TWP was funded by the Irish Cardiac Society Brian McGovern Travelling Fellowship. WTP was funded by R01 HL095712 and an AHA Established Investigator Award.

모든 기술 단계는 보스턴 아동 병원의 기관 동물 케어 및 사용위원회에 의해 승인 된 프로토콜에 따라 수행되었다. 1 준비 <ol><li> 심근 내 주입을위한 최적의 고정 센서의 위치가 해부학 및 기능 평가를 서술을위한 최적의 표준 뷰를 생성하지 수 있으므로, 이전에 주입 프로토콜을 시작으로 심 초음파에 의한베이스 라인 심장 해부학 및 기능 평가를 수행합니다. </li><li> 사출 설치는도 1a-1c에 표시됩니다. 피복 빈 바늘 주사기를 놓고, 위쪽으로 경사 배향은, 주사기 클램프 (도 1c 참조)와 검사 헤드 클램프 초음파 변환기 프로브를 고정 (도 1b 참조). 스캔 헤드 클램프 공 잠금 조인트 (도 1b)를 느슨하게하고 바늘의 축에 평행하게 정렬되도록 트랜스 듀서 방향을 조작. 스캔 - 수정스캔 머리 클램프 볼 잠금 관절을 조여 헤드 위치. 참고 : 성인 쥐에 주사를 들어, 1 / 2.5 cm 길이 30 G 바늘이 최적입니다. 기밀 주사기는 작은 볼륨의보다 정확한 제어 (5 ~ 10 μL)에 사용할 수있는 동안 한 ML의 주사기는 더 큰 볼륨을 사용할 수 있습니다. </li><li> 전체가 함께 머리를 보호하기 위해 주걱으로 트랜스 듀서 팁에 자유롭게 초음파 젤을 적용합니다. 조심스럽게 바늘을 unsheathe 바늘 직접 트랜스 듀서에서와 시각화를위한 초음파 젤 내에 바늘을 사전에 컨트롤을 장착 사용합니다. 니들은 초음파 영상에서 그 길이를 따라 명확하게 가시화되도록 바늘 제어를 탑재하여 미세 조절. 변환기가 적절 단계 1.4에서 바늘에 평행하게 정렬 된 경우는 전진 및 분사 제어 노브 (도 1C)와 같이 배출된다 후 바늘 촬상면 내에 유지한다. </li><li> 후속 단계에서, 요구를 방해하지 않는다수평 축에 하나를 이동하여 제작 / 트랜스 듀서 수평 정렬. 오히려, 바늘의 수직 (Y 축)의 위치를​​ 변경하여 장착하고 동물 플랫폼을 이동시켜 주사 중심의 특정 부분을 타겟팅. </li><li> 동물의 플랫폼에 마취 된 마우스의 후속 배치를 할 수 있도록 스캔 높이 제어 (그림 1B)를 사용하여 동물의 플랫폼에서 우량 변환기를 이동합니다. 이 바늘의 긴 축에 진동자의 X 축 배향을 방해하지 않을 것이다. </li><li> 주사기 클램프에서 정렬을 위해 사용 조심스럽게 폐기 된 주사기를 제거합니다. 새 바늘을 넣고 주사기 팁 죽은 공간을 허용, 최종 목표 볼륨으로 injectate 주사기. 공기 방울을 제거주의하십시오. 그 x 축 정렬을 조정하지 않고 주사기 클램프에 주사기를 놓습니다. 완전 분사 제어를 사용하여 주사기를 후퇴. 참고 : 초기 교육 목적을 위해, 에반의 파란색의 사용염료 (1 %)은, 트리 판 블루 스테인 (0.4 %) 또는 injectate 같은 형광 미립자의 현탁액을 주입 능력과 표적의 확인에 성공 연산자를 어시스트 할 수있다. </li></ol> 2 사출 <ol><li> 가열 플랫폼의 통합 따뜻한 켜고 37 ° C &#39;의 설정합니다. 운영자에 가장 가까운 동물의 마취 호스 클램프 및 머리, 일반적인 영상 방향에서 동물의 플랫폼 180 °를 놓습니다. 이것은 (가슴의 왼쪽에) 마음이 주사기 클램프와 바늘 동측 될 수 있습니다. 주 : 동물 플랫폼에 추가 작은 시계 방향으로 회전 조정 분사 (그림 2A)에 사용되는 흉골 단축 영상 평면에 대해 올바르게 마음을 방향을해야 할 수도 있습니다. </li><li> 이전에 구를보고 심장 초음파 검사 생쥐를 준비합니다. 2 % 이소 플루 란으로 유도 챔버에 마우스를 마취. 머리 제거 크림과 함께 가슴 털을 제거공막의 건조를 방지하기 위해 두 눈에 젤 윤활 적용 거라고. </li><li> 스캔 헤드 높이 제어 (도 1b)을 사용하여 트랜스 듀서를 올린다. 1-3%의 이소 플루 란 (그림 2A-2B)를 제공하는 코 콘 내 주둥이로 가열 된 동물 플랫폼 꼭대기 마취 마우스 부정사를 놓습니다. 부드럽게 직장 프로브를 삽입하고 전기 접촉 전극 젤을 적용, ECG 전극에 네 발을 테이프. NOTE : 마취 적정 수준이 동물의 치료를 위해 인간을 확보해야한다. 심장 박동의 변화와 가슴 벽을 통해 바늘의 위치에 대한 응답이 없어야합니다. 저체온증은 상대적 서맥, 심실 팽창, 가능한 불편을 초래할 것 같이 동물 플랫폼의 통합 항상성 온도 컨트롤은 정상 체온 (37 ± 0.5 ° C)를 유지하는 데 사용되어야한다. </li><li> 마우스 동물 플랫폼 보안되면 depila 상에 트랜스 듀서를 낮출스캔 헤드 높이 조절 (그림 1B)를 사용하여 테드의 가슴. 심장이 표준 심 초음파 기법에 따라, 흉골 단축 방향으로 가시화 될 때까지 주입을위한 최적의 초음파 설정입니다. 단축 영상 평면 (그림 2A-2B) 주입을위한 최적의 음향 창을 얻기 위해 동물 플랫폼에게 20 ~ 30 ° 시계 방향으로 돌립니다. 주 : 대안 적으로, 주입은 동물 플랫폼 (도 2c)의 반 시계 방향 회전에 의해 흉골 장축 방향에서 수행 될 수있다. </li><li> 시야를 조정하고 좌심실의 심근에서 원하는 주사 부위를 대상으로 동물 플랫폼 조절 제어를 사용합니다. 좌심실 심근 (도 3A-3C)에서 원하는 주입 위치를 대상으로 심장의베이스에 정점으로부터 전후 패닝. 참고 : midpapillary 흉골 짧은 축보기 (그림 3a는) reproducibl을 제공합니다주사 부위의 후속 영상을 허용 전자의 랜드 마크. 참고 : 또는 흉골 장축이 미리 정의 된 주입 부위 (도 3D)을 대상으로 사용될 수있다. </li><li> 완전히 수축 된 주사기 클램프 주사기를 시작으로 천천히 분사 제어를 시계 방향으로보기 (그림 4A)를 돌려 동물의 가슴쪽으로 주사기를 진행합니다. 그 가슴에 접근 심장 및 바늘 끝 모두 명확한 초음파 시각화를 허용하기 위해, 가슴의 왼쪽 위에 초음파 겔을 충분히 사용하고 심장 초음파 제어에 넓은 시야를 설정하여 음향 창을 최적화. 주사에 대한 대상 사이트에 초점 / 시간대를 설정합니다. 바늘에 약간의 조정은 제어가 그 길이를 따라 니들의 이미지를 최적화 할 수 마운트. NOTE : 일부 초음파 시스템은 디지털 목표 심근까지 바늘의 장축을 따라 선을 연장 봉침 소프트웨어 기능을 갖는다(그림 4B). 이러한 소프트웨어 도구가 도움이 될 수 있지만 필수적인 것은 아니다있다. </li><li> 동물 적절 진정으로 신중 전혀 모따 바늘 끝의 위치를​​ 관찰하고, 마우스의 흉부 벽을 통해 심근에 바늘을 전진 (1-3 %의 이소 플루 0.5-0.8 L / 분으로 100 %의 산소와 혼합) 배. 니들 팁이 목표 심근 (도 4C) 내에서 전진 할 때 멈춘다. 전체 빗면 팁 심낭 공간으로 injectate 누출을 피하기 위해, 안전하게 심근 내에 있어야한다. </li><li> 선단이 원하는 위치에있을 때, 시린지 플런저를 밀어 injectate을 제공한다. 5 ~ 30 초 동안 천천히 injectate를 제공​​ (볼륨에 따라 전달되는). injectate의 최대 50 μL은 심실 기능을 손상시키지 않고 전달 될 수있다. 주입 된 심근 지역에 과도 echobright 모양이 성공적으로 주입 후 분명있을 수 있습니다. 상대 bradycard의 개요 (초) 기간아이오와 때때로 심근에 주입 언급하고 신속하게 해결됩니다. </li><li> injectate 투여되면, 즉시 분사 제어 노브의 반 시계 방향 회전에 의해 바늘을 인출. 마우스가 보존 심실 기능없이 postprocedural 합병증을 확인하기 위해 심 초음파 관찰 몇 분 동안 마취 유지되어야한다. 표시하는 경우, 복수의 영역의 심근 직렬 동물 플랫폼의 조정에 의해 접근 니들의 각도로 재배치하여 주입 할 수있다. 심근 내 주사 후, 마우스는 그 자체 케이지에 넣고 관찰 하에서 마취에서 회복시킨다. </li><li> 이 흉골 드러 누움을 유지하기 위해 충분한 의식을 회복 할 때까지 무인 동물을 두지 마십시오. 완전히 회복 할 때까지 다른 동물의 회사에 심근 주사를 겪었다 동물을 반환하지 않습니다. 새장은 재와 온도 조절 패드에 배치해야물과 마우스 다이어트의 ADY 제공. Postprocedural 불편이 예상 적절 곧 개입 후 정상적인 동작을 재개 실패는 잠재적 인 합병증 (토론 참조) 제안하지 않습니다. </li><li> 조심스럽게 심근 내 주입의 완료가 운영자 또는 방관자로 날카로운 물건 손상의 위험을 최소화하기 위해 바늘 직후 폐기. 바늘을 재사용하면 더 도전 심근를 관통 할 수있게 팁의 둔화 및 합병증의 위험이 높은 결과가 발생할 수 있습니다. </li></ol>

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G., Suzuki, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+delivery+routes+for+stem+cell+therapy+to+the+heart:+current+and+future+approaches.">Cell delivery routes for stem cell therapy to the heart: current and future approaches.</a> J Cardiovasc Transl Res. 5 (5), 713-726 (2012).</li><li>Hou, D., 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=Radiolabeled+cell+distribution+after+intramyocardial,+intracoronary,+and+interstitial+retrograde+coronary+venous+delivery:+implications+for+current+clinical+trials.">Radiolabeled cell distribution after intramyocardial, intracoronary, and interstitial retrograde coronary venous delivery: implications for current clinical trials.</a> Circulation. 112, 150-156 (2005).</li><li>Dib, 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=Recommendations+for+successful+training+on+methods+of+delivery+of+biologics+for+cardiac+regeneration:+a+report+of+the+International+Society+for+Cardiovascular+Translational+Research).">Recommendations for successful training on methods of delivery of biologics for cardiac regeneration: a report of the International Society for Cardiovascular Translational Research).</a> JACC Cardiovasc Interv. 3 (3), 265-275 (2010).</li><li>Dib, 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=Cell+therapy+for+cardiovascular+disease:+a+comparison+of+methods+of+delivery.">Cell therapy for cardiovascular disease: a comparison of methods of delivery.</a> J Cardiovasc Transl Res. 4 (2), 177-181 (2011).</li></ol>

The authors have nothing to disclose.

생물 의약품은 혈류를 통해 직접 심근 내 주입, intrapericardial 주사, 또는 간접적으로 관리하여 심근에 전달 될 수있다. 심근 경색 모델에서 최근 세포 기반 치료 임상 시험 injectate 12-14의 전달에 개흉술 방식을 설명했다. 심근 치료 적 개입의 성공에 중요 인자는 전달 경로의 선택에 달려있다. 생물의 높은 로컬 도즈는 심근 15,16 전달함으로써 달성된다. 직접 시각화에서 심근 내 주?...

심장 질환은 매년 600,000 사망 한 회계, 미국에서 남성과 여성 모두 사망의 주요 원인이다. 심혈관 질환의 생쥐 모델은 병리 생리 학적 메커니즘을 조사 및 치료 가능성을 둘러보기에 매우 중요하다. 유전자 치료 벡터의 심근 전달, 세포 RNA를 변성하고, 심장 질환에 대한 그들의 2-7 치료 가능성의 다른 치료제 허가 조사 줄기. 현재 마우스 모델 6 치료제의 심근 전달을위한 제한된 옵션이 있습니다. 직접적인 시각화 하에서 심근 내 주입 흔히 사용하지만, 흉골 또는 개흉술을 요구하고 심장의 노출 된 영역에 한정되어있다. 이러한 LAD 결찰 같은 다른 실험 조작의 시간에 수행 될 때 편리하지만, 심근 배달 침습 절차에 대한 필요성이 전위 실험 설계를 제한하고 소개절차와 뉴 부가 효과 (예, 개흉술에 의한 섬유화). 바이러스 벡터의 경피적 심낭 전달보고되었지만 사이트 및 치료제의 분포는 균일하지 않으며 8 제어하는 것이 곤란하다. 주입 물질 있지만 효율적이고 재현 관상 전달보다 균일 분포 경피적 관상 동맥 주입 결과 뮤린 모델에 도전하고있다. 여기에, 우리는 초음파지도하에 치료제의 최소 침습, 운영자 제어 타겟팅을 할 수있는 닫힌 가슴 심근 내 주입 기술을 설명합니다. 기술은 쉽게 배울 개흉술 또는 흉골 및 부수적 인 실험 합병증의 필요성을 제거하고, 타이밍 및 심근 주사 부위에 더 큰 유연성을 제공한다. 따라서, 심장 초음파를 이용한 심근 내 주입 조작의 기술적으로 간단하고 매우 효과적인 방법을 나타낸다쥐 실험 모델에서 심근.

<table border="0" cellpadding="0" cellspacing="0" width="743">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Vevo 2100 ultrasound imaging system&#160;</td>
			<td style="width:144px;">Visualsonics</td>
			<td style="width:119px;"></td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Vevo Integrated Rail System III</td>
			<td style="width:144px;">Visualsonics</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Microscan MS400 transducer</td>
			<td style="width:144px;">Visualsonics</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Microscan MS550D transducer</td>
			<td style="width:144px;">Visualsonics</td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">PrecisionGlide needles</td>
			<td style="width:144px;">BD</td>
			<td style="width:119px;">305128</td>
			<td>30G x 1 inch</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">1cc slip tip syringe</td>
			<td style="width:144px;">Exel International&#160;</td>
			<td style="width:119px;">26048</td>
			<td>or equivalent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Gastight 50ul glass syringes</td>
			<td style="width:144px;">Hamilton&#160;</td>
			<td style="width:119px;">1705</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Trypan blue stain (0.4%)</td>
			<td style="width:144px;">Gibco&#160;</td>
			<td style="width:119px;">15250</td>
			<td>or equivalent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Isoflurane</td>
			<td style="width:144px;">Baxter</td>
			<td style="width:119px;">AHN3640</td>
			<td>or equivalent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Aquasonic 100</td>
			<td style="width:144px;">Parker Laboratories</td>
			<td style="width:119px;">(01-08)</td>
			<td>or equivalent</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:313px;">Polystyrene microspheres (red fluorescent)</td>
			<td style="width:144px;">Life Technologies</td>
			<td style="width:119px;">F-8842</td>
			<td>or equivalent</td>
		</tr>
	</tbody>
</table>

ultrasound-guided transthoracic intramyocardial injection in mice

심혈관 질환의 생쥐 모델은 병리 생리 학적 메커니즘을 조사하고 잠재적 인 재생 치료를 탐험 중요하다. 심근 주입을 포함하는 실험은 현재 개흉술을 통해 직접 수술 접근에 의해 수행된다. 관상 동맥 결찰 같은 다른 실험 조작의 시간에 수행 될 때 편리하지만, 심근 배달 침습 절차에 대한 필요성이 전위 실험 설계를 제한한다. 지금까지 초음파 해상도와 고급 비 침습적 영상 방식을 개선, 그것은 정기적으로 초음파 유도 경피적 심근 내 주입을 수행 할 지금 가능하다. 이 양상은 효율적이고 안정적​​으로 심근의 대상 지역에 에이전트를 제공합니다. 이 기술의 장점은 수술 사망률의 회피, 선택적으로 초음파지도하에 심근의 지역을 대상으로 시설, myocardiu에 injectate을 제공 할 수있는 기회를 포함복수, 소정의 시간 간격 m. 연습 기술로, 심근 내 주입의 합병증은 드문와 마우스를 신속하게 마취에서 회복 정상적인 활동으로 돌아갑니다. 이 프로토콜에 설명 된 단계에 따라 기본 심 초음파 검사 경험이있는 운전자는 신속하게이 다재 다능 한, 최소 침습 기술에서 경쟁력이 될 수 있습니다.

블루 염료 또는 형광 마이크로 스피어와 쥐의 심근 내 주입 에반의 파란색 염료를 주입 연습을 위해 유용합니다. 곧 주사 후, 중간 유두근 수준에서 심근 침윤 청색 염료, 마우스를 안락사 및 주입 청색 염료의 위치를 시각화 할 심장을 제거.도 5를 성공적으로 주입의 일례를 나타낸다 (도 5a는, 영역 밀폐 )의 점선으로. 에반스 블루 염료는 몇 시간 내에 ...

Watch this Scientific Journal Video about Ultrasound-guided Transthoracic Intramyocardial Injection in Mice at JoVE.com

Ultrasound-guided Transthoracic Intramyocardial Injection in Mice

심장 초음파 유도 경피적 심근 주입 뮤린 마음에 유전자 전달 제 또는 세포의 전달을위한 효율적인 신뢰성 있고 타겟팅 양상을 나타낸다. 이 프로토콜에 설명 된 단계에 따라, 운전자는 신속하게이 다재 다능 한, 최소 침습 기술에서 경쟁력이 될 수 있습니다.

마우스의 초음파 유도 경 흉부 심근 내 주입

Murine models of cardiovascular disease are important for investigating pathophysiological mechanisms and exploring potential regenerative therapies. ...

ultrasound-guided-transthoracic-intramyocardial-injection-in-mice

Research

JoVE Journal

Medicine

14.8K 조회수.  Boston Children's Hospital.  심장 초음파 유도 경피적 심근 주입 뮤린 마음에 유전자 전달 제 또는 세포의 전달을위한 효율적인 신뢰성 있고 타겟팅 양상을 나타낸다. 이 프로토콜에 설명 된 단계에 따라, 운전자는 신속하게이 다재 다능 한, 최소 침습 기술에서 경쟁력이 될 수 있습니다. 

비디오: Ultrasound-guided Transthoracic Intramyocardial Injection in Mice

Transthoracic Echocardiography in Mice

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene expression. Transgenic and gene targeting methods can be used to generate mice with altered cardiac size and function,1-3 and as a result, in vivo techniques are needed to evaluate their cardiac phenotype. Transthoracic echocardiography, pulse wave Doppler (PWD), and tissue Doppler imaging (TDI) can be used to provide dimensional measurements of the mouse heart and to quantify the degree of cardiac systolic and diastolic performance. Two-dimensional imaging is used to detect abnormal anatomy or movements of the left ventricle, whereas M-mode echo is used for quantification of cardiac dimensions and contractility.4,5 In addition, PWD is used to quantify localized velocity of turbulent flow,6 whereas TDI is used to measure the velocity of myocardial motion.7 Thus, transthoracic echocardiography offers a comprehensive method for the noninvasive evaluation of cardiac function in mice.

Transthoracic echocardiography offers a noninvasive method for the evaluation of cardiac function in mice. A combination of ultrasound and Doppler imaging modalities can be used to obtain dimensional measurements of the heart and intracardiac blood flow, which together provide an assessment of cardiac systolic and diastolic performance.

마우스에 Transthoracic Echocardiography

In recent years, murine models have become the primary avenue for studying the molecular mechanisms of cardiac dysfunction resulting from changes in gene ...

연구

의학

Coronary Artery Ligation and Intramyocardial Injection in a Murine Model of Infarction

Mouse models are a valuable tool for studying acute injury and chronic remodeling of the myocardium in vivo. With the advent of genetic modifications to the whole organism or the myocardium and an array of biological and/or synthetic materials, there is great potential for any combination of these to assuage the extent of acute ischemic injury and impede the onset of heart failure pursuant to myocardial remodeling. 
Here we present the methods and materials used to reliably perform this microsurgery and the modifications involved for temporary (with reperfusion) or permanent coronary artery occlusion studies as well as intramyocardial injections. The effects on the heart that can be seen during the procedure and at the termination of the experiment in addition to histological evaluation will verify efficacy. 
Briefly, surgical preparation involves anesthetizing the mice, removing the fur on the chest, and then disinfecting the surgical area. Intratracheal intubation is achieved by transesophageal illumination using a fiber optic light. The tubing is then connected to a ventilator. An incision made on the chest exposes the pectoral muscles which will be cut to view the ribs. For ischemia/reperfusion studies, a 1 cm piece of PE tubing placed over the heart is used to tie the ligature to so that occlusion/reperfusion can be customized. For intramyocardial injections, a Hamilton syringe with sterile 30gauge beveled needle is used. When the myocardial manipulations are complete, the rib cage, the pectoral muscles, and the skin are closed sequentially. Line block analgesia is effected by 0.25% marcaine in sterile saline which is applied to muscle layer prior to closure of the skin. The mice are given a subcutaneous injection of saline and placed in a warming chamber until they are sternally recumbent. They are then returned to the vivarium and housed under standard conditions until the time of tissue collection. At the time of sacrifice, the mice are anesthetized, the heart is arrested in diastole with KCl or BDM, rinsed with saline, and immersed in fixative. Subsequently, routine procedures for processing, embedding, sectioning, and histological staining are performed. 
Nonsurgical intubation of a mouse and the microsurgical manipulations described make this a technically challenging model to learn and achieve reproducibility. These procedures, combined with the difficulty in performing consistent manipulations of the ligature for timed occlusion(s) and reperfusion or intramyocardial injections, can also affect the survival rate so optimization and consistency are critical.

Numerous genetic manipulations and/or intramyocardial injections of genes, proteins, cells, and/or biomaterials are superimposed upon the dimension of time in studies of acute ischemia/ reperfusion injury and chronic remodeling in mice. This video illustrates the microsurgical procedures for ischemia/reperfusion, permanent coronary artery ligation, and intramyocardial injection studies.

경색의 Murine 모델에서 관상 동맥 내고과 Intramyocardial 사출

Mouse models are a valuable tool for studying acute injury and chronic remodeling of the myocardium in vivo.  With the advent of genetic modifications to ...

Modeling Intracerebral Hemorrhage in Mice: Injection of Autologous Blood or Bacterial Collagenase

Spontaneous intracerebral hemorrhage (ICH) defines a potentially life-threatening neurological malady that accounts for 10-15% of all stroke-related hospitalizations and for which no effective treatments are available to date1,2. Because of the heterogeneity of ICH in humans, various preclinical models are needed to thoroughly explore prospective therapeutic strategies3. Experimental ICH is commonly induced in rodents by intraparenchymal injection of either autologous blood or bacterial collagenase4. The appropriate model is selected based on the pathophysiology of hemorrhage induction and injury progression. The blood injection model mimics a rapidly progressing hemorrhage. Alternatively, bacterial collagenase enzymatically disrupts the basal lamina of brain capillaries, causing an active bleed that generally evolves over several hours5. Resultant perihematomal edema and neurofunctional deficits can be quantified from both models. In this study, we described and evaluated a modified double injection model of autologous whole blood6 as well as an ICH injection model of bacterial collagenase7, both of which target the basal ganglia (corpus striatum) of male CD-1 mice. We assessed neurofunctional deficits and brain edema at 24 and 72 hr after ICH induction. Intrastriatal injection of autologous blood (30 &mu;l) or bacterial collagenase (0.075U) caused reproducible neurofunctional deficits in mice and significantly increased brain edema at 24 and 72 hr after surgery (p&lt;0.05). In conclusion, both models yield consistent hemorrhagic infarcts and represent basic methods for preclinical ICH research.

Clinically relevant animal models of intracerebral hemorrhage (ICH) are needed to extend our knowledge of hemorrhagic stroke and to examine novel therapeutic strategies. In this study, we describe and evaluate two ICH models that implement unilateral injections of either autologous whole blood or bacterial collagenase into the basal ganglia (corpus striatum) of mice.

마우스의 모델링 Intracerebral 출혈 : Autologous 혈액이나 세균 Collagenase의 주입

Spontaneous intracerebral hemorrhage (ICH) defines a potentially life-threatening neurological malady that accounts for 10-15% of all stroke-related ...

Intramyocardial Cell Delivery: Observations in Murine Hearts

Previous studies showed that cell delivery promotes cardiac function amelioration by release of cytokines and factors that increase cardiac tissue revascularization and cell survival. In addition, further observations revealed that specific stem cells, such as cardiac stem cells, mesenchymal stem cells and cardiospheres have the ability to integrate within the surrounding myocardium by differentiating into cardiomyocytes, smooth muscle cells and endothelial cells.
Here, we present the materials and methods to reliably deliver noncontractile cells into the left ventricular wall of immunodepleted mice. The salient steps of this microsurgical procedure involve anesthesia and analgesia injection, intratracheal intubation, incision to open the chest and expose the heart and delivery of cells by a sterile 30-gauge needle and a precision microliter syringe.
Tissue processing consisting of heart harvesting, embedding, sectioning and histological staining showed that intramyocardial cell injection produced a small damage in the epicardial area, as well as in the ventricular wall. Noncontractile cells were retained into the myocardial wall of immunocompromised mice and were surrounded by a layer of fibrotic tissue, likely to protect from cardiac pressure and mechanical load.

Intramyocardial cell delivery in murine models of cardiovascular diseases, such as hypertension or myocardial infarction, is widely used to test the therapeutic potential of different cell types in regenerative studies. Therefore, a detailed description and a clear visualization of this surgical procedure will help to define the limits and advantages of cardiovascular cell therapeutic analyses in small rodents.

심근 내 휴대 납품 : 쥐과 마음의 관찰

Previous studies showed that cell delivery promotes cardiac function amelioration by release of cytokines and factors that increase cardiac tissue ...

Substernal Thyroid Biopsy Using Endobronchial Ultrasound-guided Transbronchial Needle Aspiration

Substernal thyroid goiter (STG) represents about 5.8% of all mediastinal lesions1. There is a wide variation in the published incidence rates due to the lack of a standardized definition for STG. Biopsy is often required to differentiate benign from malignant lesions. Unlike cervical thyroid, the overlying sternum precludes ultrasound-guided percutaneous fine needle aspiration of STG. Consequently, surgical mediastinoscopy is performed in the majority of cases, causing significant procedure related morbidity and cost to healthcare. Endobronchial Ultrasound-guided Transbronchial Needle Aspiration (EBUS-TBNA) is a frequently used procedure for diagnosis and staging of non-small cell lung cancer (NSCLC). Minimally invasive needle biopsy for lesions adjacent to the airways can be performed under real-time ultrasound guidance using EBUS. Its safety and efficacy is well established with over 90% sensitivity and specificity. The ability to perform EBUS as an outpatient procedure with same-day discharges offers distinct morbidity and financial advantages over surgery. As physicians performing EBUS gained procedural expertise, they have attempted to diversify its role in the diagnosis of non-lymph node thoracic pathologies. We propose here a role for EBUS-TBNA in the diagnosis of substernal thyroid lesions, along with a step-by-step protocol for the procedure.

Substernal thyroid lesions are common, and need to be differentiated from malignancy. Obtaining percutaneous fine needle biopsy is not possible due to its retrosternal location. This article proposes a protocol for biopsy of substernal thyroid lesions using Endobronchial Ultrasound-guided Transbronchial Needle Aspiration (EBUS-TBNA).

흉골 하 갑상선 조직 검사는 기관지 초음파 유도 기관지 바늘 흡인을 사용하여

Substernal thyroid goiter (STG) represents about 5.8% of all mediastinal lesions1. There is a wide variation in the published incidence rates due to the ...

Ultrasound-guided Botulinum Toxin-A Injections: A Method of Treating Sialorrhea

Neurological diseases can be complicated by sialorrhea, an excessive flow of saliva. Patients suffering from moderate to severe sialorrhea have an impaired quality of life, often worsened by correlated complications such as aspiration pneumonia, oral infections, dental caries, and maceration of the skin. Diverse therapeutic approaches have been proposed for the treatment of sialorrhea, including surgery and the use of anticholinergic agents, with limited results and the possible occurrence of serious adverse events. Recently, botulinum toxin (BoNT) injection within the major salivary glands has been proposed in patients refractory to anticholinergic therapy, with the aim of inhibiting local acetylcholine release and gland activity. 
In order to obtain a better outcome in terms of reduction of saliva production, efficacy, duration, and avoidance of major adverse events, we developed an ultrasound-guided BoNT-type A injection technique accurately described in the text. Here we present a method of treating sialorrhea with bilateral parotid and submandibular gland BoNT-type A injections under ultrasound guidance. Four quadrants of the parotid gland and two quadrants of the submandibular gland are visualized and injected using two accesses and one access, respectively. 
The ultrasound-guided procedure provides a simple, non-invasive, real-time visualization of the muscular and glandular tissues and their surrounding structures, optimizing treatment efficacy and safety.

Botulinum toxin is injected into the salivary glands for the treatment of sialorrhea. The ultrasound-guided botulinum toxin injection in both the parotid and submandibular glands demonstrates a sustained efficacy and the absence of serious side effects.

보툴리눔 독소-A 주사를 초음파 유도 : Sialorrhea을 치료하는 방법

Neurological diseases can be complicated by sialorrhea, an excessive flow of saliva. Patients suffering from moderate to severe sialorrhea have an ...

Ultrasound-guided Intracardiac Injection of Human Mesenchymal Stem Cells to Increase Homing to the Intestine for Use in Murine Models of Experimental Inflammatory Bowel Diseases

Crohn&#39;s disease (CD) is a common chronic inflammatory disease of the small and large intestines. Murine and human mesenchymal stem cells (MSCs) have immunosuppressive potential and have been shown to suppress inflammation in mouse models of intestinal inflammation, even though the route of administration can limit their homing and effectiveness 1,3,4,5. Local application of MSCs to colonic injury models has shown greater efficacy at ameliorating inflammation in the colon. However, there is paucity of data on techniques to enhance the localization of human bone marrow-derived MSCs (hMSCs) to the small intestine, the site of inflammation in the SAMP-1/YitFc (SAMP) model of experimental Crohn&#39;s disease. This work describes a novel technique for the ultrasound-guided intracardiac injection of hMSCs in SAMP mice, a well-characterized spontaneous model of chronic intestinal inflammation. Sex- and age-matched, inflammation-free AKR/J (AKR) mice were used as controls. To analyze the biodistribution and the localization, hMSCs were transduced with a lentivirus containing a triple reporter. The triple reporter consisted of firefly luciferase (fl), for bioluminescent imaging; monomeric red fluorescent protein (mrfp), for cell sorting; and truncated herpes simplex virus thymidine kinase (ttk), for positron emission tomography (PET) imaging. The results of this study show that 24 h after the intracardiac administration, hMSCs localize in the small intestine of SAMP mice as opposed to inflammation-free AKR mice. This novel, ultrasound-guided injection of hMSCs in the left ventricle of SAMP mice ensures a high success rate of cell delivery, allowing for the rapid recovery of mice with minimal morbidity and mortality. This technique could be a useful method for the enhanced localization of MSCs in other models of small-intestinal inflammation, such as TNF&#916;RE6. Future studies will determine if the increased localization of hMSCs by intra-arterial delivery can lead to increased therapeutic efficacy.

Murine studies in models of colonic inflammation have demonstrated that a small percentage (1 - 5%) of mesenchymal stem cells (MSC) injected intravenously or intraperitoneally home to the inflamed colon1,2. This study shows that ultrasound-guided intracardiac injections of MSCs result in increased localization to the intestine.

초음파 유도 Intracardiac 주입 실험 선 동적인 장 질병의 Murine 모델에 사용 하기 위해 내장을 추적 증가 인간 중간 엽 줄기 세포의

Crohn&#39;s disease (CD) is a common chronic inflammatory disease of the small and large intestines. Murine and human mesenchymal stem cells (MSCs) have ...

Percutaneous Contrast Echocardiography-guided Intramyocardial Injection and Cell Delivery in a Large Preclinical Model

Cell and gene therapy are exciting and promising strategies for the purpose of cardiac regeneration in the setting of heart failure with reduced ejection fraction (HFrEF). Before they can be considered for use, and implemented in humans, extensive preclinical studies are required in large animal models to evaluate the safety, efficacy, and fate of the injectate (e.g., stem cells) once delivered into the myocardium. Small rodent models offer advantages (e.g., cost effectiveness, amenability for genetic manipulation); however, given inherent limitations of these models, the findings in these rarely translate into the clinic. Conversely, large animal models such as rabbits, have advantages (e.g., similar cardiac electrophysiology compared to humans and other large animals), whilst retaining a good cost-effective balance. Here, we demonstrate how to perform a percutaneous contrast echocardiography-guided intramyocardial injection (IMI) technique, which is minimally invasive, safe, well tolerated, and very effective in the targeted delivery of injectates, including cells, into several locations within the myocardium of a rabbit model. For the implementation of this technique, we also have taken advantage of a widely available clinical echocardiography system. After putting in practice the protocol described here, a researcher with basic ultrasound knowledge will become competent in the performance of this versatile and minimally invasive technique for routine use in experiments, aimed at hypothesis testing of the capabilities of cardiac regenerative therapeutics in the rabbit model. Once competency is achieved, the whole procedure can be performed within 25 min after anaesthetizing the rabbit.

Novel therapeutic strategies in cardiac regenerative medicine require extensive and detailed studies in large preclinical animal models before they can be considered for use in humans. Here, we demonstrate a percutaneous contrast echocardiography-guided intramyocardial injection technique in rabbits, which is valuable for hypothesis testing the efficacy of such novel therapies.

경 피 적인 대비 심장 초음파-가이드 Intramyocardial 주입 및 큰 전 임상 모델에서 셀 배달

Cell and gene therapy are exciting and promising strategies for the purpose of cardiac regeneration in the setting of heart failure with reduced ejection ...

Transthoracic Echocardiographic Examination in the Rabbit Model

Large animal models such as the rabbit are valuable for translational preclinical research. Rabbits have a similar cardiac electrophysiology compared to that of humans and that of other large animal models such as dogs and pigs. However, the rabbit model has the additional advantage of lower maintenance costs compared to other large animal models. The longitudinal evaluation of cardiac function using echocardiography, when appropriately implemented, is a useful methodology for preclinical assessment of novel therapies for heart failure with reduced ejection fraction (e.g. cardiac regeneration). The correct use of this non-invasive tool requires the implementation of a standardized examination protocol following international guidelines. Here we describe, step by step, a detailed protocol supervised by veterinary cardiologists for performing echocardiography in the rabbit model, and demonstrate how to correctly obtain the different echocardiographic views and imaging planes, as well as the different imaging modes available in a clinical echocardiography system routinely used in human and veterinary patients.

Here we describe, step by step, a detailed protocol for performing echocardiography in the rabbit model. We show how to correctly obtain the different echocardiographic views and imaging planes, as well as the different imaging modes available in a clinical echocardiography system routinely used in human and veterinary patients.

토끼 모델의 흉부 심초음파 검사

Large animal models such as the rabbit are valuable for translational preclinical research. Rabbits have a similar cardiac electrophysiology compared to ...

International Expert Consensus and Recommendations for Neonatal Pneumothorax Ultrasound Diagnosis and Ultrasound-guided Thoracentesis Procedure

Pneumothorax (PTX) represents accumulation of the air in the pleural space. A large or tension pneumothorax can collapse the lung and cause hemodynamic compromise, a life-threatening disorder. Traditionally, neonatal pneumothorax diagnosis has been based on clinical images, auscultation, transillumination, and chest X-ray findings. This approach may potentially lead to a delay in both diagnosis and treatment. The use of lung US in diagnosis of PTX together with US-guided thoracentesis results in earlier and more precise management. The recommendations presented in this publication are aimed at improving the application of lung US in guiding neonatal PTX diagnosis and management.

Pneumothorax is a common emergency and critical disease in newborn infants that needs rapid, clear diagnosis and timely treatment. Diagnosis and treatment based on chest X-rays are associated with delayed management and radiation damage. Lung ultrasound (US) provides useful guidance for rapid, accurate diagnosis and the precise thoracentesis of pneumothorax.

신생아 기흉 초음파 진단 및 초음파 유도 흉부 시술에 대한 국제 전문가 합의 및 권장 사항

Pneumothorax (PTX) represents accumulation of the air in the pleural space. A large or tension pneumothorax can collapse the lung and cause hemodynamic ...