ويؤيد هذا العمل من قبل المؤسسة من خلال شبكة Leducq Caerus (RJH) ، عن طريق R01 NIH HL093183 ، HL088434 ، HL071763 ، HL080498 ، HL083156 وP20HL100396 (RJH). وأيد DL من قبل مؤسسة الأبحاث الألمانية. نود أن نشكر لورين Leonardson لمساعدتها التقنية في هذا المشروع.

1. احتشاء عضلة القلب خلق <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> إناث الخنازير يوركشاير (20kg) من صناعات التكنولوجيا الحيوية الحيوانية (Doylestown ، والسلطة الفلسطينية) هي مرحلة ما قبل العلاج باستخدام Telazol (تلييتامين / zolazepam) (8.0 ملغ / كلغ) والبوبرينورفين (0.6 ملغ). </li><li style=";text-align:right;direction:rtl"> وintubated الحيوانات والتهوية ثم مع الأكسجين 100 ٪. يتم الاحتفاظ التخدير العام مع البروبوفول (5-8mg/kg/hr) في جميع أنحاء الداخلي. </li><li style=";text-align:right;direction:rtl"> مستعدة للموقع الفخذ مع ايزوبروبيل 70 ٪ تليها بوفيدون اليود. ثقب عن طريق الجلد يتيح الوصول إلى الشرايين والتنسيب للغمد. بعد الإدراج غمد ، تدار 3000-5000 وحدة دولية من الهيبارين الرابع للحفاظ على الوقت لتنشيط تجلط الدم 250-300 ثواني. </li><li style=";text-align:right;direction:rtl"> تدار جرعة من الأميودارون بلعة (75 ملغ) عن طريق الحقن العضلي. بدء الحقن في الوريد من حقيبة مل 1000 المالحة مع الأميودارون (75 ملغ) ، الأتروبين (2 ملغ) ، وخلات البوتاسيوم (20 mEq) وحافظت باستخدام معدل 150 مل / ساعة في جميع أنحاء الداخلي. </li><li style=";text-align:right;direction:rtl"> A 7 الاب هوكي العصا القسطرة (القلبية ، ميامي ، فلوريدا) متقدمة على الشريان التاجي الأيسر ويتم إجراء تصوير الأوعية التاجية. متقدمة سلك دليل 0.014 بوصة (ابوت ، وبارك ، وايل) في الجزء الأمامي الأيسر الشريان التاجي النازل (LAD). و8 ملم طويلة ، 4.0 ملم VOYAGER الإفراط في سلك متقدمة بالون (ابوت ، وبارك ، وايل) والمتمركزة على LAD القريبة ، ويتم نفخ البالون التاجية إلى 3 أجهزة الصراف الآلي. ويؤكد انسداد LAD بواسطة عائية. </li><li style=";text-align:right;direction:rtl"> مع الكهربائي المستمر ومراقبة الدورة الدموية ، والحفاظ على انسداد لمدة 90 دقيقة. عدم انتظام ضربات القلب البطيني الخبيث قد يحدث أحيانا في أول 40 دقيقة ، وفي هذه الحالة التسليم الفوري للصدمات المباشرة الحالية من 200 إلى 300 J J تدار. عادة ما تكون هناك إطارين وقت الذروة من عدم انتظام ضربات القلب ، واحدة في الفترة بين 2 و 10 دقيقة والآخر بين 25 و 35 دقيقة بعد تضخم البالون. الرصد والمتابعة الحثيثة من صدمات الكهربائية الفورية هي المفتاح لاحتشاء عضلة القلب وإنشاء ناجحة ، لأن هناك حالات أكثر من 95 ٪ من الذين يدخلون الرجفان البطيني ضمن الأطر الزمنية المشار إليها. إذا كان يتم تنفيذ إزالة الرجفان فورية ومناسبة ، ومعدل البقاء على قيد الحياة مرتفعة جدا. إذا كان ضغط الدم أقل من 80 مم زئبق قطرات ، ويتم إنجاز تحميل مع حجم التسريب السريع ملحي في الوريد. </li><li style=";text-align:right;direction:rtl"> وبددت البالون وتتم إزالة كافة الأجهزة. </li><li style=";text-align:right;direction:rtl"> تتم إزالة غمد من شريان الفخذ ، هو حجب التخدير ، ويتم رصد هذا الحيوان خلال فترة الانتعاش. وانخفض ملحي في الوريد مع الأميودارون ، اتروبين والبوتاسيوم خلات التسريب في 50 مل / ساعة وتعطى له بين عشية وضحاها. يدار من الحقن العضلي من Nitroglycerline وFuroscemide. ثم سمح للحيوان لاسترداد واصلت رصد دقيق ECG ما لا يقل عن 3 ساعات. </li><li style=";text-align:right;direction:rtl"> وستعطى الاسبرين (200mg) ص لمدة خمسة أيام. </li></ol> المضاعفات شيوعا أثناء وبعد إجراء احتشاء عضلة القلب وخلق تشريح الشريان التاجي ، ثقب دموي الموقع ، عدم انتظام ضربات القلب ، وفشل القلب. وصقل تقنية ومطلوب رصد شامل لمنع وإدارة هذه المضاعفات. 2. حقن الفيروس <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> وتعد هذه الحيوانات كما هو مبين أعلاه في خلق احتشاء عضلة القلب steps1 - 3. وينبغي أن تعطى الجرعة نفسها من الهيبارين لمنع تجلط الدم خلال حقن الفيروس. </li><li style=";text-align:right;direction:rtl"> عصا الهوكي 5Fr القسطرة (القلبية ، ميامي ، فلوريدا) متقدمة على الشريان التاجي الأيسر. بعد عائية ، وهما من الأسلاك دليل 0.014 بوصة والمتقدمة ، واحدة في LAD واحدة في الشريان التاجي الأيسر المنعطف (LCX) لتحديد موقف القسطرة. </li><li style=";text-align:right;direction:rtl"> ويوجه الدم الشرياني (-20 مل) من القسطرة. لمنع نقص التروية خلال التسريب ، واستخدام نصف الدم المخفف للحل. يختلط الفيروس في الدم مع 10 مل و 20 مل مخففة لمع المالحة. كما يتم تخفيف ما تبقى من 10 مل و 20 مل و تسليم الفيروس المستخدمة المتبقية داخل تجويف القسطرة. </li><li style=";text-align:right;direction:rtl"> بدء النتروجليسرين عن طريق الوريد (20 ميكروغرام / دقيقة) عن طريق الوريد الأذن. </li><li style=";text-align:right;direction:rtl"> يتم وضع الطرف قسطرة في المسالك القريبة الرئيسي اليسار. ويتم رصد الضغط من خلال قسطرة لضمان أن لا التوتيد القسطرة. </li><li style=";text-align:right;direction:rtl"> يتم حقن الفيروس في الدم المختلط (15 مل) من خلال قسطرة خلال 15 دقيقة باستخدام مضخة التسريب في الشريان التاجي الأيسر. بعد 15 دقيقة ، يتم فيها تبادل المحقنة إلى الاحمرار ، واستمر لمدة 10 دقيقة التسريب. </li><li style=";text-align:right;direction:rtl"> يتم إصلاح سلك ومعدات قسطرة الشريان التاجي إلى الحق بنفس الطريقة التي الشريان التاجي الأيسر. </li><li style=";text-align:right;direction:rtl"> يتم حقن ما تبقى من 5 مل من الدم المختلط مع الفيروس في الشريان التاجي الأيمن لمدة 5 دقائق ثم 10 دقائق من طرد. </li></ol> 3. ممثل القضية الشكل 1 يبين تصوير البطينات اليسار في 1 الشهر بعد احتشاء عضلة القلب خلق. هو ضعف كبير في جدار البطين الأيسر الاقتراح مع جزء طرد (EF) من31 ٪. الشكل 2 يبين المقاطع العرضية من البطين الأيسر في 1 الشهر. وينظر الى ندبة كبيرة من القاعدة إلى قمة الجدار الأمامي. وEF المتوسط ​​في 1 الشهر بعد احتشاء عضلة القلب في هذا النموذج هي 36 ٪ ومتوسط ​​حجم نهاية الانبساط هو 83 مل ، في حين أن متوسط ​​هذه الحيوان في نفس الحجم من دون احتشاء عضلة القلب هو 68 ٪ و 42 مل ، على التوالي. ويبين الشكل 3 التعبير عن غال بيتا في منطقة البطين الأيسر منطقة الحدود 1 بعد شهر من حقن الغدة المرتبطة بفيروس (2 × 10 12 الجسيمات الدناز resitant من CMV rAAV1 لاك - Z). تم العثور على التعبير المعزز في مجال حقن الفيروس. <img src="/files/ftp_upload/2778/2778fig1.jpg" alt="الشكل 1" /> الشكل 1. كسر قياس الطرد من تصوير البطينات اليسار 1 شهر احتشاء عضلة القلب بعد خلق. هي متسعة البطين الأيسر إلى 92.5 مل ، وضعف ملحوظ الجدار الأمامي للحركة. <img src="/files/ftp_upload/2778/2778fig2.jpg" alt="الشكل 2" /> الشكل 2 ، والمقاطع العرضية من البطين الأيسر في 1 الشهر احتشاء عضلة القلب بعد إنشاء ملون بواسطة ثلاثي فينيل كلوريد tetrazolium لتحديد منطقة الندبة. وينظر الى ندبة كبيرة من القاعدة إلى قمة الجدار الأمامي. <img src="/files/ftp_upload/2778/2778fig3.jpg" alt="الشكل 3" /> الشكل 3. بيتا غال التعبير الناجم عن الغدة المرتبطة بفيروس (2 × 10 12 الجسيمات الدناز المقاومة للCMV rAAV1 لاك - Z). تم العثور على التعبير المعززة على الحدود بين منطقة التجارة احتشاء والأنسجة الطبيعية (1) ، بعد شهر من حقن الفيروس intracoronary. الصورة اليمنى السفلى ويبين سيطرة سلبية ، حيث تم حقن المياه المالحة فقط طبيعية في الشرايين التاجية باستخدام بروتوكول نفسه.

<ol>
	<li>Perez de Prado, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Closed-chest+experimental+porcine+model+of+acute+myocardial+infarction-reperfusion.">Closed-chest experimental porcine model of acute myocardial infarction-reperfusion.</a> J Pharmacol Toxicol Methods. 60, 301-306 (2009).</li><li>Angeli, F. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Left+ventricular+remodeling+after+myocardial+infarction:+characterization+of+a+swine+model+on+beta-blocker+therapy.">Left ventricular remodeling after myocardial infarction: characterization of a swine model on beta-blocker therapy.</a> Comp Med. 59, 272-279 (2009).</li><li>Urdaneta, F., Lobato, E. B., Kirby, D. S. &. a. m. p. ;. a. m. p., Sidi, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treating+myocardial+stunning+randomly,+with+either+propofol+or+isoflurane+following+transient+coronary+occlusion+and+reperfusion+in+pigs.">Treating myocardial stunning randomly, with either propofol or isoflurane following transient coronary occlusion and reperfusion in pigs.</a> Ann Card Anaesth. 12, 113-121 (2009).</li><li>Jaski, B. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calcium+upregulation+by+percutaneous+administration+of+gene+therapy+in+cardiac+disease+(CUPID+Trial),+a+first-in-human+phase+1/2+clinical+trial.">Calcium upregulation by percutaneous administration of gene therapy in cardiac disease (CUPID Trial), a first-in-human phase 1/2 clinical trial.</a> J Card Fail. 15, 171-181 (2009).</li></ol>

جميع المؤلفين ليس لديهم الجمعيات التجارية التي قد تشكل خطرا على النحو تضارب في المصالح فيما يتعلق المخطوطة.

من بين الأسباب المختلفة لفشل القلب ، نقص تروية هي حتى الآن الأكثر شيوعا في المسببات. للتغلب على هذا المرض الخطير ، ويجري وضع مناهج علاجية مختلفة. مطلوبة التقييمات حذرا من العلاجات قبل أن يمكن لهذه العلاجات واعدة يمكن تطبيقها على العيادات. ولذلك ، وإنشاء نموذج فشل الق?...

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		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
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<tr>
<td>Telazol</td>
<td>&nbsp;</td>
<td>Fort Dodge Animal Health</td>
<td>0856-9050-49</td>
<td>&nbsp;</td>
<tr>
<td>Propofol</td>
<td>&nbsp;</td>
<td>APP Pharmaceuticals</td>
<td>63323-270-20</td>
<td>&nbsp;</td>
<tr>
<td>Heparin</td>
<td>&nbsp;</td>
<td>APP Pharmaceuticals</td>
<td>63323-540-11</td>
<td>&nbsp;</td>
<tr>
<td>Amiodarone</td>
<td>&nbsp;</td>
<td>APP Pharmaceuticals</td>
<td>63323-616-03</td>
<td>&nbsp;</td>
<tr>
<td>Atropin Sulfate</td>
<td>&nbsp;</td>
<td>Med-Pharmex</td>
<td>54925-063-10</td>
<td>&nbsp;</td>
<tr>
<td>Potassium acetate</td>
<td>&nbsp;</td>
<td>American Regent, Inc.</td>
<td>2053-25</td>
<td>&nbsp;</td>
<tr>
<td>Nitroglycerine</td>
<td>&nbsp;</td>
<td>American Regent, Inc.</td>
<td>4810-25 </td>
<td>&nbsp;</td>
<tr>
<td>Salix (Furosemide)</td>
<td>&nbsp;</td>
<td>Intervet Inc.</td>
<td>710461</td>
<td>&nbsp;</td>
<tr>
<td>5 Fr Hockey stick catheter</td>
<td>&nbsp;</td>
<td>Cordis</td>
<td>556-278-0L</td>
<td>&nbsp;</td>
<tr>
<td>7 Fr Hockey stick catheter</td>
<td>&nbsp;</td>
<td>Cordis</td>
<td>778-278-00</td>
<td>&nbsp;</td>
<tr>
<td>Coronary wire</td>
<td>&nbsp;</td>
<td>Abbott Laboratories</td>
<td>1044588</td>
<td>&nbsp;</td>
<tr>
<td>Coronary balloon</td>
<td>&nbsp;</td>
<td>Abbott Laboratories</td>
<td>1009447-08</td>
<td>&nbsp;</td>
<tr>
<td>8Fr sheath</td>
<td>&nbsp;</td>
<td>TERUMO</td>
<td>RSS801</td>
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gene transfer for ischemic heart failure in a preclinical model

ويجري تطوير مختلف التكنولوجيات الناشئة للمرضى المصابين بقصور في القلب. راسخة التقييمات قبل السريرية ضرورية لتحديد فعاليتها وسلامتها. العلاج الجيني باستخدام نواقل الفيروسية هي واحدة من أكثر الأساليب واعدة لعلاج أمراض القلب. تسليم الفيروسية من جينات مختلفة مختلفة عن طريق تغيير الجينات الحاملة لديها امكانات لا حصر لها العلاجية. في هذا الفيديو ، وتقدم العملية الكاملة لنموذج حيواني من خلق فشل القلب تليها نقل الجينات باستخدام نموذج الخنازير. أولا ، يتم إنشاء احتشاء عضلة القلب عن طريق الاغلاق الأمامي الأيسر النازل القريبة الشريان التاجي. إعادة عرض نتائج القلب في فشل القلب المزمن. فريدة من نوعها لنموذجنا هو ندبة كبيرة نوعا ما وهو ما يعكس حقا المرضى الذين يعانون من قصور القلب الذين يحتاجون إلى العلاج شديد العدوانية من أجل تحقيق نتائج إيجابية. احتشاء عضلة القلب بعد إنشاء وتطوير ندبا ، ويتجلى حقنة intracoronary من الفيروس مع التسريب النيتروجلسرين في وقت واحد. لدينا وسيلة حقن يوفر بسيطة وفعالة نقل الجينات مع الجينات المحسنة. وقد أثبت هذا المزيج من احتشاء عضلة القلب نموذجا الخنازير مع فيروس التسليم intracoronary أن تكون منهجية متسقة وقابلة للتكرار ، والذي لا يساعد فقط على اختبار تأثير الجينات الفردية ، ولكن أيضا مقارنة فعالية العديد من الجينات العلاجية كمرشحين.

Watch this Scientific Journal Video about Gene Transfer for Ischemic Heart Failure in a Preclinical Model at JoVE.com

Gene Transfer for Ischemic Heart Failure in a Preclinical Model

ووصف طريقة نقل الجينات لعلاج قصور القلب الإقفاري باستخدام نموذج الخنازير من احتشاء عضلة القلب. لدينا وسيلة بسيطة وقابلة للتكرار تمكننا من تقييم فعالية بسهولة نقل الجينات المختلفة مع طريقة بسيطة جدا وقابلة للتكرار.

نقل الجينات لقصور القلب الدماغية في نموذج ما قبل السريرية

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine ...

gene-transfer-for-ischemic-heart-failure-in-a-preclinical-model

Research

JoVE Journal

Medicine

12.8K Views.  Mount Sinai School of Medicine. ووصف طريقة نقل الجينات لعلاج قصور القلب الإقفاري باستخدام نموذج الخنازير من احتشاء عضلة القلب. لدينا وسيلة بسيطة وقابلة للتكرار تمكننا من تقييم فعالية بسهولة نقل الجينات المختلفة مع طريقة بسيطة جدا وقابلة للتكرار.

Video: Gene Transfer for Ischemic Heart Failure in a Preclinical Model

A Simplified Technique for Producing an Ischemic Wound Model

One major obstacle in current diabetic wound research is a lack of an ischemic wound model that can be safely used in diabetic animals. Drugs that work well in non-ischemic wounds may not work in human diabetic wounds because vasculopathy is one major factor that hinders healing of these wounds. We published an article in 2007 describing a rabbit ear ischemic wound model created by a minimally invasive surgical technique. Since then, we have further simplified the procedure for easier operation. On one ear, three small skin incisions were made on the vascular pedicles, 1-2 cm from the ear base. The central artery was ligated and cut along with the nerve. The whole cranial bundle was cut and ligated, leaving only the caudal branch intact. A circumferential subcutaneous tunnel was made through the incisions, to cut subcutaneous tissues, muscles, nerves, and small vessels. The other ear was used as a non-ischemic control. Four wounds were made on the ventral side of each ear. This technique produces 4 ischemic wounds and 4 non-ischemic wounds in one animal for paired comparisons. After surgery, the ischemic ear was cool and cyanotic, and showed reduced movement and a lack of pulse in the ear artery. Skin temperature of the ischemic ear was 1-10 &deg;C lower than that on the normal ear and this difference was maintained for more than one month. Ear tissue high-energy phosphate contents were lower in the ischemic ear than the control ear. Wound healing times were longer in the ischemic ear than in the non-ischemic ear when the same treatment was used. The technique has now been used on more than 80 rabbits in which 23 were diabetic (diabetes time ranging from 2 weeks to 2 years). No single rabbit has developed any surgical complications such as bleeding, infection, or rupture in the skin incisions. The model has many advantages, such as little skin disruption, longer ischemic time, and higher success rate, when compared to many other models. It can be safely used in animals with reduced resistance, and can also be modified to meet different testing requirements.

We have developed a minimally invasive technique to create a rabbit ischemic ear wound model by dividing the central artery and nerve and the cranial neurovascular bundle. A subcutaneous tunnel then cuts all subcutaneous tissues. This procedure causes minimal skin disruption and can be safely used in diabetic animals.

أسلوب مبسط لإنتاج نموذج الجرح الدماغية

One major obstacle in current diabetic wound research is a lack of an ischemic wound model that can be safely used in diabetic animals. Drugs that work ...

Permanent Ligation of the Left Anterior Descending Coronary Artery in Mice: A Model of Post-myocardial Infarction Remodelling and Heart Failure

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It is characterized by fluid retention, shortness of breath, and fatigue, in particular on exertion. Heart failure is a growing public health problem, the leading cause of hospitalization, and a major cause of mortality. Ischemic heart disease is the main cause of heart failure.
Ventricular remodelling refers to changes in structure, size, and shape of the left ventricle. This architectural remodelling of the left ventricle is induced by injury (e.g., myocardial infarction), by pressure overload (e.g., systemic arterial hypertension or aortic stenosis), or by volume overload. Since ventricular remodelling affects wall stress, it has a profound impact on cardiac function and on the development of heart failure. A model of permanent ligation of the left anterior descending coronary artery in mice is used to investigate ventricular remodelling and cardiac function post-myocardial infarction. This model is fundamentally different in terms of objectives and pathophysiological relevance compared to the model of transient ligation of the left anterior descending coronary artery. In this latter model of ischemia/reperfusion injury, the initial extent of the infarct may be modulated by factors that affect myocardial salvage following reperfusion. In contrast, the infarct area at 24 hr after permanent ligation of the left anterior descending coronary artery is fixed. Cardiac function in this model will be affected by 1) the process of infarct expansion, infarct healing, and scar formation; and 2) the concomitant development of left ventricular dilatation, cardiac hypertrophy, and ventricular remodelling.
Besides the model of permanent ligation of the left anterior descending coronary artery, the technique of invasive hemodynamic measurements in mice is presented in detail.

Heart failure is the leading cause of hospitalization and a major cause of mortality. A model of permanent ligation of the left anterior descending coronary artery in mice is applied to investigate ventricular remodelling and cardiac dysfunction post-myocardial infarction. The technique of invasive hemodynamic measurements in mice is presented.

ربط دائم في الشريان التاجي الأيسر الأمامي تنازلي في الفئران: نموذج ما بعد إعادة تشكيل عضلة القلب احتشاء وقصور القلب

Heart failure is a syndrome in which the heart fails to pump blood at a rate commensurate with cellular oxygen requirements at rest or during stress. It ...

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves focal endothelial-selective transgenesis, thereby allowing an investigator to determine the biological roles of endothelial-expressed transgenes and to quantify the in vivo transcriptional activity of DNA sequences in large artery endothelial cells. The method uses surgical isolation of rabbit common carotid arteries and an arteriotomy to deliver a transgene-expressing viral vector into the arterial lumen. A short incubation period of the vector in the lumen, with subsequent aspiration of the lumen contents, is sufficient to achieve efficient and durable expression of the transgene in the endothelium, with no detectable transduction or expression outside of the isolated arterial segment. The method allows assessment of the biological activities of transgene products both in normal arteries and in models of human vascular disease, while avoiding systemic effects that could be caused either by targeting gene delivery to other sites (e.g. the liver) or by the alternative approach of delivering genetic constructs to the endothelium by germ line transgenesis. Application of the method is limited by the need for a skilled surgeon and anesthetist, a well-equipped operating room, the costs of purchasing and housing rabbits, and the need for expertise in gene-transfer vector construction and use. Results obtained with this method include: transgene-related alterations in arterial structure, cellularity, extracellular matrix, or vasomotor function; increases or reductions in arterial inflammation; alterations in vascular cell apoptosis; and progression, retardation, or regression of diseases such as intimal hyperplasia or atherosclerosis. The method also allows measurement of the ability of native and synthetic DNA regulatory sequences to alter transgene expression in endothelial cells, providing results that include: levels of transgene mRNA, levels of transgene protein, and levels of transgene enzymatic activity.

In Vivo Gene Transfer to the Rabbit Common Carotid Artery Endothelium

This method is to introduce a transgene into the endothelium of rabbit carotid arteries. Introduction of the transgene allows the assessment of the biological role of the transgene product either in normal arteries or disease models. The method is also useful for measuring activity of DNA regulatory sequences.

في فيفو نقل الجينات ببطانة الشريان السباتي المشترك الأرنب

The goal of this method is to introduce a transgene into the endothelium of isolated segments of both rabbit common carotid arteries. The method achieves ...

Tachycardia-Induced Cardiomyopathy As a Chronic Heart Failure Model in Swine

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment methods. Here, we present such a model by tachycardia-induced cardiomyopathy, which can be produced by rapid cardiac pacing in swine.
A single pacing lead is introduced transvenously into fully anaesthetized healthy swine, to the apex of the right ventricle, and fixated. Its other end is then tunneled dorsally to the paravertebral region. There, it is connected to an in-house modified heart pacemaker unit that is then implanted in a subcutaneous pocket. 
After 4 - 8 weeks of rapid ventricular pacing at rates of 200 - 240 beats/min, physical examination revealed signs of severe heart failure - tachypnea, spontaneous sinus tachycardia, and fatigue. Echocardiography and X-ray showed dilation of all heart chambers, effusions, and severe systolic dysfunction. These findings correspond well to decompensated dilated cardiomyopathy and are also preserved after the cessation of pacing.
This model of tachycardia-induced cardiomyopathy can be used for studying the pathophysiology of progressive chronic heart failure, especially hemodynamic changes caused by new treatment modalities like mechanical circulatory supports. This methodology is easy to perform and the results are robust and reproducible.

Here, we present a protocol to produce tachycardia-induced cardiomyopathy in swine. This model represents a potent way to study the hemodynamics of progressive chronic heart failure and the effects of applied treatment.

اعتلال عضلة القلب الناجم عن عدم انتظام دقات القلب كنموذج فشل القلب مزمن في الخنازير

A stable and reliable model of chronic heart failure is required for many experiments to understand hemodynamics or to test effects of new treatment ...

Isolation of Atrial Cardiomyocytes from a Rat Model of Metabolic Syndrome-related Heart Failure with Preserved Ejection Fraction

In this article, we describe an optimized, Langendorff-based procedure for the isolation of single-cell atrial cardiomyocytes (ACMs) from a rat model of metabolic syndrome (MetS)-related heart failure with preserved ejection fraction (HFpEF). The prevalence of MetS-related HFpEF is rising, and atrial cardiomyopathies associated with atrial remodeling and atrial fibrillation are clinically highly relevant as atrial remodeling is an independent predictor of mortality. Studies with isolated single-cell cardiomyocytes are frequently used to corroborate and complement in vivo findings. Circulatory vessel rarefication and interstitial tissue fibrosis pose a potentially limiting factor for the successful single-cell isolation of ACMs from animal models of this disease.
We have addressed this issue by employing a device capable of manually regulating the intraluminal pressure of cardiac cavities during the isolation procedure, substantially increasing the yield of morphologically and functionally intact ACMs. The acquired cells can be used in a variety of different experiments, such as cell culture and functional Calcium imaging (i.e., excitation-contraction-coupling).
We provide the researcher with a step-by-step protocol, a list of optimized solutions, thorough instructions to prepare the necessary equipment, and a comprehensive troubleshooting guide. While the initial implementation of the procedure might be rather difficult, a successful adaptation will allow the reader to perform state-of-the-art ACM isolations in a rat model of MetS-related HFpEF for a broad spectrum of experiments.

Here, we describe an optimized, Langendorff-based procedure for the isolation of single-cell atrial cardiomyocytes from a rat model of metabolic syndrome-related heart failure with preserved ejection fraction. A manual regulation of intraluminal pressure of cardiac cavities is implemented to yield functionally intact myocytes suitable for excitation-contraction-coupling studies.

عزل كارديوميوسيتيس خوارج من نموذج الفئران من فشل القلب المتصلة بالمتلازمة الأيضية مع كسر قذفي الحفاظ على

In this article, we describe an optimized, Langendorff-based procedure for the isolation of single-cell atrial cardiomyocytes (ACMs) from a rat model of ...

Induction and Phenotyping of Acute Right Heart Failure in a Large Animal Model of Chronic Thromboembolic Pulmonary Hypertension

The development of acute right heart failure (ARHF) in the context of chronic pulmonary hypertension (PH) is associated with poor short-term outcomes. The morphological and functional phenotyping of the right ventricle is of particular importance in the context of hemodynamic compromise in patients with ARHF. Here, we describe a method to induce ARHF in a previously described large animal model of chronic PH, and to phenotype, dynamically, right ventricular function using the gold standard method (i.e., pressure-volume PV loops) and with a non-invasive clinically available method (i.e., echocardiography). Chronic PH is first induced in pigs by left pulmonary artery ligation and right lower lobe embolism with biological glue once a week for 5 weeks. After 16 weeks, ARHF is induced by successive volume loading using saline followed by iterative pulmonary embolism until the ratio of the systolic pulmonary pressure over systemic pressure reaches 0.9 or until the systolic systemic pressure decreases below 90 mmHg. Hemodynamics are restored with dobutamine infusion (from 2.5 &#181;g/kg/min to 7.5 &#181;g/kg/min). PV-loops and echocardiography are performed during each condition. Each condition requires around 40 minutes for induction, hemodynamic stabilization and data acquisition. Out of 9 animals, 2 died immediately after pulmonary embolism and 7 completed the protocol, which illustrates the learning curve of the model. The model induced a 3-fold increase in mean pulmonary artery pressure. The PV-loop analysis showed that ventriculo-arterial coupling was preserved after volume loading, decreased after acute pulmonary embolism and was restored with dobutamine. Echocardiographic acquisitions allowed to quantify right ventricular parameters of morphology and function with good quality. We identified right ventricular ischemic lesions in the model. The model can be used to compare different treatments or to validate non-invasive parameters of right ventricular morphology and function in the context of ARHF.

We present a protocol to induce and phenotype an acute right heart failure in a large animal model with chronic pulmonary hypertension. This model can be used to test therapeutic interventions, to develop right heart metrics&#160;or to improve the understanding of acute right heart failure pathophysiology.

تحريض وتوصيف ظاهري لقصور القلب الأيمن الحاد في نموذج حيواني كبير لارتفاع ضغط الدم الرئوي الخثاري المزمن

The development of acute right heart failure (ARHF) in the context of chronic pulmonary hypertension (PH) is associated with poor short-term outcomes. The ...

Preclinical Model of Hind Limb Ischemia in Diabetic Rabbits

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular disease is the development of ischemia. In severe cases, patients can develop critical limb ischemia in which they experience constant pain and an increased risk of limb amputation. Current therapies for peripheral ischemia include bypass surgery or percutaneous interventions such as angioplasty with stenting or atherectomy to restore blood flow. However, these treatments often fail to the continued progression of vascular disease or restenosis or are contraindicated due to the overall poor health of the patient. A promising potential approach to treat peripheral ischemia involves the induction of therapeutic neovascularization to allow the patient to develop collateral vasculature. This newly formed network alleviates peripheral ischemia by restoring perfusion to the affected area. The most frequently employed preclinical model for peripheral ischemia utilizes the creation of hind limb ischemia in healthy rabbits through femoral artery ligation. In the past, however, there has been a strong disconnect between the success of preclinical studies and the failure of clinical trials regarding treatments for peripheral ischemia. Healthy animals typically have robust vascular regeneration in response to surgically induced ischemia, in contrast to the reduced vascularity and regeneration in patients with chronic peripheral ischemia. Here, we describe an optimized animal model for peripheral ischemia in rabbits that includes hyperlipidemia and diabetes. This model has reduced collateral formation and blood pressure recovery in comparison to a model with a higher cholesterol diet. Thus, the model may provide better correlation with human patients with compromised angiogenesis from the common co-morbidities that accompany peripheral vascular disease.

We describe a surgical procedure used to induce peripheral ischemia in rabbits with hyperlipidemia and diabetes. This surgery acts as a preclinical model for conditions experienced in peripheral artery disease in patients. Angiography is also described as a means to measure the extent of introduced ischemia and recovery of perfusion.

نموذج قبل السريرية من نقص التروية الأطراف هند في الأرانب السكري

Peripheral vascular disease is a widespread clinical problem that affects millions of patients worldwide. A major consequence of peripheral vascular ...

Establishing a Swine Model of Post-myocardial Infarction Heart Failure for Stem Cell Treatment

Although advances have been achieved in the treatment of heart failure (HF) following myocardial infarction (MI), HF following MI remains one of the major causes of mortality and morbidity around the world. Cell-based therapies for cardiac repair and improvement of left ventricular function after MI have attracted considerable attention. Accordingly, the safety and efficacy of these cell transplantations should be tested in a preclinical large animal model of HF prior to clinical use. Pigs are widely used for cardiovascular disease research due to their similarity to humans in terms of heart size and coronary anatomy. Therefore, we sought to present an effective protocol for the establishment of a porcine chronic HF model using closed-chest coronary balloon occlusion of the left circumflex artery (LCX), followed by rapid ventricular pacing induced with pacemaker implantation. Eight weeks later, the stem cells were administered by intramyocardial injection in the peri-infarct area. Then the infarct size, cell survival, and left ventricular function (including echocardiography, hemodynamic parameters, and electrophysiology) were evaluated. This study helps establish a stable preclinical large animal HF model for stem cell treatment.

We sought to establish a swine model of heart failure induced by left circumflex artery blockage and rapid pacing to test the effect and safety of intramyocardial administration of stem cells for cell-based therapies.

إنشاء نموذج الخنازير من فشل القلب احتشاء ما بعد عضلة القلب لعلاج الخلايا الجذعية

Although advances have been achieved in the treatment of heart failure (HF) following myocardial infarction (MI), HF following MI remains one of the major ...

A Preclinical Model of Exertional Heat Stroke in Mice

Heat stroke is the most severe manifestation of heat-related illnesses. Classic heat stroke (CHS), also known as passive heat stroke, occurs at rest, whereas exertional heat stroke (EHS) occurs during physical activity. EHS differs from CHS in etiology, clinical presentation, and sequelae of multi-organ dysfunction. Until recently, only models of CHS have been well established. This protocol aims to provide guidelines for a refined preclinical mouse model of EHS that is free from major limiting factors such as the use of anesthesia, restraint, rectal probes, or electric shock. Male and female C57Bl/6 mice, instrumented with core temperature (Tc) telemetric probes were utilized in this model. For familiarization with the running mode, mice undergo 3 weeks of training using both voluntary and forced running wheels. Thereafter, mice run on a forced wheel inside a climatic chamber set at 37.5 &#176;C and 40%-50% relative humidity (RH) until displaying symptom limitation (e.g., loss of consciousness) at Tc of 42.1-42.5 &#176;C, although suitable results can be obtained at chamber temperatures between 34.5-39.5 &#176;C and humidity between 30%-90%. Depending on the desired severity, mice are removed from the chamber immediately for recovery in ambient temperature or remain in the heated chamber for a longer duration, inducing a more severe exposure and a higher incidence of mortality. Results are compared with sham-matched exercise controls (EXC) and/or na&#239;ve controls (NC). The model mirrors many of the pathophysiological outcomes observed in human EHS, including loss of consciousness, severe hyperthermia, multi-organ damage as well as inflammatory cytokine release, and acute phase responses of the immune system. This model is ideal for hypothesis-driven research to test preventative and therapeutic strategies that may delay the onset of EHS or reduce the multi-organ damage that characterizes this manifestation.

The protocol describes the development of a standardized, repeatable, preclinical model of exertional heat stroke (EHS) in mice free from adverse external stimuli such as electric shock. The model provides a platform for mechanistic, preventative, and therapeutic studies.

نموذج ما قبل السريرية من السكتة الدماغية الحرارة مجهود في الفئران

Heat stroke is the most severe manifestation of heat-related illnesses. Classic heat stroke (CHS), also known as passive heat stroke, occurs at rest, ...

Large Animal Model for Evaluating the Efficacy of the Gene Therapy in Ischemic Heart

Coronary artery disease is one of the significant causes of mortality and morbidity worldwide. Despite the progression of current therapeutics, a considerable proportion of coronary artery disease patients remain symptomatic. Gene therapy-mediated therapeutic angiogenesis offers a novel therapeutic method for improving myocardial perfusion and relieving symptoms. Gene therapy with different angiogenic factors has been studied in few clinical trials. Due to the novelty of the method, the progress of myocardial gene therapy is a continuous path from bench to bedside. Therefore, large animal models are needed for evaluating the safety and efficacy. The more the large animal model identifies the original disease and the endpoints used in clinics, the more predictable outcomes are from clinical trials. Here, we introduce a large animal model for evaluating the efficacy of the gene therapy in the ischemic porcine heart. We use clinically relevant imaging methods such as ultrasound imaging and 15H2O-PET. For targeting the gene transfers into the desired area, electroanatomical mapping is used. The aim of this method is: (1) to mimic chronic coronary artery disease, (2) to induce therapeutic angiogenesis at hypoxic areas of the heart, and (3) to evaluate the safety and efficacy of the gene therapy by using relevant endpoints.

Myocardial gene therapy for ischemic heart disease holds great promise for future therapeutics. Here, we introduce a large animal model for evaluating the efficacy of gene therapy in the ischemic heart.