This work was supported in part by the United States Department of Veterans Affairs Biomedical Laboratory Research and Development Program Merit Review Award I01-BX002336, the National Institute of Health grant R56-HL095498, as well as with the resources and the use of facilities at the VA Connecticut Healthcare System, West Haven, CT.

해당 기관 동물 관리 및 사용위원회의 승인을 얻을 수있다. 1. 마취 및 수술 전 절차 <ol><li> 아크릴 유도 챔버에 투여 기화 3 % 이소 플루 란과 0.8 L / 분 산소와 남성 C57BL / 6 얼음, 세 팔주를 마취. </li><li> 발가락 핀치에 반응의 부족으로 적절한 마취를 확인합니다. 연속 흡입에 의해 3 %의 이소 플루 란 - 작업 테이블에 마우스 부정사를 놓고 실리콘 마스크를 위치 2 기화 제공한다. </li><li> 화학적 제모 크림을 사용하여 복부 이하로 목에서 복부 머리를 제거합니다. </li><li> 관심의 영역에서 동맥과 정맥의 흐름과 혈관 직경의 기본 특성을 기록하기 전에 AVF 수술에 도플러 초음파 검사를 수행합니다. 1,2 </li><li> 25 G 바늘로 1 ML의 주사기를 연결하고 원하는 약물을 주사기를로드합니다. 벤드 요구로부터 60도 각도로 바늘 약 4mm제작 팁. 곡선 바늘 홀더로 25 G 바늘을 잡고. </li></ol> 2. 수술 절차 <ol><li> 국소 방부제와 절개 사이트를 준비하고 외과 드레이프를 적용합니다. 수술 전반에 걸쳐 무균 기술을 유지하기 위해 멸균 장갑과 악기를 사용합니다. </li><li> 메스 그냥 치골 위를 하단 간 날의 수준에서 연장 중간 선 복부 절개를합니다. </li><li> 트랙터를 삽입하고 오른쪽 방향으로 복강에서 모든 창자를 내용 만. 식염수로 적신 거즈에 창자를 감싸. 대동맥과 IVC의 전체보기를 얻기 위해 역 복막 및​​ 하부 대장을 연결하는 멤브레인을 해부하다. </li><li> 근위 및 원위 체결을 준비하고, 주변 조직에서 적외선 신장 대동맥과 IVC를 해부하다. </li><li> 근위 대동맥 바로 왼쪽 신장 정맥 아래 수준에서 근위 IVC 모두에서 하나의 미세 클립을 배치합니다. D 모두에서 두 번째 미세 클립을 배치istal 대동맥 및 말초 IVC. </li><li> 대동맥 주위의 결합 조직을 잡고 대동맥의 등 표면이 약간 동맥 천자에 대한 노출되도록 전술 한 바와 같이, 내측 회전합니다. (1) </li><li> 빠르게 펑크 사이트를 노출합니다. 천자 사이트는 혈관, 대동맥 분기에 왼쪽 신장 정맥으로부터의 거리의 약 3 분기 꼬리 측면에있을 것입니다. 오른손 천자 있도록 충분한 노출이되도록 왼쪽으로 회전 된 위치에 유지 대동맥, 대동맥의 왼쪽 측면 마진을 해부. 대동맥과 IVC 사이 해부하지 않도록주의하십시오. </li><li> 회전 위치를 유지하고 대동맥 부착 1ml를 주사기 함유 약액에 25 G 바늘을 사용을 통해 IVC에 대동맥 천공. (도 1a) </li><li> 왼손 사용 - (200 μL, 100) 약액을 주입. 바늘 팽창 얇은 통해 알 수있는투명 약액이 IVC (도 1B, C)에서 정맥혈 변위 IVC 벽. 남아 15 분 동안 위치에 바늘을 유지한다. </li><li> 만 원위부 대동맥 및 말초 IVC를 클램프를 해제하기 위해 말초 미세 클립을 제거합니다. </li><li> 바늘을 제거한 후 인접 역 복막 조직을 당겨 대동맥의 천자 사이트를 다룹니다. </li><li> 근위 대동맥과 근위 IVC를 해제 클램프 근위 미세 클립을 제거합니다. 디 클램핑시, 동맥 혈액은 IVC 대신 어두운 정맥 혈류로 유입 관찰된다. 1 분 동안 구멍 구멍을 덮고 보관하십시오. </li><li> 압축없이 30 초 동안 관찰하여 지혈의 확인 후, 그들의 자연적인 위치로 창자를 반환하고 승인 된 동물 프로토콜에 따라 실행 봉합과 복부를 닫습니다. </li></ol> 3. 수술 후 절차 <ol><li> 복부의 폐쇄 후, ANEST을 중지hesia. 기관 동물 관리 및 사용위원회에서 권장하는 지침에 따라 진통과 상처 치료를 포함한 수술 후 치료를 적용합니다. 진통제를 위해 우리는 / kg이 intrasmuscularly 24 시간마다 12 시간이 수술 절차에 따라 0.1 밀리그램에서 프레 노르 핀을 사용합니다. </li><li> 수술 후 첫 날, AVF의 개통을 확인하는 도플러 초음파를 수행합니다. 또한, 다른 선박을 측정하고 연속적으로 유동 특성 및 수술 전 기본 값의 변화를 비교한다. 1,2 </li></ol><img alt="그림 1" src="/files/ftp_upload/53905/53905fig1.jpg" /> AVF 수술시 관내 배달을보기 그림 1. (A) 수술 사진. 근위 및 말초 동맥뿐만 아니라, 미세 클립을 적용하여 IVC 클램프. 연결된 주사기 함유 약액에 25 G 바늘을 사용을 통해 IVC에 대동맥 천공. (B)를 HIG그녀의 전원 사진 주입하기 전에 구멍 IVC의 (4 배 확대). 바늘 끝은 어두운 색의 정맥혈에 의해 가려한다. 노란 화살촉은 주입 후 천공 IVC의 IVC. (C) 높은 전원 사진 (4 배 배율)의 벽에 벽 직경을 나타낸다. 니들 팁 (흑색 화살표) 정맥혈 변위 투명 약액 등 가볍게 팽창 IVC 얇은 벽 (노란 화살표 머리)를 통해 볼 수있다.

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The authors have nothing to disclose.

AVF 뮤린 모델이 수정 AVF 생성시 정맥 내피 관내 약물 전달을 포함한다. AVF는 25 G 바늘 적외선 신장 대동맥을 천공과 같은 바늘을 통해 약물 용액을 주입 한 다음 IVC에 반대 대동맥 벽을 통해 구멍을 확장하여 만들어졌습니다. 용액 내 대정맥 유지된다, 즉., AVF의 정맥 상지에서, 디 클램핑까지. 어떤 다른 쥐 AVF 모델 17-19에서이 모델을 구별하는 분자 분석 협착, 급성 혈전증, 가능한 ?...

쥐 aortovenous 루 (AVF) 대동맥과 하대 정맥 (IVC) 사이의 구멍 모델은 지금에 나오는이 모델에서 확립 된 기술. 1, 25 G 바늘을 천공하는 적외선 신장 대동맥의 양쪽 벽이다 적외선 신장 대정맥 인접; 전방 대동맥 입구 구멍은 간단한 압축 수리 및 봉합 수리를 필요로하지 않습니다. 고해상도 도플러 초음파 조직 학적 분석에 의해 일련 후속 검사 성숙 단계 후 실패한 단계 인간 AVF의 알려진 기전을 recapitulating을 가지고 AVF를 나타낸다. 2 AVF의 성숙을 조절하는 메커니즘을 탐구하기 위해, 성숙 AVF의 내피 세포로 치료제의 전달을위한 개선 된 방법이 필요하다. 혈관 치료제의 배달 중 혈관 내 전달을 통해 루멘, 또는 외막에 외부 배달을 통해 할 수있다. 외부 납품의 일례이다 COMmonly는 플루로 닉 겔의 외막 응용 프로그램을 사용했다. 체온으로 가온 될 때이 공중 합체는, 열 가역성 겔 및 고체에서 액체로 변환된다. 플루 겔 혼합 약물이 생체 내에서 국소 적으로 적용될 때 종래 연구 도시 한 지속적인 약물 전달이 달성된다. 바이러스 벡터 또는 플루 겔의 siRNA 3,4- 외막 애플리케이션 혈관 주위 전달체로서 효과적인 것으로보고되었다. 5,6-에게 우리를 또한 펩타이드에 의해 외막 자극과 외식 인간 복재 정맥의 치료가 내피 세포 수용체 단백질의 인산화 결과보고 있습니다. (7) 반면에, 연구자들은 또한 정맥 이식편의 8 ~ 10 개 토끼 11, 12 모델 모두 바이러스 성 및 비 바이러스 벡터의 관내 배달을 사용하고 있습니다. 이 보고서에서 유전자 전달 수확을 정맥 후 생체 하였다. Eslami 등은. 혈관 내 바이러스 성 유전자 전달보고y를 바이 패스를 생성하지 않고 현장에서 혈관을 경동맥합니다. 13 Gloverman 등. 쥐 대퇴 동맥 - 표면 상복부 정맥 루에서 벌거 벗은 DNA의보고 관내 및 외막 배달. 14 메이요 그룹은 마우스 경동맥 동맥 - 경정맥 정맥 루의 외막 약물 전달을 보도했다. (15, 16) 그러나,이 이전에보고 된 모델을 만들 봉합 문합이 필요 AVF. 이 보고서에서 쥐에서 동시 AVF 생성과 관내 약물 전달은 AVF 창조의 봉합이없는 모델을 사용하여 설명한다. 이 변형 AVF 뮤린 모델을 사용하여 누공 정맥 사지 관내 약물 전달을위한 간단한 방법이 수행 될 수있다.

<table><tbody><tr><td>Pluronic F-127</td><td>Sigma-Aldrich</td><td>P2443-250G</td><td>Used as 30% solution in d-water</td></tr><tr><td>GFP antibody</td><td>NOVUS BIOLOGICALS INC</td><td>NB100-1770</td><td></td></tr><tr><td>Ad-CMV-GFP</td><td>VECTOR BIOLABS</td><td>1060</td><td></td></tr><tr><td>0.9% Sodium Chloride Irrigation, USP</td><td>Baxter</td><td>2F7122</td><td></td></tr><tr><td>BD PrecisionGlide Needle 25 G x 5/8</td><td>BD</td><td>305122</td><td></td></tr><tr><td>BD 1 ml Syringe Tuberculin Slip Tip</td><td>BD</td><td>309659</td><td></td></tr><tr><td>Scalpel</td><td>Surgical Design Inc</td><td>22079707</td><td></td></tr><tr><td>6-0 ETHILON P-1 11 mm 3/8c Reverse Cutting</td><td>ETHICON INC</td><td>697G</td><td></td></tr><tr><td>Vevo 770 ultrasound machine&amp;nbsp;</td><td>Visualsonics&amp;nbsp;</td><td>20 - 60 Mhz scan head; RMV-704</td><td></td></tr><tr><td>Vascular clamp&amp;nbsp;</td><td>Roboz Surgical Instrument Co.</td><td>RS-5424</td><td></td></tr><tr><td>Clamp applying forceps&amp;nbsp;</td><td>Roboz Surgical Instrument Co.</td><td>RS-5410</td><td></td></tr></tbody></table>

intraluminal drug delivery to the mouse arteriovenous fistula endothelium

치료제의 배달 동정맥루 (AVF) 성숙이 중 관내 또는 외부 경로를 통해 투여 될 수있다 강화한다. 간단한 AVF 뮤린 모델 누공 생성과 동시에 정맥 내피 약액 관내 투여와 결합시켰다. 이 모델의 기술적 인 측면을 설명합니다. 전신 마취하에 복부 절개가 이루어지고 대동맥과 하대 정맥 (IVC)가 노출된다. 적외선 신장 대동맥과 IVC는 클램핑에 대한 해부한다. 근위 및 원위 클램프 후에 천자 부위가 노출되고 25 G 바늘은 대동맥 및 IVC로 양쪽 벽을 천공하는데 사용된다. 즉시 천자 후, 리포터 유전자 발현 바이러스 벡터는 15 분 인큐베이션 한 후, 동일한 니들을 통해 IVC에 주입 하였다. 관내 투여 방법은 외부 경로에 의한 투여에 비해 정맥 내피 더 강력한 바이러스 성 유전자 전달을 활성화. 이 노브배달 방법은 L AVF 성숙에서 내피 세포의 역할을 살펴 수술시의 관내 약물 전달을 활성화 연구를 촉진 할 것이다.

마우스 (33)의 연속으로, 제 수술 일에 생존은 97.0 %였다; 초음파에 의해 결정 AVF 개통, 84.9 %였다. 기존의 외부 노선이 혈관 내 전달 경로의 유전자 전달 효율을 비교 하​​였다. 관내 배달 (ILD), 즉시 천자 후, 아데노 바이러스-GFP의 200 μL (AD-GFP) 벡터 솔루션 (1 × 10 9 PFU / ml)에 15 분 배양 시간 뒤에, 천공 바늘을 통해 IVC?...

Watch this Scientific Journal Video about Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium at JoVE.com

Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium

After puncturing the aorta through the inferior vena cava (IVC) to create an aorto-caval fistula in the mouse, solution containing a drug is infused into the IVC via the same needle, followed by incubation. This method enables more robust drug delivery to the venous endothelium compared to the external route.

마우스 동정맥루 내피에 관내 약물 전달

Delivery of therapeutic agents to enhance arteriovenous fistula (AVF) maturation can be administered either via intraluminal or external routes. The ...

intraluminal-drug-delivery-to-mouse-arteriovenous-fistula

Research

JoVE Journal

Medicine

9.3K 조회수.  Yale University. After puncturing the aorta through the inferior vena cava (IVC) to create an aorto-caval fistula in the mouse, solution containing a drug is infused into the IVC via the same needle, followed by incubation. This method enables more robust drug delivery to the venous endothelium compared to the external route.

비디오: Intraluminal Drug Delivery to the Mouse Arteriovenous Fistula Endothelium

Osmotic Drug Delivery to Ischemic Hindlimbs and Perfusion of Vasculature with Microfil for Micro-Computed Tomography Imaging

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to stimulate microcirculation. The choice of delivery method for these agents is important because the route of administration profoundly affects the bioactivity and efficacy of these agents1,2. In this article, we demonstrate how to locally administer a substance in ischemic hindlimbs by using a catheterized osmotic pump. This pump can deliver a fixed volume of aqueous solution continuously for an allotted period of time. We also present our mouse model of unilateral hindlimb ischemia induced by ligation of the common femoral artery proximal to the origin of profunda femoris and epigastrica arteries in the left hindlimb. Lastly, we describe the in vivo cannulation and ligation of the infrarenal abdominal aorta and perfusion of the hindlimb vasculature with Microfil, a silicone radiopaque casting agent. Microfil can perfuse and fill the entire vascular bed (arterial and venous), and because we have ligated the major vascular conduit for exit, the agent can be retained in the vasculature for future ex vivo imaging with the use of small specimen micro-CT3.

We show here the in vivo insertion of an osmotic pump for constant local drug delivery and the creation of hindlimb ischemia in a mouse model. Moreover, the hindlimb vasculature is perfused with Microfil, a silicone radiopaque agent, to prepare for micro-computed tomography (micro-CT) imaging.

마이크로 컴퓨터 단층 촬영 영상에 대한 Microfil와 허혈성 뒷발과 혈관의 관류에 삼투의 약물 전달 (Drug Delivery)

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to ...

연구

의학

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 ...

Adult Mouse Venous Hypertension Model: Common Carotid Artery to External Jugular Vein Anastomosis.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model creating an artificial fistula between the common carotid artery (CCA) and the external jugular vein (EJV) has been widely described and proved technically feasible. This construct provokes a consistent cerebral venous hypertension (CVH), and therefore has helped studying the contribution of venous hypertension to formation, clinical symptoms, and prognosis of brain AVMs and dural AVFs. Equivalent mice models have been only scarcely described and have shown trouble with stenosis of the fistula. An established murine model would allow the study of not only pathophysiology but also potential genetic therapies for these cerebrovascular diseases.
We present a model of arteriovenous fistula that produces a durable intracranial venous hypertension in the mouse. Microsurgical anastomosis of the murine CCA and EJV can be difficult due to diminutive anatomy and frequently result in a non-patent fistula. In this step-by-step protocol we address all the important challenges encountered during this procedure. Avoiding excessive retraction of the vein during the exposure, using 11-0 sutures instead of 10-0, and making a carefully planned end-to-side anastomosis are some of the critical steps. Although this method requires advanced microsurgical skills and a longer learning curve that the equivalent in the rat, it can be consistently developed.
This novel model has been designed to integrate transgenic mouse techniques with a previously well-established experimental system that has proved useful to study brain AVMs and dural AVFs. By opening the possibility of using transgenic mice, a broader spectrum of valid models can be achieved and genetic treatments can also be tested. The experimental construct could also be further adapted to the study of other cerebrovascular diseases related with venous hypertension such as migraine, transient global amnesia, transient monocular blindness, etc.

We describe a method for creating a reliable model of cerebral venous hypertension in the adult mouse. This model has been widely described and tested in the rat. This new counterpart in the mice opens the possibility of using genetic modified animals and thereby broadens the applications of the model.

성인 마우스 정맥 ​​고혈압 모델 : 외부 경정맥 정맥 문합에 경동맥.

The understanding of the pathophysiology of brain arteriovenous malformations and arteriovenous fistulas has improved thanks to animal models. A rat model ...

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. ...

A Novel Murine Model of Arteriovenous Fistula Failure: The Surgical Procedure in Detail

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the exact pathophysiology remains unknown. In order to unravel the pathophysiology a murine model of AVF-failure was developed in which the configuration of the anastomosis resembles the preferred situation in the clinical setting. A model was described in which an AVF is created by connecting the venous end of the branch of the external jugular vein to the side of the common carotid artery using interrupted sutures. At a histological level, we observed progressive stenotic intimal lesions in the venous outflow tract that is also seen in failed human AVFs. Although this procedure can be technically challenging due to the small dimensions of the animal, we were able to achieve a surgical success rate of 97% after sufficient training. The key advantage of a murine model is the availability of transgenic animals. In view of the different proposed mechanisms that are responsible for AVF failure, disabling genes that might play a role in vascular remodeling can help us to unravel the complex pathophysiology of AVF failure.

Here we present a murine model of arteriovenous fistula (AVF) failure in which a clinically relevant anastomotic configuration is incorporated. This model can be used to study the pathophysiology and to test possible therapeutic interventions.

동정맥루 실패의 소설 쥐 모델 : 세부 수술

The arteriovenous fistula (AVF) still suffers from a high number of failures caused by insufficient remodeling and intimal hyperplasia from which the ...

Murine Model of Central Venous Stenosis using Aortocaval Fistula with an Outflow Stenosis

Central venous stenosis is an important entity contributing to arteriovenous fistula (AVF) failure. A murine AVF model was modified to create a partial ligation of the inferior vena cava (IVC) in the outflow of the fistula, mimicking central venous stenosis. Technical aspects of this model are introduced. The aorta and IVC are exposed, following an abdominal incision. The infra-renal aorta and IVC are dissected for proximal clamping, and the distal aorta is exposed for puncture. The IVC at the midpoint between the left renal vein and the aortic bifurcation is carefully dissected to place an 8-0 suture beneath the IVC. After clamping the aorta and IVC, an AVF is created by puncturing the infra-renal aorta through both walls into the IVC with a 25 G needle, followed by ligating a 22 G intra-venous (IV) catheter and IVC together. The catheter is then removed, creating a reproducible venous stenosis without occlusion. The aorta and IVC are unclamped after confirming primary hemostasis. This novel model of central vein stenosis is easy to perform, reproducible, and will facilitate studies on AVF failure.

An aortocaval fistula was created by puncturing the murine infra-renal aorta through both walls into the inferior vena cava and was followed by creation of a stenosis in its outflow via partial ligation of the inferior vena cava. This reproducible model can be used to study central venous stenosis.

유출 협착증을 가진 Aortocaval 누공을 사용하여 중앙 정맥 협착증의 뮤린 모형

Central venous stenosis is an important entity contributing to arteriovenous fistula (AVF) failure. A murine AVF model was modified to create a partial ...

Multiphoton Intravital Imaging for Monitoring Leukocyte Recruitment during Arteriogenesis in a Murine Hindlimb Model

Arteriogenesis strongly depends on leukocyte and platelet recruitment to the perivascular space of growing collateral vessels. The standard approach for analyzing collateral arteries and leukocytes in arteriogenesis is ex vivo (immuno-) histological methodology. However, this technique does not allow the measurement of dynamic processes such as blood flow, shear stress, cell-cell interactions, and particle velocity. This paper presents a protocol to monitor in vivo processes in growing collateral arteries during arteriogenesis utilizing intravital imaging. The method described here is a reliable tool for dynamics measurement and offers a high-contrast analysis with minimal photo-cytotoxicity, provided by multiphoton excitation microscopy. Prior to analyzing growing collateral arteries, arteriogenesis was induced in the adductor muscle of mice by unilateral ligation of the femoral artery.
After the ligation, the preexisting collateral arteries started to grow due to increased shear stress. Twenty-four hours after surgery, the skin and subcutaneous fat above the collateral arteries were removed, constructing a pocket for further analyses. To visualize blood flow and immune cells during in vivo imaging, CD41-fluorescein isothiocyanate (FITC) (platelets) and CD45-phycoerythrin (PE) (leukocytes) antibodies were injected intravenously (i.v.) via a catheter placed in the tail vein of a mouse. This article introduces intravital multiphoton imaging as an alternative or in vivo complementation to the commonly used static ex vivo (immuno-) histological analyses to study processes relevant for arteriogenesis. In summary, this paper describes a novel and dynamic in vivo method to investigate immune cell trafficking, blood flow, and shear stress in a hindlimb model of arteriogenesis, which enhances evaluation possibilities notably.

The recruitment of leukocytes and platelets constitutes an essential component necessary for the effective growth of collateral arteries during arteriogenesis. Multiphoton microscopy is an efficient tool for tracking cell dynamics with high spatio-temporal resolution in vivo and less photo-toxicity to study leukocyte recruitment and extravasation during arteriogenesis.

쥐 뒷다리 모델에서 동맥 형성 중 백혈구 모집을 모니터링하기 위한 다광자 생체 내 이미징

Arteriogenesis strongly depends on leukocyte and platelet recruitment to the perivascular space of growing collateral vessels. The standard approach for ...

A Murine Model of Hemodialysis Access-Related Hand Dysfunction

Chronic kidney disease is a major public health problem, and the prevalence of end-stage renal disease (ESRD) requiring chronic renal replacement therapies such as hemodialysis continues to increase. Autogenous arteriovenous fistula (AVF) placement remains a primary vascular access option for ESRD patients. Unfortunately, approximately half of the hemodialysis patients experience dialysis access-related hand dysfunction (ARHD), ranging from subtle paresthesia to digital gangrene. Notably, the underlying biologic drivers responsible for ARHD are poorly understood, and no adequate animal model exists to elucidate the mechanisms and/or develop novel therapeutics for the prevention/treatment of ARHD. Herein, we describe a new mouse model in which an AVF is created between the left common iliac artery and vein, thereby facilitating the assessment of limb pathophysiology. The microsurgery includes vessel isolation, longitudinal venotomy, creation of arteriovenous anastomosis, and venous reconstruction. Sham surgeries include all the critical steps except for AVF creation. Iliac AVF placement results in clinically relevant alterations in central hemodynamics, peripheral ischemia, and impairments in hindlimb neuromotor performance. This novel preclinical AVF model provides a useful platform that recapitulates common neuromotor perturbations reported by hemodialysis patients, allowing researchers to investigate the mechanisms of ARHD pathophysiology and test potential therapeutics.

This protocol details the surgical steps of murine common iliac arteriovenous fistula creation. We developed this model to study hemodialysis access-related limb pathophysiology.

혈액 투석 접근 관련 손 기능 장애의 쥐 모델

Chronic kidney disease is a major public health problem, and the prevalence of end-stage renal disease (ESRD) requiring chronic renal replacement ...

Drug Treatment by Central Venous Catheter in a Mouse Model of Angiotensin II Induced Abdominal Aortic Aneurysm and Monitoring by 3D Ultrasound

Since pharmaceutical treatment options are lacking in the clinical management of abdominal aortic aneurysm (AAA), animal models, in particular mouse models, are applied to advance the understanding of the disease pathogenesis and to identify potential therapeutic targets. Testing novel drug candidates to block AAA growth in these models generally requires repeated drug administration during the time course of the experiment. Here, we describe a compiled protocol for AAA induction, insertion of an intravenous catheter to facilitate prolonged therapy, and serial AAA monitoring by 3D ultrasound. Aneurysms are induced in apolipoprotein E (ApoE) deficient mice by angiotensin II release over 28 days from osmotic mini-pumps implanted subcutaneously into the mouse back. Subsequently, the surgical procedure for external jugular vein catheterization is conducted to allow for daily intravenous drug treatment or repeated blood sampling via a subcutaneous vascular access button. Despite the two dorsal implants, the monitoring of AAA development is readily facilitated by sequential semi-automated 3D ultrasound analysis, which yields comprehensive information on the expansion of aortic diameter and volume and on aneurysm morphology, as illustrated by experimental examples.

This protocol describes the consecutive implantation of an osmotic pump to induce abdominal aortic aneurysm by angiotensin II release in apolipoprotein E (ApoE) deficient mice and of a vascular access port with a jugular vein catheter for repeated drug treatment. Monitoring of aneurysm development by 3D ultrasound is effectively conducted despite dorsal implants.

안지오텐신 II의 마우스 모델에서 중앙 정맥 카테터에 의한 약물 치료 유도 복부 대동맥류 및 3D 초음파에 의한 모니터링

Since pharmaceutical treatment options are lacking in the clinical management of abdominal aortic aneurysm (AAA), animal models, in particular mouse ...

A Modified Technique for Arteriovenous Fistula Construction in Rabbits

Juxta-anastomotic stenosis is a challenging problem that often causes non-maturation and decreases the patency of an arteriovenous fistula (AVF). Injury to the veins and arteries during the operation and hemodynamic changes can lead to intimal hyperplasia, leading to juxta-anastomotic stenosis. To reduce injury to the veins and arteries during the operation, this study proposes a new modified no-touch technique (MNTT) for AVF construction that can decrease the rate of juxta-anastomotic stenosis and improve the AVF patency. To unravel the hemodynamic changes and mechanisms of the MNTT, this study presented an AVF procedure using this technique. Although this procedure is technically challenging, 94.4% procedural success was achieved after adequate training. Ultimately, 13 out of 34 rabbits had a functional AVF 4 weeks after the surgery, leading to a 38.2% AVF patency rate. However, at 4 weeks, the survival rate was 86.1%. Ultrasonography showed active blood flow through AVF anastomosis. Furthermore, the spiral laminar flow was observed in the vein and artery near the anastomosis, suggesting that this technique may improve the hemodynamics of the AVF. On histological observation, significant venous intimal hyperplasia was observed at the AVF anastomosis, whereas no significant intimal hyperplasia was observed at the proximal external jugular vein (EJV) of the anastomosis. This technique will improve the understanding of the mechanisms underlying the use of MNTT for AVF construction and provide technical support for the further optimization of the surgical approach in AVF construction.

The present protocol proposes the creation of an arteriovenous fistula in rabbits using a modified no-touch technique. The technique involves the side-to-side anastomosis of the common carotid artery and external jugular vein without the dissection of the perivenous tissues or cutting off the artery.

토끼의 동정맥루 건설을위한 수정 된 기술

Juxta-anastomotic stenosis is a challenging problem that often causes non-maturation and decreases the patency of an arteriovenous fistula (AVF). Injury ...

마우스 동정맥루 내피에 관내 약물 전달

요약

더 많은 비디오 탐색

이 비디오의 챕터

마이크로 컴퓨터 단층 촬영 영상에 대한 Microfil와 허혈성 뒷발과 혈관의 관류에 삼투의 약물 전달 (Drug Delivery)

마우스 Aortocaval 누관의 기술적 측면

성인 마우스 정맥 고혈압 모델 : 외부 경정맥 정맥 문합에 경동맥.

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

동정맥루 실패의 소설 쥐 모델 : 세부 수술

유출 협착증을 가진 Aortocaval 누공을 사용하여 중앙 정맥 협착증의 뮤린 모형

쥐 뒷다리 모델에서 동맥 형성 중 백혈구 모집을 모니터링하기 위한 다광자 생체 내 이미징

혈액 투석 접근 관련 손 기능 장애의 쥐 모델

안지오텐신 II의 마우스 모델에서 중앙 정맥 카테터에 의한 약물 치료 유도 복부 대동맥류 및 3D 초음파에 의한 모니터링

토끼의 동정맥루 건설을위한 수정 된 기술

마우스 동정맥루 내피에 관내 약물 전달

요약

더 많은 비디오 탐색

이 비디오의 챕터

마이크로 컴퓨터 단층 촬영 영상에 대한 Microfil와 허혈성 뒷발과 혈관의 관류에 삼투의 약물 전달 (Drug Delivery)

마우스 Aortocaval 누관의 기술적 측면

성인 마우스 정맥 ​​고혈압 모델 : 외부 경정맥 정맥 문합에 경동맥.

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

동정맥루 실패의 소설 쥐 모델 : 세부 수술

유출 협착증을 가진 Aortocaval 누공을 사용하여 중앙 정맥 협착증의 뮤린 모형

쥐 뒷다리 모델에서 동맥 형성 중 백혈구 모집을 모니터링하기 위한 다광자 생체 내 이미징

혈액 투석 접근 관련 손 기능 장애의 쥐 모델

안지오텐신 II의 마우스 모델에서 중앙 정맥 카테터에 의한 약물 치료 유도 복부 대동맥류 및 3D 초음파에 의한 모니터링

토끼의 동정맥루 건설을위한 수정 된 기술

성인 마우스 정맥 고혈압 모델 : 외부 경정맥 정맥 문합에 경동맥.