Name: a rat carotid balloon injury model to test anti-vascular remodeling therapeutics
Uploaded: 2016-09-19T12:30:00
Description: Watch this Scientific Journal Video about A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics at JoVE.com

The work was supported by the intramural research program of the NIH, National Institute on Aging, and by a Priority Research Centers Program grant from the National Research Foundation (NRF-2009-0093812) funded by the Ministry of Science, Information and Communication Technology, &amp; Future Planning, the Republic of Korea (H.T.). We thank Dr. Han-sol Park for putting "material" part together for the manuscript.

참고 : 쥐 풍선 손상 모델과 재조합 sRAGE 초음파 초음파 연구의 주입을 포함하여 관련 절차의 사용은 노화에 국립 연구소의 동물 관리 및 사용위원회 (ACUC), NIH에 의해 승인되었습니다. 1. 사전 수술 준비 <ol><li> 악기, 외과 플랫폼 및 개인 보호 장비. <ol><li> 이 절차에 사용 된 모든 수술기구 및 시약에 대한 재료 및 장비의 표를 참조하십시오. 수술하?...

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경동맥 동맥 내강의 벽화 내피 세포를 제거하는 부상을 생성하기 위해 풍선을 팽창하는 데 사용되는 두 가지 방법이 있었다. 하나는 액체 (20)에 부착 된 주사기를 채우기 위하여, 그리고 다른 하나는 공기 압력 (21)을 사용하는 것이다. 우리는 정확한 액량 (0.02 mL)을 각각의 방법에 사용되기 때문에, 액체로 채워진 주사기를 사용하는 것을 선호한다. 이 절차를 진행 각 동물의 부상 ?...

혈관 성형술은 동맥 경화증과 같은 병리학 적 조건에서 결과 좁아진 또는 방해 동맥을 확대하는 데 사용되는 혈관 절차입니다. 혈관 성형술 중 하나 흔한 합병증은 수술 부상과 이후의 염증에 의한 혈관 리모델링으로 인해 발생 후 운영 신생 내막의 증식, 또는 재 협착이다. 이러한 조건은 혈관 부드러운 세포 및 여러 병태 생리 학적 결과 1-3의 확산으로 이어질. 신생 내막의 증식 용기를 재 - 두껍게하고, 제 1 년 이내에 60 % 후 혈관 성형술 환자까지에서 발생합니다. 따라서, 재 협착은 널리 사용되는 혈관 성형 절차 (4)의 큰 방해가된다. 약물 용출 스텐트의 주입이 재 협착을 방지하는 데 도움이 될 수 있지만, 선택된 후보는이 비용이 많이 드는 절차 (5)를받을 수 있습니다. 동물과 임상 연구 모두 만성 염증이 vascu에 의해 생성 된 것을 설립LAR 부상 및 / 또는 수술 상처 후 혈관 신생 내막의 성장 2,4-의 주요 자극 역할을합니다. 따라서, 쥐 풍선 경동맥 손상 모델을 모방 임상 적 상황과 혈관 재 형성 및 혈관 증식 6-9에 관련된 세포 계수를 식별하기위한 유용한 모델 시스템으로서 기능한다. 이 모델 시스템은 전임상 연구 병진 10-14에서 신생 내막 성장을 억제 약물 및 치료 시약 매우 유용한 평가 도구 및 / 또는 스크린이다. 뮤린 경동맥 와이어 손상 모델 (15) 및 쥐의 대퇴 동맥 와이어 손상 모델 (16), 쥐 경동맥 풍선 손상 모델이 입은 부상의 재현을 용이하게 시술의 용이성을 위해 크기가 충분히 큰 있다는 장점을 가진다 비교. 그것은 되어온 대 일차 전지의 더 큰 개수 (예, 혈관 평활근 세포, 내피 세포)를 제공 할 수있다하는 ional는 시험 관내 연구 혈관 재 형성을 지배하는 분자 메커니즘을 묘사하기. 중요한 것은, 마우스에 비해 래트는 생리 학적 연구 (17)을위한 더 나은 모델이 알려져있다. 래트 모델의 단점 또는 제한 유전자 변형 유전자 녹아웃 모델의 부족이지만, 이러한 단점은 래트 게놈 시퀀스의 가용성 및 렌더링 CRISPR-CAS 기술 강력한 게놈 편집 도구의 최근 발전에 의해 극복 될 수있다 다른 모델 시스템 (18, 19)의 게놈 서열의 광대 한 범위의 수 조작. 쥐 풍선 손상 모델이 여러 연구소 및 다양한 포괄적 인 프로토콜에 의해 사용되었지만 (20, 21)을 발표 한,이 프로토콜은 사전 수술 준비에 대한 자세한 내용을 제공하는 것을 목적으로이 수술 연습을 설정하려면이 절차에 새로운 연구를 안내 할 수 있습니다. 우리는 또한 t의 수술 후 관리를 강조살아있는 동물 13,22에서뿐만 아니라 동맥 리모델링에 대한 치료 효과의 사후 병리 및 histomorphological 분석뿐만 아니라, 초음파 초음파 연구 허용 그는 동물.

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			<td height="21" style="height:21px;width:287px;">2 F Fogarty balloon embolectomy catheter&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
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			<td height="21" style="height:21px;width:287px;">Standard scalpel</td>
			<td style="width:163px;">Fine Science Tools</td>
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			<td height="40" style="height:40px;width:287px;">Small&#160; curved forceps (Large radius Dumont#7shanks curved)&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:163px;">Fine Science Tools&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
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			<td height="21" style="height:21px;width:287px;">Large, medium and small micro-scissors</td>
			<td style="width:163px;">Roboz</td>
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			<td height="21" style="height:21px;width:287px;">Needles (20 G)&#160;&#160;</td>
			<td style="width:163px;">TycoHealthcare</td>
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			<td height="41" style="height:41px;width:287px;">Micro-surgery forceps with micro-blunted atraumatic tips</td>
			<td style="width:163px;">Fine Science Tools&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
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			<td height="21" style="height:21px;width:287px;">Atraumatic straight small arterial clamps&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:163px;">Fine Science Tools&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
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			<td height="41" style="height:41px;width:287px;">Retractor&#160; with maximum spread 5.5 cm long blunt teeth</td>
			<td style="width:163px;">Fine Science Tools&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
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			<td height="21" style="height:21px;width:287px;">Silk suture (4.0 and&#160; 6.0 )</td>
			<td style="width:163px;">Fine Science Tools&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
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			<td height="21" style="height:21px;width:287px;">Syringe (1.0 ml)&#160;</td>
			<td style="width:163px;">BD&#160;</td>
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			<td height="21" style="height:21px;width:287px;">Curity gauze sponges</td>
			<td style="width:163px;">AllegroMedical</td>
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			<td height="21" style="height:21px;width:287px;">Cotton tip applicators sterile and non-sterile</td>
			<td style="width:163px;">Puritan Medical Products</td>
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			<td height="21" style="height:21px;width:287px;">Compact hot bead sterilizer</td>
			<td style="width:163px;">Fine Science Tools</td>
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			<td height="21" style="height:21px;width:287px;">Self-regulating heating pad</td>
			<td style="width:163px;">Fine Science Tools&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
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			<td height="21" style="height:21px;width:287px;">ADS200 anesthesia system/ventilator</td>
			<td style="width:163px;">Paragon Medical</td>
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			<td height="21" style="height:21px;width:287px;">Isoflurane (forane), liquid form</td>
			<td style="width:163px;">Baxter</td>
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			<td height="21" style="height:21px;width:287px;">Sodium chloride 0.9% (Saline)&#160;</td>
			<td style="width:163px;">Hospira</td>
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			<td height="41" style="height:41px;width:287px;">Buprenex (buprenorphine)&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;</td>
			<td style="width:163px;">Reckitt Benckiser Healthcare (UK) Ltd.&#160;</td>
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			<td height="21" style="height:21px;width:287px;">70% alcohol</td>
			<td style="width:163px;">Fisher</td>
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			<td height="21" style="height:21px;width:287px;">1: 10 Betadine</td>
			<td style="width:163px;">Fisher</td>
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	</tbody>
</table>

a rat carotid balloon injury model to test anti-vascular remodeling therapeutics

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is also a valuable model system to test, and to evaluate therapeutics and drugs that negate maladaptive remodeling in the vessel. The injury, or barotrauma, in the vessel lumen caused by an inflated balloon via an inserted catheter induces subsequent neointimal growth, often leading to hyperplasia or thickening of the vessel wall that narrows, or obstructs the lumen. The method described here is sufficiently sensitive, and the results can be obtained in relatively short time (2 weeks after the surgery). The efficacy of the drug or therapeutic against the induced-remodeling can be evaluated either by the post-mortem pathological and histomorphological analysis, or by ultrasound sonography in live animals. In addition, this model system has also been used to determine the therapeutic window or the time course of the administered drug. These studies can leadto the development of a better administrative strategy and a better therapeutic outcome. The procedure described here provides a tool for translational studies that bring drug and therapeutic candidates from bench research to clinical applications.

2 주 풍선 손상 후, 래트를 안락사되고 경동맥은 히스 - 형태 학적 분석을 위해 분리된다. 모두 왼쪽 경동맥 및 취소 운영 오른쪽 동맥 처리, 단면 크로스, 파라핀 내장되어 운영했다. 파라핀 샘플은 더 얇은 단면 및 헤 마톡 실린 - 에오신 (H &amp; E) 염색을합니다. 히스 - 형태소 분석은 디지털 이미지 분석 시스템을 이용하여 수행된다. 동맥 수확 히스 - 형태소 분석의 상세 <s...

Watch this Scientific Journal Video about A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics at JoVE.com

A Rat Carotid Balloon Injury Model to Test Anti-vascular Remodeling Therapeutics

The rat carotid balloon injury model described below allows researchers to evaluate drugs or therapeutics that negate injury-induced arterial hyperplasia. Detailed pre-surgical preparation, surgical procedure, and post-surgical cares of the animal are described.

생쥐 경동맥 풍선 손상 모델은 반대로 혈관 리모델링의 치료를 테스트하는

The rat carotid balloon injury is a well-established surgical model that has been used to study arterial remodeling and vascular cell proliferation. It is ...

The overall goal of this Rat Carotid Balloon Injury procedure is to mimic clinical angioplasty and its potential consequences. This message can help answer key questions in the cardiovascular research field, about the cardiovascular surgical injury elicited neointimal hyperplasia which can significantly compromise the benefits of the surgery. The main advantage of this technique is that the result can be obtained within two weeks, allowing a quick evaluation of the therapeutic reagents administered during the treatment period.Demonstrating the procedure will be Dr.Natalia Petrasheskatya from the laboratory of cardiovascular sciences and at the National Institute on Aging. Before beginning the surgery, autoclave all of the surgical instruments and use 70%ethanol to disinfect the operating platform surface. Put on the proper personal protective equipment, placing a box of sterilized gloves near the surgical platform for additional use.Next, fill a 1 milliliter syringe with sterile water and attach the syringe to a sterile, water-filled, two-way stopcock. Gently push the water through the stopcock to fill the lure-lock portion of the balloon catheter, and remove any air bubbled from the opening of the catheter. Then, test the balloon inflation to confirm that the balloon can be inflated with 0.02 milliliters of water.Now, check the pedal withdrawal reflex of an anesthetized 400 to 450 gram male Wistar rat. If the rat does not withdraw its foot, transfer the animal to the surgical platform. Cover the rat with a sterile surgical sheet, and cut the sheet to expose the neck region.Then adjust the animal into the supine position on a headed pad with the head toward the surgeon and tape the limbs into place. To isolate the left carotid artery, us a number 14 scalpel to make a straight incision from below the chin to the top of the sternum, just below the ribcage. Using a 7S forceps, blunt dissect under the skin and through the salivary gland tissues to expose the underlying muscle layer.Separate the muscle tissues to gain access to the carotid vasculature and the vagus nerve in the neck. Continue to blunt dissect the tissues surrounding the carotid artery, carefully separating the vagus nerve and the vascular fascia without damaging either tissue, until the bifurcation of the common carotid artery and the internal and external carotid branches are clearly visible. When the left common carotid artery has been exposed all the way to the sternum, place a 4-0 silk stay suture behind the artery as close to the sternum as possible.Permanently ligate the smaller arteries with the sutures, along the external carotid artery, including the ascending pharyngeal, the occipital, and the superior thyroid to avoid arterial leakage. Place another 4-0 silk stay suture immediately distal to the bifurcation, behind the external carotid artery as far from the bifurcation as possible. Then gently retract the overlying carotid artery to the right to visualize the internal carotid artery and place a 4-0 silk stay suture around the artery to avoid significant retrograde blood-loss.To introduce the balloon catheter, retract the proximal suture on the common carotid artery, and temporarily stop the blood flow with an arterial clamp. Make a small incision on the external carotid artery as distal to the suture as possible, and gently insert the uninflated balloon catheter into the incision. Advance the catheter into the lumen until it is close to the arterial clamp on the common carotid artery.Then remove the clamp and advance the catheter to the aortic arch, approximately 35 to 40 millimeters from the incision. Now use the syringe of sterile water to manually inflate the catheter to a 0.02 milliliter volume, and turn the stopcock to the locked position. Slowly withdraw the catheter while rotating to produce the injury.When the instrument is close to the arterial incision, deflate the balloon and retract the catheter back to its original position. After repeating the balloon injury procedure three times, carefully remove the catheter from the arterial lumen and close the arteriotomy with size 5-0 silk stay sutures. Release the internal carotid artery to restore the blood flow and check for arterial leakage.Finally, remove the stay sutures and clamps and close the glandular tissue with 6-0 silk sutures. Then, close the skin and monitor the animal until it is fully recovered. Two weeks after the balloon injury, the carotid arteries can be isolated for histomorphological analysis.The balloon injury elicits neointimal growth, or thickening of the vessel wall. If left untreated or treated with a saline control, compared to a non-operated vessel section from the same animal. By contrast, the vessel section from a rat that received a therapeutic reagent that blocks neointimal growth demonstrates a significantly reduced thickening in the vessel wall.The effects of the balloon injury and therapeutic treatment can also be evaluated in vivo, by ultrasound sonography. And these data collaborate well with the results from histomorphological analysis. When mastered, this techniques can be completed in 30 minutes, if it is performed properly.While attempting this procedure, it is important to remember to prepare and test the balloon catheter for leaks prior to the surgery. After watching this video, you should have a good understanding of how to perform the rat carotid balloon injury procedure, for evaluating tracks and therapeutic reagents that block neointimal growth and to study the mag-ni-sen of injury elicited neointimal growth. Don't forget that working with animals requires an approved protocol by your institutional AICUC.You should also consult with your institutional laboratory animal veterinarians to establish the best practices for post-surgical care, and monitoring of the animals.

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