우리는 스캐닝 전자 현미경 샘플 처리에 대한 그녀의 기술 지원으로 Maryland School of Medicine의 Electron Microscopy Core Facility에서 Hsia Ru-ching 박사에게 감사드립니다.

윤리 성명서 : 동물 취급을위한 프로토콜은 메릴랜드 대학의 기관 동물 관리 및 사용위원회 (IACUC) (프로토콜 번호 0416009)의 승인을 받았고 AAALAC-International 표준에 따라 시행되었습니다. 1. 수술 절차 <ol><li> 마취 기법 <ol><li> 생존 수술에 사용 된 모든기구를 121 ° C에서 30 분 동안 증기 멸균하여 소독하십시오. </li><li> 정밀 증발기 를 통해 전달 되는 100 % O 2 및 2.5 % 이소 플루 란 을 포함 하는 유도 탱크를 통해 마취를 유도합니다. 사후 유도, isoflurane을 중단하고 O 2로 챔버를 플러시하십시오. 안면 마스크 를 통한 1.5-2 % isoflurane과 흡입에 의한 1 L / min O2로 마취를 유지하십시오. </li><li> 인원을 보호하기 위해 배기 가스 흡착을위한 숯 제거제에 유도 챔버와 안면 마스크를 부착하십시오. 시위로 적절한 마취기를 확보하십시오.유해한 자극 (발가락 핀치)에 대한 반응이 없다는 것. </li><li> 등온 패드가있는 수술 트레이로 구성된 수술장을 만들어 수술 중에 열을 받치도록하십시오. 추가적인 등온 패드는 회복 케이지에서 동물에게 열적지지를 제공합니다. </li></ol></li><li> 동물 준비 <ol><li> 10-12 주령의 수컷 C57BL / 6 마우스에 대해 다음 조사를 수행하십시오. </li><li> 탈모 제 또는 40 번 칼날이 달린 전기 클리퍼로 흉골에서 사타구니 부위까지 동물 복부 복부 표면의 머리카락을 제거합니다. <ol><li> 탈모제의 경우 2-3 분 동안이 화합물을 수술 부위에 바른 후면으로 제거하십시오. 우리는 상업적으로 이용 가능한 수산화칼슘과 수산화 나트륨 화합물을 사용합니다. </li></ol></li><li> 70 % 알콜로 어깨 부분을 준비하고 25 게이지 또는 더 작은 보어 바늘로 carprofen (5 mg / kg)을 피하 주사하십시오. 이치료는 동물을위한 수술 후 진통제를 제공 할 것입니다. </li><li> 외과 분야에 동물을 전송하고 지느러미 응급실에서 위치. </li><li> 깨끗한 코튼 어플리케이터 또는면 거즈로 8-12 %의 섭취 된 요오드를 문질러 수술 부위를 준비하십시오. 그런 다음 70 % 알코올로 피부를 두 번 헹굽니다. </li><li> 각막 건조의 발병률을 줄이기 위해 안구 윤활제를 양안에 넣으십시오. 멸균 드레이프로 수술 부위를 덮으십시오. </li></ol></li><li> 수술 기법 <ol><li> 중간 복부 개복술 절개를 대략 2-2.5 cm 길이로 만들고, 칼피스를 즉시 꼬리뼈 골반쪽으로 가져오고 골반쪽으로 확장합니다. </li><li> 소장과 십이지장을 동원하여 오른쪽 옆으로 반사시킵니다. 노출을 향상시키기 위해 내장을 포장 할 수 있도록 주사 용 멸균 식염수로 닦고 닦아 낸 멸균 된면을 겹쳐서 감습니다. </li><li> 소장이 오른쪽에 동원되어복강 내막을 노출시키고 복부 대동맥을 좌측 신장 정맥에서 대동맥 분기로 노출시킨다 ( 그림 1 ). </li><li> 멸균 된 면봉으로 된 애플리케이터로 각 5 초 동안 30 회 연속 분쇄합니다. </li><li> 패킹을 제거하고 내장이 원래 위치로 돌아가도록하십시오. </li><li> 실행중인 4-0 흡수 monofilament 봉합사 (polydioxanone)로 근막을 닫으십시오. 피부는 6-0 비 흡착성 모노 필라멘트 봉합사 (나일론)로 막습니다. </li></ol></li><li> 복구 및 사후 절차 관리 <ol><li> 절차가 끝나면 등온 패드에 깨끗한 침구가있는 복구 케이지에 동물을 넣어 동물이 정상적으로 보행 할 수있을 때까지 열 지원을 계속하십시오. 흉골의 머리맡을 유지할 수있을 때까지 동물을 방치하지 마십시오. </li><li> 수술 후 처음 4 시간 동안 매 시간마다 동물을 관찰하십시오. 동물이 정상적으로 돌아 오는 동안 나는<html> 할당 된 축산 실로 그것이 완전히 회복 될 때까지 동물은 다른 동물의 회사에 수용되지 않을 것입니다. </li><li> 수술 후 첫 72 시간 동안은 동물을 하루 두 번, 그 후 일주일에 적어도 3 번은 동물을 관찰하십시오. 모니터링은 일주일에 세 번 동물의 무게를 측정하는 것을 포함합니다. </li><li> 수술 후 첫 72 시간 동안 carpofen (5 mg / kg)을 매일 두 번 피하 투여하십시오. </li></ol></li></ol> 2. 조직 조달 <ol><li> 안락사의 방법 <ol><li> 미리 정해진 시간에 동물을 안락사하십시오. </li><li> 정밀 증발기를 통해 전달되는 100 % O 2 및 2.5 % 이소 플루 란을 포함하는 유도 탱크를 통해 마취를 유도합니다. <ol><li> 내피의 완전성을 연구하기 위해 Evans Blue 염료를 동물에게 투여하십시오. 참고 : Evans Blue는 알부민에 대한 결합 친화력이 높은 음전하를 띤 아조 염료이며 손상되지 않은 내피가없는 경우에만 혈관을 염색 할 수 있습니다s = &quot;xref&quot;&gt; 6. </li><li> 내피 완전성 연구를 위해, 안락사시, 0.3 % 에반스 블루 (Evans Blue) 염색약 5 mL와 생리 학적 압력 5 mL PBS로 5 분간 동물을 관류시킨다. 동물은 재관류 과정 중에 마취 외과 수술 평면에 유지됩니다. </li></ol></li><li> 깊은 마취기를 마친 후 흉골 절개로 가슴을 엽니 다. 오른쪽 아트리움에서 열혈을하여 동물의 혈액을 배수시키고 21G 주사 바늘로 왼쪽 심실에 액세스하고 오른쪽 심방의 유출액이 깨끗해질 때까지 인산염 완충 생리 식염수 (PBS)를 주입하십시오. </li><li> PBS로 재관류 한 후 정중선 절개를 통해 복부로 들어갑니다. </li><li> 다시 한번, 소장을 신장 우측 대동맥을 드러내는 복부의 오른쪽으로 동원시키고, 인접한 조직으로부터 대동맥을 급히 절개하여 왼쪽 신장 혈관에서 대동맥 분기로 절제합니다. </li><li> excised 대동맥을 4 % paraformald에 보관하십시오.조직 처리가 발생할 때까지 ehyde 용액. <ol><li> 내피 완전성 연구를 위해, 대동맥을 세로로 열고 내강 표면 전체를 노출시키는 왁스 시트에 핀을 고정시킵니다. Evans Blue dye 6로 염색 정도에 따라 혈관 내피의 정성 평가를 수행하십시오. </li><li> 다른 조직학 검사의 경우, 대동맥을 횡 방향으로 절개하고 조직 학적 방법에 따라 최적 절개 온도 (OCT) 화합물에 삽입하십시오 7 . </li></ol></li></ol></li></ol>

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이 작품은 Veterans Affairs Career Development Award (1IK2BX001553-01) (TSM) 및 Vascular Cures EJ Wylie 장학금 (TSM)에 의해 지원되었습니다.

우리는 내측 과형성 및 내피 장벽 기능 장애를 초래하는 쥐 대동맥 손상 모델의 효과를 특성화했다. 대동맥 내막을 따라 부분적인 EC 분리는 세포 - 세포 접촉의 손실과 세포 돌출의 증가를 수반했다. 따라서, 내피 장벽 기능이 현저히 손상되어 분열 촉진 물질 민감성 신호 전달 경로가 자극되어 VSMC가 증식되고 혈관벽이 두꺼워진다. 이 모델의 강점은 다른 대동맥 질환 모델보다 배우고 실행하는 ?...

재 협착 심혈관 수술 (바이 패스 수술, 혈관 성형술 또는 스텐트 삽입)은 이러한 과정의 내구성을 감소시키는 중요한 문제입니다. 모든 재 혈관 화 절차는 재협착에 시달립니다. 재 협착 (약물 방출 스텐트 및 약물 코팅 된 풍선)을 방지하는 현재의 전략은 혈관 평활근 세포 (VSMC) 및 내피 세포 증식 (EC)을 억제한다. 결과적으로, 이러한 중재는 VSMC가 중재하는 재 협착을 예방할뿐만 아니라 내피 세포의 재생을 막습니다. 손상되지 않은 내피없이 환자는 출혈 합병증의 위험이있는 현장 혈전증의 위험을 줄이기 위해 강력한 항 혈소판제를 사용해야합니다. 이상적인 치료법은 VSMC의 증식을 억제하면서 내피 세포의 재생을 촉진합니다. 따라서, 생체 내에서 VSMC 증식과 내피 장벽 기능을 동시에 연구 할 필요가있다. 현재,재 협착의 알 마우스 모델 1 . 이 모델에는 경동맥 결찰 및 대퇴 동맥 와이어 손상 2가 포함 됩니다. 대동맥 모델은 스텐트 배치 3 , 풍선 손상 4 및 대동맥 동종 이식 5를 포함 합니다. 현재 모델은 모두 제한되어 있습니다. 경동맥 연결은 혈류가 매개되는 신생 내막 병변을 유발하고 내피 손상을 일으키지 않습니다. 또한 경동맥과 대퇴 동맥은 인간 혈관보다 세포 배양이 훨씬 적어서 병독성이 제한됩니다. 직경 약 1.3 mm 인 마우스 대동맥은 임상 관련 (관상 동맥) 인간 동맥 (3)과 근사하는 유일한 혈관입니다. 질병의 쥐 대동맥 모델의 번역 가능성에도 불구하고, 현재 모델에는 한계가 있습니다. 이러한 모델은 첨단 미세 수술 기술과 혈관 성형 풍선 및 스텐트와 같은 전문 장비가 필요합니다. 여기서, 우리는VSMC의 증식을 동시에 유도하고 내피 장벽 기능을 파괴하는 새로운 재현 가능 기술.

<table><tbody><tr><td>Ocular lubricant</td><td>Dechra</td><td>17033-211-38</td><td>Pharmaceutical agents</td></tr><tr><td>Isoflurane</td><td>VetOne</td><td>502017</td><td>Pharmaceutical agents</td></tr><tr><td>Carprofen</td><td>Zoetis</td><td>26357</td><td>Pharmaceutical agents</td></tr><tr><td>Precision vaporizer</td><td>Summit Medical</td><td>10675</td><td>Surgical supplies</td></tr><tr><td>Charcoal scavenger</td><td>Bickford Inc.</td><td>80120</td><td>Surgical supplies</td></tr><tr><td>Isothermal pad</td><td>Harvard Apparatus</td><td>50-7053-R</td><td>Surgical supplies</td></tr><tr><td>Sterile cotton-tipped applicator</td><td>Fisher Scientific</td><td>23-400-124</td><td>Surgical supplies</td></tr><tr><td>4-0 absorbable monofilament suture&amp;nbsp;</td><td>Ethicon, Inc</td><td>J310</td><td>Surgical supplies</td></tr><tr><td>5-0 non-absorbable monofilament suture</td><td>Ethicon,Inc</td><td>1666</td><td>Surgical supplies</td></tr><tr><td>21-gauge x 1 inch needle</td><td>BD Biosciences</td><td>305165</td><td>Surgical supplies</td></tr><tr><td>25-gauge x 1 inch&amp;nbsp; needle</td><td>BD Biosciences</td><td>305125</td><td>Surgical supplies</td></tr><tr><td>Dry sterilizer</td><td>Cellpoint&amp;nbsp;</td><td>7770</td><td>Surgical supplies</td></tr><tr><td>Fine scissors</td><td>Fine Science Tools</td><td>14058-09</td><td>Surgical instruments</td></tr><tr><td>Adson forceps</td><td>Fine Science Tools</td><td>11006-12</td><td>Surgical instruments</td></tr><tr><td>Dumont #5 fine forceps</td><td>Fine Science Tools</td><td>11254-20</td><td>Surgical instruments</td></tr><tr><td>Vannas Spring Scissors 3 mm cutting edge</td><td>Fine Science Tools</td><td>15000-00</td><td>Surgical instruments</td></tr><tr><td>Needle driver</td><td>Fine Science Tools</td><td>91201-13</td><td>Surgical instruments</td></tr><tr><td>Scalpel handle #4</td><td>Fine Science Tools</td><td>10004-13</td><td>Surgical instruments</td></tr><tr><td>Scalpel blades #10</td><td>Fine Science Tools</td><td>10010-00</td><td>Surgical instruments</td></tr><tr><td>PBS&amp;nbsp;</td><td>Lonza</td><td>17-516F</td><td>Reagents for tissue processing</td></tr><tr><td>Evans Blue</td><td>Sigma-Aldrich</td><td>E2129</td><td>Reagents for tissue processing</td></tr><tr><td>Paraformaldehyde</td><td>Sigma-Aldrich</td><td>P6148</td><td>Reagents for tissue processing</td></tr><tr><td>Modeling wax</td><td>Bego</td><td>40001</td><td>Reagents for tissue processing</td></tr><tr><td>OCT compound</td><td>Tissue-Tek Sakura</td><td>4583</td><td>Reagents for tissue processing</td></tr><tr><td>Mayer&#039;s hematoxylin solution</td><td>Sigma-Aldrich</td><td>MHS16</td><td>Reagents for immunohistological analysis</td></tr><tr><td>Eosin Y solution alcoholic&amp;nbsp;</td><td>Sigma-Aldrich</td><td>HT110316</td><td>Reagents for immunohistological analysis</td></tr><tr><td>Elastin stain kit</td><td>Sigma-Aldrich</td><td>HT25A</td><td>Reagents for immunohistological analysis</td></tr><tr><td>Click-it Edu Alexa-488 Imaging Kit</td><td>Invitrogen</td><td>C10337</td><td>Reagents for immunohistological analysis</td></tr><tr><td>Anti-Erk1/2 antibody</td><td>Cell Signaling Technology</td><td>4695</td><td>Reagents for immunohistological analysis</td></tr><tr><td>Anti-phospho-Erk1/2 antibody</td><td>Cell Signaling Technology</td><td>4370</td><td>Reagents for immunohistological analysis</td></tr><tr><td>Anti-p27kip1 antibody</td><td>Cell Signaling Technology</td><td>3698</td><td>Reagents for immunohistological analysis</td></tr><tr><td>Trichloroacetic acid</td><td>Sigma-Aldrich</td><td>T9159</td><td>Reagents for immunohistological analysis</td></tr></tbody></table>

murine aortic crush injury: an efficient in vivo model of smooth muscle cell proliferation and endothelial function

혈관 성형술이든 우회 수술이든 동맥 재건술은 내피 세포 손상 및 혈관 평활근 세포 증식을 유발하는 의원 성 외상을 포함합니다. 일반적인 쥐 모델은 경동맥과 대퇴 동맥과 같은 작은 혈관을 연구합니다. 여기서 우리는 VSMC 증식과 내피 장벽 기능이 대형 혈관에서 동시에 평가 될 수 있는 생체 내 시스템을 설명한다. 우리는 C57BL / 6 마우스의 손상에 대한 신장 하 대동맥 반응을 연구했습니다. 대동맥은 좌측 신장 정맥에서 대퇴 동맥 분 지로 손상되었는데, 코튼 팁 도포기로 5 초간 30 회 변형시켰다. 형태 학적 변화는 전통적인 조직학으로 평가되었다. 대동맥 벽 두께는 내강 표면에서 외막까지 측정되었다. VSMC 증식을 입증하기 위해 EdU 통합 및 DAPI 및 알파 - 액틴에 의한 카운터 염색을 사용 하였다. ERK1 / 2 (내막 과형성 형성의 알려진 조절 인자)의 활성화가 억제되었다Western blot 분석으로 채취 하였다. 염증의 효과는 B 세포, T 세포 및 대 식세포에 대한 면역 조직 화학 법에 의해 결정되었다 . 내피 세포의 각면을 주사 전자 현미경 (SEM)으로 시각화 하였다. Evans Blue 염색으로 내피 장벽 기능을 측정 하였다. Transmural 손상으로 대동맥 벽이 두꺼워졌다. 이 손상은 손상 후 3 일째에 가장 현저하게 VSMC 증식을 유도하고 ERK1 / 2의 조기 활성화 및 p27 kip1 발현을 감소 시켰다. 부상으로 혈관벽에 B 세포, T 세포 또는 대 식세포 침윤이 증가하지 않았습니다. 상해로 인해 부분 내피 세포 누출과 세포 - 세포 접촉의 손실이 발생했습니다. 손상으로 인해 7 일 후에 기준선으로 돌아온 내피 장벽 기능이 크게 상실되었습니다. 횡격막 횡격막 둔 상동맥 손상 모델은 대형 선박에서 VSMC 증식과 내피 장벽 기능을 동시에 연구 할 수있는 효율적인 시스템을 제공합니다.

OCT에 내장 된 횡단 절제 된 대동맥을 절단하고 hematoxylin과 eosin으로 염색 한 후 Verhoeff-Van Gieson (VVG) 염색으로 반으로 염색하여 내부 및 외부 신축성 층을 확인 하였다. 가식 과정 (개복술과 작은 창자 동원)으로 치료 한 동물의 대동맥에 비해 크러쉬 부상으로 대동맥 벽 비후가 유발되었습니다. 외막에서 내강까지의 거리로 평가 한 벽 두께는 손상 3 일 후 가장 컸다 (42.2 ± 1....

Watch this Scientific Journal Video about Murine Aortic Crush Injury: An Efficient In Vivo Model of Smooth Muscle Cell Proliferation and Endothelial Function at JoVE.com

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

심혈관 수술 (바이 패스 수술, 혈관 성형술 또는 스텐트 삽입) 후의 재 협착은 이러한 과정의 내구성을 감소시키는 중요한 문제입니다. 이상적인 치료법은 내피 세포의 재생을 촉진시키면서 평활근 세포 (VSMC)의 증식을 억제합니다. 우리는 생체 내에서 VSMC의 증식과 내피 기능의 동시 평가를위한 모델을 설명합니다.

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 ...

murine-aortic-crush-injury-an-efficient-vivo-model-smooth-muscle-cell

Universidad de Cartagena UDEC

Research

JoVE Journal

Biology

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

8.3K 조회수.  Baltimore Veterans Affairs Medical Center.  심혈관 수술 (바이 패스 수술, 혈관 성형술 또는 스텐트 삽입) 후의 재 협착은 이러한 과정의 내구성을 감소시키는 중요한 문제입니다. 이상적인 치료법은 내피 세포의 재생을 촉진시키면서 평활근 세포 (VSMC)의 증식을 억제합니다. 우리는 생체 내에서 VSMC의 증식과 내피 기능의 동시 평가를위한 모델을 설명합니다. 

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

Using In Vivo and Tissue and Cell Explant Approaches to Study the Morphogenesis and Pathogenesis of the Embryonic and Perinatal Aorta

The aorta is the largest artery in the body. The aortic wall is composed of an inner layer of endothelial cells, a middle layer of alternating elastic lamellae and smooth muscle cells (SMCs), and an outer layer of fibroblasts and extracellular matrix. In contrast to the widespread study of pathological models (e.g., atherosclerosis) in the adult aorta, much less is known about the embryonic and perinatal aorta. Here, we focus on SMCs and provide protocols for the analysis of the morphogenesis and pathogenesis of embryonic and perinatal aortic SMCs in normal development and disease. Specifically, the four protocols included are: i) in vivo embryonic fate mapping and clonal analysis; ii) explant embryonic aorta culture; iii) SMC isolation from the perinatal aorta; and iv) subcutaneous osmotic mini-pump placement in pregnant (or non-pregnant) mice. Thus, these approaches facilitate the investigation of the origin(s), fate, and clonal architecture of SMCs in the aorta in vivo. They allow for modulating embryonic aorta morphogenesis in utero by continuous exposure to pharmacological agents. In addition, isolated aortic tissue explants or aortic SMCs can be used to gain insights into the role of specific gene targets during fundamental processes such as muscularization, proliferation, and migration. These hypothesis-generating experiments on isolated SMCs and the explanted aorta can then be assessed in the in vivo context through pharmacological and genetic approaches.

Using In Vivo and Tissue and Cell Explant Approaches to Study the Morphogenesis and Pathogenesis of the Embryonic and Perinatal Aorta

Protocols for studying the embryonic and perinatal murine aorta using in vivo clonal analysis and fate mapping, aortic explants, and isolated smooth muscle cells are detailed here. These diverse approaches facilitate the investigation of the morphogenesis of the embryonic and perinatal aorta in normal development and the pathogenesis in disease.

Morphogenesis 배아 및 Perinatal 대동맥의 병 인을 공부 하 고 Vivo에서 , 조직 및 세포 Explant 접근을 사용 하 여

The aorta is the largest artery in the body. The aortic wall is composed of an inner layer of endothelial cells, a middle layer of alternating elastic ...

연구

발생학

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. ...

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 ...

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 ...

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 ...

Balloon-based Injury to Induce Myointimal Hyperplasia in the Mouse Abdominal Aorta

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine balloon denudation model, which is comparable with established vessel injury models in large animals. The aorta denudation model with balloon catheters mimics the clinical setting and leads to comparable pathobiological and physiological changes. Briefly, after performing a horizontal incision in the aorta abdominalis, a balloon catheter will be inserted into the vessel, inflated, and introduced retrogradely. Inflation of the balloon will lead to intima injury and overdistension of the vessel. After removing the catheter, the aortic incision will be closed with single stiches. The model shown in this article is reproducible, easy to perform, and can be established quickly and reliably. It is especially suitable for evaluating expensive experimental therapeutic agents, which can be applied in an economical fashion. By using different knockout-mouse strains, the impact of different genes on MH development can be assessed.

This article demonstrates a murine model to study the development of myointimal hyperplasia (MH) after aortic balloon injury.

풍선-기반 부상 마우스 복 부 대동맥에서 Myointimal 증식을 유도 하

The use of animal models is essential for a better understanding of MH, one major cause for arterial stenosis.In this article, we demonstrate a murine ...

Construction of a Human Aorta Smooth Muscle Cell Organ-On-A-Chip Model for Recapitulating Biomechanical Strain in the Aortic Wall

Conventional two-dimensional cell culture techniques and animal models have been used in the study of human thoracic aortic aneurysm and dissection (TAAD). However, human TAAD sometimes cannot be characterized by animal models. There is an apparent species gap between clinical human studies and animal experiments that may hinder the discovery of therapeutic drugs. In contrast, the conventional cell culture model is unable to simulate in vivo biomechanical stimuli. To this end, microfabrication and microfluidic techniques have developed greatly in recent years, providing novel techniques for establishing organoids-on-a-chip models that replicate the biomechanical microenvironment. In this study, a human aorta smooth muscle cell organ-on-a-chip (HASMC-OOC) model was developed to simulate the pathophysiological parameters of aortic biomechanics, including the amplitude and frequency of cyclic strain experienced by human aortic smooth muscle cells (HASMCs) that play a vital role in TAAD. In this model, the morphology of HASMCs became elongated in shape, aligned perpendicularly to the strain direction, and presented a more contractile phenotype under strain conditions than under static conventional conditions. This was consistent with the cell orientation and phenotype in native human aortic walls. Additionally, using bicuspid aortic valve-related TAAD (BAV-TAAD) and tricuspid aortic valve-related TAAD (TAV-TAAD) patient-derived primary HASMCs, we established BAV-TAAD and TAV-TAAD disease models, which replicate HASMC characteristics in TAAD. The HASMC-OOC model provides a novel in vitro platform that is complementary to animal models for further exploring the pathogenesis of TAAD and discovering therapeutic targets.

Here, we developed a human aorta smooth muscle cell organ-on-a-chip model to replicate the in vivo biomechanical strain of smooth muscle cells in the human aortic wall.

대동맥 벽의 생체 역학적 변형을 요약하기 위한 인간 대동맥 평활근 세포 장기 온 칩 모델 구축

Conventional two-dimensional cell culture techniques and animal models have been used in the study of human thoracic aortic aneurysm and dissection ...

생체공학

Quantitative Micro-CT Analysis of Aortopathy in a Mouse Model of &#946;-aminopropionitrile-induced Aortic Aneurysm and Dissection

Aortic aneurysm and dissection is associated with significant morbidity and mortality in the population and can be highly lethal. While animal models of aortic disease exist, in vivo imaging of the vasculature has been limited. In recent years, micro-computerized tomography (micro-CT) has emerged as a preferred modality for imaging both large and small vessels both in vivo and ex vivo. In conjunction with a method of vascular casting, we have successfully used micro-CT to characterize the frequency and distribution of aortic pathology in &#946;-aminopropionitrile-treated C57/Bl6 mice. Technical limitations of this method include variations in the quality of the perfusion introduced by poor animal preparation, the application of proper methodologies for vessel size quantification, and the non-survivability of this procedure. This article details a methodology for the intravascular perfusion of a lead-based radiopaque silicone rubber for the quantitative characterization of aortopathy in a mouse model of aneurysm and dissection. In addition to visualizing aortic pathology, this method may be used for examining other vascular beds in vivo or vascular beds removed post-mortem.

This article describes a detailed methodology of using a radiopaque lead-based silicone rubber to perfuse the murine vasculature for aortic diameter quantification in a mouse model of aortic aneurysm and dissection.

Β-aminopropionitrile-유도 대동맥 동맥 및 해 부의 마우스 모델에서 Aortopathy의 양적 마이크로 CT 분석

Aortic aneurysm and dissection is associated with significant morbidity and mortality in the population and can be highly lethal. While animal models of ...

Murine Cervical Aortic Transplantation Model using a Modified Non-Suture Cuff Technique

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. However, only minor improvement in the long-term results of transplanted solid organs could be observed over the past decades. In this context, chronic allograft vasculopathy (CAV) still represents the leading cause of late organ failure in cardiac, renal and pulmonary transplantation.
Thus far, the underlying pathogenesis of CAV development remains unclear, explaining why effective treatment strategies are presently missing and emphasizing a need for relevant experimental models in order to study the underlying pathophysiology leading to CAV formation. The following protocol describes a murine heterotopic cervical aortic transplantation model using a modified non-suture cuff technique. In this technique, a segment of the thoracic aorta is interpositioned in the right common carotid artery. With the use of the non-suture cuff technique, an easy to learn and reproducible model can be established, minimizing the possible heterogeneity of sutured vascular micro anastomoses.

Here, we present a protocol of heterotopic aortic transplantation in mice using the non-suture cuff technique in a cervical murine model. This model can be used to study the underlying pathology of chronic allograft vasculopathy (CAV) and can help evaluate new therapeutic agents in order to prevent its formation.

수정된 비봉합사 커프 기술을 사용한 뮤린 자궁 경부 대동맥 이식 모델

With the introduction of powerful immunosuppressive protocols, distinct advances are possible in the prevention and therapy of acute rejection episodes. ...

Isolation of Primary Patient-specific Aortic Smooth Muscle Cells and Semiquantitative Real-time Contraction Measurements In Vitro

Smooth muscle cells (SMCs) are the predominant cell type in the aortic media. Their contractile machinery is important for the transmission of force in the aorta and regulates vasoconstriction and vasodilation. Mutations in genes encoding for the SMC contractile apparatus proteins are associated with aortic diseases, such as thoracic aortic aneurysms. Measuring SMC contraction in vitro is challenging, especially in a high-throughput manner, which is essential for screening patient material. Currently available methods are not suitable for this purpose. This paper presents a novel method based on electric cell-substrate impedance sensing (ECIS). First, an explant protocol is described to isolate patient-specific human primary SMCs from aortic biopsies and patient-specific human primary dermal fibroblasts for the study of aortic aneurysms. Next, a detailed description of a new contraction method is given to measure the contractile response of these cells, including the subsequent analysis and suggestion for comparing different groups. This method can be used to study the contraction of adherent cells in the context of translational (cardiovascular) studies and patient and drug screening studies.

Isolation of Primary Patient-specific Aortic Smooth Muscle Cells and Semiquantitative Real-time Contraction Measurements In Vitro

This paper describes an explant culture-based method for the isolation and culturing of primary, patient-specific human aortic smooth muscle cells and dermal fibroblasts. Furthermore, a novel method is presented for measuring cell contraction and subsequent analysis, which can be used to study patient-specific differences in these cells.

일차 환자 특이적 대동맥 평활근 세포 및 체외에서 반정량적 실시간 수축 측정의 분리

Smooth muscle cells (SMCs) are the predominant cell type in the aortic media. Their contractile machinery is important for the transmission of force in ...

소뇌 대동맥 파열 상해 : 효율적

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