Name: an in vivo mouse model of total intravenous anesthesia during cancer resection surgery
Uploaded: 2021-06-08T14:30:00
Description: Watch this Scientific Journal Video about An In Vivo Mouse Model of Total Intravenous Anesthesia During Cancer Resection Surgery at JoVE.com

作者希望感谢癌症神经免疫实验室的成员和帕克维尔莫纳什大学莫纳什药学研究所的Cameron Nowell博士。这项工作得到了国家健康和医学研究委员会1147498，国家乳腺癌基金会IIRS-20-025，澳大利亚和新西兰麻醉师学院（ANZCA），永久和CTC癌症治疗的资助。

所有动物研究均在莫纳什大学机构动物护理和使用委员会的批准下进行。在这项研究中，使用了6-8周龄的雌性Balb / c小鼠。
1. 准备癌细胞
<ol><li>在培养基中培养肿瘤细胞。在含有10%FBS和200mM谷氨酰胺的α-MEM中培养66cl4小鼠乳腺癌细胞。细胞的支原体文本应为阴性。可选：使用稳定转导的细胞表达萤火虫荧光素酶进行生物发光成像，以便在切除手术后进行癌症复发监测<sup class="...

<ol>
	<li>Sullivan, 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=Global+cancer+surgery:+delivering+safe,+affordable,+and+timely+cancer+surgery.">Global cancer surgery: delivering safe, affordable, and timely cancer surgery.</a> Lancet Oncology. 16 (11), 1193-1224 (2015).</li><li>Lim, J. 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=The+effect+of+propofol+and+sevoflurane+on+cancer+cell,+natural+killer+cell,+and+cytotoxic+T+lymphocyte+function+in+patients+undergoing+breast+cancer+surgery:+an+in+vitro+analysis.">The effect of propofol and sevoflurane on cancer cell, natural killer cell, and cytotoxic T lymphocyte function in patients undergoing breast cancer surgery: an in vitro analysis.</a> BMC Cancer. 18 (1), 159 (2018).</li><li>Pandit, J. 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A., Dubowitz, J., Hiller, J., Riedel, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anesthetic+technique+and+cancer+outcomes:+a+meta-analysis+of+total+intravenous+versus+volatile+anesthesia.">Anesthetic technique and cancer outcomes: a meta-analysis of total intravenous versus volatile anesthesia.</a> Canadian Journal of Anesthesia. 66 (5), 546-561 (2019).</li><li>Makito, K., Matsui, H., Fushimi, K., Yasunaga, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Volatile+versus+total+intravenous+anesthesia+for+cancer+prognosis+in+patients+having+digestive+cancer+surgery.">Volatile versus total intravenous anesthesia for cancer prognosis in patients having digestive cancer surgery.</a> Anesthesiology. 133 (4), 764-773 (2020).</li><li>Oh, T. K., Kim, H. H., Jeon, Y. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Retrospective+analysis+of+1-year+mortality+after+gastric+cancer+surgery:+Total+intravenous+anesthesia+versus+volatile+anesthesia.">Retrospective analysis of 1-year mortality after gastric cancer surgery: Total intravenous anesthesia versus volatile anesthesia.</a> Acta Anaesthesiologica Scandinavica. 63 (9), 1169-1177 (2019).</li><li>Lai, H. 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=Propofol-based+total+intravenous+anesthesia+is+associated+with+better+survival+than+desflurane+anesthesia+in+hepatectomy+for+hepatocellular+carcinoma:+a+retrospective+cohort+study.">Propofol-based total intravenous anesthesia is associated with better survival than desflurane anesthesia in hepatectomy for hepatocellular carcinoma: a retrospective cohort study.</a> British Journal of Anaesthesia. 123 (2), 151-160 (2019).</li><li>Hong, B., 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=Anesthetics+and+long-term+survival+after+cancer+surgery-total+intravenous+versus+volatile+anesthesia:+a+retrospective+study.">Anesthetics and long-term survival after cancer surgery-total intravenous versus volatile anesthesia: a retrospective study.</a> BMC Anesthesiology. 19 (1), 233 (2019).</li><li>Flecknell, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Special+Techniques.">Special Techniques.</a> Laboratory Animal Anaesthesia. Fourth edition. , (2015).</li><li>Cicero, L., Fazzotta, S., Palumbo, V. D., Cassata, G., Lo Monte, A. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Anesthesia+protocols+in+laboratory+animals+used+for+scientific+purposes.">Anesthesia protocols in laboratory animals used for scientific purposes.</a> Acta Biomedica. 89 (3), 337-342 (2018).</li><li>Sloan, E. K., 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+sympathetic+nervous+system+induces+a+metastatic+switch+in+primary+breast+cancer.">The sympathetic nervous system induces a metastatic switch in primary breast cancer.</a> Cancer Research. 70 (18), 7042-7052 (2010).</li><li>Al-Hashimi, M., Scott, S. W. M., Thompson, J. P., Lambert, D. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Opioids+and+immune+modulation:+more+questions+than+answers.">Opioids and immune modulation: more questions than answers.</a> British Journal of Anaesthesia. 111 (1), 80-88 (2013).</li><li>DeMarco, G. J., Nunamaker, E. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Review+of+the+effects+of+pain+and+analgesia+on+immune+system+function+and+inflammation:+relevance+for+preclinical+studies.">A Review of the effects of pain and analgesia on immune system function and inflammation: relevance for preclinical studies.</a> Comparative Medicine. 69 (6), 520-534 (2019).</li><li>Hiller, J. G., Perry, N. J., Poulogiannis, G., Riedel, B., Sloan, E. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perioperative+events+influence+cancer+recurrence+risk+after+surgery.">Perioperative events influence cancer recurrence risk after surgery.</a> Nature Reviews Clinical Oncology. 15 (4), 205-218 (2018).</li></ol>

本研究报告了在乳腺癌小鼠模型中使用丙泊酚进行全静脉麻醉 （TIVA） 的方案，该方案复制了需要癌症手术的患者 TIVA 临床实践的关键方面。该协议允许在癌症进展小鼠模型中研究癌症手术后的短期和长期临床相关结果，包括细胞因子水平和癌症复发的测量（图2和 图3）。该方法将有助于评估TIVA对癌症相关结局的影响，并将其与其他麻醉技术（如挥发...

超过60%的癌症患者接受麻醉进行手术切除1。目前，尚无具体的临床指南来确定癌症患者使用的麻醉选择。对麻醉师的调查表明，他们更喜欢挥发性麻醉，包括在癌症手术期间2，3。然而，越来越多的证据表明，与挥发性麻醉相比，在癌症手术中使用基于丙泊酚的全静脉麻醉 （TIVA） 可能与术后结局（无进展生存期、总生存期）的改善有关4.随后的临床研究继续报告相互矛盾的结果5，6，7，8。这些发现支持需要临床前研究，以更好地了解不同麻醉剂对癌症相关结果的机制影响。
然而，在模拟癌症手术的 体内 研究中，麻醉通常是手术的附带部分。选择麻醉的基本原理通常不是实验设计的重点，并且可能无法评估其对癌症相关终点的影响。例如，需要维持癌症手术麻醉的 体内 研究最常使用吸入挥发性麻醉9。在 体内 研究中使用异丙酚的地方，它通过单次推注给药和腹腔内给药给药，这不复制临床肿瘤麻醉方案10。丙泊酚给药的方法诱导适合快速手术的轻度麻醉。但是，它不允许维持癌症切除手术所需的麻醉，这可能会延长。此外，腹膜内递送的吸收动力学与临床给药方法不同。
开发了一种用于癌症切除手术的基于丙泊酚的TIVA模型，以满足这一需求。开发了一种持续维持麻醉的方案，通过滴定麻醉剂以允许对手术刺激做出反应，以将麻醉剂输送的关键方面复制到接受癌症手术的患者。所得方案与癌症小鼠模型一起使用，以在癌症切除手术期间提供TIVA。评估对短期和长期癌症相关结局的影响。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.9% saline</td>
			<td>Fresnius Kabi</td>
			<td>AUST R 197198</td>
			<td></td>
		</tr>
		<tr>
			<td>Artery forceps</td>
			<td>Proscitech</td>
			<td>TS1322-140</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>Temgesic</td>
			<td>TEMG I</td>
			<td></td>
		</tr>
		<tr>
			<td>Heated surgical mat</td>
			<td>Custom</td>
			<td>-</td>
			<td></td>
		</tr>
		<tr>
			<td>Hypodermic needle (30 G, 1 mL insulin syringe)</td>
			<td>Terumo</td>
			<td>NN3013R</td>
			<td></td>
		</tr>
		<tr>
			<td>IVIS Lumina</td>
			<td>PerkinElmer</td>
			<td>126274</td>
			<td></td>
		</tr>
		<tr>
			<td>Luciferin</td>
			<td>Promega</td>
			<td>P1041/2/3</td>
			<td></td>
		</tr>
		<tr>
			<td>Polyurethane catheter</td>
			<td>Intramedic</td>
			<td>427401</td>
			<td></td>
		</tr>
		<tr>
			<td>Povidone Iodine</td>
			<td>Betadine</td>
			<td>AUST R 29562</td>
			<td></td>
		</tr>
		<tr>
			<td>Propofol Lipuro, 2%</td>
			<td>Braun</td>
			<td>3521490</td>
			<td></td>
		</tr>
		<tr>
			<td>Sevoflurane</td>
			<td>Baxter</td>
			<td>ANZ2L9117</td>
			<td></td>
		</tr>
		<tr>
			<td>Sevoflurane vaporiser</td>
			<td>Vetquip</td>
			<td>VQ1334</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile gauze</td>
			<td>Multigate Medical Products</td>
			<td>11-600A</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical scissors</td>
			<td>Proscitech</td>
			<td>TS1044</td>
			<td></td>
		</tr>
		<tr>
			<td>Sutures, 5-0 nylon</td>
			<td>Dynek</td>
			<td>V504</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe pump</td>
			<td>Harvard Apparatus</td>
			<td>70-4500</td>
			<td></td>
		</tr>
		<tr>
			<td>Syringes (1 mL)</td>
			<td>Terumo</td>
			<td>SS+01T</td>
			<td></td>
		</tr>
	</tbody>
</table>

an in vivo mouse model of total intravenous anesthesia during cancer resection surgery

麻醉是癌症护理的常规组成部分，用于诊断和治疗程序。麻醉技术最近被认为会影响长期癌症结果，可能是通过调节影响癌细胞行为和免疫细胞功能的肾上腺素能炎症反应。新出现的证据表明，与吸入挥发性麻醉相比，基于丙泊酚的全静脉麻醉（TIVA）可能对长期癌症结局有益。然而，现有的临床表现并不一致。迫切需要确定所涉及的潜在机制的临床前研究，以指导临床研究的设计，从而加快洞察力。大多数麻醉的临床前模型都是从 体内 麻醉的使用中推断出来的，并且没有最佳设计来研究麻醉本身作为主要终点的影响。本文描述了一种在乳腺癌切除小鼠模型中提供丙泊酚-TIVA麻醉的方法，该方法复制了癌症患者临床递送的关键方面。该模型可用于研究麻醉对不同癌症类型癌症结果的作用机制，并可以外推到临床前麻醉研究的其他非癌症领域。

该方法描述了在小鼠癌症切除手术期间使用丙泊酚进行全静脉麻醉（TIVA）的模型。丙泊酚在该小鼠模型中通过使用注射泵的静脉导管递送（图1A，B），以在癌症手术麻醉的临床环境中复制TIVA的递送。注射泵的使用允许从吸入麻醉的初始诱导快速转换为静脉内给药，从而最大限度地减少挥发性麻醉的暴露。
丙泊酚基TIVA实现稳定麻醉后，切除...

Watch this Scientific Journal Video about An In Vivo Mouse Model of Total Intravenous Anesthesia During Cancer Resection Surgery at JoVE.com

An In Vivo Mouse Model of Total Intravenous Anesthesia During Cancer Resection Surgery

本文描述了一种模拟小鼠癌症切除手术期间全静脉麻醉（TIVA）的方法。目标是向癌症患者复制麻醉输送的关键特征。该方法允许研究麻醉技术如何影响切除手术后的癌症复发。

癌症切除手术中全静脉麻醉的 体内 小鼠模型

Anesthesia is a routine component of cancer care that is used for diagnostic and therapeutic procedures. The anesthetic technique has recently been ...

该新方案描述了使用丙泊酚麻醉的小鼠手术麻醉。该方法能够研究疾病小鼠模型中的丙泊酚麻醉。该方案允许评估全静脉麻醉，避免使用挥发性麻醉。该方案可用于探索丙泊酚麻醉如何影响疾病相关结果。例如，在这里，我们评估手术切除肿瘤后的短期和长期癌症结果。首先，在含有10%FBS和200毫摩尔谷氨酰胺的α MEM培养基中稳定转导以表达萤火虫荧光素的66个CL4 Mirai乳腺癌细胞。将细胞在 37 摄氏度但 5% 二氧化碳下孵育。当细胞达到约80%汇合度时传代培养细胞。当细胞处于对数生长期，在EDTA溶液中滴有两毫升时，提起贴壁细胞并使用血细胞计数器计数细胞。然后在PBS中稀释细胞，并将稀释的细胞悬液保持在冰上，直到小鼠准备好注射。小鼠完全麻醉后，用一次性酒精棉签擦拭左第四个乳腺脂肪垫区域以准备注射部位。用癌细胞悬浮液填充连接到无菌 27 号皮下注射针头的 25 微升汉密尔顿注射器。使用镊子提拉并固定皮肤。将1次10次注射到第五个细胞和20微升PBS到距离约1毫米的第四个左乳腺脂肪垫中。如果细胞用荧光素酶标记，则使用0.5毫升胰岛素注射器和30号皮下注射针头在麻醉小鼠的侧尾静脉中注射100微升D-荧光素。注射两分钟后，将小鼠置于生物发光成像系统中，乳腺脂肪垫朝上并捕获图像10秒钟。成像后，将鼠标放在干净的笼子中，让它从麻醉中恢复。使用卡尺监测原发性肿瘤生长的生长。计算肿瘤体积并在原发肿瘤达到80至90立方毫米体积时切除肿瘤。使用含有2%丙泊酚制剂的30号1毫升胰岛素注射器设置自动注射泵。在含有百分之三氟烷或异氟烷的诱导室中诱导麻醉。然后将麻醉的小鼠转移到37摄氏度的加热垫上，并使用鼻锥用2%至3%的七氟烷或异氟烷维持麻醉。在静脉插管期间根据需要调整七氟醚的输送，以保持稳定的麻醉深度，表现为踏板反射丧失和呼吸频率低于每分钟 100 次。在眼睛上涂抹水性润滑剂以防止角膜干燥，然后皮下注射每公斤丁丙诺啡0.05毫克。为了提供基于丙泊酚的全静脉麻醉，请使用连接到无菌聚氨酯导管的无菌 30 号皮下注射针插管外侧尾静脉。通过血液回气进入导管确认正确放置。在一分钟内给予 2% 的丙泊酚作为每公斤 27 毫克的初始推注，以开始全静脉麻醉。停止七氟醚给药。将丙泊酚输注保持在每分钟 2.2 至 4.0 毫克/千克的速度，以在手术过程中保持稳定的麻醉深度。剃除腹部区域，并用聚维酮碘溶液对手术区域进行消毒。在左第四乳腺脂肪垫中做一个低于肿瘤的一厘米切口。使用镊子和剪刀解剖肿瘤和左腹股沟淋巴结。如果使用荧光素酶标记的肿瘤细胞，如图所示在侧尾静脉中注射d-荧光素，并捕获图像60秒以获得清晰的边缘。如果发现残留肿瘤，请从乳腺脂肪垫上切除其他组织。一旦在手术部位达到止血，使用5-0尼龙缝合线关闭皮肤。用聚维酮碘溶液擦拭伤口。手术完成后，停止麻醉并将鼠标放在加热垫上的干净笼子中，使其从麻醉中恢复。每 15 分钟监控一次鼠标，直到鼠标恢复正常警觉状态。然后在手术后 48 小时内每 12 小时监测并提供镇痛。对于原发性肿瘤复发或远处复发评估，每周监测一次小鼠，使用生物发光成像系统在手术后一周开始。采用生物发光成像技术鉴定原发性肿瘤切除术后肺部远处肿瘤复发情况。该模型可用于评估围手术期发生的事件。在丙泊酚下进行癌症手术后24小时，评估循环血浆细胞因子。快速插管尾静脉和最小化七氟醚暴露持续时间所需的做法。七氟醚应逐渐降低滴定，同时推注丙泊酚以防止过度镇静。该方法可用于研究丙泊酚麻醉对心脏手术或危重疾病（如败血症）小鼠模型的影响。

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An In Vivo Mouse Model of Total Intravenous Anesthesia During Cancer Resection Surgery

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医学

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

多光子成像在口腔鳞状细胞癌原位小鼠模型的肿瘤细胞的侵袭

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

Combination Radiotherapy in an Orthotopic Mouse Brain Tumor Model

在原位小鼠脑肿瘤模型结合放疗

Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the ...

An Ex vivo Model to Study Hormone Action in the Human Breast

An Ex vivo Model to Study Hormone Action in the Human Breast

一个<em&gt;体外</em&gt;模型在乳腺癌研究激素的作用

The study of hormone action in the human breast has been hampered by lack of adequate model systems. Upon in vitro culture, primary mammary epithelial ...

An Orthotopic Mouse Model of Spontaneous Breast Cancer Metastasis

自发性乳腺癌转移的原位小鼠模型

Metastasis is the primary cause of mortality of breast cancer patients. The mechanism underlying cancer cell metastasis, including breast cancer ...

Depletion of Mouse Cells from Human Tumor Xenografts Significantly Improves Downstream Analysis of Target Cells

从人类肿瘤异种移植的小鼠细胞的消耗显着改善靶细胞下游分析

The use of in vitro cell line models for cancer research has been a useful tool. However, it has been shown that these models fail to reliably mimic ...

An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model

An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model

用于评估椎弓根螺钉的麻醉，手术和收获方法<em&gt;体内</em&gt;猪腰椎模型

Pedicle screw fixation is the gold standard for the treatment of spinal diseases. However, many studies have reported the issue of loosening pedicle ...

In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur

In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur

在体内MRI 对小鼠股骨骨愈合过程中骨折愈伤组织发育的评价

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The ...

Isometric Contractility Measurement of the Mouse Mesenteric Artery Using Wire Myography

用线 Myography 测定小鼠肠系膜动脉的等轴收缩力

The wire myograph technique is used to assess the contractility of vascular smooth muscles in response to depolarization, GPCR agonists/inhibitors and ...

Noninvasive Determination of Vortex Formation Time Using Transesophageal Echocardiography During Cardiac Surgery

经食管超声心动图在心脏手术中无创地测定涡旋形成时间

Trans-mitral blood flow produces a three-dimensional rotational body of fluid, known as a vortex ring, that enhances the efficiency of left ventricular ...

癌症切除手术中全静脉麻醉的 体内 小鼠模型

癌症切除手术中全静脉麻醉的体内小鼠模型