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>

<ol>
	<li>Dzau, V. J., Braun-Dullaeus, R. C., Sedding, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+proliferation+and+atherosclerosis:+new+perspectives+and+therapeutic+strategies.">Vascular proliferation and atherosclerosis: new perspectives and therapeutic strategies.</a> Nat Med. 8 (11), 1249-1256 (2002).</li><li>Hui, D. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intimal+hyperplasia+in+murine+models.">Intimal hyperplasia in murine models.</a> Curr Drug Targets. 9 (3), 251-260 (2008).</li><li>Ferns, G. A., Avades, T. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+mechanisms+of+coronary+restenosis:+insights+from+experimental+models.">The mechanisms of coronary restenosis: insights from experimental models.</a> Int J Exp Pathol. 81 (2), 63-88 (2000).</li><li>Sata, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+mouse+model+of+vascular+injury+that+induces+rapid+onset+of+medial+cell+apoptosis+followed+by+reproducible+neointimal+hyperplasia.">A mouse model of vascular injury that induces rapid onset of medial cell apoptosis followed by reproducible neointimal hyperplasia.</a> J Mol Cell Cardiol. 32 (11), 2097-2104 (2000).</li><li>Xu, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+models+of+arteriosclerosis:+from+arterial+injuries+to+vascular+grafts.">Mouse models of arteriosclerosis: from arterial injuries to vascular grafts.</a> Am J Pathol. 165 (1), 1-10 (2004).</li><li>Lv, L., Meng, Q., Ye, M., Wang, P., Xue, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=STAT4+deficiency+protects+against+neointima+formation+following+arterial+injury+in+mice.">STAT4 deficiency protects against neointima formation following arterial injury in mice.</a> J Mol Cell Cardiol. 74C, 284-294 (2014).</li><li>Zhu, B., Kuhel, D. G., Witte, D. P., Hui, D. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Apolipoprotein+E+inhibits+neointimal+hyperplasia+after+arterial+injury+in+mice.">Apolipoprotein E inhibits neointimal hyperplasia after arterial injury in mice.</a> Am J Pathol. 157 (6), 1839-1848 (2000).</li><li>Fu, Y., Duru, E. A., Davies, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+metabolic+syndrome+on+the+response+to+arterial+injury.">Effect of metabolic syndrome on the response to arterial injury.</a> J Surg Res. , (2014).</li><li>Kuhel, D. G., Zhu, B., Witte, D. P., Hui, D. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Distinction+in+genetic+determinants+for+injury-induced+neointimal+hyperplasia+and+diet-induced+atherosclerosis+in+inbred+mice.">Distinction in genetic determinants for injury-induced neointimal hyperplasia and diet-induced atherosclerosis in inbred mice.</a> Arterioscler Thromb Vasc Biol. 22 (6), 955-960 (2002).</li><li>Cooley, B. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+strain+differential+neointimal+response+in+vein+grafts+and+wire-injured+arteries.">Mouse strain differential neointimal response in vein grafts and wire-injured arteries.</a> Circ J. 71 (10), 1649-1652 (2007).</li><li>Ishigami, N., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deficiency+of+CuZn+superoxide+dismutase+promotes+inflammation+and+alters+medial+structure+following+vascular+injury.">Deficiency of CuZn superoxide dismutase promotes inflammation and alters medial structure following vascular injury.</a> J Atheroscler Thromb. 18 (11), 1009-1017 (2011).</li></ol>

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 border="0" cellpadding="0" cellspacing="0" width="644">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Name of Reagent/ Equipment</td>
			<td style="width:101px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:208px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Fixed Core Wire Guide</td>
			<td style="width:101px;">Cook</td>
			<td style="width:119px;">G02426</td>
			<td>Diameter 0.015 inch, Straight</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Dilation Forceps</td>
			<td style="width:101px;">Roboz</td>
			<td style="width:119px;">RS-4927</td>
			<td>Curved, blunt tips</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Dumont Tweesers #5</td>
			<td style="width:101px;">World Precision Instruments</td>
			<td style="width:119px;">14095</td>
			<td>Straight, sharp tips</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Dumont Vessel Cannulation Forceps</td>
			<td style="width:101px;">World Precision Instruments</td>
			<td style="width:119px;">503373</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">McPherson-Vannas Scissors</td>
			<td style="width:101px;">World Precision Instruments</td>
			<td style="width:119px;">501234</td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Mosquito Forceps</td>
			<td style="width:101px;">World Precision Instruments</td>
			<td style="width:119px;">501291</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Ethilon Nylon Suture 9-0</td>
			<td style="width:101px;">Ethicon</td>
			<td style="width:119px;">7717G</td>
			<td>9-0, Black Nylon Monofilament</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Micro AROSuture, Sterile 11-0, 70 Microns, MET Point</td>
			<td style="width:101px;">AROSurgical</td>
			<td style="width:119px;">VT4A00N07</td>
			<td>11-0, Black Nylon Monofilament</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">3M Precise Multi-Shot DS Disposable Skin Stapler</td>
			<td style="width:101px;">3M</td>
			<td style="width:119px;">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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JoVE Journal

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19.9K 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

Microsurgical Venous Pouch Arterial-Bifurcation Aneurysms in the Rabbit Model: Technical Aspects

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials have shown disappointing long-term results with unacceptable high rates of aneurysm recanalization and delayed aneurysm rupture.2 To overcome these problems, animal experimental studies are crucial for the development of better endovascular devices.3-5
Several animal models in rats, rabbits, canines and swine are available.6-8 Comparisons of the different animal models showed the superiority of the rabbit model with regard to hemodynamics and comparability of the coagulation system and cost-effectiveness.9-11 
The venous pouch arterial bifurcation model in rabbits is formed by a venous pouch sutured into an artificially created true bifurcation of both common carotid arteries (CCA). The main advantage of this model are true bifurcational hemodynamics.12 The major drawbacks are the sofar high microsurgical technical demands and high morbidity and mortality rates of up to 50%.13 These limitations have resulted in less frequent use of this aneurysm model in the recent years. These shortcomings could be overcome with improved surgical procedures and modified peri- and postoperative analgetic management and anticoagulation.14-16 Our techniques reported in this paper demonstrate this optimized technique for microsurgical creation of arterial bifurcation aneurysms.

An optimized technique for the microsurgical creation of arterial bifurcation aneurysms mimicking bifurcation human cerebral aneurysms is described. A venous pouch is sutured into an artificially created true bifurcation of both common carotid arteries. Facilitated microsurgical techniques and aggressive postoperative anticoagulation and analgesia lead to minimized morbidity rates and high aneurysm patency rates.

토끼 모델에서 Microsurgical 정맥 주머니 아티 - 분기점의 동맥류 : 기술적인 관점

For ruptured human cerebral aneurysms endovascular embolization has become an equivalent alternative to aneurysm clipping.1 However, large clinical trials ...

연구

의학

Femoral Arterial and Venous Catheterization for Blood Sampling, Drug Administration and Conscious Blood Pressure and Heart Rate Measurements

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically implanted catheters permit a researcher to effectively and humanely administer substances, perform repeated blood sampling and assists in conscious direct measurements of blood pressure and heart rate. Once the catheter is implanted long-term sampling is possible. Patency and catheter life depends on multiple factors including the lock solution used, flushing regimen and catheter material. This video will demonstrate the methodology of femoral artery and venous catheterization of the rat. In addition the video will demonstrate the use of the femoral venous and arterial catheters for blood sampling, drug administration and use of the arterial catheter in taking measurements of blood pressure and heart rate in a conscious freely-moving rat. A tether and harness attached to a swivel system will allow the animal to be housed and have samples taken by the researcher with minimal disruption to the animal. To maintain patency of the catheter, careful daily maintenance of the catheter is required using lock solution (100 U/ml heparinized saline), machine-ground blunt tip syringe needles and the use of syringe filters to minimize potential contamination. With careful aseptic surgical techniques, proper catheter materials and careful catheter maintenance techniques, it is possible to sustain patent catheters and healthy animals for long periods of time (several weeks).

Chronic catheterization of blood vessels in the rat is often required for administration of substances, obtain blood sample over a period of time or for direct conscious blood pressure measurements. Femoral arterial catheterization of the rat and corresponding measurements of blood pressure in the conscious animal will be demonstrated.

혈액 샘플링, 마약 관리 및 의식 검사 중 혈압과 심장 박동을 측정 대퇴 동맥혈과 정맥 Catheterization

In multiple fields of study, access to the circulatory system in laboratory studies is necessary. Pharmacological studies in rats using chronically ...

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 ...

Technical Aspects of the Mouse Aortocaval Fistula

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta and inferior vena cava (IVC) are exposed. The proximal infrarenal aorta and the distal aorta are dissected for clamp placement and needle puncture, respectively. Special attention is paid to avoid dissection between the aorta and the IVC. After clamping the aorta, a 25 G needle is used to puncture both walls of the aorta into the IVC. The surrounding connective tissue is used for hemostatic compression. Successful creation of the AVF will show pulsatile arterial blood flow in the IVC. Further confirmation of successful AVF can be achieved by post-operative Doppler ultrasound.

The goal is to produce an arteriovenous fistula that is simple and reproducible. This method does not use sutures or glue adhesive. Therefore the samples can be used with the least amount of foreign materials for analysis.

마우스 Aortocaval 누관의 기술적 측면

Technical aspects of creating an arteriovenous fistula in the mouse are discussed. Under general anesthesia, an abdominal incision is made, and the aorta ...

The Rabbit Blood-shunt Model for the Study of Acute and Late Sequelae of Subarachnoid Hemorrhage: Technical Aspects

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and controllable animal models that simulate both conditions are presently uncommon. Therefore, new models are needed in order to mimic human pathophysiological conditions resulting from SAH.
This report describes the technical nuances of a rabbit blood-shunt SAH model that enables control of intracerebral pressure (ICP). An extracorporeal shunt is placed between the arterial system and the subarachnoid space, which enables examiner-independent SAH in a closed cranium. Step-by-step procedural instructions and necessary equipment are described, as well as technical considerations to produce the model with minimal mortality and morbidity. Important details required for successful surgical creation of this robust, simple and consistent ICP-controlled SAH rabbit model are described.

The experimental intracranial pressure-controlled blood shunt subarachnoid hemorrhage (SAH) model in the rabbit combines the standard procedures &#8212; subclavian artery cannulation and transcutaneous cisterna magna puncture, which enables close mimicking of human pathophysiological conditions after SAH. We present step-by-step instructions and discuss key surgical points for successful experimental SAH creation.

지주막 하 출혈의 급성과 늦은 후유증의 연구에 대한 토끼 피 션트 모델 : 기술적 측면

Early brain injury and delayed cerebral vasospasm both contribute to unfavorable outcomes after subarachnoid hemorrhage (SAH). Reproducible and ...

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 ...

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. ...

Murine Model of Intestinal Ischemia-reperfusion Injury

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, hypotension, necrotizing enterocolitis, bowel transplantation, trauma and chronic inflammation. Intestinal ischemia-reperfusion (IR) injury is a consequence of acute mesenteric ischemia, caused by inadequate blood flow through the mesenteric vessels, resulting in intestinal damage. Reperfusion following ischemia can further exacerbate damage of the intestine. The mechanisms of IR injury are complex and poorly understood. Therefore, experimental small animal models are critical for understanding the pathophysiology of IR injury and the development of novel therapies.
Here we describe a mouse model of acute intestinal IR injury that provides reproducible injury of the small intestine without mortality. This is achieved by inducing ischemia in the region of the distal ileum by temporally occluding the peripheral and terminal collateral branches of the superior mesenteric artery for 60 min using microvascular clips. Reperfusion for 1 hr, or 2 hr after injury results in reproducible injury of the intestine examined by histological analysis. Proper position of the microvascular clips is critical for the procedure. Therefore the video clip provides a detailed visual step-by-step description of this technique. This model of intestinal IR injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

Here we describe the detailed procedure of intestinal ischemia-reperfusion in mice which results in reproducible injury without mortality to encourage the standardization of this technique across the field. This model of intestinal ischemia-reperfusion injury can be utilized to study the cellular and molecular mechanisms of injury and regeneration.

창자 허혈 - 재관류 손상의 쥐 모델

Intestinal ischemia is a life-threatening condition associated with a broad range of clinical conditions including atherosclerosis, thrombosis, ...

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.

Induction of Accelerated Atherosclerosis in Mice: The &quot;Wire-Injury&quot; Model

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. ...

Open Tracheostomy Gastric Acid Aspiration Murine Model of Acute Lung Injury Results in Maximal Acute Nonlethal Lung Injury

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute respiratory distress syndrome (ARDS), characterized by neutrophilic alveolitis, and injury to both alveolar epithelium and vascular endothelium. Clinically, ARDS is defined by acute onset of hypoxemia, bilateral patchy pulmonary infiltrates and non-cardiogenic pulmonary edema. Human studies have provided us with valuable information about the physiological and inflammatory changes in the lung caused by ARDS, which has led to various hypotheses about the underling mechanisms. Unfortunately, difficulties determining the etiology of ARDS, as well as a wide range of pathophysiology have resulted in a lack of critical information that could be useful in developing therapeutic strategies.
Translational animal models are valuable when their pathogenesis and pathophysiology accurately reproduce a concept proven in both in vitro and clinical settings. Although large animal models (e.g., sheep) share characteristics of the anatomy of human trachea-bronchial tree, murine models provide a host of other advantages including: low cost; short reproductive cycle lending itself to greater data acquisition; a well understood immunologic system; and a well characterized genome leading to the availability of a variety of gene deletion and transgenic strains. A robust model of low pH induced ARDS requires a murine ALI that targets mainly the alveolar epithelium, secondarily the vascular endothelium, as well as the small airways leading to the alveoli. Furthermore, a reproducible injury with wide differences between different injurious and non-injurious insults is important.
The murine gastric acid aspiration model presented here using hydrochloric acid employs an open tracheostomy and recreates a pathogenic scenario that reproduces the low pH pneumonitis injury in humans. Additionally, this model can be used to examine interaction of a low pH insult with other pulmonary injurious entities (e.g., food particles, pathogenic bacteria).

This protocol induces acute lung injury in a mouse that has close fidelity to the pathogenesis of acid pneumonitis observed in humans. We generate a maximal acute nonlethal low pH lung injury and account for differences in rodent-human anatomic respiratory structure using an open tracheostomy coupled with circumferential pressure release.

급성 폐 손상의 열기 기관 절개 위산 흡인 쥐 모델은 최대한 급성 치명적 폐 손상의 결과

Acid pneumonitis is a major cause of sterile acute lung injury (ALI) in humans. Acid pneumonitis spans the clinical spectrum from asymptomatic to acute ...