Name: autologous microfractured and purified adipose tissue for arthroscopic management of osteochondral lesions of the talus
Uploaded: 2018-01-23T13:00:00.000+00:00
Duration: 7 min 24 s
Description: Watch this Scientific Journal Video about Autologous Microfractured and Purified Adipose Tissue for Arthroscopic Management of Osteochondral Lesions of the Talus at JoVE.com

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=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&amp;deg; TIP / 60&amp;deg; TIP</td><td>ARTHREX</td><td>AR-8655-05&lt;br/&gt; 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

Repair of a Critical-sized Calvarial Defect Model Using Adipose-derived Stromal Cells Harvested from Lipoaspirate

Craniofacial skeletal repair and regeneration offers the promise of de novo tissue formation through a cell-based approach utilizing stem cells. Adipose-derived stromal cells (ASCs) have proven to be an abundant source of multipotent stem cells capable of undergoing osteogenic, chondrogenic, adipogenic, and myogenic differentiation. Many studies have explored the osteogenic potential of these cells in vivo with the use of various scaffolding biomaterials for cellular delivery. It has been demonstrated that by utilizing an osteoconductive, hydroxyapatite-coated poly(lactic-co-glycolic acid) (HA-PLGA) scaffold seeded with ASCs, a critical-sized calvarial defect, a defect that is defined by its inability to undergo spontaneous healing over the lifetime of the animal, can be effectively show robust osseous regeneration. This in vivo model demonstrates the basis of translational approaches aimed to regenerate the bone tissue - the cellular component and biological matrix. This method serves as a model for the ultimate clinical application of a progenitor cell towards the repair of a specific tissue defect.

This protocol describes the isolation of adipose-derived stromal cells from lipoaspirate and the creation of a 4 mm critical-sized calvarial defect to evaluate skeletal regeneration.

修复的临界大小的颅骨缺损模型脂肪间充质干细胞的收获Lipoaspirate

Craniofacial skeletal repair and regeneration offers the promise of de novo tissue formation through a cell-based approach utilizing stem cells. ...

科研

医学

Treatment of Ankle Osteoarthritis with Total Ankle Replacement Through a Lateral Transfibular Approach

Total ankle replacement (TAR) is a valid option for the treatment of ankle osteoarthritis. The traditional surgical approach for TAR is the anterior approach. Recently, the lateral transfibular approach to the ankle has gained popularity since a new TAR implant was designed to be performed via this approach that results in an ideal visualization of the center of rotation of the ankle and curved resections that allow for sparing bone cuts. The aim of the present paper is to present our preoperative, operative, and postoperative protocols for the treatment of the osteoarthritis of the ankle with TAR via lateral approach. We present our preoperative clinical and radiographic protocol. In addition, we describe our surgical technique with some technical tips. Finally, we report our follow-up schedule that includes the collection of clinical, functional, and radiographic data. The results of this procedure are encouraging: TAR through a lateral transfibular approach provides reliable pain relief and improvements in functional outcomes in patients with ankle osteoarthritis.

We present our preoperative, operative, and postoperative protocols for the treatment of osteoarthritis of the ankle with total ankle replacement via lateral transfibular approach.

外侧 Transfibular 法治疗踝关节骨性关节炎

Total ankle replacement (TAR) is a valid option for the treatment of ankle osteoarthritis. The traditional surgical approach for TAR is the anterior ...

行为学

Matrix-assisted Autologous Chondrocyte Transplantation for Remodeling and Repair of Chondral Defects in a Rabbit Model

Articular cartilage defects are considered a major health problem because articular cartilage has a limited capacity for self-regeneration 1. Untreated cartilage lesions lead to ongoing pain, negatively affect the quality of life and predispose for osteoarthritis. During the last decades, several surgical techniques have been developed to treat such lesions. However, until now it was not possible to achieve a full repair in terms of covering the defect with hyaline articular cartilage or of providing satisfactory long-term recovery 2-4. Therefore, articular cartilage injuries remain a prime target for regenerative techniques such as Tissue Engineering. In contrast to other surgical techniques, which often lead to the formation of fibrous or fibrocartilaginous tissue, Tissue Engineering aims at fully restoring the complex structure and properties of the original articular cartilage by using the chondrogenic potential of transplanted cells. Recent developments opened up promising possibilities for regenerative cartilage therapies.
The first cell based approach for the treatment of full-thickness cartilage or osteochondral lesions was performed in 1994 by Lars Peterson and Mats Brittberg who pioneered clinical autologous chondrocyte implantation (ACI) 5. Today, the technique is clinically well-established for the treatment of large hyaline cartilage defects of the knee, maintaining good clinical results even 10 to 20 years after implantation 6. In recent years, the implantation of autologous chondrocytes underwent a rapid progression. The use of an artificial three-dimensional collagen-matrix on which cells are subsequently replanted became more and more popular 7-9.
MACT comprises of two surgical procedures: First, in order to collect chondrocytes, a cartilage biopsy needs to be performed from a non weight-bearing cartilage area of the knee joint. Then, chondrocytes are being extracted, purified and expanded to a sufficient cell number in vitro. Chondrocytes are then seeded onto a three-dimensional matrix and can subsequently be re-implanted. When preparing a tissue-engineered implant, proliferation rate and differentiation capacity are crucial for a successful tissue regeneration 10. The use of a three-dimensional matrix as a cell carrier is thought to support these cellular characteristics 11.
The following protocol will summarize and demonstrate a technique for the isolation of chondrocytes from cartilage biopsies, their proliferation in vitro and their seeding onto a 3D-matrix (Chondro-Gide, Geistlich Biomaterials, Wollhusen, Switzerland). Finally, the implantation of the cell-matrix-constructs into artificially created chondral defects of a rabbit's knee joint will be described. This technique can be used as an experimental setting for further experiments of cartilage repair.

An experimental technique for the treatment of chondral defects in the rabbit's knee joint is described. The implantation of autologous chondrocytes seeded on a matrix is a well-accepted method for the remodeling and repair of articular cartilage lesions providing satisfying long-term results. Matrix-assisted autologous chondrocyte transplantation (MACT) offers a standardized and clinically established implantation method.

基质辅助重塑和修复软骨缺损的自体软骨细胞移植在兔模型

Articular cartilage defects are considered a major health problem because articular cartilage has a limited capacity for self-regeneration 1. Untreated ...

Treatment of Osteochondral Defects in the Rabbit's Knee Joint by Implantation of Allogeneic Mesenchymal Stem Cells in Fibrin Clots

The treatment of osteochondral articular defects has been challenging physicians for many years. The better understanding of interactions of articular cartilage and subchondral bone in recent years led to increased attention to restoration of the entire osteochondral unit. In comparison to chondral lesions the regeneration of osteochondral defects is much more complex and a far greater surgical and therapeutic challenge. The damaged tissue does not only include the superficial cartilage layer but also the subchondral bone. For deep, osteochondral damage, as it occurs for example with osteochondrosis dissecans, the full thickness of the defect needs to be replaced to restore the joint surface 1. Eligible therapeutic procedures have to consider these two different tissues with their different intrinsic healing potential 2. In the last decades, several surgical treatment options have emerged and have already been clinically established 3-6.
Autologous or allogeneic osteochondral transplants consist of articular cartilage and subchondral bone and allow the replacement of the entire osteochondral unit. The defects are filled with cylindrical osteochondral grafts that aim to provide a congruent hyaline cartilage covered surface 3,7,8. Disadvantages are the limited amount of available grafts, donor site morbidity (for autologous transplants) and the incongruence of the surface; thereby the application of this method is especially limited for large defects.
New approaches in the field of tissue engineering opened up promising possibilities for regenerative osteochondral therapy. The implantation of autologous chondrocytes marked the first cell based biological approach for the treatment of full-thickness cartilage lesions and is now worldwide established with good clinical results even 10 to 20 years after implantation 9,10. However, to date, this technique is not suitable for the treatment of all types of lesions such as deep defects involving the subchondral bone 11.
The sandwich-technique combines bone grafting with current approaches in Tissue Engineering 5,6. This combination seems to be able to overcome the limitations seen in osteochondral grafts alone. After autologous bone grafting to the subchondral defect area, a membrane seeded with autologous chondrocytes is sutured above and facilitates to match the topology of the graft with the injured site. Of course, the previous bone reconstruction needs additional surgical time and often even an additional surgery. Moreover, to date, long-term data is missing 12.
Tissue Engineering without additional bone grafting aims to restore the complex structure and properties of native articular cartilage by chondrogenic and osteogenic potential of the transplanted cells. However, again, it is usually only the cartilage tissue that is more or less regenerated. Additional osteochondral damage needs a specific further treatment. In order to achieve a regeneration of the multilayered structure of osteochondral defects, three-dimensional tissue engineered products seeded with autologous/allogeneic cells might provide a good regeneration capacity 11.
Beside autologous chondrocytes, mesenchymal stem cells (MSC) seem to be an attractive alternative for the development of a full-thickness cartilage tissue. In numerous preclinical in vitro and in vivo studies, mesenchymal stem cells have displayed excellent tissue regeneration potential 13,14. The important advantage of mesenchymal stem cells especially for the treatment of osteochondral defects is that they have the capacity to differentiate in osteocytes as well as chondrocytes. Therefore, they potentially allow a multilayered regeneration of the defect.
In recent years, several scaffolds with osteochondral regenerative potential have therefore been developed and evaluated with promising preliminary results 1,15-18. Furthermore, fibrin glue as a cell carrier became one of the preferred techniques in experimental cartilage repair and has already successfully been used in several animal studies 19-21 and even first human trials 22.
The following protocol will demonstrate an experimental technique for isolating mesenchymal stem cells from a rabbit's bone marrow, for subsequent proliferation in cell culture and for preparing a standardized in vitro-model for fibrin-cell-clots. Finally, a technique for the implantation of pre-established fibrin-cell-clots into artificial osteochondral defects of the rabbit's knee joint will be described.

An experimental technique for the treatment of osteochondral defects in the rabbit's knee joint is described. The implantation of allogeneic mesenchymal stem cells into osteochondral defects provides a promising development in the field of tissue engineering. The preparation of fibrin-cell-clots in vitro offers a standardized method for implantation.

异基因骨髓间充质干细胞植入兔子的膝关节软骨缺损治疗纤维蛋白凝块

The treatment of osteochondral articular defects has been challenging physicians for many years. The better understanding of interactions of articular ...

Application of Atomic Force Microscopy to Detect Early Osteoarthritis

Biomechanical properties of cells and tissues not only regulate their shape and function but are also crucial for maintaining their vitality. Changes in elasticity can propagate or trigger the onset of major diseases like cancer or osteoarthritis (OA). Atomic force microscopy (AFM) has emerged as a strong tool to qualitatively and quantitatively characterize the biomechanical properties of specific biological target structures on a microscopic scale, measuring forces in a range from as small as the piconewton to the micronewton. Biomechanical properties are of special importance in musculoskeletal tissues, which are subjected to high levels of strain. OA as a degenerative disease of the cartilage results in the disruption of the pericellular matrix (PCM) and the spatial rearrangement of the chondrocytes embedded in their extracellular matrix (ECM). Disruption in PCM and ECM has been associated with changes in the biomechanical properties of cartilage. In the present study we used AFM to quantify these changes in relation to the specific spatial pattern changes of the chondrocytes. With each pattern change, significant changes in elasticity were observed for both the PCM and ECM. Measuring the local elasticity thus allows for drawing direct conclusions about the degree of local tissue degeneration in OA.

We present a method to investigate early osteoarthritic changes at the cellular level in articular cartilage by using atomic force microscopy (AFM).

原子力显微镜在早期骨关节炎检测中的应用

Biomechanical properties of cells and tissues not only regulate their shape and function but are also crucial for maintaining their vitality. Changes in ...

生物工程

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 ...

<meta charset="utf8"></head><body>使用外植体模型对骨形成进行多模态分析

Comprehensive Characterization of Tissue Mineralization in an Ex Vivo Model

The extensive characterization of tissue mineralization in the context of bone regeneration represents a significant challenge, given the numerous modalities that are currently available for analysis. Here, we propose a workflow for a comprehensive evaluation of new bone formation using a relevant large animal osseous ex vivo explant. A bone defect (diameter = 3.75 mm; depth = 5.0 mm) is created in an explanted sheep femoral head and injected with a macroporous bone substitute loaded with a pro-osteogenic growth factor (bone morphogenetic protein 2 - BMP2). Subsequently, the explant is maintained in culture for a 28-day period, allowing cellular colonization and subsequent bone formation. To evaluate the quality and structure of newly mineralized tissue, the following successive methods are set up: (i) Characterization and high-resolution 3D images of the entire explant using micro-CT, followed by deep learning image analyses to enhance the discrimination of mineralized tissues; (ii) Nano-indentation to determine the mechanical properties of the newly formed tissue; (iii) Histological examinations, such as Hematoxylin/Eosin/Saffron (HES), Goldner's trichrome, and Movat's pentachrome to provide a qualitative assessment of mineralized tissue, particularly with regard to the visualization of the osteoid barrier and the presence of bone cells; (iv) Back-scattering scanning electron microscopy (SEM) mapping with internal reference to quantify the degree of mineralization and provide detailed insights into surface morphology, mineral composition, and bone-biomaterial interface; (v) Raman spectroscopy to characterize the molecular composition of the mineralized matrix and to provide insights into the persistence of BMP2 within the cement through the detection of peptide bonds. This multimodal analysis will provide an effective assessment of newly formed bone and comprehensive qualitative and quantitative insights into mineralized tissues. Through the standardization of these protocols, we aim to facilitate interstudy comparisons and improve the validity and reliability of research findings.

<meta charset="utf8"></head><body>作者聚焦:在骨再生研究中表征组织矿化的高级技术

The proposed protocol entails a global approach to assess bone formation in the context of bone regeneration using multimodal analyses. It aims to provide qualitative and quantitative information on new bone formation, enhancing the rigor and validity of basic and pre-clinical investigations.

在离体模型中全面表征组织矿化

The extensive characterization of tissue mineralization in the context of bone regeneration represents a significant challenge, given the numerous ...

Destabilization of the Medial Meniscus and Cartilage Scratch Murine Model of Accelerated Osteoarthritis

Osteoarthritis is the most prevalent musculoskeletal disease in people over 45, leading to an increasing economic and societal cost. Animal models are used to mimic many aspects of the disease. The present protocol describes the destabilization and cartilage scratch model (DCS) of post-traumatic osteoarthritis. Based on the widely used destabilization of the medial meniscus (DMM) model, DCS introduces three scratches on the cartilage surface. The current article highlights the steps to destabilize the knee by transecting the medial meniscotibial ligament followed by three intentional superficial scratches on the articular cartilage. The possible analysis methods by dynamic weight-bearing, microcomputed tomography, and histology are also demonstrated. While the DCS model is not recommended for studies that focus on the effect of osteoarthritis on the cartilage, it enables the study of osteoarthritis development in a shorter time window, with special focus on (1) osteophyte formation, (2) osteoarthritic and injury pain, and (3) the effect of cartilage damage in the whole joint.

The present protocol describes the controlled microblade scratches on the surface of the articular cartilage after destabilizing the mouse knee by cutting the medial miniscotibial ligament. This animal model presents an accelerated form of osteoarthritis (OA) suitable for studying osteophyte formation, osteosclerosis, and early-stage pain.

加速性骨关节炎内侧半月板和软骨划痕鼠模型的不稳定

Osteoarthritis is the most prevalent musculoskeletal disease in people over 45, leading to an increasing economic and societal cost. Animal models are ...

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

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修复的临界大小的颅骨缺损模型脂肪间充质干细胞的收获Lipoaspirate

外侧 Transfibular 法治疗踝关节骨性关节炎

基质辅助重塑和修复软骨缺损的自体软骨细胞移植在兔模型

异基因骨髓间充质干细胞植入兔子的膝关节软骨缺损治疗纤维蛋白凝块

原子力显微镜在早期骨关节炎检测中的应用

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

在离体模型中全面表征组织矿化

在离体模型中全面表征组织矿化

在离体模型中全面表征组织矿化

加速性骨关节炎内侧半月板和软骨划痕鼠模型的不稳定