This study was supported and funded by the Avenir Foundation (Fondation de l'Avenir, 255 rue de Vaugirard, 75719 Paris cedex 15, Paris, France). This study would never have been conducted without the precious help of the veterinary team of the Laboratory of Biosurgical Research from the Alain Carpentier Foundation.

所有的动物都根据动物研究伦理委员会（法国农业部的指导方针）处理。该协议获得在巴黎第五大学授权的动物experimentations当地伦理委员会的批准（注册号MESR 2035.02;医学学部巴黎笛卡尔，法国巴黎）。 1. ADSC文化和标签<ol><li>获得一个私人机构证实脂肪干细胞。在37℃和5％的CO 2与含10％胎儿小牛血清和1％抗生素（青霉素和链霉素）的α极限必需培养基培养同种异体脂肪干细胞。 </li><li>使用标准的细胞扩增程序（37℃和5％CO 2）得到每只动物约24×10 6个脂肪干细胞。收获使用标准的胰蛋白酶溶液（10毫升的T150皿）的脂肪干细胞。用血球进行细胞计数。 </li><li>执行小区使用PKH67染色PR标记移植前一天ocedure如下所述。 注:在染色过程的目标是允许移植后细胞跟踪。 <ol><li>为了标注2×10 6的脂肪干细胞，准备溶液A（2×10 6细胞用100微升稀释的C）和溶液B（PKH67 4μL用1毫升稀释液的C）。 </li><li>孵育溶液A和B为2分钟。然后，添加100μL胎儿小牛血清1分钟。 </li><li>冲洗以1500 xg离心与柠檬酸的RPMI 300微升的溶液（RPMI的298.5微升和柠檬酸1.5μL），离心5分钟。重复此操作2次。保持标记细胞，并保护他们免受光，直到板结构。 </li></ol></li></ol> 2.双击ADSC表构建<ol><li>一天移植前，准备与每细胞培养基的阱4毫升，如上所述，PKH标记之前12孔温度响应细胞培养皿。种子菜1.5×10 6 PKH-labele每孔D细胞孵育12小时（37℃和5％CO 2）。 </li><li>上移植的当天，通过温育在室温下将培养皿30分钟分离汇合细胞片。 </li><li>轻轻吸了吸取1 ADSC表和图层它的疏水纸（1.5厘米直径）。使用这种纸张作为支撑膜。 </li><li>掌握另一ADSC片，轻轻地沉积其上的另一个的顶部，以获得一个双层构建体。 </li><li>重复这些程序根据需要（4每只动物），得到尽可能多的ADSC表结构。 </li></ol> 3.食管内镜黏膜下剥离术（ESD） <ol><li>上移植的当天，用10毫克/千克肌内氯胺酮预先用药的动物，并以8毫克/公斤静脉内异丙酚诱导它们。然后，执行气管插管并保持与异氟烷2.5％吸入麻醉。 </li><li>一旦动物是在全身麻醉下，进行ESD与gastroscOPE，一个视频内窥镜，以及电外科单元。 <ol><li>使用内窥镜刀和软凝固，以获得半周背侧标记范围从40厘米到从牙弓45厘米。 </li><li>注入含有靛蓝胭脂红染料成用于粘膜层从肌肉层分离的粘膜下层甘油溶液。 </li><li>使用内窥镜刀endocut I模式来获得圆周切口。 </li><li>使用的内窥镜刀强迫凝固模式来执行粘膜解剖，从近端切口移动到远端切口。 </li></ol></li><li>在静电放电过程结束时，观察半周，5-厘米长的食管瘢痕露出肌层。 </li></ol> 4.移植内镜<ol><li>静电放电后，立即内镜移植动物4双ADSC表结构。 <ol><li>掌握内镜钳和ADSC表结构通过使用大量的，透明的内窥镜盖保护它运输到创伤部位中。 </li><li>的ADSC表构建体的整个表面施加到溃疡床。 </li><li>轻轻地应用ADSC表结构为10分钟获得一个完整的，稳定的植入。为应用程序，使用内窥镜钳子或内窥镜帽并应用ADSC的整个表面构造到食道粘膜下层。 </li></ol></li><li>重复此步骤，对每个动物四次，以获得4移栽双贴壁细胞表结构。 注:我们的目标是覆盖近似一半的受伤面积。 </li></ol> 5.术后评估及随访<ol><li>对于每个动物，确保后的ESD镇痛为0.2毫克/千克吗啡为第一天并用埃索美拉唑的28天的抗酸处理（40毫克/天），每天三次肌内注射。执行与amoxicill抗生素预防在（1克/天）7天。授权于翌日第1天的液体和固体。 </li><li>跟进动物28天。获得使用圆润平卡斯吞咽困难得分16，体重变化的测量日常临床评估。 </li><li>多式联运狭窄的评价:全身麻醉，在预定的3天，14和28下，担负狭窄发生多式联运评价。 <ol><li>执行使用瘢痕描述，狭窄测量和纸张支撑膜检测内窥镜评价。使用胃镜通过狭窄跨越测量狭窄（胃镜的直径为8毫米）。观察疤痕，关注炎性方面（粘膜红斑，粘膜水肿，和粘膜自发性出血）和瘢痕内不存在或纸张支撑膜的存在。 </li><li>通过胃镜操作者运河直接申请pCLE绿色探头插入疤痕床。搜索一个自发并组织与PKH67标记ADSC表结构植入兼容绿色信号。 </li><li>执行重晶石食管造影的2正交发生率与放射性箍（正面和左路的发生率）。保持为狭窄评价最紧密的发生率（的狭窄（％度）= [1-（狭窄下短轴长度/狭窄下正常轴）的长度×100）]）17，18。 </li></ol></li><li>动物牺牲。 <ol><li>在第28天，在随访期结束时，执行动物牺牲。虽然动物仍然是在全身麻醉下，注入100毫克/公斤静脉注射苯巴比妥。确保动物死亡利用临床检查为缺乏心跳和呼吸运动。 </li><li>胸骨切开术和器官曝光后，取出食道从每个动物处死（用10％福尔马林固定，石蜡包埋，4微米厚的部分）。染色幻灯片用苏木和藏红花15。对于数字化计算机分析幻灯片和比较动物组15。 </li></ol></li></ol>

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作者什么都没有透露。

在这种猪模型中，ADSC表移植在技术上是成功的，并允许细胞植入监测体内 pCLE评价。临床，内窥镜，放射性，和组织学评价证明增强的ESD后在食管狭窄预防的内窥镜ADSC片的有效性。 含有靛蓝胭脂红染料片的ADSC甘油溶液的内窥镜移植是在再生医学的创新方法。大木等。第一次描述了内镜移植过程。在他们的研究中，细胞表构建体组成层叠在聚偏二氟乙烯的薄片口腔粘膜上皮细胞片，用作载体膜10，11，19的。支持膜与内镜钳抓住并小心地放在床上溃疡。切尔温和应用的10分钟后，获得的L-片的粘附性。这个过程还没有被再现。在这项研究中，双ADSC表构建物移植多种原因。 ADSC的选择是基于几个研究，证明了其抗炎能力，特别是在皮肤愈合过程。脂肪干细胞是公知的调节角质形成细胞和成纤维细胞的相互作用7。纤维化发展的机制是复杂的，仅部分的理解。的旁分泌效应似乎发挥ADSC动作的主要作用，包括抗炎因子的生产，如TGFβ和促血管生成因子20。 内镜ADSC表移植有几个限制。首先，被应用细胞片10分钟以食管瘢痕获得稳定的粘合性。在这部分程序是具有挑战性的，技术上的困难，不得不由有经验的内镜医师进行。然而，所有的动物都成功地移植，确认了内窥镜检查过程的可行性。最近，以改善细胞片移植的成功使用3D印刷装置的报告21。第二，这种技术具有未知存活率有关。食管瘢痕的细胞一个积极的环境。温度响应盘允许生产的细胞片的构造通过细胞外基质和细胞的连接的形成。因此，为了提高生存率和ADSC相互作用，一个双ADSC表构建体被选择22,23。这项研究并不是用来评估两种细胞存活率或移植对食管狭窄预防剂量效应，这两者都是重要的问题。第三，如在大木等。 ，使用支撑膜的必要携带ADSC表构建体。前前后后呃移植，纸支撑膜不能没有构造破坏拆除。此帧内食管异物负责增加炎症。一个优化的技术将允许移除纸张支撑膜。 与pCLE移植的监测也具有挑战性。这个新的内窥镜成像技术可以在体内的组织学分析，包括腺图案的可视化，微血管和细胞的改变。 pCLE在许多消化系统疾病，如炎性肠疾病，巴雷特食管，和粘液性胰腺囊肿24,25进行评价。所述pCLE兼容PKH67荧光团被用于细胞标记。与ADSC片的存在相容的荧光信号在4只动物中观察到第3天这肯定结果反映的ADSC表移植的成功。然而，贝科使用PKH67具有降低的信号随着时间的推移和细胞分裂后，pCLE评价移植26后仅几天是可能的。大木等。和金井等人。通过免疫染色监测细胞片移植，和他们证实细胞的存在，直到移植后8一天。成熟细胞在这项研究中，具有特定的免疫染色的优点，使用。 食管狭窄是以下的ESD的最严重的不良事件，从而限制了技术发展中的一个。最佳的技术来防止狭窄形成仍然缺乏。在临床实践中，患者具有扩展食管癌的ESD被规定不同的预防性治疗方法，如皮质类固醇，内窥镜球囊扩张术，或食道支架置入27。生物材料的局部应用已经进行，结果各不相同28,29 </SUP&gt;。再生医学的目的是要重建使用各种方法，如组织替代或局部免疫调节的组织或器官。对组织替代，使用具有或不具有上皮细胞的生物支架在文献30,31进行评价。在食管瘢痕观察到早熟，提高再上皮化。对于局部免疫调节，小数据是在大约ADSC注射或羊膜应用32,33的文献报道。一个延迟的，狭窄的形成和降低狭窄率进行观察。 在这种猪模型，成功地进行了内镜细胞表移植过程。在双细胞片移植组织的脂肪干细胞表现出了对防止超期ESD后食管狭窄形成能力。参与ADSC愈合机制过程知之甚少，必须在今后的研究进一步调查。因此，鉴于这些有前景的结果的，临床试验应进行评估扩展后的ESD患者ADSC表构建体的效力。

浅表性食管肿瘤的管理层已与新的内镜技术的发展变化。如今，内窥镜切除是一线治疗。事实上，它是具有较低的发病率和死亡率比等于肿瘤结果1，2，3一手术相关。内窥镜粘膜切除术（EMR）和内镜黏膜下切除术（ESD）是最广泛使用的技术。在扩展浅表肿瘤的情况下，静电放电是优选的。相比EMR，ESD允许整块切除，无论病灶大小和形状4，5，6。 ESD的主要延迟并发症是食道狭窄的形成，这通常发生一周和两周之间切除后。最近公布的研究表明，狭窄形成是相关的切除的大小。日本内镜学会建议避免ESD尺寸大于¾食管周长，因为它们是在病例的90％以上与狭窄发展相关联，并且负责严重馈送麻烦和生活质量严重恶化的要大。 食管狭窄的预防仍然是具有挑战性的。参与狭窄形成机制只知道部分。狭窄的地层仿佛来自两个不同的机制，该协会导致:（1）促炎症细胞招聘和（2）过度纤维化发展7。几个预防性治疗方法已被提出。然而，结果并不令人满意，有什么好处和严重的副作用8,9。最近，日本队，大木等人。 ，提出了自体口腔粘膜细胞的单层细胞片移植到esophageal疤痕。在ESD 10,11后立即进行移植。他们证明这种创新方法的有效性，首先在犬模型，然后在病人。 脂肪组织来源的基质细胞（脂肪干细胞）是有希望在再生医学。其在几个领域中的应用已表明有趣的结果，特别是在伤口愈合过程。 ADSC疗法提供了几个优点，因为细胞是容易分离的，并具有抗炎特性，局部免疫调节作用，血管新生诱导，分化能力为间充质和非间充质系12，13，14相关联。 在先前的研究中，我们的团队表现出双ADSC表镜下移植治疗食管狭窄上一页成效在猪模型15增强的ESD后ention。在这篇文章中，ADSC板结构和内窥镜移植技术的报告介绍。

<table border="0" cellpadding="0" cellspacing="0" width="1071">
	<colgroup>
		<col />
		<col />
		<col />
		<col />
	</colgroup>
	<tbody>
		<tr height="20">
			<td height="20" style="height:20px;width:313px;">Transparent endoscopic cap Q180 compatible</td>
			<td style="width:159px;">Olympus Optical Co</td>
			<td style="width:107px;"></td>
			<td style="width:492px;"></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">GIF-Q180 gastroscope&#160;</td>
			<td>Olympus Optical Co</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Videoscope System Exera II&#160;</td>
			<td>Olympus Optical Co</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Injection needle 18 G</td>
			<td>Olympus Optical Co</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Electrosurgery unit ERBE ICC 350&#160;</td>
			<td>ERBE Technology</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Indigo carmin 1%</td>
			<td>Life</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Endoscopic hybrid knife</td>
			<td>Life</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Minisonde Z pCLE green probe</td>
			<td>Mauna Kea Technology</td>
			<td></td>
			<td>You must learn how tu use the probe. The manipulation could be difficult</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Fetal bovine serum</td>
			<td>Sigma Aldrich</td>
			<td>12105C</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Trypsin</td>
			<td>Sigma Aldrich</td>
			<td>T146</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Alpha minimum essential medium</td>
			<td>Thermo Fisher</td>
			<td>22561-021</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Phosphate-Buffered Salines</td>
			<td>Thermo Fisher</td>
			<td>10010-023</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">PKH67 dye kit</td>
			<td>Sigma Aldrich</td>
			<td>Mini67-KT</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">12-well temperature responsive cell culture dish</td>
			<td>Upcell Thermo Scientific</td>
			<td align="right">174900</td>
			<td>Feel the weel with 4 mL standard medium culture 30 minutes before seeding cells</td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Esomeprazole 40 mg&#160;</td>
			<td>Biogaran</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Moprhine sulfate 50 mg/ mL</td>
			<td>Lavoisier</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Amoxicilline 1 g</td>
			<td>Biogaran</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Ketamine 250 mg/5 mL</td>
			<td>Panpharma</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Propofol 10 mg/ mL</td>
			<td>Fresenius</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;">Hydrophobic paper</td>
			<td>Carrefour</td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

adsc-sheet transplantation to prevent stricture after extended esophageal endoscopic submucosal dissection

在过去几年中，细胞的表结构刺激了再生医学的广泛兴趣，尤其是对重建手术程序。多样化的技术脂肪组织衍生的基质细胞（脂肪干细胞）与各种生物材料相结合的发展导致了多种类型的组织工程替代品，如骨，软骨，和从啮齿动物，猪或人脂肪干细胞的脂肪组织的结构。扩展食管内镜黏膜下剥离术（ESD）负责食管狭窄的形成。狭窄预防仍然是具有挑战性的，没有可用的有效的治疗方法。先前的研究报道了犬模型和人类中的粘膜细胞片移植的有效性。脂肪干细胞归因抗炎特性，局部免疫调节作用，血管新生诱导，分化能力为间充质和非间充质细胞谱系。这个原始的研究描述了TRA内镜一个ADSC组织工程构建的nsplantation防止食管狭窄的猪模型。在ADSC结构是由一纸支撑膜在彼此分层2同种异体ADSC表。将脂肪干细胞标记的PKH67荧光允许基于探针的共聚焦激光显微内镜（pCLE）监控。上移植当天，进行5厘米和已知诱导食管狭窄半周的ESD。动物立即用胃镜ADSC 4移植结构。温和应用的10分钟后，获得的ADSC构建体的完整的粘合。动物处死28天所有动物均成功移植。移植证实第3天以积极的pCLE的评价。相比于移植的动物，对照组动物出现严重狭窄，与主要的纤维化组织的发展，越来越频繁的消化道麻烦，减少体重增加。在我们的模型中，allogeni移植Ç脂肪干细胞，双细胞片组织，增强的ESD后成功并以较低的食管狭窄率密切相关。

脂肪干细胞并获得ADSC片的过程的培养示于图1。 图2示出了移植物的结构，在其纸载体膜在彼此层叠2 ADSC片组成。脂肪干细胞以前与PKH67荧光团标记，以便与 pCLE 体内移植监控。 图3示出了延长的食管内窥镜黏膜下层剥离术的不同步骤，从而产生一个5厘米和半周食管疤痕。 图4示出的内窥镜移植过程。移植是成功的所有动物平缓应用的10分钟后。 图5示出在第3天的正pCLE评价，证实了移植过程的成功。 允许多狭窄analys随访期<html>是。不同的评估被显示在下面的附图和表格。与对照动物相比，移植的动物组显示出较不频繁的消化道麻烦和更大的增益的日28.一个动物体重从移植组相比对照组中的所有动物发展为严重的食管狭窄。相比于移植的动物，动物控制开发显著纤维化组织。 表1示出在随访期临床和内窥镜的发现。 图6示出在移植和非移植动物随访期结束时的内窥镜和放射学发现。 图7显示了移植和控制动物之间的不同组织的调查结果。这些数据证实ADSC表移植治疗食管狭窄预防的效果。 018fig1.jpg"/&gt; 图1. ADSC表建设。 A和B早通道脂肪干细胞在不同的放大倍率标准的培养皿。 ADSC张由商业温度反应培养皿培养PKH标记的脂肪干细胞获得。 C.当培养温度降低至20℃（室温）下，所有的细胞从培养皿表面作为完整的ADSC片自发地分离，而不需要酶处理。利用转移膜移植施工前的四片ADSC收获。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55018/55018fig1large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图2" src="/files/ftp_upload/55018/55018fig2.jpg" /> 图2.双ADSC表建设。 A，B和C. transpla前一天ntation，PKH标记脂肪干细胞收获在早期传代次数（P4 - P5），接种在12孔温度响应细胞培养，并在37℃和5％的CO 2用α-最低必需培养基中培养的，包括10％胎小牛肉血清和1％抗生素（青霉素和链霉素）。各孔涂敷有聚N-异丙基膜（在每个孔的底部）中，用1.5×10 6个细胞接种。当培养温度降低至20℃（室温）下，所有的细胞自发地脱离如无需酶处理完整ADSC片。 D. 2 ADSC片构成的双细胞表构建体的理论架构层叠在彼此并施加到纸载体膜。一个完整的双单元表结构的E.宏观视图。从参考15.修改<a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55018/55018fig2large.jpg" target="_blank">请点击这里Ť</a><html> Ø查看此图的放大版本。 <img alt="图3" src="/files/ftp_upload/55018/55018fig3.jpg" /> 图3.内镜黏膜下剥离过程。 答内窥镜标记创建划定切除区域，范围从40到45厘米的牙弓和涉及一半圆周。 B.含有靛蓝染料的甘油溶液的黏膜下注射。这种喷射的目的是粘膜层从肌肉层，以露出粘膜下层分离。 C.周围切口被外部执行的内窥镜标记，揭示注入粘膜下层。 D.在黏膜下层剥离术的端部的内窥镜视图，示出5厘米长和半周食管疤痕。等="_空白"&gt;点击此处查看该图的放大版本。 <img alt="图4" src="/files/ftp_upload/55018/55018fig4.jpg" /> 图4.双击ADSC表内镜移植过程。 A.四ADSC构建准备移栽。 B.在ADSC保护移植前的大帽内窥镜下施工。动物咽部的交叉，显示了内窥镜下盖保护的ADSC构造C.内镜视图。 D.应用的10分钟是足以获得片粘附。使用内窥镜帽或内镜钳进行应用。 <a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55018/55018fig4large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图5" srC ="/文件/ ftp_upload / 55018 / 55018fig5.jpg"/&gt; 图5. pCLE结果。在第3天pCLE示出了类似于那些在体外获得，以成功的细胞片移植兼容自发和强烈的信号。无信号于14日和28 A.悬浮脂肪干细胞的体外 pCLE信号找到。 B.在细胞片组织脂肪干细胞的体外 pCLE信号。 C. 体内 pCLE探针应用到食管疤痕。 ð。脂肪干细胞的体内 pCLE信号移植后第3天显示。从参考15.修改<a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55018/55018fig5large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图6" src="/files/ftp_upload/55018/55018fig6.jpg" /> 图6.结果形态分析在28天相比于对照组，内镜与第28天放射学研究结果显示短而轻狭窄而不上游扩张和完全再上皮黏膜。从参考15.修改<a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55018/55018fig6large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="图7" src="/files/ftp_upload/55018/55018fig7.jpg" /> 图7.狭窄区域的组织学评价。后HES-标签和幻灯片数字化进行的组织学分析。 在移植的动物（A）中 ，愈合过程进行了改进，以增加再上皮和减少纤维化的发展，相比 非移植动物（B）。激烈的纤维化组织的发展是由于观察到粘膜肌层破坏，纤维变性内肌层浸润和大的粘膜缺损。从参考15.修改<a href="https://ecsource-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/55018/55018fig7large.jpg" target="_blank">请点击此处查看该图的放大版本。</a> <img alt="表格1" src="/files/ftp_upload/55018/55018table1.jpg" /> 在随访期间表1.临床评估。每天使用临床评估根据麦罗平卡斯得分和权重的变化进行临床评估。 （*）醇厚平卡斯得分16。从参考15修改。

Watch this Scientific Journal Video about ADSC-sheet Transplantation to Prevent Stricture after Extended Esophageal Endoscopic Submucosal Dissection at JoVE.com

ADSC-sheet Transplantation to Prevent Stricture after Extended Esophageal Endoscopic Submucosal Dissection

这项研究报告内窥镜脂肪组织的基质细胞（ADSC）的成功的方法折叠移植食管狭窄预防猪模型的扩展内镜黏膜下剥离术（ESD）之后。

ADSC表移植到扩展食管内镜黏膜下剥离术之后防止狭窄

In past years, the cell-sheet construct has spurred wide interest in regenerative medicine, especially for reconstructive surgery procedures. The ...

adsc-sheet-transplantation-to-prevent-stricture-after-extended

Nanyang Technological University

Research

JoVE Journal

Medicine

8.4K 次观看.  Université Paris Descartes Sorbonne Paris Cité. 这项研究报告内窥镜脂肪组织的基质细胞（ADSC）的成功的方法折叠移植食管狭窄预防猪模型的扩展内镜黏膜下剥离术（ESD）之后。 

视频: ADSC-sheet Transplantation to Prevent Stricture after Extended Esophageal Endoscopic Submucosal Dissection

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

The number of patients with end-stage renal disease, and the number of kidney allograft recipients continuously increases. Episodes of acute cellular allograft rejection (AR) are a negative prognostic factor for long-term allograft survival, and its timely diagnosis is crucial for allograft function 1. At present, AR can only be definitely diagnosed by core-needle biopsy, which, as an invasive method, bares significant risk of graft injury or even loss. Moreover, biopsies are not feasible in patients taking anticoagulant drugs and the limited sampling site of this technique may result in false negative results if the AR is focal or patchy. As a consequence, this gave rise to an ongoing search for new AR detection methods, which often has to be done in animals including the use of various transplantation models.
Since the early 60s rat renal transplantation is a well-established experimental method for the examination and analysis of AR 2. We herein present in addition small animal positron emission tomography (PET) using 18F-fluorodeoxyglucose (FDG) to assess AR in an allogeneic uninephrectomized rat renal transplantation model and propose graft FDG-PET imaging as a new option for a non-invasive, specific and early diagnosis of AR also for the human situation 3. Further, this method can be applied for follow-up to improve monitoring of transplant rejection 4.

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

We herein present a rat renal transplantation model to non-invasively assess acute allograft rejection using positron emission tomography with 18F-fluorodeoxyglucose.

大鼠肾移植后急性排斥反应的非侵入性的成像使用<sup&gt; 18</sup&gt; F-FDG PET

The number of patients with end-stage renal  disease, and the number of kidney allograft recipients continuously increases.  Episodes of acute cellular ...

科研

医学

Intrasplenic Transplantation of Hepatocytes After Partial Hepatectomy in NOD.SCID Mice

Partial hepatectomy is a versatile and reproducible method to study liver regeneration and the effect of cell based therapeutics in various pathological conditions. Partial hepatectomy also facilitates the increased engraftment and proliferation of transplanted cells by accelerating neovascularization and cell migration towards the liver. Here, we describe a simple protocol for performing 30% hepatectomy and transplantation of cells in the spleen of a non-obese diabetic/severe combined immunodeficient NOD.SCID (NOD.CB17-Prkdcscid/J) mouse. 
In this procedure, two small incisions are made. The first incision is to expose and resect the left lobe of the liver, and another small incision is made to expose the spleen for the intrasplenic transplantation of cells. This procedure does not require any specialized surgical skills, and it can be completed in 5-7 minutes with less stress and pain, faster recovery, and better survival. We have demonstrated the transplantation of hepatocytes isolated from a green fluorescent protein (GFP) expressing mouse (Transgenic C57BL/6-Tg (UBC-GFP) 30Scha/J), as well as hepatocyte like cells of human origin (NeoHep) in partially hepatectomized NOD.SCID mice.

We have described a protocol for performing partial hepatectomy (PHx) and cell transplantation via spleen in NOD.SCID (NOD.CB17-Prkdcscid/J) mice. In this protocol, an incision is made to expose and resect the left lobe of the liver followed by another incision for the intrasplenic transplantation of cells.

肝细胞部分肝切除术后脾移植。小鼠

Partial hepatectomy is a versatile and reproducible method to study liver regeneration and the effect of cell based therapeutics in various pathological ...

Discovering Middle Ear Anatomy by Transcanal Endoscopic Ear Surgery: A Dissection Manual

The middle ear is located in the center of the temporal bone and bears a highly complex anatomy. The recently introduced exclusively endoscopic transcanal approach to the middle ear is a minimally invasive technique sparing the bone and mucosa of the mastoid bone, since the middle ear is accessed through the external auditory canal. This emerging method has several advantages over the traditional (microscopic) approaches to the middle ear such as the panoramic wide-angle views of the anatomy, the possibility to approach and magnify tiny structures, and the possibility of looking around the corner using angled endoscopes.
The cadaveric dissection method presented here consists of an overview on the technical requirements and a precise description of a step-by-step protocol to discover the anatomy of the middle ear. Each step and anatomical structure is carefully described in order to provide a comprehensive guide to endoscopic ear anatomy. In our opinion, this is particularly important to any novice in endoscopic ear surgery as it provides thorough anatomical knowledge and may improve surgical skills.

The aim of this article is to describe the methodology of exclusively endoscopic cadaveric middle ear dissection. Moreover, we aim to provide a comprehensive guide to endoscopic middle ear anatomy.

Transcanal 内窥镜下耳部手术发现中耳解剖: 解剖手册

The middle ear is located in the center of the temporal bone and bears a highly complex anatomy. The recently introduced exclusively endoscopic transcanal ...

Transplantation of Adipose Tissue-Derived Stem Cell Sheet to Reduce Leakage After Partial Colectomy in A Rat Model

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of clinical efficacy, they are until now imperfect solutions. Stem cell therapy using ASC sheets could provide a solution to this problem. ASCs are considered as promising candidates for promoting tissue healing because of their trophic and immunomodulatory properties. Here, we provide methods to produce high-density ASC sheets, that are transplanted onto a colorectal anastomosis in a rat model to reduce the leakage. ASCs formed cell sheets in thermo-responsive culture dishes that could be easily detached. On the day of the transplantation, a partial colectomy with a 5-suture colorectal anastomosis was performed. Animals were immediately transplanted with 1 ASC sheet per rat. ASC sheets adhered spontaneously to the anastomosis without any glue, suture, or any biomaterial. Animal groups were sacrificed 3 and 7 days postoperatively. Compared to transplanted animals, the incidence of anastomotic abscesses and leakage was higher in control animals. In our model, the transplantation of ASC sheets after colorectal anastomosis was successful and associated with a lower leakage rate.

The preparation and transplantation of adipose tissue derived stem cell (ASC) sheets onto insufficiently sutured colorectal anastomoses in a rat model is presented. This novel application shows that ASC sheets can reduce colorectal anastomosis leakage.

脂肪组织干细胞移植对大鼠部分结肠切除术后渗漏的减少

Anastomotic leakage is a disastrous complication after colorectal surgery. Although current methods for leakage prevention have different levels of ...

Reduced Complications after Arterial Reconnection in a Rat Model of Orthotopic Liver Transplantation

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation of human liver transplantation due to the absence of arterial reconnection. Described here is a modified transplantation procedure that includes the incorporation of hepatic artery (HA) reconnection, leading to a marked improvement in transplant outcomes. With a mean anhepatic time of 12 min and 14 s, HA reconnection results in improved perfusion of the transplanted liver and an increase in long-term recipient survival from 37.5% to 88.2%. This protocol includes the use of 3D-printed cuffs and holders to connect the portal vein and infrahepatic inferior vena cava. It can be implemented for studying multiple aspects of liver transplantation, from immune response and infection to technical aspects of the procedure. By incorporating a simple and practical method for arterial reconnection using a microvascular technique, this modified rat OLT protocol closely mimics aspects of human liver transplantation and will serve as a valuable and clinically relevant research model.

The goal of this study is to modify the rat orthotopic liver transplant model to better represent human liver transplantation and improve recipient survival. The presented method reestablishes hepatic arterial inflow by connecting the donor liver&#39;s common hepatic artery to the recipient liver&#39;s proper hepatic artery.

矫形肝移植大鼠模型中动脉重联后并发症减少

The rat orthotopic liver transplantation (OLT) model is a powerful tool to study acute and chronic rejection. However, it is not a complete representation ...

Simultaneous Laryngopharyngeal and Conventional Esophageal pH Monitoring

In addition to typical reflux symptoms, many patients with gastroesophageal reflux disease (GERD) present with extraesophageal symptoms such as cough, hoarseness or asthma, which can be caused by laryngopharyngeal reflux (LPR). Due to their multifactorial origin, those symptoms can be a great diagnostic and therapeutic challenge. Esophageal pH-monitoring is commonly used to determine abnormal esophageal acid exposure and confirm the diagnosis of GERD. However, for better evaluation of acid exposure above the upper esophageal sphincter, a new laryngopharyngeal pH measurement system is now available and may lead to more reliable results in patients with predominantly extraesophageal symptoms. This article aims to present a standardized protocol for simultaneous pH measurement using esophageal and laryngopharyngeal pH probes in order to obtain acid exposure scores from both measurements.

Laryngopharyngeal pH monitoring has been specifically designed to measure acid exposure above the upper esophageal sphincter and complements diagnostic evaluation in patients that present with mainly extraesophageal reflux symptoms. Patients with suspected laryngopharyngeal reflux (LPR) were evaluated using distal esophageal and laryngopharyngeal pH testing simultaneously.

同时监测喉咽和常规食管 pH 值

In addition to typical reflux symptoms, many patients with gastroesophageal reflux disease (GERD) present with extraesophageal symptoms such as cough, ...

Intramyocardial Transplantation of MSC-Loading Injectable Hydrogels after Myocardial Infarction in a Murine Model

One of the major issues facing current cardiac stem cell therapies for preventing postinfarct heart failure is the low retention and survival rates of transplanted cells within the injured myocardium, limiting their therapeutic efficacy. Recently, the use of scaffolding biomaterials has gained attention for improving and maximizing stem cell therapy. The objective of this protocol is to introduce a simple and straightforward technique to transplant bone marrow-derived mesenchymal stem cells (MSCs) using injectable hydroxyphenyl propionic acid (GH) hydrogels; the hydrogels are favorable as a cell delivery platform for cardiac tissue engineering applications due to their ability to be cross-linked in situ and high biocompatibility. We present a simple method to fabricate MSC-loading GH hydrogels (MSC/hydrogels) and evaluate their survival and proliferation in three-dimensional (3D) in vitro culture. In addition, we demonstrate a technique for intramyocardial transplantation of MSC/hydrogels in mice, describing a surgical procedure to induce myocardial infarction (MI) via left anterior descending (LAD) coronary artery ligation and subsequent MSC/hydrogels transplantation.

Stem cell-based therapy has emerged as an efficient strategy to repair injured cardiac tissues after myocardial infarction. We provide an optimal in vivo application for stem cell transplantation using gelatin hydrogels that are able to be enzymatically cross-linked.

Murine 模型心肌梗塞后 MSC 加载注射水凝胶的心内移植

One of the major issues facing current cardiac stem cell therapies for preventing postinfarct heart failure is the low retention and survival rates of ...

Digital Home-Monitoring of Patients after Kidney Transplantation: The MACCS Platform

The MACCS (Medical Assistant for Chronic Care Service) platform enables secure sharing of key medical information between patients after kidney transplantation and physicians. Patients provide information such as vital signs, well-being, and medication intake via smartphone apps. The information is transferred directly into a database and electronic health record at the kidney transplant center, which is used for routine patient care and research. Physicians can send an updated medication plan and laboratory data directly to the patient app via this secure platform. Other features of the app are medical messages and video consultations. Consequently, the patient is better-informed, and self-management is facilitated. In addition, the transplant center and the patient's local nephrologist automatically exchange notes, medical reports, laboratory values, and medication data via the platform. A telemedicine team reviews all incoming data on a dashboard and takes action, if necessary. Tools to identify patients at risk for complications are under development. The platform exchanges data via a standardized secure interface (Health Level 7 (HL7), Fast Healthcare Interoperability Resources (FHIR)). The standardized data exchange based on HL7 FHIR guarantees interoperability with other eHealth solutions and allows rapid scalability to other chronic diseases. The underlying data protection concept is in concordance with the latest European General Data Protection Regulation. Enrollment started in February 2020, and 131 kidney transplant recipients are actively participating as of July 2020. Two large German health insurance companies are currently funding the telemedicine services of the project. The deployment for other chronic kidney diseases and solid organ transplant recipients is planned. In conclusion, the platform is designed to enable home monitoring and automatic data exchange, empower patients, reduce hospitalizations, and improve adherence, and outcomes after kidney transplantation.

The MACCS platform&#160;is a comprehensive telemedicine concept aiming at better outcomes after kidney transplantation by sharing key medical information between patients and physicians. A telemedicine team reviews incoming data to detect potential complications and to improve adherence in kidney transplant recipients to achieve better long-term outcomes.

数字家庭-肾移植后患者的监测:MACCS平台

The MACCS (Medical Assistant for Chronic Care Service) platform enables secure sharing of key medical information between patients after kidney ...

Endoscopic Cholesteatoma Surgery

Implementation of endoscopes in cholesteatoma surgery resulted in considerable changes in the management of cholesteatoma in the last two decades. Compared to the microscopic approach with an excellent but straight-line view and limited illumination, the introduction of endoscopes provides a wide-angled panoramic view. Moreover, angled lenses allow the surgeon to visualize the middle ear and its hidden recesses through a transcanal, minimally-invasive approach. The endoscope enables the surgeon to remove limited cholesteatoma of the middle ear and its recesses using an exclusive endoscopic technique by taking advantage of these benefits. This reduces the rate of residual disease and sparing external incisions and excessive temporal bone drilling as in a transmastoid approach. Since transcanal endoscopic access is mainly a one-handed technique, it implies the need for specific procedures and technical refinements. This article describes a step-by-step guide as a surgical manual for endoscopic removal of epitympanic cholesteatoma. Different techniques for cholesteatoma dissection and bone removal for epitympanectomy, including curettage and powered instruments such as drills and ultrasonic devices with their outcomes, are discussed. This may offer ear surgeons insight into technical refinements and the latest technological developments and open the horizon for different techniques.

The present protocol describes a step-by-step guide for the complete endoscopic removal of epitympanic cholesteatoma with different techniques for cholesteatoma dissection and bone removal for epitympanectomy.

内窥镜胆脂瘤手术

Implementation of endoscopes in cholesteatoma surgery resulted in considerable changes in the management of cholesteatoma in the last two decades. ...

Porcine Liver Transplantation Without Veno-Venous Bypass As an Extended Criteria Donor Model

Liver transplantation is regarded as the gold standard for the treatment of a variety of fatal hepatic diseases. However, unsolved issues of chronic graft failure, ongoing organ donor shortages, and the increased use of marginal grafts call for the improvement of current concepts, such as the implementation of organ machine perfusion. In order to evaluate new methods of graft reconditioning and modulation, translational models are required. With respect to anatomical and physiological similarities to humans and recent progress in the field of xenotransplantation, pigs have become the main large animal species used in transplantation models. After the initial introduction of a porcine orthotopic liver transplant model by Garnier et al. in 1965, several modifications have been published over the past 60 years.
Due to specifies-specific anatomical traits, a veno-venous bypass during the anhepatic phase is regarded as a necessity to reduce intestinal congestion and ischemia resulting in hemodynamic instability and perioperative mortality. However, the implementation of a bypass increases the technical and logistical complexity of the procedure. Furthermore, associated complications such as air embolism, hemorrhage, and the need for a simultaneous splenectomy have been reported previously.
In this protocol, we describe a model of porcine orthotopic liver transplantation without the use of a veno-venous bypass. The engraftment of donor livers after static cold storage of 20 h - simulating extended criteria donor conditions - demonstrates that this simplified approach can be performed without significant hemodynamic alterations or intraoperative mortality and with regular uptake of liver function (as defined by bile production and liver-specific CYP1A2 metabolism). The success of this approach is ensured by an optimized surgical technique and a sophisticated anesthesiologic volume and vasopressor management.
This model should be of special interest for workgroups focusing on the immediate postoperative course, ischemia-reperfusion injury, associated immunological mechanisms, and the reconditioning of extended criteria donor organs.

In this protocol, a model of porcine orthotopic liver transplantation after static cold storage of donor organs for 20 h without the use of a veno-venous bypass during engraftment is described. The approach uses a simplified surgical technique with minimization of the anhepatic phase and sophisticated volume and vasopressor management.

无静脉-静脉旁路术的猪肝移植作为扩展标准供体模型

Liver transplantation is regarded as the gold standard for the treatment of a variety of fatal hepatic diseases. However, unsolved issues of chronic graft ...