Name: normothermic negative pressure ventilation ex situ lung perfusion: evaluation of lung function and metabolism
Uploaded: 2022-02-14T14:00:00
Description: Watch this Scientific Journal Video about Normothermic Negative Pressure Ventilation Ex Situ Lung Perfusion: Evaluation of Lung Function and Metabolism at JoVE.com

这项研究是由医院研究基金会资助的。

本手稿中执行的程序符合加拿大动物护理理事会的指南以及实验动物护理和使用指南。阿尔伯塔大学的机构动物护理委员会批准了这些协议。仅使用35-50公斤的雌性幼年约克郡猪。参与ESLP程序的所有个人都需要进行适当的生物安全培训。整个NPV-ESLP实验的示意图如图 1所示。
1. 术前准备
<ol><li>将风琴室放在ESLP推车上，并将硅支撑膜（见 材料表</st...

<ol>
	<li>Chambers, D. C., 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=The+international+thoracic+organ+transplant+registry+of+the+international+society+for+heart+and+lung+transplantation:+Thirty-fifth+adult+lung+and+heart-lung+transplant+report-2018;+focus+theme:+Multiorgan+transplantation.">The international thoracic organ transplant registry of the international society for heart and lung transplantation: Thirty-fifth adult lung and heart-lung transplant report-2018; focus theme: Multiorgan transplantation.</a> The Journal of Heart and Lung Transplantation. 37 (10), 1169-1183 (2018).</li><li>Valapour, M., 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=OPTN/SRTR+2017+annual+data+report:+Lung.">OPTN/SRTR 2017 annual data report: Lung.</a> American Journal of Transplantation. 19, 404-484 (2019).</li><li>Chambers, D. C., 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=The+registry+of+the+international+society+for+heart+and+lung+transplantation:+Thirty-fourth+adult+lung+and+heart-lung+transplantation+report-2017;+focus+theme:+Allograft+ischemic+time.">The registry of the international society for heart and lung transplantation: Thirty-fourth adult lung and heart-lung transplantation report-2017; focus theme: Allograft ischemic time.</a> The Journal of Heart and Lung Transplantation. 36 (10), 1047-1059 (2017).</li><li>Klein, A. S., 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=Organ+donation+and+utilization+in+the+united+states,+1999-2008.">Organ donation and utilization in the united states, 1999-2008.</a> American Journal of Transplantation. 10 (4), 973-986 (2010).</li><li>Singh, E., 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=Sequence+of+refusals+for+donor+quality,+organ+utilization,+and+survival+after+lung+transplantation.">Sequence of refusals for donor quality, organ utilization, and survival after lung transplantation.</a> The Journal of Heart and Lung Transplantation. 38 (1), 35-42 (2019).</li><li>Bhorade, S. M., Vigneswaran, W., McCabe, M. A., Garrity, E. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liberalization+of+donor+criteria+may+expand+the+donor+pool+without+adverse+consequence+in+lung+transplantation.">Liberalization of donor criteria may expand the donor pool without adverse consequence in lung transplantation.</a> The Journal of Heart and Lung Transplantation. 19 (12), 1199-1204 (2000).</li><li>Snell, G. I., Griffiths, A., Levvey, B. J., Oto, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Availability+of+lungs+for+transplantation:+Exploring+the+real+potential+of+the+donor+pool.">Availability of lungs for transplantation: Exploring the real potential of the donor pool.</a> The Journal of Heart and Lung Transplantation. 27 (6), 662-667 (2008).</li><li>Cypel, M., 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=Normothermic+ex+vivo+lung+perfusion+in+clinical+lung+transplantation.">Normothermic ex vivo lung perfusion in clinical lung transplantation.</a> The New England Journal of Medicine. 364 (15), 1431-1440 (2011).</li><li>Wallinder, A., 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=Early+results+in+transplantation+of+initially+rejected+donor+lungs+after+ex+vivo+lung+perfusion:+A+case-control+study.">Early results in transplantation of initially rejected donor lungs after ex vivo lung perfusion: A case-control study.</a> European Journal of Cardio-Thoracic Surgery. 45 (1), 40-45 (2014).</li><li>Cypel, M., 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=Experience+with+the+first+50+ex+vivo+lung+perfusions+in+clinical+transplantation.">Experience with the first 50 ex vivo lung perfusions in clinical transplantation.</a> The Journal of Thoracic and Cardiovascular Surgery. 144 (1), 1200-1206 (2012).</li><li>Buchko, M. T., 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=Total+parenteral+nutrition+in+ex+vivo+lung+perfusion:+Addressing+metabolism+improves+both+inflammation+and+oxygenation.">Total parenteral nutrition in ex vivo lung perfusion: Addressing metabolism improves both inflammation and oxygenation.</a> American Journal of Transplantation. 19 (12), 3390-3397 (2019).</li><li>Andreasson, A. S. I., 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=Profiling+inflammation+and+tissue+injury+markers+in+perfusate+and+bronchoalveolar+lavage+fluid+during+human+ex+vivo+lung+perfusion.">Profiling inflammation and tissue injury markers in perfusate and bronchoalveolar lavage fluid during human ex vivo lung perfusion.</a> European Journal of Cardio-Thoracic Surgery. 51 (3), 577-586 (2017).</li><li>Sadaria, M. R., 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=Cytokine+expression+profile+in+human+lungs+undergoing+normothermic+ex-vivo+lung+perfusion.">Cytokine expression profile in human lungs undergoing normothermic ex-vivo lung perfusion.</a> The Annals of Thoracic Surgery. 92 (2), 478-484 (2011).</li><li>Ricard, J. D., Dreyfuss, D., Saumon, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ventilator-induced+lung+injury.">Ventilator-induced lung injury.</a> European Respiratory Journal. 42, 2-9 (2003).</li><li>Aboelnazar, N. S., 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=Negative+pressure+ventilation+decreases+inflammation+and+lung+edema+during+normothermic+ex-vivo+lung+perfusion.">Negative pressure ventilation decreases inflammation and lung edema during normothermic ex-vivo lung perfusion.</a> The Journal of Heart and Lung Transplantation. 37 (4), 520-530 (2018).</li><li>Lai-Fook, S. J., Rodarte, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pleural+pressure+distribution+and+its+relationship+to+lung+volume+and+interstitial+pressure.">Pleural pressure distribution and its relationship to lung volume and interstitial pressure.</a> Journal of Applied Physiology. 70 (3), 967-978 (1991).</li><li>Buchko, M. T., 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=Clinical+transplantation+using+negative+pressure+ventilation+ex+situ+lung+perfusion+with+extended+criteria+donor+lungs.">Clinical transplantation using negative pressure ventilation ex situ lung perfusion with extended criteria donor lungs.</a> Nature Communications. 11 (1), 5765 (2020).</li><li>Buchko, M. T., 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=A+low-cost+perfusate+alternative+for+ex+vivo+perfusion.">A low-cost perfusate alternative for ex vivo perfusion.</a> Transplantation Proceedings. 52 (10), 2941-2946 (2020).</li><li>Forgie, K. A., 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=Left+lung+orthotopic+transplantation+in+a+juvenile+porcine+model+for+ESLP.">Left lung orthotopic transplantation in a juvenile porcine model for ESLP.</a> The Journal of Visualized Experiments. , (2021).</li></ol>

有几个关键的手术步骤以及故障排除需要确保成功的ESLP运行。与成人肺相比，幼年猪肺非常脆弱，因此采购外科医生在处理猪肺时必须小心。在解剖肺部时，尝试“非接触”技术以避免引起创伤和肺不张至关重要。“非接触式”是指在采购过程中使用最低限度的手动操作肺部。手术期间使用呼吸机进行肺复张操作在猪肺中远不如人肺有效。不建议像人类肺部那样手动将空气重定向到肺泡，因为这...

肺移植 （LTx） 仍然是终末期肺病的标准护理1;然而，LTx具有显着的局限性，包括供体器官利用率不足2 和40%3的候补名单死亡率，高于任何其他实体器官移植4，5。由于器官质量问题，供体器官利用率较低（20-30%）。过长的地理运输距离加上严格的供体器官接受标准加剧了这些质量问题。LTx在长期移植和患者预后方面也落后于其他实体器官移植2。原发性移植物功能障碍 （PGD） 最常由缺血性再灌注损伤 （IRI） 引起，是 LTx 后 30 天死亡率和发病率的主要原因，并增加了慢性移植物功能障碍的风险6，7。努力降低IRI和延长安全运输时间对于改善患者预后至关重要。
非原位肺灌注（ESLP）是一项创新技术，在减轻这些限制方面显示出希望。ESLP有助于移植前供体肺的保存，评估和修复。在移植扩展标准供体（ECD）肺后，它显示出令人满意的短期和长期结果，有助于增加适合LTx的供体肺的数量，一些中心的器官利用率提高了20%8，9，10。与目前LTx的临床标准冷静态保存（CSP）相比，ESLP具有以下几个优点：器官保存时间不限于6小时，可以在植入前评估器官功能，并且由于连续的器官灌注，可以对优化器官功能的灌注物进行修改11。
目前绝大多数为人类使用而设计的ESLP设备都使用正压通气（PPV）;然而，最近的文献表明，这种通气策略不如负压通气（NPV）ESLP，PPV诱发更显着的呼吸机诱导的肺损伤12，13，14，15。在人和猪肺中，与正压非原位肺灌注 （PPV-ESLP） 相比，NPV-ESLP 在各种生理领域表现出卓越的器官功能，包括促炎细胞因子产生、肺水肿和大疱形成15。NPV-ESLP中整个肺表面胸内压的均匀分布被认为是这一优势的重要因素15，16。除了临床前益处外，NPV-ESLP的临床安全性和可行性已在最近的一项临床试验中得到证实17。利用一种新型NPV-ESLP装置，成功保存、评估了12个扩展标准供体人肺，随后移植了100%的30天和1年生存率。
本手稿的目的是展示我们实验室的NPV-ESLP装置的工作方案，在正常温度条件下使用幼年猪肺12小时。详细介绍了手术检索，还描述了我们的定制软件平台的启动、管理和终止。还解释了组织收集和样本管理的策略。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0 ETHIBOND Green 1 x 36&#34; Endo Loop 0</td>
			<td>ETHICON</td>
			<td>D8573</td>
			<td></td>
		</tr>
		<tr>
			<td>2-0 SILK Black 12&#34; x 18&#34; Strands</td>
			<td>ETHICON</td>
			<td>SA77G</td>
			<td></td>
		</tr>
		<tr>
			<td>ABL 800 FLEX Blood Gas Analyzer</td>
			<td>Radiometer</td>
			<td>989-963</td>
			<td></td>
		</tr>
		<tr>
			<td>Adult-Pediatric Electrostatic Filter HME - Small</td>
			<td>Covidien</td>
			<td>352/5877</td>
			<td></td>
		</tr>
		<tr>
			<td>Arterial Filter</td>
			<td>SORIN GROUP</td>
			<td>01706/03</td>
			<td></td>
		</tr>
		<tr>
			<td>Backhaus Towel Clamp</td>
			<td>Pilling</td>
			<td>454300</td>
			<td></td>
		</tr>
		<tr>
			<td>Biomedicus Pump</td>
			<td>Maquet</td>
			<td>BPX-80</td>
			<td></td>
		</tr>
		<tr>
			<td>Cable Ties &#8211; White 12&#8221;</td>
			<td>HUASU International</td>
			<td>HS4830001</td>
			<td></td>
		</tr>
		<tr>
			<td>Calcium Chloride</td>
			<td>Fisher Scientific</td>
			<td>C69-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Cooley Sternal Retractor</td>
			<td>Pilling</td>
			<td>341162</td>
			<td></td>
		</tr>
		<tr>
			<td>CUSHING Gutschdressing Forceps</td>
			<td>Pilling</td>
			<td>466200</td>
			<td></td>
		</tr>
		<tr>
			<td>D-glucose</td>
			<td>Sigma-Aldrich</td>
			<td>G5767-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Deep Deaver Retractor</td>
			<td>Pilling</td>
			<td>481826</td>
			<td></td>
		</tr>
		<tr>
			<td>Debakey Straight Vascular Tissue Forceps</td>
			<td>Pilling</td>
			<td>351808</td>
			<td></td>
		</tr>
		<tr>
			<td>Debakey-Metzenbaum Dissecting</td>
			<td>Pilling</td>
			<td>342202</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td>Pilling</td>
			<td>342202</td>
			<td></td>
		</tr>
		<tr>
			<td>Endotracheal Tube 9.0mm CUFD</td>
			<td>Mallinckrodt</td>
			<td>9590E</td>
			<td>Cuff removed for ESLP apparatus</td>
		</tr>
		<tr>
			<td>Flow Transducer</td>
			<td>BIO-PROBE</td>
			<td>TX 40</td>
			<td></td>
		</tr>
		<tr>
			<td>Human Albumin Serum</td>
			<td>Grifols Therapeutics</td>
			<td>2223708</td>
			<td></td>
		</tr>
		<tr>
			<td>Infusion Pump</td>
			<td>Baxter</td>
			<td>AS50</td>
			<td></td>
		</tr>
		<tr>
			<td>Inspire 7 M Hollow Fiber Membrane Oxygenator</td>
			<td>SORIN GROUP</td>
			<td>K190690</td>
			<td></td>
		</tr>
		<tr>
			<td>Intercept Tubing 1/4&#34; x 1/16&#34; x 8&#39;</td>
			<td>Medtronic</td>
			<td>3108</td>
			<td></td>
		</tr>
		<tr>
			<td>Intercept Tubing 3/8&#34; x 3/32&#34; x 6&#39;</td>
			<td>Medtronic</td>
			<td>3506</td>
			<td></td>
		</tr>
		<tr>
			<td>Intercept Tubing Connector 3/8&#34; x 1/2&#34;</td>
			<td>Medtronic</td>
			<td>6013</td>
			<td></td>
		</tr>
		<tr>
			<td>MAYO Dissecting Scissors</td>
			<td>Pilling</td>
			<td>460420</td>
			<td></td>
		</tr>
		<tr>
			<td>Medical Carbon Dioxide Tank</td>
			<td>Praxair</td>
			<td>5823115</td>
			<td></td>
		</tr>
		<tr>
			<td>Medical Nitrogen Tank</td>
			<td>Praxair</td>
			<td>NI M-K</td>
			<td></td>
		</tr>
		<tr>
			<td>Medical Oxygen Tank</td>
			<td>Praxair</td>
			<td>2014408</td>
			<td></td>
		</tr>
		<tr>
			<td>Organ Chamber</td>
			<td>Tevosol</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>PlasmaLyte A</td>
			<td>Baxter</td>
			<td>TB2544</td>
			<td></td>
		</tr>
		<tr>
			<td>Poole Suction Tube</td>
			<td>Pilling</td>
			<td>162212</td>
			<td></td>
		</tr>
		<tr>
			<td>Potassium Phosphate</td>
			<td>Fischer Scientific</td>
			<td>P285-500G</td>
			<td></td>
		</tr>
		<tr>
			<td>Scale</td>
			<td>TANITA</td>
			<td>KD4063611</td>
			<td></td>
		</tr>
		<tr>
			<td>Silicon Support Membrane</td>
			<td>Tevosol</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Bicarbonate</td>
			<td>Sigma-Aldrich</td>
			<td>792519-1KG</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Chloride 0.9%</td>
			<td>Baxter</td>
			<td>JB1324</td>
			<td></td>
		</tr>
		<tr>
			<td>Sorin XTRA Cell Saver</td>
			<td>SORIN GROUP</td>
			<td>75221</td>
			<td></td>
		</tr>
		<tr>
			<td>Sternal Saw</td>
			<td>Stryker</td>
			<td>6207</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical Electrocautery Device</td>
			<td>Kls Martin</td>
			<td>ME411</td>
			<td></td>
		</tr>
		<tr>
			<td>Temperature Sensor probe</td>
			<td>Omniacell Tertia Srl</td>
			<td>1777288F</td>
			<td></td>
		</tr>
		<tr>
			<td>THAM Buffer</td>
			<td>Thermo Fisher Scientific</td>
			<td>15504020</td>
			<td>made from UltraPureTM Tris</td>
		</tr>
		<tr>
			<td>TruWave Pressure Transducer</td>
			<td>Edwards</td>
			<td>VSYPX272</td>
			<td></td>
		</tr>
		<tr>
			<td>Two-Lumen Central Venous Catheter 7fr</td>
			<td>Arrowg+ard</td>
			<td>CS-12702-E</td>
			<td></td>
		</tr>
		<tr>
			<td>Vorse Tubing Clamp</td>
			<td>Pilling</td>
			<td>351377</td>
			<td></td>
		</tr>
		<tr>
			<td>Willauer-Deaver Retractor</td>
			<td>Pilling</td>
			<td>341720</td>
			<td></td>
		</tr>
		<tr>
			<td>Yankauer Suction Tube</td>
			<td>Pilling</td>
			<td>162300</td>
			<td></td>
		</tr>
	</tbody>
</table>

normothermic negative pressure ventilation ex situ lung perfusion: evaluation of lung function and metabolism

肺移植（LTx）仍然是终末期肺病的标准护理。合适的供体器官的短缺以及对供体器官质量的担忧因地理运输距离过长和严格的供体器官接受标准而加剧，限制了当前的LTx工作。 非原位 肺灌注（ESLP）是一项创新技术，在减轻这些限制方面显示出希望。在供体炎症环境之外的肺部生理通气和灌注为ESLP提供了优于传统冷静态保存（CSP）的几个优势。有证据表明，负压通气 （NPV） ESLP 优于正压通气 （PPV） ESLP，PPV 可诱发更显著的呼吸机诱发的肺损伤、促炎细胞因子产生、肺水肿和大疱形成。NPV的优势可能是由于胸内压在整个肺表面的均匀分布。定制NPV-ESLP设备的临床安全性和可行性已在最近一项涉及延长标准供体（ECD）人肺的临床试验中得到证实。本文在12小时持续时间的正温NPV-ESLP幼猪模型中描述了该定制设备的使用，特别注意管理技术。指定术前准备，包括ESLP软件初始化，ESLP回路的启动和脱气，以及抗血栓，抗微生物和抗炎剂的添加。描述了术中中心插管、肺活检、放血、采血、心脏切除术和肺切除术的技术。此外，还特别关注麻醉注意事项，概述了麻醉诱导、维持和动态修改。该协议还指定了自定义设备的灌注和通气的初始化、维护和终止。详细描述了动态器官管理技术，包括改变通气和代谢参数以优化器官功能。最后，对肺功能的生理代谢评估进行表征，并在代表性结果中有所体现。

在肺灌注和通气开始时（保存模式），随着灌注液升温至正常体温，肺部通常具有低肺动脉压 （< 10 mmHg） 和低动态顺应性 （< 10 mL/mmHg）。体重 35-50 公斤的约克郡猪通常会产生重达 350-500 克的肺。在NPV-ESLP的第一个小时内，测量的呼气潮气量（TVe）为0-2mL/kg，吸气潮气量（TVi）为100-200mL。TVe通常在3-6小时内达到4-6毫升/千克，之后可能会继续增加，但自然稳定在6-8毫升/千克范围内。TVi 将始终超过 T...

Watch this Scientific Journal Video about Normothermic Negative Pressure Ventilation Ex Situ Lung Perfusion: Evaluation of Lung Function and Metabolism at JoVE.com

Normothermic Negative Pressure Ventilation Ex Situ Lung Perfusion: Evaluation of Lung Function and Metabolism

本文描述了猪负压通气 异位 肺灌注模型，包括定制平台上的采购、附着和管理。重点是麻醉和手术技术，以及故障排除。

非原位负压通气肺灌注：肺功能和代谢的评估

Lung transplantation (LTx) remains the standard of care for end-stage lung disease. A shortage of suitable donor organs and concerns over donor organ ...

Darren Freed博士创造了这种ESLP设备，它使用专利的负压通气技术和软件，这在ESLP研究领域的其他任何地方都看不到。我们的实验室之前已经证明，与正压通气相比，负压通气在ESLP期间引起的肺损伤较少。这可能是由于负压通气的生理机制更多。该装置和方案旨在延长供体肺的安全保存时间，并将边缘肺修复为可接受的移植标准。设备本身操作非常简单。最大的挑战是在不造成任何伤害的情况下通过手术切除幼猪肺。与成年人的肺部相比，它们非常脆弱。首先通过增加峰值压力来维持每公斤 10 毫升的呼气潮气量，以获得最大的肺泡募集。进行中线胸骨切开术。使用电烙术切开皮肤和皮下组织。从胸骨下部的后表直接解剖软组织附件。使用胸骨锯分开胸骨。插入胸骨牵开器并张开。进行心脏切除术后，双侧打开并取出纵隔胸膜以露出肺部。然后将胸膜附件从横膈膜向左下肺叶分开，使用 Deaver 牵开器将横膈膜向上固定。现在，将左侧的下肺韧带分开，然后继续向上向肺门方向移动。接下来，将下腔静脉和胸膜附件与横膈膜分开，然后使用 Deaver 牵开器向上缩回横膈膜。将右侧的下肺韧带分开，然后继续向上朝肺门。然后分开无名静脉和拱形血管以暴露气管。钝化解剖气管周围组织后，在最大吸气时使用管夹夹住气管，确保呼气潮量为每公斤 10 毫升。横断气管并向上提起夹具部分以提供手术牵引。然后用沉重的Metzenbaum剪刀和空出的手使用钝性夹层从食道上解剖后气管。分割任何剩余的胸膜附件。横断左支气管上方和下方的主动脉，并用一段降主动脉从胸部取出肺。用夹子称量肺部后，迅速将它们存放在装满冰块的冷却器中。首先向灌注回路中加入 500 毫升浓缩红细胞，以达到 1.5 升灌注液的最终体积。拍摄肺部照片以进行数据记录。对于肺活检，用零丝线包围一立方厘米的部分并系好。然后按照文本手稿中的描述，使用剪刀进行组织分析，切除这部分绑扎的部分，并分割活检段。接下来，要将 3/8 半英寸管接头固定到主肺动脉上，请使用镊子或按扣抓住主肺动脉的对侧。然后，将带有半英寸部分的适配器插入主肺动脉并将其固定到位，同时使用零丝或 Ethibond 扎带将适配器固定到位。现在将肺置于硅胶支撑膜上的仰卧位置，并将它们连接到ESLP设备。将第二个管夹放在气管支气管位置附近的气管上。取下更远端的钳夹后，用气管插管插管气管，并使用两条扎带将其固定到位。然后使用管夹夹住通气管，然后从气管中松开近端夹具。将肺动脉适配器连接到肺动脉管线。一旦主肺动脉脱气，开始灌注时间。在设置页面上，单击启动加热器并将温度设置为 38 摄氏度。输入猪的体重以计算心输出量。在主页上，将持续气道正压通气设置为 20 厘米水，然后单击开始持续气道正压通气。通气开始时，松开通气管。将动脉压力传感器归零后，用管夹将肺动脉线夹在压力传感器上方。打开传感器，单击设置页面上的零肺动脉压和零血流，并在主页上确认读数归零。关闭压力传感器旋塞阀以读取管路压力。打开肺动脉插管的线。在主页上选择10%心输出量。单击返回 PA 手册并松开肺管。在将灌注液固定到机器上之前，请将其固定在顶盖中，以防止在ESLP期间形成冷凝。抽取10毫升灌注液进行离心分析，并抽取时间零动脉血气。离心前将样品放在冰上。一旦肺部灌注 10 分钟，将流量增加到心输出量的 20%。当灌注液温度达到 32 摄氏度时，用夹子将腔室盖固定到位以形成气密密封。确保肺部处于最佳位置并修复漏气。盖好盖子后，在关闭持续气道正压通气之前夹紧通气管。在设置页面上，选择零胸内压、气道压力和气流。然后确认读数在主页上归零。在松开通气管之前单击启动 CPAP。设置文稿中提到的通气参数，点击按下开始通风，激活负压通气。将侧端口通风管连接到腔室。在接下来的几次呼吸中，保持文本手稿中提到的通风参数。灌注10分钟后，增加流量至20%，灌注20分钟后，增加流量至30%心输出量。向系统中添加葡萄糖和胰岛素。根据手稿说明，使用 50% 心输出量和脱氧混合扫描气体在 5 分钟内评估肺功能。在保存模式下的每个奇数小时，抽取 10 毫升灌注液样品以供将来分析。在评估模式五分钟后，每小时从脱氧前和脱氧后端口抽取一毫升脱氧前动脉血气样本。灌注和通气 12 小时内平均肺动脉压、动态顺应性和肺血管阻力的趋势可能受所采用的特定 ESLP 实验方案的影响。本文展示了在整个负压通气异位肺灌注期间，在 5 小时和 11 小时的评估时间点动脉氧分压与吸入氧分数的比例的典型趋势。水肿形成是与内皮通透性相关的炎症的另一个重要指标，在负压通气异位肺灌注 12 小时结束时，典型体重增加 30%。灌注液可作为整体肺状态的替代指标。因此，灌注液的血气分析提供了有关肺部代谢状态的广泛信息。在ESLP的评估模型中，在12小时的运行中发生在3，5，7，9和11小时，观察到左动脉氧分压呈上升趋势。在手术切除肺部时，尽可能轻柔并快速有效地移动非常重要。该装置，方案和实验室的建立导致了成功的临床试验和一些延长供体肺保存和修复的发现。

Research

JoVE Journal

Medicine

Normothermic Negative Pressure Ventilation Ex Situ Lung Perfusion: Evaluation of Lung Function and Metabolism

Pressure Controlled Ventilation to Induce Acute Lung Injury in Mice

压力控制通气诱导小鼠急性肺损伤

Murine models are extensively used to investigate acute injuries of different organs systems (1-34). Acute lung injury (ALI), which occurs with prolonged ...

科研

医学

Noninvasive Intratracheal Intubation to Study the Pathology and Physiology of Mouse Lung

无创性气管内插管，以研究小鼠肺的病理生理

The use of a model that mimics the condition of lung diseases in humans is critical for studying the pathophysiology and/or etiology of a particular ...

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program

的隔离方法<em&gt;离体</em&gt;肺灌注的大鼠模型：教训制定鼠EVLP课程中学

The number of acceptable donor lungs available for lung transplantation is severely limited due to poor quality. Ex-Vivo Lung Perfusion (EVLP) has allowed ...

Ex Situ Normothermic Machine Perfusion of Donor Livers

Ex Situ Normothermic Machine Perfusion of Donor Livers

供肝易地常温灌注机

In contrast to conventional static cold preservation (0-4 &#176;C), ex situ machine perfusion may provide better preservation of donor livers. Continuous ...

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

A Small Animal Model of Ex Vivo Normothermic Liver Perfusion

体外常温肝灌注的小动物模型

There is a significant shortage of liver allografts available for transplantation, and in response the donor criteria have been expanded. As a result, ...

Lung Fixation under Constant Pressure for Evaluation of Emphysema in Mice

小鼠肺气肿评价持续压力下的肺固定

Emphysema is a significant feature of chronic obstructive pulmonary disease (COPD). Studies involving an emphysematous mouse model require optimal lung ...

Normothermic Ex Situ Heart Perfusion in Working Mode: Assessment of Cardiac Function and Metabolism

工作模式下的心功能和代谢评估

The current standard method for organ preservation (cold storage, CS), exposes the heart to a period of cold ischemia that limits the safe preservation ...

Normothermic Ex Vivo Pancreas Perfusion for the Preservation of Pancreas Allografts before Transplantation

Normothermic Ex Vivo Pancreas Perfusion for the Preservation of Pancreas Allografts before Transplantation

非原生离体胰腺灌注用于移植前胰腺同种异 体 移植物的保存

Pancreas transplantation (PTx) is a curative treatment for people who live with the burden of a diagnosis of diabetes mellitus (DM). However, due to organ ...

Lung Rapid Recovery Procurement Combined with Abdominal Normothermic Regional Perfusion in Controlled Donation after Circulatory Death

肺快速恢复采购联合腹部正常区域灌注在循环死亡后受控捐献中的应用

Controlled donation after circulatory death (cDCD) has contributed to increasing donor numbers all over the world. Experiences published in the last years ...

Functional Assessment of the Donor Heart During Ex Situ Perfusion: Insights from Pressure-Volume Loops and Surface Echocardiography

Functional Assessment of the Donor Heart During Ex Situ Perfusion: Insights from Pressure-Volume Loops and Surface Echocardiography

非原位灌注期间供体心脏的功能评估：来自压力-容量环和表面超声心动图的见解

Heart transplantation remains the gold standard treatment for advanced heart failure. However, the current critical organ shortage has resulted in the ...