这些过程是使用 Lipogems 系统执行的。

在涉及人类参加者的研究中所执行的所有程序都符合机构和/或国家研究委员会的道德标准, 以及1964《赫尔辛基宣言》及其后的修正案或可比较的道德标准.
1. 病史
<ol><li>开始临床检查与详细的患者历史。 注: 在踝关节不稳定的情况下, 必须始终怀疑其是否伴有肿胀、僵硬、疼痛和关节阻塞等反复扭伤。此外, 在许多情况下, 肝移植可能与系统性疾病史有关, 如炎症或血管疾病、神经病变或神经系统疾病和糖尿病。必须评估和考虑使用可能影响愈合的药物或医疗问题。</li>
</ol>2. 临床检查
<ol><li>评估病人在直立位置, 以突出脚踝或后足畸形。评估肌肉和肌腱的功能和踝关节的运动范围 (ROM)。弥漫性压痛经常存在, 尤其是在最大屈曲和伸展时, 在关节的高度接触敏感区域并不少见。</li>
	<li>进行前和后抽屉测试, 以确定伴随的侧踝关节不稳定。</li>
	<li>在术前会诊中, 记录以下临床和功能评分: 美国骨科足 & 踝关节 (AOFAS) 踝部和后足评分15, 视觉模拟量表 (VAS) 疼痛评分16, 和12项短格式健康调查 (SF-12)17。</li>
</ol>3. 放射性评估
<ol><li>执行双侧负重的脚部和脚踝的 x 片。这包括传统的负重前后 (AP) 榫, 以及踝关节侧面负重的观点。在中性位置执行 AP 榫, 并与15度的内部旋转, 以更好地可视化距骨。 注: 只有50% 的肝移植可以诊断为 x 射线;在大病灶的情况下, radiolucency 周围的分离骨区域可以被注意到18。</li>
	<li>执行常规的计算机断层扫描 (ct 扫描) 的脚踝。CT 扫描允许准确的位置和大小的病变, 也确定骨碎片的情况下脱离。CT 的薄弱点是显示软骨状态的能力。前一项研究显示了0.81 和0.99 的灵敏度和特异性, 分别用于在 CT19,20上检测 OLTs。</li>
	<li>执行踝关节磁共振成像 (MRI)。MRI 是软骨和下骨评估的基础。此外, MRI 不涉及电离辐射, 并允许更好的可视化的软组织。文献报告的敏感性和特异性96% 在检测 OLTs21,22。</li>
</ol>4. 外科技术
<ol><li>脂肪组织的采集和处理
	<ol>
<li>准备克莱因溶液: 1 升9克/升生理盐水, 50 毫升1% 利多卡因, 1 毫升1:1000 肾上腺素, 10 毫升8.4% 碳酸氢钠, 0.1 毫升的10毫克/毫升的曲安奈德。</li>
		<li>用手术刀刀片创建两个约0.5 厘米的脐切口。使用60毫升注射器和18G 针 (图 1), 向腹部皮下脂肪组织注入大约300毫升的 Klein 溶液。</li>
		<li>收获 40-45 毫升干细胞使用13G 钝套管连接到一个20毫升注射器和引进的处理套件 (图 2)。通常, 在围脐区进行收割。</li>
		<li>在闭合系统中插入 100-130 毫升的 lipoaspirate。通过一个大的过滤器将 lipoaspirate 推入设备, 以获得第一簇复位;与此同时, 相应数量的盐水溶液出口到浪费袋。一个关键的作用是发挥不锈钢大理石, 以获得一个临时乳液之间的油, 血液和盐水。用盐水溶液的重力反流去除油残渣和被污染的血液。</li>
		<li>经过这个洗涤步骤 (流动的解决方案显得清晰和 lipoaspirate 黄色), 停止盐水通量和反转装置 (灰色帽), 导致第二次脂肪簇减少。通过将漂浮的脂肪簇推到第二个切割六角形过滤器上, 以10毫升的注射器将液体从下面推入, 从而获得减少。将最终产品收集到连接到设备上部开口的10毫升注射器中。 注: 脂肪组织处理试剂盒改进了标准的 lipofilling 技术: 实际上, 该系统由封闭、全浸入、低压圆柱系统组成, 以获得含有大量周的流体和均匀产品。该程序允许通过轻度机械力来处理脂肪细胞, 并保持基质血管龛的完整性。这一过程是最不受创伤的可能, 并使最终产品在短时间 (15-20 分钟), 没有酶治疗或扩展。未损坏的 vasculostromal 龛有助于愈合过程。</li>
		<li>一旦干细胞已经收割, 在腹部敷上绷带。</li>
	</ol></li>
	<li>手术方法与脂肪组织注射
	<ol>
<li>将患者置于脊柱麻醉下的仰卧位, 用止血带, 在250柱的压力下, 在大腿的水平上, 以减少出血, 并允许一个更好的关节镜可视化1。</li>
		<li>用 dermographic 笔标记皮肤上的解剖地标。地标是避免医源性损伤的关键。 标记以下内容 (图 3): 足 (LM) 和内侧 (MM) 前关节线 (刘), 与踝关节的肌痛和跖屈进行鉴别 胫骨前肌腱 (达) 和跟腱 大隐静脉, 只跑在内侧踝前 腓浅神经 (SPN)</li>
		<li>首先, 在胫骨前肌腱内侧执行内侧门, 与软点重合。此门户代表方法门户。在大多数情况下, 在背的脚踝的抑郁症是可见的和可触及的。<ol>
<li>用刀片切开皮肤, 然后用钝夹层刺穿胶囊。注意避免隐神经和大隐静脉。当神经是大约7.4 毫米侧向门户时, 隐静脉位于侧向门户的9毫米。这构成了两个主要查看门户之一。</li>
		</ol>
</li>
		<li>检查关节线, 放置前外侧门, 内侧至侧面踝, 侧向伸肌腱肌腱。 注意: 在执行前外侧门时, 防止对中间背皮神经 (腓浅神经侧支) 的任何损伤;由于这个原因, 切割后, 皮肤必须采取钝解剖。</li>
		<li>检查关节软骨以评估其大小和位置。用 palpator 评价软骨的状况和质量。软骨病变的距骨通常位于 posteromedially 或 anterolaterally。<ol>
<li>使用广角2.7 毫米 arthroscopes, 以30°视角进行关节镜下治疗, 虽然一些外科医生将使用更大的4毫米关节镜, 并保持仪器在关节前休会。无创关节牵引技术和超足底屈曲可以用来访问大部分的距圆顶。</li>
			<li>在后病变的情况下, 经放置一个 Hintermann, 以分散关节, 并允许暴露的损害。根据病灶侧的 Hintermann, 在胫骨和距骨内侧或侧面的两根 K 线上有一个开口杠杆臂。在侧位病变的情况下, 注意在胫骨骨内插入近端的 K 线, 避免腓骨, 以更好地分散关节的注意力。</li>
		</ol>
</li>
		<li>用刮准备病变, 去除受损和不稳定的软骨, 钙化层和坏死和硬化骨, 以形成一个有肩边界的规则型的包含病变。对于此步骤, 请使用标准的关节镜刮 (图 4)。</li>
		<li>刺激骨髓干细胞从下骨 microperforations 全面从外到内的病变。<ol>
<li>进行穿孔间隔约3毫米之间。诱导微使用软骨选择在健康的下骨的缺陷下面 (图 5)。 注: 出血导致纤维蛋白凝块的形成。血液凝固产物, 通过炎症级联活化, 释放血管活性介质, 生长因子和细胞因子。这些因素具有刺激血管浸润和骨髓间充质干细胞迁移到病变软骨部分的能力。这些多能细胞被刺激分化成成纤维细胞, 软骨细胞, 和骨细胞, 并发挥重要作用, 刺激修复病变。分泌生长因子在关节环境中促进细胞外基质的形成和软骨的产生。同时, 病变的骨骼部分的细胞产生一个未成熟的骨组织, 逐渐被成熟的骨骼所取代。</li>
		</ol>
</li>
		<li>使用抽吸器中的剃须刀取出关节内的水, 用棉花海绵将剩余的液体除去, 直到关节完全干燥为止。</li>
	</ol></li>
	<li>脂肪源性干细胞注射
	<ol>
<li>注入 5-7 毫升脂肪来源的干细胞, 以前准备在步骤 4.1.3, 进入踝关节使用两个门户 (内侧或前外侧) 之一。</li>
		<li>松开止血带</li>
	</ol></li>
</ol>5. 术后护理
<ol><li>避免踝关节运动15天, 与操作肢体完全放电。</li>
	<li>手术后15天, 允许主动和被动运动的脚踝, 直到完全恢复和运动范围达到。</li>
	<li>手术后30天允许充电。</li>
</ol>6. 临床和放射学随访
<ol><li>在六和十二月以及此后每年对临床和影像患者进行评估。后续协议包括 AOFAS 脚踝和后足评分, VAS 疼痛评分, 和 SF-12 在每个端点15,16,17。影像学检查包括对手术踝关节的 MRI 和 CT 扫描。</li>
</ol>

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G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Osteochondral+lesions+of+the+talus:+a+new+magnetic+resonance+grading+system+with+arthroscopic+correlation.">Osteochondral lesions of the talus: a new magnetic resonance grading system with arthroscopic correlation.</a> Arthroscopy. 19 (4), 353-359 (2003).</li><li>Leumann, 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=A+novel+imaging+method+for+osteochondral+lesions+of+the+talus--comparison+of+SPECT-CT+with+MRI.">A novel imaging method for osteochondral lesions of the talus--comparison of SPECT-CT with MRI.</a> Am. J. Sports Med. 39 (5), 1095-1101 (2011).</li><li>Kim, Y. S., Park, E. H., Kim, Y. C., Koh, Y. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+outcomes+of+mesenchymal+stem+cell+injection+with+arthroscopic+treatment+in+older+patients+with+osteochondral+lesions+of+the+talus.">Clinical outcomes of mesenchymal stem cell injection with arthroscopic treatment in older patients with osteochondral lesions of the talus.</a> Am. J. Sports Med. 41 (5), 1090-1099 (2013).</li><li>Kim, Y. S., Lee, H. J., Choi, Y. J., Kim, Y. I., Koh, Y. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Does+an+injection+of+a+stromal+vascular+fraction+containing+adipose-derived+mesenchymal+stem+cells+influence+the+outcomes+of+marrow+stimulation+in+osteochondral+lesions+of+the+talus?+A+clinical+and+magnetic+resonance+imaging+study.">Does an injection of a stromal vascular fraction containing adipose-derived mesenchymal stem cells influence the outcomes of marrow stimulation in osteochondral lesions of the talus? A clinical and magnetic resonance imaging study.</a> Am. J. Sports Med. 42 (10), 2424-2434 (2014).</li><li>Kim, Y. S., Koh, Y. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Injection+of+Mesenchymal+Stem+Cells+as+a+Supplementary+Strategy+of+Marrow+Stimulation+Improves+Cartilage+Regeneration+After+Lateral+Sliding+Calcaneal+Osteotomy+for+Varus+Ankle+Osteoarthritis:+Clinical+and+Second-Look+Arthroscopic+Results.">Injection of Mesenchymal Stem Cells as a Supplementary Strategy of Marrow Stimulation Improves Cartilage Regeneration After Lateral Sliding Calcaneal Osteotomy for Varus Ankle Osteoarthritis: Clinical and Second-Look Arthroscopic Results.</a> Arthroscopy. 32 (5), 878-889 (2016).</li><li>Kim, Y. S., Lee, M., Koh, Y. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Additional+mesenchymal+stem+cell+injection+improves+the+outcomes+of+marrow+stimulation+combined+with+supramalleolar+osteotomy+in+varus+ankle+osteoarthritis:+short-term+clinical+results+with+second-look+arthroscopic+evaluation.">Additional mesenchymal stem cell injection improves the outcomes of marrow stimulation combined with supramalleolar osteotomy in varus ankle osteoarthritis: short-term clinical results with second-look arthroscopic evaluation.</a> J. Exp. Orthop. 3 (1), 12 (2016).</li><li>Hanke, C. W., Bernstein, G., Bullock, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Safety+of+tumescent+liposuction+in+15,336+patients.+National+survey+results.">Safety of tumescent liposuction in 15,336 patients. National survey results.</a> Dermatol Surg. 21 (5), 459-462 (1995).</li><li>Illouz, Y. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complications+of+liposuction.">Complications of liposuction.</a> Clin Plast Surg. 33 (1), 129-163 (2006).</li><li>Dixit, V. V., Wagh, M. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Unfavourable+outcomes+of+liposuction+and+their+management.">Unfavourable outcomes of liposuction and their management.</a> Indian J Plast Surg. 46 (2), 377-392 (2013).</li><li>Lehnhardt, M., Homann, H. H., Daigeler, A., Hauser, J., Palka, P., Steinau, H. U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Major+and+lethal+complications+of+liposuction:+review+of+72+cases+in+Germany+between+1998+and+2002.">Major and lethal complications of liposuction: review of 72 cases in Germany between 1998 and 2002.</a> Plast Reconstr Surg. 121 (6), 396e-403e (2008).</li><li>Usuelli, F. G., de Girolamo, L., Grassi, M., D'Ambrosi, R., Montrasio, U. A., Boga, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=All-Arthroscopic+Autologous+Matrix-Induced+Chondrogenesis+for+the+Treatment+of+Osteochondral+Lesions+of+the+Talus.">All-Arthroscopic Autologous Matrix-Induced Chondrogenesis for the Treatment of Osteochondral Lesions of the Talus.</a> Arthrosc Tech. 4 (3), e255-e259 (2015).</li><li>Simonson, D. C., Roukis, T. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Safety+of+ankle+arthroscopy+for+the+treatment+of+anterolateral+soft-tissue+impingement.">Safety of ankle arthroscopy for the treatment of anterolateral soft-tissue impingement.</a> Arthroscopy. 30 (2), 256-259 (2014).</li><li>Suzangar, M., Rosenfeld, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ankle+arthroscopy:+is+preoperative+marking+of+the+superficial+peroneal+nerve+important?.">Ankle arthroscopy: is preoperative marking of the superficial peroneal nerve important?.</a> J. Foot. Ankle Surg. 51 (2), 179-181 (2012).</li><li>Kraeutler, M. J., 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=Current+Concepts+Review+Update:+Osteochondral+Lesions+of+the+Talus.">Current Concepts Review Update: Osteochondral Lesions of the Talus.</a> Foot Ankle Int. 38 (3), 331-342 (2017).</li><li>Looze, C. 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=Evaluation+and+Management+of+Osteochondral+Lesions+of+the+Talus.">Evaluation and Management of Osteochondral Lesions of the Talus.</a> Cartilage. 8 (1), 19-30 (2017).</li><li>Dragoo, J. L., 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=Healing+full-thickness+cartilage+defects+using+adipose-derived+stem+cells.">Healing full-thickness cartilage defects using adipose-derived stem cells.</a> Tissue Eng. 13 (7), 1615-1621 (2007).</li><li>Lee, S. Y., Kim, W., Lim, C., Chung, S. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Treatment+of+Lateral+Epicondylosis+by+Using+Allogeneic+Adipose-Derived+Mesenchymal+Stem+Cells:+A+Pilot+Study.">Treatment of Lateral Epicondylosis by Using Allogeneic Adipose-Derived Mesenchymal Stem Cells: A Pilot Study.</a> Stem Cells. 33 (10), 2995-3005 (2015).</li><li>Feisst, V., Meidinger, S., Locke, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+bench+to+bedside:+use+of+human+adipose-derived+stem+cells.">From bench to bedside: use of human adipose-derived stem cells.</a> Stem Cells Cloning. 8, 149-162 (2015).</li></ol>

费德里克二世 Usuelli, MD, 报告个人费用从 Integra 和 Geistlich, 以及赠款和个人的费用, 从在提交的工作以外的。

近年来, 临床和基础研究已经将注意力集中在干细胞治疗不同肌肉骨骼疾病的作用上。本文的目的是描述使用 microfractured 和纯化脂肪组织与关节镜 microperforations 治疗骨距软骨病变的协议。该协议涉及几个关键步骤, 具有高风险的并发症。在脂肪收获过程中, 并发症可分为局部或全身并发症。
术后最常见的并发症是轮廓不规则, 发病率为2.7%。这可以避免使用小套管, 不进行肤浅?...

关节镜是治疗距骨 (OLTs) 软骨病变的金标准, 目的是减轻疼痛, 恢复功能, 改善生活质量, 特别是在年轻和积极的患者。
目前, 关节镜技术可分为三种。修复技术通过病灶清除和 microperforations, 刺激骨髓细胞获得。重建技术用自体或异源 ostechondral 移植取代病变。再生技术利用多能细胞分化和复制的能力来重建受损组织1,2,3,4,5,6.
近年来, 再生技术已经成为众多的体外和体内研究的主题, 用于治疗 OLTs, 特别是从脂肪组织中提取的间充质干细胞 (干细胞)7,8,9. 这些间充质干细胞表现出与其他组织分离的其他多能细胞相似的形态学和功能特征;他们也有能力区分成几个和不同的细胞线都在体外和在体内10,11,12,13.对这些细胞的研究主要是由于它们的局部化, 事实上它们代表了正常体重的10% 到 30%, 每克组织的浓度为5000细胞,13,14。另一方面, 限制使用这些细胞的因素与它们在实验室过程中的处理有关。lipoaspirate 含有脂肪细胞、胶原纤维和正常血管成分的聚合体, 酶 i 型胶原处理, 在培养前受到溶血。目的是描述使用 microfractured 和纯化脂肪组织治疗距骨软骨病变的方案。

<table>
	<tbody>
		<tr>
			<td>PROCESS KIT - PROCESSING KIT FOR FAT TISSUE</td>
			<td>LIPOGEMS</td>
			<td>LG PK 60</td>
			<td>Lipogems Kit to obtain microfractured and purified ADSCs</td>
		</tr>
		<tr>
			<td>HINTERMANN SPREADER</td>
			<td>INTEGRA</td>
			<td>119654</td>
			<td>The spreader allow to access most of the talar dome, in particular in case of posterior lesion</td>
		</tr>
		<tr>
			<td>CUP CURETTE</td>
			<td>ARTHREX</td>
			<td>AR-8655-02</td>
			<td>To remove the damaged cartilage and necrotic and sclerotic bone</td>
		</tr>
		<tr>
			<td>CHONDRAL PICK 30&#176; TIP / 60&#176; TIP</td>
			<td>ARTHREX</td>
			<td>AR-8655-05 
			AR-8655-06</td>
			<td>To perfrom microperforation at the level of the lesion, stimulating bleeding from the subchondral bone</td>
		</tr>
		<tr>
			<td>SHAVER</td>
			<td>ARTHREX</td>
			<td>AR-7300SR</td>
			<td>To clean the joint and aspirate water</td>
		</tr>
	</tbody>
</table>

autologous microfractured and purified adipose tissue for arthroscopic management of osteochondral lesions of the talus

近年来, 再生技术已被越来越多的研究和使用, 以治疗软骨病变的距骨。特别是, 有几项研究把注意力集中在脂肪组织的间充质干细胞上。脂肪来源的干细胞 (干细胞) 表现出与其他间充质细胞相似的形态学特征和性质, 并能分化成多种细胞株。此外, 这些细胞也广泛适用于皮下组织, 代表 10-30% 的正常体重, 每克组织5000细胞浓度。
在所提出的技术, 第一步是从腹部收获干细胞和一个过程的微和纯化;其次, 手术的过程是完全关节, 较少软组织剥离, 更好的关节可视化, 和更快的恢复相比, 标准开放程序。关节镜的特点是第一阶段的病灶是确定, 孤立, 并准备与 microperforations;第二步, 执行干, 涉及注射脂肪组织在病变的水平。
2016年1月至2016年9月期间, 四例患者经关节镜治疗, microfractured 和纯化的脂肪组织在距骨软骨病变。所有患者报告在手术后六月的临床改善没有报告的并发症。在最新的随访中, 功能评分是令人鼓舞的, 并证实该技术提供了可靠的疼痛缓解和改善的软骨病变的距骨。

2016年1月至2016年9月期间, 四例患者经关节镜治疗, microfractured 和纯化的脂肪组织在距骨软骨病变。所有患者在手术后六月报告临床改善。在表 1中报告了初步临床结果。无并发症发生。
近年来, 使用干细胞治疗足踝部病变的情况有所增加。在 2013年, Kim et al.23治疗65老年患者, 50 岁以上的症状?...

Watch this Scientific Journal Video about Autologous Microfractured and Purified Adipose Tissue for Arthroscopic Management of Osteochondral Lesions of the Talus at JoVE.com

Autologous Microfractured and Purified Adipose Tissue for Arthroscopic Management of Osteochondral Lesions of the Talus

本研究的目的是报告一个协议的关节镜治疗软骨病变的距骨使用 microfractured 和纯化脂肪干细胞。

自体 Microfractured 与纯化脂肪组织在关节镜下治疗距骨软骨病变

In recent years, regenerative techniques have been increasingly studied and used to treat osteochondral lesions of the talus. In particular, several ...

autologous-microfractured-purified-adipose-tissue-for-arthroscopic

Research

JoVE Journal

Medicine

10.2K 次观看.  IRCCS Istituto Ortopedico Galeazzi - C.A.S.C.O. Piede e Caviglia. 本研究的目的是报告一个协议的关节镜治疗软骨病变的距骨使用 microfractured 和纯化脂肪干细胞。

视频: Autologous Microfractured and Purified Adipose Tissue for Arthroscopic Management of Osteochondral Lesions of the Talus

Breathing-controlled Electrical Stimulation (BreEStim) for Management of Neuropathic Pain and Spasticity

Electrical stimulation (EStim) refers to the application of electrical current to muscles or nerves in order to achieve functional and therapeutic goals. It has been extensively used in various clinical settings. Based upon recent discoveries related to the systemic effects of voluntary breathing and intrinsic physiological interactions among systems during voluntary breathing, a new EStim protocol, Breathing-controlled Electrical Stimulation (BreEStim), has been developed to augment the effects of electrical stimulation. In BreEStim, a single-pulse electrical stimulus is triggered and delivered to the target area when the airflow rate of an isolated voluntary inspiration reaches the threshold. BreEStim integrates intrinsic physiological interactions that are activated during voluntary breathing and has demonstrated excellent clinical efficacy. Two representative applications of BreEStim are reported with detailed protocols: management of post-stroke finger flexor spasticity and neuropathic pain in spinal cord injury.

Breathing-controlled Electrical Stimulation BreEStim for Management of Neuropathic Pain and Spasticity

The purpose is to present a new method, breathing-control electrical stimulation (BreEStim) for management of neuropathic pain and spasticity.

呼吸控制电气的刺激（BreEStim）的神经性疼痛和痉挛的管理

Electrical stimulation (EStim) refers to the application of electrical current to muscles or nerves in order to achieve functional and therapeutic goals. ...

科研

医学

Steps for the Autologous Ex vivo Perfused Porcine Liver-kidney Experiment

The use of ex vivo perfused models can mimic the physiological conditions of the liver for short periods, but to maintain normal homeostasis for an extended perfusion period is challenging. We have added the kidney to our previous ex vivo perfused liver experiment model to reproduce a more accurate physiological state for prolonged experiments without using live animals. Five intact livers and kidneys were retrieved post-mortem from sacrificed pigs on different days and perfused for a minimum of 6 hr. Hourly arterial blood gases were obtained to analyze pH, lactate, glucose and renal parameters. The primary endpoint was to investigate the effect of adding one kidney to the model on the acid base balance, glucose, and electrolyte levels. The result of this liver-kidney experiment was compared to the results of five previous liver only perfusion models. In summary, with the addition of one kidney to the ex vivo liver circuit, hyperglycemia and metabolic acidosis were improved. In addition this model reproduces the physiological and metabolic responses of the liver sufficiently accurately to obviate the need for the use of live animals. The ex vivo liver-kidney perfusion model can be used as an alternative method in organ specific studies. It provides a disconnection from numerous systemic influences and allows specific and accurate adjustments of arterial and venous pressures and flow.

Steps for the Autologous Ex vivo Perfused Porcine Liver-kidney Experiment

Study of the animal organ physiology can be achieved by performing an experimental ex vivo perfusion system. The addition of a porcine kidney, as a homeostatic organ to our previously developed ex vivo liver perfusion model can be a principal step to achieve a better physiological environment.

The use of ex vivo perfused models can mimic the physiological conditions of the liver for short periods, but to maintain normal homeostasis for an ...

Generation and 3-Dimensional Quantitation of Arterial Lesions in Mice Using Optical Projection Tomography

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating important information on the pathogenesis of lesion formation and the action of novel therapies. Use of atherosclerosis-prone mice, surgical methods of lesion induction, and dietary modification has dramatically improved understanding of the mechanisms that contribute to disease development and the potential of new treatments.
Classically, analysis of lesions is performed ex vivo using 2-dimensional histological techniques. This article describes application of optical projection tomography (OPT) to 3-dimensional quantitation of arterial lesions. As this technique is non-destructive, it can be used as an adjunct to standard histological and immunohistochemical analyses.
Neointimal lesions were induced by wire-insertion or ligation of the mouse femoral artery whilst atherosclerotic lesions were generated by administration of an atherogenic diet to apoE-deficient mice.
Lesions were examined using OPT imaging of autofluorescent emission followed by complementary histological and immunohistochemical analysis. OPT clearly distinguished lesions from the underlying vascular wall. Lesion size was calculated in 2-dimensional sections using planimetry, enabling calculation of lesion volume and maximal cross-sectional area. Data generated using OPT were consistent with measurements obtained using histology, confirming the accuracy of the technique and its potential as a complement (rather than alternative) to traditional methods of analysis.
This work demonstrates the potential of OPT for imaging atherosclerotic and neointimal lesions. It provides a rapid, much needed ex vivo technique for the routine 3-dimensional quantification of vascular remodelling.

Ex vivo analysis of arterial lesions from animal models of cardiovascular disease classically relies on histological and immunohistochemical techniques. These provide 2-dimensional measurements in 3-dimensional lesions. This manuscript describes the generation of arterial lesions for quantitative analysis in 3-dimensions using optical projection tomography.

生成和动脉病变的小鼠的三维定量使用光学投影层析成像

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating ...

Cerenkov Luminescence Imaging of Interscapular Brown Adipose Tissue

Brown adipose tissue (BAT), widely known as a &#8220;good fat&#8221; plays pivotal roles for thermogenesis in mammals. This special tissue is closely related to metabolism and energy expenditure, and its dysfunction is one important contributor for obesity and diabetes. Contrary to previous belief, recent PET/CT imaging studies indicated the BAT depots are still present in human adults. PET imaging clearly shows that BAT has considerably high uptake of 18F-FDG under certain conditions. In this video report, we demonstrate that Cerenkov luminescence imaging (CLI) with 18F-FDG can be used to optically image BAT in small animals. BAT activation is observed after intraperitoneal injection of norepinephrine (NE) and cold treatment, and depression of BAT is induced by long anesthesia. Using multiple-filter Cerenkov luminescence imaging, spectral unmixing and 3D imaging reconstruction are demonstrated. Our results suggest that CLI with 18F-FDG is a practical technique for imaging BAT in small animals, and this technique can be used as a cheap, fast, and alternative imaging tool for BAT research.

In this video report, we show the application of Cerenkov Luminescence Imaging (CLI) for interscapular brown adipose tissue in mice under activated and depressed conditions.

肩胛间褐色脂肪组织的切伦科夫发光成像

Brown adipose tissue (BAT), widely known as a &#8220;good fat&#8221; plays pivotal roles for thermogenesis in mammals. This special tissue is closely ...

Encapsulation Thermogenic Preadipocytes for Transplantation into Adipose Tissue Depots

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular weight metabolites in tissues of the treated host to achieve long-term survival. The semipermeable membrane allows engrafted encapsulated cells to avoid rejection by the immune system. The encapsulation procedure was designed to enable a controlled release of bioactive compounds, such as insulin, other hormones, and cytokines. Here we describe a method for encapsulation of catabolic cells, which consume lipids for heat production and energy dissipation (thermogenesis) in the intra-abdominal adipose tissue of obese mice. Encapsulation of thermogenic catabolic cells may be potentially applicable to the prevention and treatment of obesity and type 2 diabetes. Another potential application of catabolic cells may include detoxification from alcohols or other toxic metabolites and environmental pollutants.

Here, we present a protocol for encapsulation of catabolic cells, which consume lipids for heat production in intra-abdominal adipose tissue and increase energy dissipation in obese mice.

封装产热前脂肪细胞移植到脂肪组织仓库

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular ...

A Technique for Subcutaneous Abdominal Adipose Tissue Biopsy via a Non-diathermy Method

Adipose tissue biopsies offer tissue samples that, upon analysis, may provide insightful overviews of mechanisms relating to metabolism and disease. To obtain subcutaneous adipose tissue biopsies in the abdominal area, researchers and physicians use either a surgical or a needle-based technique. However, surgical subcutaneous fat biopsies can offer tissue samples that may provide a more comprehensive overview of the complexities of biological indices in white adipose tissue. Usually, a surgical adipose tissue biopsy includes a diathermy treatment for cauterizing blood vessels to prevent excessive bleeding. Nevertheless, side effects, such as flash fires and skin lesions in the tissue, have been reported after diathermy. Therefore, we aimed to standardize a surgical abdominal adipose tissue biopsy performed under local anesthesia using a non-diathermy method. We conducted 115 subcutaneous adipose tissue biopsies in healthy men using a non-diathermy abdominal surgical biopsy method. Our results showed three cases of excessive post-operation bleeding out of 115 operations (2.61%).In conclusion, our standardized subcutaneous abdominal adipose tissue surgical biopsy using a non-diathermy method can be safely applied to healthy men at the bedside, with minimal side effects.

We standardized an abdominal adipose tissue biopsy using a non-diathermy method performed under local anesthesia. Three cases of excessive post-operation bleeding out of 115 operations (2.61%) occurred.We conclude that an abdominal adipose tissue surgical biopsy using a non-diathermy method can be safely applied to healthy men.

经非透法皮下腹脂肪组织活检技术

Adipose tissue biopsies offer tissue samples that, upon analysis, may provide insightful overviews of mechanisms relating to metabolism and disease. To ...

Assessment of Human Adipose Tissue Microvascular Function Using Videomicroscopy

While obesity is closely linked to the development of metabolic and cardiovascular disease, little is known about mechanisms that govern these processes. It is hypothesized that pro-atherogenic mediators released from fat tissues particularly in association with central/visceral adiposity may promote pathogenic vascular changes locally and systemically, and the notion that cardiovascular disease may be the consequence of adipose tissue dysfunction continues to evolve. Here, we describe a unique method of videomicroscopy that involves analysis of vasodilator and vasoconstrictor responses of intact small human arterioles removed from the adipose depot of living human subjects. Videomicroscopy is used to examine functional properties of isolated microvessels in response to pharmacological or physiological stimuli using a pressured system that mimics in vivo conditions. The technique is a useful approach to gain understanding of the pathophysiology and molecular mechanisms that contribute to vascular dysfunction locally within the adipose tissue milieu. Moreover, abnormalities in the adipose tissue microvasculature have also been linked with systemic diseases. We applied this technique to examine depot-specific vascular responses in obese subjects. We assessed endothelium-dependent vasodilation to both increased flow and acetylcholine in adipose arterioles (50 - 350 &#181;m internal diameter, 2 - 3 mm in length) isolated from two different adipose depots during bariatric surgery from the same individual. We demonstrated that arterioles from visceral fat exhibit impaired endothelium-dependent vasodilation compared to vessels isolated from the subcutaneous depot. The findings suggest that the visceral microenvironment is associated with vascular endothelial dysfunction which may be relevant to clinical observation linking increased visceral adiposity to systemic disease mechanisms. The videomicroscopy technique can be used to examine vascular phenotypes from different fat depots as well as compare findings across individuals with different degrees of obesity and metabolic dysfunction. The method can also be used to examine vascular responses longitudinally in response to clinical interventions.

Videomicroscopy systems are used to examine functional properties of isolated adipose tissue arterioles in response to physiological and pharmacological stimuli. This technique can be used to examine microvascular phenotypes in different adipose tissue domains in obese humans.

应用 Videomicroscopy 评价人脂肪组织微血管功能

While obesity is closely linked to the development of metabolic and cardiovascular disease, little is known about mechanisms that govern these processes. ...

Clinical Protocol of Producing Adipose Tissue-Derived Stromal Vascular Fraction for Potential Cartilage Regeneration

Osteoarthritis (OA) is one of the most common debilitating disorders.&#160;Recently, numerous attempts have been made to improve the functions of the knees by using different forms of mesenchymal stem cells (MSCs). In Korea, bone marrow concentrates and cord blood-derived stem cells have been approved by the Korean Food and Drug Administration (KFDA) for cartilage regeneration. In addition, an adipose tissue-derived stromal vascular fraction (SVF) has been allowed by the KFDA for joint injections in human patients. Autologous adipose tissue-derived SVF contains extracellular matrix (ECM) in addition to mesenchymal stem cells. ECM excretes various cytokines that, along with hyaluronic acid (HA) and platelet-rich plasma (PRP) activated by calcium chloride, may help MSCs to regenerate cartilage and improve knee functions. In this article, we presented a protocol to improve knee functions by regenerating cartilage-like tissue in human patients with OA. The result of the protocol was first reported in 2011 followed by a few additional publications. The protocol involves liposuction to obtain autologous lipoaspirates that are mixed with collagenase. This lipoaspirates-collagenase mixture is then cut and homogenized to remove large fibrous tissue that may clog up the needle during the injection. Afterwards, the mixture is incubated to obtain adipose tissue-derived SVF. The resulting adipose tissue-derived SVF, containing both adipose tissue-derived MSCs and remnants of ECM, is injected into knees of patients, combined with HA and calcium chloride activated PRP. Included are three cases of patients who were treated with our protocol resulting in improvement of knee pain, swelling, and range of motion along with MRI evidence of hyaline cartilage-like tissue.

Here, we present a protocol to produce an adipose tissue-derived stromal vascular fraction and its application to improve knee functions by regenerating cartilage-like tissue in human patients with osteoarthritis.

制备脂肪组织基质血管部分用于潜在软骨再生的临床协议

Osteoarthritis (OA) is one of the most common debilitating disorders.&#160;Recently, numerous attempts have been made to improve the functions of the ...

Inguinal Subcutaneous White Adipose Tissue (ISWAT) Transplantation Model of Murine Islets

Pancreatic islet transplantation is a well-established therapeutic treatment for type 1 diabetes. The kidney capsule is the most commonly used site for islet transplantation in rodent models. However, the tight kidney capsule limits the transplantation of sufficient islets in large animals and humans. The inguinal subcutaneous white adipose tissue (ISWAT), a new subcutaneous space, was found to be a potentially valuable site for islet transplantation. This site has better blood supply than other subcutaneous spaces. Moreover, the ISWAT accommodates a larger islet mass than the kidney capsule, and transplantation into it is simple. This manuscript describes the procedure of mouse islet isolation and transplantation in the ISWAT site of syngeneic diabetic mouse recipients. Using this protocol, murine pancreatic islets were isolated by standard collagenase digestion and a basement membrane matrix hydrogel was used for fixing the purified islets in the ISWAT site. The blood glucose levels of the recipient mice were monitored for more than 100 days. Islet grafts were retrieved at day 100 after transplantation for histological analysis. The protocol for islet transplantation in the ISWAT site described in this manuscript is simple and effective.

Inguinal Subcutaneous White Adipose Tissue ISWAT Transplantation Model of Murine Islets

In this protocol, a method of murine islet isolation and transplantation into the inguinal subcutaneous white adipose tissue is described. Isolated syngeneic murine islets are transplanted into a murine recipient using a basement membrane hydrogel. The blood glucose level of the recipients is monitored, and histology analysis of the islet grafts is performed.

毛里岛皮下白脂肪组织（ISWAT）移植模型

Pancreatic islet transplantation is a well-established therapeutic treatment for type 1 diabetes. The kidney capsule is the most commonly used site for ...

Fat-Covered Islet Transplantation Using Epididymal White Adipose Tissue

Islet transplantation is a cellular replacement therapy for severe diabetes mellitus. The intraperitoneal cavity is typically the transplant site for this procedure. However, intraperitoneal islet transplantation has some limitations, including poor transplant efficacy, difficult graft detection ability, and a lack of graftectomy capability for post-transplant analysis. In this paper, "fat-covered islet transplantation", an intraperitoneal islet transplantation method that utilizes epididymal white adipose tissue, is used to assess the therapeutic effects of bioengineered islets. The simplicity of the method lies in the seeding of islets onto epididymal white adipose tissue and using the tissue to cover the islets. While this method can be categorized as an intraperitoneal islet transplantation technique, it shares characteristics with intra-adipose tissue islet transplantation. The fat-covered islet transplantation method demonstrates more robust therapeutic effects than intra-adipose tissue islet transplantation, however, including the improvement of blood glucose and plasma insulin levels and the potential for graft removal. We recommend the adoption of this method for assessing the mechanisms of islet engraftment into white adipose tissue and the therapeutic effects of bioengineered islets.

This fat-covered islet transplantation method is suitable for the detection of engrafted islets in the intraperitoneal cavity. Notably, it does not require the use of biobinding agents or suturing.

使用附睾白色脂肪组织进行脂肪覆盖的胰岛移植

Islet transplantation is a cellular replacement therapy for severe diabetes mellitus. The intraperitoneal cavity is typically the transplant site for this ...