficient mice.
Lesions were examined using OPT imaging of autofluorescent emission followed by complementary histological and immunohistochemical analysis. OPT clearly distinguished lesions from the underlying vascular wall. Lesion size was calculated in 2-dimensional sections using planimetry, enabling calculation of lesion volume and maximal cross-sectional area. Data generated using OPT were consistent with measurements obtained using histology, confirming the accuracy of the technique and its potential as a complement (rather than alternative) to traditional methods of analysis.
This work demonstrates the potential of OPT for imaging atherosclerotic and neointimal lesions. It provides a rapid, much needed ex vivo technique for the routine 3-dimensional quantification of vascular remodelling.

Ex vivo analysis of arterial lesions from animal models of cardiovascular disease classically relies on histological and immunohistochemical techniques. These provide 2-dimensional measurements in 3-dimensional lesions. This manuscript describes the generation of arterial lesions for quantitative analysis in 3-dimensions using optical projection tomography.

생성 및 마우스의 동맥 병변의 3 차원 정량 광학 프로젝션 단층 촬영을 사용하여

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating ...

A New Murine Model of Endovascular Aortic Aneurysm Repair

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of Abdominal aortic aneurysm (AAA) pathophysiology. There is no abdominal aortic aneurysm endograft exclusion model cheap and reproducible, which would allow deep investigations of AAA before and after treatment. We hereby describe how to induce, and then to exclude with a covered coronary stentgraft an abdominal aortic aneurysm in a rat. The well known elastase induced AAA model was first reported in 19901 in a rat, then described in mice2. Elastin degradation leads to dilation of the aorta with inflammatory infiltration of the abdominal wall and intra luminal thrombus, matching with human AAA. Endovascular exclusion with small covered stentgraft is then performed, excluding any interactions between circulating blood and the aneurysm thrombus. Appropriate exclusion and stentgraft patency is confirmed before euthanasia by an angiography thought the left carotid artery. Partial control of elastase diffusion makes aneurysm shape different for each animal. It is difficult to create an aneurysm, which will allow an appropriate length of aorta below the aneurysm for an easy stentgraft introduction, and with adequate proximal and distal neck to prevent endoleaks. Lots of failure can result to stentgraft introduction which sometimes lead to aorta tear with pain and troubles to stitch it, and endothelial damage with post op aorta thrombosis. Giving aspirin to rats before stentgraft implantation decreases failure rate without major hemorrhage. Clamping time activates neutrophils, endothelium and platelets, and may interfere with biological analysis.

Histological and biochemical modifications after aneurysm endograft exclusion are still unclear. We describe a new model of endograft implantation on a murine aneurysm. Through thrombus analysis with and without persistant circulating blood stream interface, and new endovascular biomaterials evaluation, this model will help to better understand endovascular aneurysm exclusion pathobiology.

혈관 동맥류 수리의 새로운 쥐과 모델

Endovascular aneurysm exclusion is a validated technique to prevent aneurysm rupture. Long-term results highlight technique limitations and new aspects of ...

Murine Model of Femoral Artery Wire Injury with Implantation of a Perivascular Drug Delivery Patch

Percutaneous interventions including balloon angioplasty and stenting have been used to restore blood flow in vessels with occlusive vascular disease. While these therapies lead to the rapid restoration of blood flow, these technologies remain limited by restenosis in the case of bare metal stents and angioplasty, or reduced healing and possibly enhanced risk of thrombosis in the case of drug eluting stents. A key pathophysiological mechanism in the formation of restenosis is intimal hyperplasia caused by the activation of vascular smooth muscle cells and inflammation due to arterial stretch and injury. Surgeries that induce arterial injury in genetically modified mice are useful for the mechanistic study of the vascular response to injury but are often technically challenging to perform in mouse models due to the their small size and lack of appropriate sized devices. We describe two approaches for a surgical technique that induces endothelial denudation and arterial stretch in the femoral artery of mice to produce robust neointimal hyperplasia. The first approach creates an arteriotomy in the muscular branch of the femoral artery to obtain vascular access. Following wire injury this arterial branch is ligated to close the arteriotomy. A second approach creates an arteriotomy in the main femoral artery that is later closed through localized cautery. This method allows for vascular access through a larger vessel and, consequently, provides a less technically demanding procedure that can be used in smaller mice. Following either method of arterial injury, a degradable drug delivery patch can be placed over or around the injured artery to deliver therapeutic agents.

We describe a surgical technique that produces wire injury in the femoral artery of mice to induce neointimal hyperplasia to serve as a model testing system for the perivascular delivery of therapeutic compounds for the inhibition of restenosis.

혈관 주위 약물 전달 패치의 주입과 대퇴 동맥 와이어 부상의 쥐 모델

Percutaneous interventions including balloon angioplasty and stenting have been used to restore blood flow in vessels with occlusive vascular disease. ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. Moreover, by rupture of the defective vascular wall, atherosclerosis induces occlusive thrombus formation, which represents the main cause of myocardial infarction or stroke and the most frequent cause of death. Despite the advances in the cardiovascular field, many questions remain unanswered, and additional basic research is essential to improve our understanding of the molecular mechanisms during atherosclerosis and its effects. Due to limited clinical studies, there is a need for representative animal models recreating atherosclerotic conditions such as neointima formation after stent implantation, balloon angioplasty, or endarterectomy. Since the mouse presents many advantages and is the most frequently used model for studying molecular processes, the current study proposes an invasive procedure of endothelial denudation, also known as the wire-injury model, which is representative of the human condition of neointima formation in arteries after revascularization procedures.

This study describes an invasive procedure for the induction of accelerated atherosclerosis in mice. In comparison to other methods using electric- or cryo-induced injury, mechanical-induced injury mimics the human condition of restenosis after revascularization therapies and is ideal for the study of the molecular mechanisms involved.

마우스에서 가속 죽상 경화증의 유도: "와이어 부상" 모델

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

Reproducible Arterial Denudation Injury by Infrarenal Abdominal Aortic Clamping in a Murine Model

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These complications can be attributed to impairments in re-endothelialization within the wound margins. Yet, the cellular and molecular mechanisms of re-endothelialization remain to be defined. While several animal models to study re-endothelialization after arterial denudation are available, few are performed in the mouse because of surgical limitations. This undermines the opportunity to exploit transgenic mouse lines and investigate the contribution of specific genes to the process of re-endothelialization. Here, we present a step-by-step protocol for creating a highly reproducible murine model of arterial denudation injury in the infrarenal abdominal aorta using external vascular clamping. Immunocytochemical staining of injured aortas for fibrinogen and &#946;-catenin demonstrate the exposure of a pro-thrombotic surface and the border of intact endothelium, respectively. The method presented here has the advantages of speed, excellent overall survival rate, and relative technical ease, creating a uniquely practical tool for imposing arterial denudation injury in transgenic mouse models. Using this method, investigators may elucidate the mechanisms of re-endothelialization under normal or pathological conditions.

Understanding the cellular and molecular mechanisms of re-endothelialization following arterial denudation injury is of paramount importance in preventing thrombosis and restenosis of arteries. Here we describe a protocol for reproducible arterial denudation injury of the infrarenal abdominal aorta. The procedure was developed to investigate the underlying mechanisms that regulate endothelial regeneration using mouse models.

쥐 모델에서 Infrarenal 복부 대동맥 클램핑에 의해 재현 동맥 발가 벗김 부상

Percutaneous vascular interventions uniformly result in arterial denudation injuries that subsequently lead to thrombosis and restenosis. These ...

A Rat Carotid Artery Pressure-Controlled Segmental Balloon Injury with Periadventitial Therapeutic Application

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the luminal surface area in arteries thereby reducing adequate blood flow to organs and distal tissues. Clinically, revascularization procedures such as balloon angioplasty with or without stent placement aim to restore blood flow. Although these procedures reestablish blood flow by reducing plaque burden, they damage the vessel wall, which initiates the arterial healing response. The prolonged healing response causes arterial restenosis, or re-narrowing, ultimately limiting the long-term success of these revascularization procedures. Therefore, preclinical animal models are integral for analyzing the pathophysiological mechanisms driving restenosis, and provide the opportunity to test novel therapeutic strategies. Murine models are cheaper and easier to operate on than large animal models. Balloon or wire injury are the two commonly accepted injury modalities used in murine models. Balloon injury models in particular mimic the clinical angioplasty procedure and cause adequate damage to the artery for the development of restenosis. Herein we describe the surgical details for performing and histologically analyzing the modified, pressure-controlled rat carotid artery balloon injury model. Additionally, this protocol highlights how local periadventitial application of therapeutics can be used to inhibit neointimal hyperplasia. Lastly, we present light sheet fluorescence microscopy as a novel approach for imaging and visualizing the arterial injury in three-dimensions.

The rat carotid artery balloon injury mimics the clinical angioplasty procedure performed to restore blood flow in atherosclerotic vessels. This model induces the arterial injury response by distending the arterial wall, and denuding the intimal layer of endothelial cells, ultimately causing remodeling and an intimal hyperplastic response.

쥐 경동맥 압력 제어 세그먼트 풍선 부상 과식 치료 응용 프로그램

Cardiovascular disease remains the leading cause of death and disability worldwide, in part due to atherosclerosis. Atherosclerotic plaque narrows the ...

High-Resolution Three-Dimensional Imaging of the Footpad Vasculature in a Murine Hindlimb Gangrene Model

Peripheral arterial disease (PAD) is a significant cause of morbidity resulting from chronic exposure to atherosclerotic risk factors. Patients suffering from its most severe form, chronic limb-threatening ischemia (CLTI), face substantial impairments to daily living, including chronic pain, limited walking distance without pain, and nonhealing wounds. Preclinical models have been developed in various animals to study PAD, but mouse hindlimb ischemia remains the most widely used. There can be significant variation in response to ischemic insult in these models depending on the mouse strain used and the site, number, and means of arterial disruption. This protocol describes a unique method combining femoral artery and vein electrocoagulation with the administration of a nitric oxide synthase (NOS) inhibitor to reliably induce footpad gangrene in Friend Virus B (FVB) mice that resembles the tissue loss of CLTI. While traditional means of assessing reperfusion such as laser Doppler perfusion imaging (LDPI) are still recommended, intracardiac perfusion of the lipophilic dye 1,1'-dioctadecyl-3,3,3',3'-tetramethylindocarbocyanine perchlorate (DiI) is used to label the vasculature. Subsequent whole-mount confocal laser scanning microscopy allows for high-resolution, three-dimensional (3D) reconstruction of footpad vascular networks that complements traditional means of assessing reperfusion in hindlimb ischemia models.

The present protocol describes a unique, clinically relevant model of peripheral arterial disease that combines femoral artery and vein electrocoagulation with the administration of a nitric oxide synthase inhibitor to induce hindlimb gangrene in FVB mice. Intracardiac DiI perfusion is then used for high-resolution, three-dimensional imaging of the footpad vasculature.

Murine Hindlimb Gangrene 모델에서 발판 혈관 구조의 고해상도 입체 이미징

Peripheral arterial disease (PAD) is a significant cause of morbidity resulting from chronic exposure to atherosclerotic risk factors. Patients suffering ...

Murine Aortic Crush Injury: An Efficient In Vivo Model of Smooth Muscle Cell Proliferation and Endothelial Function

Arterial reconstruction, whether angioplasty or bypass surgery, involves iatrogenic trauma causing endothelial disruption and vascular smooth muscle cell (VSMC) proliferation. Common murine models study small vessels such as the carotid and femoral arteries. Herein we describe an in vivo system in which both VSMC proliferation and endothelial barrier function can be simultaneously assessed in a large vessel. We studied the infrarenal aortic response to injury in C57BL/6 mice. The aorta was injured from the left renal vein to the aortic bifurcation by 30 transmural crushes of 5-seconds duration with a cotton-tipped applicator. Morphological changes were assessed with conventional histology. Aorta wall thickness was measured from the luminal surface to the adventitia. EdU integration and counter staining with DAPI and alpha-actin was used to demonstrate VSMC proliferation. Activation of ERK1/2, a known moderator of intimal hyperplasia formation, was determined by Western Blot analysis. The effect of inflammation was determined by immunohistochemistry for B-cells, T-cells, and macrophages. En face sections of endothelium were visualized with scanning electron microscopy (SEM). Endothelial barrier function was determined with Evans Blue staining. Transmural injury resulted in aortic wall thickening. This injury induced VSMC proliferation, most prominently at 3 days after injury, and early activation of ERK1/2 and decreased p27kip1 expression. Injury did not result in increased B-cells, T-cells, or macrophages infiltration in the vessel wall. Injury caused partial endothelial cell denudation and loss of cell-cell contact. Injury resulted in a significant loss of endothelial barrier function, which returned to baseline after seven days. The murine transmural blunt aortic injury model provides an efficient system to simultaneously study both VSMC proliferation and endothelial barrier function in a large vessel.

Murine Aortic Crush Injury: An Efficient In Vivo Model of Smooth Muscle Cell Proliferation and Endothelial Function

Restenosis following cardiovascular procedures (bypass surgery, angioplasty, or stenting) is a significant problem reducing the durability of these procedures. An ideal therapy would inhibit smooth muscle cell (VSMC) proliferation while promoting regeneration of the endothelium. We describe a model for simultaneous assessment of VSMC proliferation and endothelial function in vivo.

소뇌 대동맥 파열 상해 : 효율적<em&gt; In Vivo</em&gt; 평활근 세포 증식과 내피 기능 모델

Arterial reconstruction, whether angioplasty or bypass surgery, involves iatrogenic trauma causing endothelial disruption and vascular smooth muscle cell ...