Name: stromal vascular fraction-enriched fat grafting for the treatment of symptomatic end-neuromata
Uploaded: 2017-11-23T13:00:00.000+00:00
Duration: 7 min 58 s
Description: Watch this Scientific Journal Video about Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata at JoVE.com

作者没有致谢。

该议定书遵循了大学医院苏黎世人类研究伦理委员会的指导方针。
1. Neuromata 和患者选择的诊断
<ol><li>执行术前神经阻滞的近端, 以确定诊断。评估前臂中间三分之一的块, 排除 SBRN 交叉支配区的潜在诱因, 如侧前臂皮神经 (域)、骨间后神经 (针) 和尺神经背支,10。</li>
	<li>仅在减轻疼痛的患者中进行手术 (在疼痛的视觉模拟量表 (VAS) 上至少有5点)11。</li>
</ol>2. 手术准备
<ol><li>在手术前禁止病人进食6小时的食物或液体。</li>
	<li>将病人放在手术室的中央, 在一个沙滩椅位置的通用手术台上。</li>
	<li>将受影响的手臂/手放在手臂上休息。</li>
	<li>要求麻醉进行气管内麻醉 (芬太尼2µg/千克, 罗库0.6 毫克/千克, 异丙酚 150-250 毫克)。</li>
	<li>用聚维酮碘在吸脂术 (如内大腿和/或上/下腹) 和神经瘤切除部位对皮肤进行消毒。</li>
	<li>用不育的手术窗帘盖住病人, 留下暴露在抽脂部位和手术部位。</li>
</ol>3. 肿胀吸脂
<ol><li>在抽脂领域用 #11 刀片手术刀做几处直径约 3-5 毫米的小刺 (穿刺) 切口。切割到皮下层的距离。</li>
	<li>注射含有稀释利多卡因和肾上腺素 (500 毫升林格氏乳酸溶液, 20 毫升1% 利多卡因, 1 毫克肾上腺素) 的肿胀溶液, 通过14口径者 (浸润套管) 进入皮下脂肪层 (3 毫米深度)脂肪收获区。</li>
	<li>等待至少20分钟, 使肿胀的解决方案生效。</li>
	<li>钩3毫米收集套管 (外径3毫米, 长度26厘米), 以60毫升图米注射器和开始收割脂肪通过刺切口。 注: nondominant 手持续控制套管的放置和过程。</li>
	<li>拉回图米注射器的柱塞到60毫升的标记, 并放置一个蚊子钳沿撤回柱塞, 以保持负压在注射器和避免持续真空创造的主导手。</li>
	<li>在所提供的图米注射器支架中, 将图米注射器放置在垂直位置, 以获得至少220毫升的脂肪组织;这使得脂肪沉淀和分离的液体。</li>
	<li>在没有大量血液的情况下, 要特别注意获得高质量的脂肪移植。 注意: 当增强血液含量被注意时, 改变抽脂的区域。</li>
	<li>用简单的缝线缝合切口, 并按要求施加压力 (如压缩带)。</li>
	<li>将 lipoaspirate 转化为一次性无菌隔离系统。</li>
	<li>保持一个图米注射器与 lipoaspirate 在提供的立场为了得到脂肪分数为以后 lipofilling。</li>
</ol>4. 脂肪组织加工
<ol><li>关闭无菌耗材集。 注意: 隔离系统 (参见材料表) 通过执行湿测试来确认完整性。</li>
	<li>按照显示屏上的说明进行操作。 注意: 该过程主要是由软件控制的, 只需在控制面板上按下 "下一步" 或 "后退" 按钮, 就可对列出的步骤进行手动确认。lipoaspirate 被转移到组织收集罐。在组织收集罐中, 用林格氏乳酸溶液对 lipoaspirate 进行称量和自动清洗, 以去除残余的血细胞和润湿液。</li>
	<li>如果需要, 增加更多的脂肪。 注: 根据所收获的脂肪组织的重量, 隔离系统要求用户添加更多的脂肪或计算额外的酶试剂添加量。</li>
	<li>在隔离系统提示时添加所需的酶试剂量。 注意: 在酶消化过程中, 油脂和特别的分离得到了实现。因此, 组织处于摆动运动中。酶消化完成后, 特别自动转移到离心室中进行细胞浓度测定。在离心机室中, 特别被集中到一个细胞颗粒 (离心机速度大约 3000 x g)。最后, 细胞颗粒经历了一系列的冲洗与林格的乳酸溶液, 以减少残留酶水平的输出。当工艺完成时, 获得5毫升的清水 (特别)。</li>
</ol>5. 手术程序
<ol><li>要接近的神经瘤, 做一个皮肤切口在以前被诊断的神经瘤站点在前臂的远端三分之一。</li>
	<li>在外科手术双目放大镜放大镜 (2.5-3.5X) 的使用下, 继续解剖皮下组织, 并确定累及神经近端的听神经瘤。<ol>
<li>随后, 跟踪神经远端, 并确定了听神经瘤 (基于视觉检查)。</li>
	</ol>
</li>
	<li>一旦完全延长的神经瘤是明确的, 开始通过一个直接的反式神经元剪刀切除。如果没有发现任何神经瘤, 只会进行松。</li>
	<li>继续与其余的神经残肢的松, 直到3厘米近的手腕。 注意: 神经残端应位于主切口正中。</li>
	<li>将两到四小穿刺 (刺伤) 切口 (直径1毫米) 与手术刀一起通过表皮和真皮皮肤在主要方法周围。</li>
	<li>将二到四钝套管 (16 口径, 长度9厘米) 通过刺切口神经到树桩。 注意: 套管的尖端应该指向神经残端。</li>
	<li>用胶粘敷料固定套管的位置。</li>
	<li>将缝线周围的套管, 并留在原位缝合材料没有固定的节点。</li>
	<li>缝合主切口, 以防止移植的流出。 注意: 确保套管保持原位。</li>
</ol>6. 特别脂肪移植的应用
注: 经过大约1.5 小时的加工, 5 毫升的加工特别已准备好进一步使用。
<ol><li>在10毫升的交流注射器中抽取5毫升的特别。</li>
	<li>以同样的方式, 在对立的10毫升注射器中抽取2毫升的沉淀脂质分数。</li>
	<li>混合5毫升的特别与2毫升的沉淀脂质分数通过交替拉回一个注射器的柱塞和拉下来的沟通, 反对注射器的柱塞, 直到充分的混合得到。</li>
	<li>在两到四10毫升的注射器中, 将特别的脂肪移植物均分。</li>
	<li>通过将注射器连接到钝套管, 在神经残端周围均匀地分布准备好的特别-富脂接枝。 注: 在应用移植前, 确保套管在正确的位置, 并注意不要脱臼套管, 而应用脂肪移植。</li>
	<li>小心地取下套管。</li>
	<li>然后, 用柜绷带将小的穿刺切口带上。</li>
	<li>固定的手通过应用夹板在内在的加号位置 (掌 (MCP) 关节屈曲在 60-70, 间 (IP) 关节充分延长, 拇指在拳头投射, 手腕在背屈°)。如果只有松的神经被执行, 不需要夹板, 手臂和手腕包扎10天。</li>
</ol>7. 术后病人的处理
<ol><li>将病人转移至 post-anesthesia 恢复后, 并在病房内进行足够的监测。</li>
	<li>手术后10天继续固定手腕。</li>
	<li>在48小时后后进行绷带的第一次更换, 并按固定间隔继续 (即每隔一天直到出院)。</li>
	<li>切除缝合术后 12-14 天。</li>
	<li>在随访2、6、12和36月时对疼痛进行临床评估。</li>
</ol>

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作者没有什么可透露的。

本研究的目的是有条不紊地说明的技术方面, 特别脂肪移植作为一种新的治疗症状性端 neuromata。
Neuromata 的 SBRN 与外科手术成功率只有 33%12。瓦伦et al.提出了一系列有利的结果, 早期随访使用神经脂肪移植 (框架) 没有添加特别8患者的痛苦 neuromata 的上肢5。然而, 疼痛减少在6月后疼痛复发在12月随访5。
虽然疼痛减少没有证明是统计学意义上的特别丰富脂肪移植的 SBRN, 一个明确的趋势, 以恒定的疼痛减少被注意到所有疼痛方式10。此外, 疼痛减少持续2月至36月后无疼痛复发。因此, 2 月后的临床评估似乎是未来远期预后的可靠预测指标。然而, 一个更大的随机序列是必要的检测潜在的统计意义。这一有利的结果可能是在更大的剩余体积脂肪移植的背景下看到的。以往的研究表明, ADSC 脂肪吸收率降低时, 应用与 non-enriched 嫁接13比较。此外, 特别脂肪移植的再生潜能由细胞水平上的许多机制推动, 主要是改善血管生成、炎症和纤维化。ADSC 作为特别的一个组成部分, 通过表达生长因子如 VEGF 和 bFGF 在细胞水平上的14来触发血管生成。免疫系统受造血细胞的影响, 如巨噬细胞的 M2 表型或 T 辅助单元和自然杀伤细胞15。细胞信号可以受到减少的炎细胞因子的影响, 如 TNF α和 IL-616。它对神经组织本身也有有益的作用, 它可以减少无序的神经生长和随后痛苦的皮肤疤痕。白et al.建议每200µL 脂肪组织的理想浓度为1万细胞, 以获得最佳的移植成活和作用机制。高剂量的特别与显著增加的脂肪吸收率17相关。
多项手术技术在治疗痛苦的 neuromata SBRN 已广泛发表。肱肌的神经瘤切除术和肌内移位术仍然是流行和广泛的表现。特别脂肪移植与肌内移位相比, 不需要肱肌的手术准备, 并与较短的手腕固定。此外, 该技术不限于所呈现的神经群体, 并可以适用于任何在上肢或下肢的任何瘤。
尽管手术前进行了有效的神经阻滞来确认诊断并排除了重叠神经的疼痛, 但不能绝对排除相互干扰。SBRN 和域的解剖重叠可能导致术后神经支配区域的无条理的生长。此外, 目前的研究是有限的, 由于少数病人。虽然这项研究是迄今为止最大的调查长期结果和说明特别脂肪的技术方面的一个单一的神经人口, 进一步的随机试验与更多的参与者是需要确认的有利这种新技术的发展趋势。

对有症状的末端 neuromata 的管理 SBRN 仍然是挑战与许多现有的外科治疗。主要地, 神经瘤切除并且剩余的神经末梢被安置入邻居结构例如肌肉1,2, 骨头3, 或静脉4。这被假设通过模仿末端器官的气氛防止再生神经束的 out-sprouting。然而, 这些技术都没有被证明是优越的, 因为高的疼痛复发率和不足的疼痛减少往往被观察到。
最近, 脂肪移植周围剩余的神经残端产生了可喜的结果, 并主张作为一个新的治疗方案的痛苦结束 neuromata 的上肢5。除了作为机械屏障, 以防止继续运动和轻拍疼痛, 脂肪移植的有益方面是相当归因于细胞水平。人体脂肪组织包含多种细胞类型, 包括脂肪、肥大细胞、周、内皮细胞、血管平滑肌细胞、成纤维细胞和脂肪来源的干细胞 (干细胞) 以及造血干细胞6。这种异质细胞群被称为脂肪衍生再生细胞 (ADRCs) 或特别, 并可从 lipoaspirate 获得一个隔离系统 (见材料表) 和应用酶试剂 (见材料表)7。确切的行动机制还没有完全理解;然而, 特别已与改善血管生成, 免疫, 分化和分泌细胞外基质8。
特别的浓缩细胞数量增加到脂肪移植, 以减少吸收率和增加再生潜力的脂肪移植, 最近的研究显示疼痛复发后12月的脂肪移植没有添加特别5. 此外, 生物作用还通过减少残端的轴突发芽和随后的瘢痕坚持对神经本身产生有益的影响, 这可能导致神经卡压.
第一次临床研究描述特别脂肪移植作为一种新的手术选择治疗症状性端 neuromata SBRN 最近发表了10。一般来说, 该技术可以应用于任何发生在意外或医源性神经损伤后的听神经瘤。本次后续研究的目的是有条不紊地说明这一程序的技术方面。

<table><tbody><tr><td>Cytori Celution System</td><td>Cytori</td><td></td><td>isolation system</td></tr><tr><td>Celase</td><td>Cytori</td><td></td><td>enyzme reagent</td></tr><tr><td>liposuction system</td><td></td><td></td><td></td></tr><tr><td>tumescent solution (500 ml Lactated Ringers, 20 ml of 1% lidocaine and 1 mg epinephrine)</td><td></td><td></td><td></td></tr><tr><td>14-gauge infiltrator</td><td></td><td></td><td></td></tr><tr><td>3 mm-collecting cannula (15, 26, 32 cm)</td><td></td><td></td><td></td></tr><tr><td>60 mL Tommey syringes</td><td></td><td></td><td></td></tr><tr><td>mosquito clamp</td><td></td><td></td><td></td></tr><tr><td>Toomey syringe stand</td><td></td><td></td><td></td></tr><tr><td>4 cannulas (fat injecting cannulas 1.5 mm, syringe NO.2, 90 mm)</td><td>Chalybs</td><td></td><td></td></tr><tr><td>10 mL syringes</td><td></td><td></td><td></td></tr><tr><td>connecting device for syringes</td><td></td><td></td><td></td></tr><tr><td>#11- and #15-blade scalpel</td><td></td><td></td><td></td></tr><tr><td>stevens scissors</td><td></td><td></td><td></td></tr><tr><td>pins pincette</td><td></td><td></td><td></td></tr><tr><td>bipolar electrocautery</td><td></td><td></td><td></td></tr><tr><td>suture material</td><td></td><td></td><td></td></tr><tr><td>adhesive dressing</td><td></td><td></td><td></td></tr><tr><td>binocular loupes magnifier glasses (2.5-3.5x)</td><td>Carl Zeiss</td><td></td><td></td></tr><tr><td>Statistical Package for Social Sciences, version 22</td><td>IBM</td><td></td><td></td></tr></tbody></table>

stromal vascular fraction-enriched fat grafting for the treatment of symptomatic end-neuromata

本研究的目的是有条不紊地说明和突出的关键步骤, 基质血管分数 (特别)-丰富脂肪移植作为一种新的治疗症状端 neuromata 外周感觉神经, 并在本研究, 特别是桡神经 (SBRN) 的浅支。尽管有许多现有的治疗方法, 持续的术后疼痛和常见的疼痛复发仍然是非常普遍的, 独立的程序评估。神经瘤被 microsurgically 切除相应地规范化的协议。而不是在邻近的解剖结构, 如肌肉或骨骼的再生神经残肢的迁移, 脂肪移植应用 perifocally 和作为机械屏障。为了降低脂肪的吸收率, 提高移植物的再生潜能, 将高浓度的特别集成在嫁接中。特别是从皮下脂肪分离的酶和机械 lipoaspirate 由一个特定的商业隔离系统。富含特别的脂肪移植在细胞水平上提供了机械屏障和各种生物效应, 包括改善血管生成、炎症和纤维化。在神经再生过程中, 机械和生物效应都有助于减少神经残端的无条理的轴突突起, 从而防止了痛苦的端 neuromata 的复发。

利用统计软件对数据进行分析, 并以标准偏差表示。应用科尔莫戈罗夫斯米尔诺夫试验验证了研究人群的非参数分布, 证实了负经验分布。根据研究人群的非参数性质, 采用魏氏对相关样本进行测试, 比较术前和术后结局。统计学意义被认为在 p 值小于 0.0510。
五例平均年龄为49.8 ±16.6 岁的患者包括在该研究中。在手术前和术后疼痛评分的比较是在表 1中的每一个疼痛模式和整体疼痛。特别脂肪移植的治疗不能明显减轻疼痛。然而, 所有疼痛模式的相关减少可从术后2月开始, 并显示为持续36月随访后无疼痛复发 (图 1)。
自发性疼痛可从术后1.6 ±0.55 降至手术后1.2 ± 1.1 (p = 0.414), 峰值从2.2 ±1.3 到1.4 ± 1.34 (p = 0.180), 感觉从1.6 ±1.14 到1.2 ± 1.64 (p = 0.713), 从2.8 ±0.45 到1.8 ± 1.3 (p = 0.197) 和运动 pa 的轻拍痛苦在从2.8 ±0.45 到1.4 ± 1.34 (p = 0.066)。从术前2.2 ±0.97 到术后1.4 ±1.26 的疼痛总体减轻, 手术后36月 (p = 0.104)。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/55962/55962fig1.jpg"> 图 1: 疼痛模式的趋势.术前和2、6、12、36月均进行疼痛模式评估 (平均± SD)。此图已从以前的出版物中修改过10.<a href="http://cloudflare.jove.com/files/ftp_upload/55962/55962fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td colspan="6">表1。平均疼痛评分±标准偏差 (p 值) 特别富脂接枝前后</td>
			<td></td>
		</tr>
<tr>
<td></td>
			<td>自发性疼痛</td>
			<td>穗</td>
			<td>感觉</td>
			<td>轻拍疼痛</td>
			<td>运动疼痛</td>
			<td>整体疼痛评分</td>
		</tr>
<tr>
<td>术前评分</td>
			<td>1.6 ±0.55</td>
			<td>2.2 ±1。3</td>
			<td>1.6 ±1.14</td>
			<td>2.8 ±0.45</td>
			<td>2.8 ±0.45</td>
			<td>2.2 ±0.97</td>
		</tr>
<tr>
<td>2月</td>
			<td>0.8 ± 0.84 (0.102)</td>
			<td>1.4 ± 1.52 (0.285)</td>
			<td>1.2 ± 1.64 (0.713)</td>
			<td>1.6 ± 0.89 (0.083)</td>
			<td>1.4 ± 1.52 (0.102)</td>
			<td>1.28 ± 1.24 (0.225)</td>
		</tr>
<tr>
<td>6月</td>
			<td>1.6 ± 1.52 (1.0)</td>
			<td>1.8 ± 1.64 (0.655)</td>
			<td>1.4 ± 1.52 (0.713)</td>
			<td>1.6 ± 1.52 (0.157)</td>
			<td>1.2 ± 1.64 (0.102)</td>
			<td>1.52 ± 1.45 (0.345)</td>
		</tr>
<tr>
<td>12月</td>
			<td>1.6 ± 1.52 (1.0)</td>
			<td>1.8 ± 1.64 (0.655)</td>
			<td>1.4 ± 1.52 (0.713)</td>
			<td>1.8 ± 1.3 (0.180)</td>
			<td>1.6 ± 1.52 (0.109)</td>
			<td>1.64 ± 1.38 (0.345)</td>
		</tr>
<tr>
<td>36月</td>
			<td>1.2 ± 1.1 (0.414)</td>
			<td>1.4 ± 1.34 (0.180)</td>
			<td>1.2 ± 1.64 (0.713)</td>
			<td>1.8 ± 1.3 (0.197)</td>
			<td>1.4 ± 1.34 (0.066)</td>
			<td>1.4 ± 1.26 (0.104)</td>
		</tr>
<tr>
<td colspan="4">p 值显示术前和手术前后评分的差异</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
</tbody></table>表 1: 影响神经瘤皮肤的多种疼痛方式.不同的疼痛模式影响的神经瘤的皮肤, 我们的病人评价为每一个疼痛的方式 [无 (0), 轻度 (1), 中度 (2), 和严重疼痛 (3), 0 至 3)。临床检查是在手术前和随访时进行的2、6、12和36月后。术后疼痛评分的评估, 在最初的位置疼痛, 以及在现场的特别丰富脂肪移植。已从以前的出版物10中修改了此项。

Watch this Scientific Journal Video about Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata at JoVE.com

Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata

基质血管部分富集脂肪移植治疗症状性端 neuromata

本研究的目的是为了说明基质血管部分 (特别)-富集脂肪移植的技术过程, 作为一种新的治疗症状性端 neuromata。这种技术提供了机械屏障和生物作用的优势, 在细胞水平的加工和集中特别。

The purpose of this study was to methodically illustrate and highlight the crucial steps of stromal vascular fraction (SVF)-enriched fat grafting as a ...

stromal-vascular-fraction-enriched-fat-grafting-for-treatment

Nanyang Technological University

Research

JoVE Journal

Medicine

7.6K 次观看.  University Hospital Zurich. 本研究的目的是为了说明基质血管部分 (特别)-富集脂肪移植的技术过程, 作为一种新的治疗症状性端 neuromata。这种技术提供了机械屏障和生物作用的优势, 在细胞水平的加工和集中特别。

视频: Stromal Vascular Fraction-enriched Fat Grafting for the Treatment of Symptomatic End-neuromata

A Novel Mammary Fat Pad Transplantation Technique to Visualize the Vessel Generation of Vascular Endothelial Stem Cells

Endothelial cells (ECs) are the fundamental building blocks of the vascular architecture and mediate vascular growth and remodeling to ensure proper vessel development and homeostasis. However, studies on endothelial lineage hierarchy remain elusive due to the lack of tools to gain access as well as to directly evaluate their behavior in vivo. To address this shortcoming, a new tissue model to study angiogenesis using the mammary fat pad has been developed. The mammary gland develops mostly in the postnatal stages, including puberty and pregnancy, during which robust epithelium proliferation is accompanied by extensive vascular remodeling. Mammary fat pads provide space, matrix, and rich angiogenic stimuli from the growing mammary epithelium. Furthermore, mammary fat pads are located outside the peritoneal cavity, making them an easily accessible grafting site for assessing the angiogenic potential of exogenous cells. This work also describes an efficient tracing approach using fluorescent reporter mice to specifically label the targeted population of vascular endothelial stem cells (VESCs) in vivo. This lineage tracing method, coupled with subsequent tissue whole-mount microscopy, enable the direct visualization of targeted cells and their descendants, through which the proliferation capability can be quantified and the differentiation commitment can be fate-mapped. Using these methods, a population of bipotent protein C receptor (Procr) expressing VESCs has recently been identified in multiple vascular systems. Procr+ VESCs, giving rise to both new ECs and pericytes, actively contribute to angiogenesis during development, homeostasis, and injury repair. Overall, this manuscript describes a new mammary fat pad transplantation and in vivo lineage tracing techniques that can be used to evaluate the stem cell properties of VESCs.

This work demonstrates a novel approach to assess the proliferation, differentiation, and vessel-forming potential of vascular endothelial stem cells (VESCs) through mammary fat pad transplantation followed by whole-mount tissue preparation for microscopic observation. A lineage tracing strategy to investigate the behavior of VESCs in vivo is also presented.

新型乳腺脂肪移植技术可视化血管内皮干细胞的血管生成

Endothelial cells (ECs) are the fundamental building blocks of the vascular architecture and mediate vascular growth and remodeling to ensure proper ...

科研

发育生物学

A Modified Heterotopic Swine Hind Limb Transplant Model for Translational Vascularized Composite Allotransplantation (VCA) Research

Vascularized Composite Allotransplantation (VCA) such as hand and face transplants represent a viable treatment option for complex musculoskeletal trauma and devastating tissue loss. Despite favorable and highly encouraging early and intermediate functional outcomes, rejection of the highly immunogenic skin component of a VCA and potential adverse effects of chronic multi-drug immunosuppression continue to hamper widespread clinical application of VCA. Therefore, research in this novel field needs to focus on translational studies related to unique immunologic features of VCA and to develop novel immunomodulatory strategies for immunomodulation and tolerance induction following VCA without the need for long term immunosuppression.
This article describes a reliable and reproducible translational large animal model of VCA that is comprised of an osteomyocutaneous flap in a MHC-defined swine heterotopic hind limb allotransplantation. Briefly, a well-vascularized skin paddle is identified in the anteromedial thigh region using near infrared laser angiography. The underlying muscles, knee joint, distal femur, and proximal tibia are harvested on a femoral vascular pedicle. This allograft can be considered both a VCA and a vascularized bone marrow transplant with its unique immune privileged features. The graft is transplanted to a subcutaneous abdominal pocket in the recipient animal with a skin component exteriorized to the dorsolateral region for immune monitoring.
Three surgical teams work simultaneously in a well-coordinated manner to reduce anesthesia and ischemia times, thereby improving efficiency of this model and reducing potential confounders in experimental protocols. This model serves as the groundwork for future therapeutic strategies aimed at reducing and potentially eliminating the need for chronic multi-drug immunosuppression in VCA.

A Modified Heterotopic Swine Hind Limb Transplant Model for Translational Vascularized Composite Allotransplantation VCA Research

Vascularized Composite Allotransplantations (VCA) have become a clinical reality. However, broad clinical application of VCA is limited by chronic multi-drug immunosuppression. The authors present a reliable and reproducible large animal model to translate novel immunomodulatory strategies that can minimize or potentially eliminate the need of immunosuppression in VCA.

平移血管复合同种异体移植（VCA）研究一种改进的异位猪后肢移植模型

Vascularized Composite Allotransplantation (VCA) such as hand and face transplants represent a viable treatment option for complex musculoskeletal trauma ...

医学

Assessment of Viability of Human Fat Injection into Nude Mice with Micro-Computed Tomography

Lipotransfer is a vital tool in the surgeon&#8217;s armamentarium for the treatment of soft tissue deficits of throughout the body. Fat is the ideal soft tissue filler as it is readily available, easily obtained, inexpensive, and inherently biocompatible.1 However, despite its burgeoning popularity, fat grafting is hampered by unpredictable results and variable graft survival, with published retention rates ranging anywhere from 10-80%. 1-3
To facilitate investigations on fat grafting, we have therefore developed an animal model that allows for real-time analysis of injected fat volume retention. Briefly, a small cut is made in the scalp of a CD-1 nude mouse and 200-400 &#181;l&#160;of processed lipoaspirate is placed over the skull. The scalp is chosen as the recipient site because of its absence of native subcutaneous fat, and because of the excellent background contrast provided by the calvarium, which aids in the analysis process. Micro-computed tomography (micro-CT) is used to scan the graft at baseline and every two weeks thereafter. The CT images are reconstructed, and an imaging software is used to quantify graft volumes.
Traditionally, techniques to assess fat graft volume have necessitated euthanizing the study animal to provide just a single assessment of graft weight and volume by physical measurement ex vivo. Biochemical and histological comparisons have likewise required the study animal to be euthanized. This described imaging technique offers the advantage of visualizing and objectively quantifying volume at multiple time points after initial grafting without having to sacrifice the study animal. The technique is limited by the size of the graft able to be injected as larger grafts risk skin and fat necrosis. This method has utility for all studies evaluating fat graft viability and volume retention. It is particularly well-suited to providing a visual representation of fat grafts and following changes in volume over time.

Fat grafting is an essential technique for reconstructing soft tissue deficits. However, it remains an unpredictable procedure characterized by variable graft survival. Our goal was to devise a mouse model that utilizes a novel imaging method to compare volume retention between differing techniques of fat graft preparation and delivery.

评估人体脂肪注射生存能力的裸鼠与微型计算机断层扫描

Lipotransfer is a vital tool in the surgeon&#8217;s armamentarium for the treatment of soft tissue deficits of throughout the body. Fat is the ideal soft ...

A Patient-Derived Xenograft Model for Venous Malformation

Venous malformation (VM) is a vascular anomaly that arises from impaired development of the venous network resulting in dilated and often dysfunctional veins. The purpose of this article is to carefully describe the establishment of a murine xenograft model that mimics human VM and is able to reflect patient heterogeneity. Hyper-activating non-inherited (somatic) TEK (TIE2) and PIK3CA mutations in endothelial cells (EC) have been identified as the main drivers of pathological vessel enlargement in VM. The following protocol describes the isolation, purification and expansion of patient-derived EC expressing mutant TIE2 and/or PIK3CA. These EC are injected subcutaneously into the back of immunodeficient athymic mice to generate ectatic vascular channels. Lesions generated with TIE2 or PIK3CA-mutant EC are visibly vascularized within 7&#8210;9 days of injection and recapitulate histopathological features of VM patient tissue. This VM xenograft model provides a reliable platform to investigate the cellular and molecular mechanisms driving VM formation and expansion. In addition, this model will be instrumental for translational studies testing the efficacy of novel drug candidates in preventing the abnormal vessel enlargement seen in human VM.

We present a detailed protocol to generate a murine xenograft model of venous malformation. This model is based on the subcutaneous injection of patient-derived endothelial cells containing hyper-activating TIE2 and/or PIK3CA gene mutations. Xenograft lesions closely recapitulate the histopathological features of VM patient tissue.

静脉畸形的患者衍生异种移植模型

Venous malformation (VM) is a vascular anomaly that arises from impaired development of the venous network resulting in dilated and often dysfunctional ...

Isolation of Murine Adipose Tissue-derived Microvascular Fragments as Vascularization Units for Tissue Engineering

A functional microvascular network is of pivotal importance for the survival and integration of engineered tissue constructs. For this purpose, several angiogenic and prevascularization strategies have been established. However, most cell-based approaches include time-consuming in vitro steps for the formation of a microvascular network. Hence, they are not suitable for intraoperative one-step procedures. Adipose tissue-derived microvascular fragments (ad-MVF) represent promising vascularization units. They can be easily isolated from fat tissue and exhibit a functional microvessel morphology. Moreover, they rapidly reassemble into new microvascular networks after in vivo implantation. In addition, ad-MVF have been shown to induce lymphangiogenesis. Finally, they are a rich source of mesenchymal stem cells, which may further contribute to their high vascularization potential. In previous studies we have demonstrated the remarkable vascularization capacity of ad-MVF in engineered bone and skin substitutes. In the present study, we report on a standardized protocol for the enzymatic isolation of ad-MVF from murine fat tissue.

We present a protocol to isolate adipose tissue-derived microvascular fragments that represent promising vascularization units. They can be rapidly isolated, do not require in vitro processing and, thus, may be used for one-step prevascularization in different fields of tissue engineering.

小鼠脂肪组织来源的微血管片段作为血管化单元的分离组织工程

A functional microvascular network is of pivotal importance for the survival and integration of engineered tissue constructs. For this purpose, several ...

生物工程

Transplantation of Schwann Cells Inside PVDF-TrFE Conduits to Bridge Transected Rat Spinal Cord Stumps to Promote Axon Regeneration Across the Gap

Among various models for spinal cord injury in rats, the contusion model is the most often used because it is the most common type of human spinal cord injury. The complete transection model, although not as clinically relevant as the contusion model, is the most rigorous method to evaluate axon regeneration. In the contusion model, it is difficult to distinguish regenerated from sprouted or spared axons due to the presence of remaining tissue post injury. In the complete transection model, a bridging method is necessary to fill the gap and create continuity from the rostral to the caudal stumps in order to evaluate the effectiveness of the treatments. A reliable bridging surgery is essential to test outcome measures by reducing the variability due to the surgical method. The protocols described here are used to prepare Schwann cells (SCs) and conduits prior to transplantation, complete transection of the spinal cord at thoracic level 8 (T8), insert the conduit, and transplant SCs into the conduit. This approach also uses in situ gelling of an injectable basement membrane matrix with SC transplantation that allows improved axon growth across the rostral and caudal interfaces with the host tissue.

This article describes a technique to insert a hollow conduit between the spinal cord stumps after complete transection and fill with Schwann cells (SCs) and injectable basement membrane matrix in order to bridge and promote axon regeneration across the gap.

氟 TrFE 导管内许旺细胞移植桥断大鼠脊髓残肢促进轴突再生跨越间隙

Among various models for spinal cord injury in rats, the contusion model is the most often used because it is the most common type of human spinal cord ...

Mechanical Micronization of Lipoaspirates for Regenerative Therapy

Stromal vascular fraction (SVF) has become a regenerative tool for various diseases; however, legislation strictly regulates the clinical application of cell products using collagenase. Here, we present a protocol to generate an injectable mixture of SVF cells and native extracellular matrix from adipose tissue by a purely mechanical process. Lipoaspirates are put into a centrifuge and spun at 1,200 x g for 3 min. The middle layer is collected and separated into two layers (high-density fat at the bottom and low-density fat on the top). The upper layer is directly emulsified by intersyringe shifting, at a rate of 20 mL/s for 6x to 8x. The emulsified fat is centrifuged at 2,000 x g for 3 min, and the sticky substance under the oil layer is collected and defined as the extracellular matrix (ECM)/SVF-gel. The oil on the top layer is collected. Approximately 5 mL of oil is added to 15 mL of high-density fat and emulsified by intersyringe shifting, at a rate of 20 mL/s for 6x to 8x. The emulsified fat is centrifuged at 2,000 x g for 3 min, and the sticky substance is also ECM/SVF-gel. After the transplantation of the ECM/SVF-gel into nude mice, the graft is harvested and assessed by histologic examination. The result shows that this product has the potential to regenerate into normal adipose tissue. This procedure is a simple, effective mechanical dissociation procedure to condense the SVF cells embedded in their natural supportive ECM for regenerative purposes.

Here, we present a protocol to obtain stromal vascular fraction from adipose tissue through a series of mechanical processes, which include emulsification and multiple centrifugations.

再生疗法用脂质吸入剂的机械微化

Stromal vascular fraction (SVF) has become a regenerative tool for various diseases; however, legislation strictly regulates the clinical application of ...

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.

糖尿病家兔欣德肢体缺血的临床前模型

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

<meta charset="utf8"></head><body>用于再生医学的纤维蛋白水凝胶中的机械基质血管组分

The Combination of Mechanically Isolated Stromal Vascular Fraction and Fibrin Hydrogel: A Processing Protocol

The regenerative potential of adipose-derived stromal cells (ASCs) has gained significant attention in regenerative and translational research. In the past, the extraction of these cells from adipose tissue required a multistep enzyme-based process, resulting in a heterogenous cell mix consisting of ACSs and other cells, which are jointly termed the stromal vascular fraction (SVF). More recently introduced mechanical SVF (mSVF) isolation protocols are less time-consuming and bypass regulatory concerns. We recently proposed a protocol that generates mSVF rich in stromal cells based on a combination of emulsification and centrifugation. One current issue in mSVF application for wound therapy application is the lack of a scaffold providing protection from mechanical manipulation and desiccation. Fibrin hydrogels have been shown to be a useful adjunct in cell transfer for wound healing purposes in the past. In the work herein, we delineate the preparation steps of an mSVF-fibrin hydrogel construct as a novel approach for translational research and clinical application.

<meta charset="utf8"></head><body>作者聚焦：探索脂肪衍生基质血管成分对伤口愈合的潜力

Mechanically isolated stromal vascular fraction (SVF) in combination with a fibrin hydrogel offers an easy and efficient carrier for viable adipose-derived stromal cells for various indications, including tissue engineering and or wound healing purposes. Here, we present the preparation of a mechanical SVF (mSVF)-fibrin hydrogel construct for translational research and clinical application.

机械分离的基质血管组分和纤维蛋白水凝胶的组合：一种加工方案

The regenerative potential of adipose-derived stromal cells (ASCs) has gained significant attention in regenerative and translational research. In the ...

基质血管部分富集脂肪移植治疗症状性端 neuromata

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再生疗法用脂质吸入剂的机械微化

糖尿病家兔欣德肢体缺血的临床前模型

机械分离的基质血管组分和纤维蛋白水凝胶的组合：一种加工方案

机械分离的基质血管组分和纤维蛋白水凝胶的组合：一种加工方案