部分支持这项研究是由NIH资助1RO1DK74566和1RO1AM52984。笔者想感谢她的帮助杰奎琳·麦卡锡女士在手术过程中，在RRC的DRS。剑谱王和Harshini的萨罗吉尼为帮助他们在一些兔子，荣丸为她组织学工作，博士和她的高效液相色谱法测量高能磷酸盐的李明女士的手术和术后护理。

1。解剖学基础兔耳伤口护理的最惠国待遇研究材料，自20世纪初。耳提供三个维管束（中央，颅，尾）;中央束是最大的，而大小不等的船只在头部和尾部束。偶尔，尾动脉可能缺席。1所有这些船只很容易确定背侧后，耳朵被剃（ 图1）。远端皮肤上腹部，不接受从近端血管的血液供应。背耳上的三个血管束有许多分支，穿透软骨提供血液供应，远端腹侧皮肤（ 图2）。在一些兔子，一个非常小的皮下血管也发生在腹侧方，供应血液不超过1/3的腹侧皮肤。在一个开放的环状皮肤切口，有埃森tially从腹部切口无出血。 2。外科手术<ol><li>兔子称重，随着颈后，耳朵被剃光。 </li><li>丁丙诺啡（0.01毫克/千克）的术前给出IM手术前一小时，。 </li><li>麻醉诱导氯胺酮（50毫克/公斤）和甲苯噻嗪（5毫克/千克）IM的混合物。 </li><li>兔子在手术过程中，用加热垫保护眼睛的保护与软膏，并在手术过程中监测其生命体征。 </li><li>无菌手术区域准备与手术擦洗，并披上使两个耳朵暴露。 </li><li>一只耳朵呈现缺血，而另一只耳朵配对的非缺血性控制供应。 </li><li>由动脉和静脉血管蒂确定。 </li><li>使用15号刀片，三个小（1-1.5厘米）垂直切口上的血管蒂，从耳朵基地约1-2厘米。 </li><li>中央静脉从周围组织分离，缝合用于包围它的回缩。 4-0缝线结扎中央动脉和分歧。 </li><li>随行的中央神经也被削减。 </li><li>整个颅蒂结扎和划分。 </li><li>尾蒂被保存下来。 </li><li>通过3个切口皮下隧道。这是通过与钝性剥离钳和用刀子和剪刀的锐利解剖。所有的皮下组织，肌肉，神经，和小血管分支已停产。下方的真皮小vasculatures完全分歧。所有的皮下组织切割刀片软骨水平。 </li><li>进一步尖锐和生硬的夹层是用来清理的基础，留下裸露的软骨关于5-10毫米宽。 </li><li>此过程中断所有的皮下血管分支，以防止他们从迅速建立抵押品。 </li><li>出血最小的。如果有任何，它很容易被控制的纱布压缩，而不需要为电灼。 </li><li>耳内的皮肤不携带大量的血液供应，从而不切开。 </li><li>使用4-0或5-0缝合皮肤切口封闭，并用无菌纱布覆盖。 </li><li> 3小的皮肤切口，并会导致皮肤损伤远远低于开放手术过程的结果。 </li><li>四个圆形全层皮肤伤口上创建的每个耳腹面，用6毫米的不锈钢冲。伤口之间的距离至少有20-30毫米。软骨内冲伤口的皮肤被删除。 ，与皮肤或单独的骨膜也将被删除。造粒和上皮发生的基础是软骨，但软骨是不穿孔（ 图3）。 </li><li>当两个耳朵上的伤口已经完成，测试药物被应用到两个伤口的一侧，而对照药物是APPL灭蝇灯对对方的伤口。一个闭塞敷料（TegaDerm，3M）是用来覆盖伤口。这可防止伤口变得干燥。 </li><li>在底部的耳朵的伤口通常比伤口愈合得更快，在耳朵上方。建议对伤口愈合的对比研究，比较一方的两个伤口，另一侧的两个伤口，或单独比较的顶部和底部的伤口。 </li></ol> 3。术后护理<ol><li>一个芬太尼透皮贴剂的补丁连接到背部皮肤释放芬太尼（25微克/小时）为2-3天，以减少可能出现的疼痛。 </li><li>兔子被允许自由进入食品和饮料。 </li><li>每天进行换药。拆除旧敷料，并用无菌棉签清洗伤口。数码照片进行比较。应用新的敷料，伤口再次由TegaDerm覆盖。没有麻醉需要在更衣室通道安格斯。 </li><li>皮肤温度测量每日每只耳朵上进行比较。 </li><li>与经皮氧张力仪或激光多普勒流量计可以测量额外的耳朵的血液循环。 </li><li>一周后缝线可能会被删除。 </li><li>对于非糖尿病的动物，最通常的耳朵伤口愈合2-3个星期内。 </li></ol> 4。潜在的并发症，以及如何避免这种微创手术是安全的，并发症是罕见的。除了一个过渡性的术后轻度耳肿胀，没有我们的兔子有任何严重的手术并发症。然而，也有一些潜在的并发症，此过程: <ol><li>出血。出血可能发生在切口止血是不完整的或损坏，如果一个较大的血管（如主要蒂船只之一）。小心剥离，这是很容易避免。除了三大维管束，有重新皮下许多小血管分支。皮下隧道时，这些船只都被切断，但没有搭售是必要的。可以实现简单止血纱布按压。插入几分钟隧道纱布的一角，这是很容易做到。关闭切口前，确保无活动性出血存在。 </li><li>如果被污染的切口，切口感染，裂开:这些并发症可能发生。这可避免使用严格的无菌技术和无抗生素是必要的。然而，如果感染发生，适当引流和换药是必要的。 </li><li>耳水肿:由于血管结扎及淋巴循环中断，可能会出现轻度耳肿胀几天。然而，它本身消失，没有治疗是必要的。 </li></ol> 5。代表结果自2007年以来，我们在此过程中超过70只，其中包括23糖尿病1imals。在糖尿病组中，手术是在不同的糖尿病时间:7兔有两个星期，对糖尿病有四个，其中有两个月，十有12个月，2例糖尿病时间24个月。 后立即手术，缺血性耳出现凉爽和紫绀，轻度水肿。最重要的耳动脉中央动脉有一个强脉冲在正常的耳朵，但这种脉冲是在缺血性耳不存在。耳运动减少，但并未完全消失。皮肤表明明确的洼地，在隧道发现任何动物，但没有缺血或坏死（ 图4）。 正常和缺血耳朵的皮肤温度测量与数字温度计（欧姆龙保健，班诺克本，白细胞介素，或一个K型红外点温度计​​尔科技，宾夕法尼亚州匹兹堡欧姆龙）。非缺血性和缺血性耳之间的温度差异范围从1到10℃，W2-6°C。 图范围内的第i个最兔下降。 5显示了30只兔的平均气温差异。温差在初期较高，手术后15天之后逐渐下降，但仍维持在一个月结束。 在一些动物处死后，他们组织高能磷酸含量测定。 6个组织样本取自每个耳朵和高能量的磷酸盐含量测定高效液相色谱法。 ATP和总能量浓度较高，在正常的耳朵比缺血耳朵图。 6显示了一个例子在手术后23天牺牲一只兔子。平均组织ATP含量分别为0.349±0.047μmol/克的（平均值±SD）在正常耳与0.237±0.059μmol/克的缺血耳（P &lt;0.005）。平均总能量（ATP + ADP + AMP）的为（0.882±0.137）μmol/克的正常耳与0.556±0.115μmol/克的"ische<html>麦克风耳（P &lt;0.001）。 伤口愈合时间永远不再为缺血性耳比在各组的非缺血性耳（非糖尿病或糖尿病）。 图。 7显示了对缺血性和非缺血性耳朵之间的26对伤口愈合的比较。愈合时间为13至18天（平均14.9±1.6天），与17至27天的正常耳（平均20.5±3.4天）对缺血性耳（P = 0.001）。老年人和糖尿病兔伤口愈合时间更长，尤其是在糖尿病的时间超过12个月。 在这些兔子，所有的切口愈合，无任何并发症通常包括与糖尿病时间24个月的兔子。 图8是一个组织学样品展示愈合的皮下隧道，在肌肉的缺席而被纤维组织取代。然而，皮肤完好。 / 3341fig1.jpg"/&gt; 图1。兔耳提供三个维管束。中央是最大的，颅较小，尾包是最小的。这些船只很容易看到表面上被剃​​得耳后。 <img alt="图2" src="/files/ftp_upload/3341/3341fig2.jpg" /> 图2。背耳供血不仅要背的皮肤，而且还通过穿透血管的腹部皮肤上的三个血管束。箭头穿透血管通过软骨。插图显示越过这些船只的部分。 <img alt="图3" src="/files/ftp_upload/3341/3341fig3.jpg" /> 图3。四个6毫米的伤口上创建腹耳侧，骨膜也将被删除。造粒和上皮发生的基础是软骨，但软骨是不穿孔。 3 SMaL公司<html>升皮肤切口封闭，用很少的皮肤破坏。 <img alt="图4" src="/files/ftp_upload/3341/3341fig4.jpg" /> 图4。关闭后皮肤切口，皮下隧道缩进清楚地看到。 <img alt="图5" src="/files/ftp_upload/3341/3341fig5.jpg" /> 图5。正常和缺血性之间的30只兔耳皮肤的平均温度差。早期的差异范围从0°C至10°C后手术。差异逐渐减小，但仍维持在一个月结束。 <img alt="图6" src="/files/ftp_upload/3341/3341fig6.jpg" /> 图6。耳组织高能磷酸含量减少缺血耳相比，在手术后23天（P &lt;0.001），非缺血性耳。 41/3341fig7.jpg"/&gt; 图7。创面愈合时间较长比非缺血性耳）（26伤口缺血耳朵。愈合被定义为诚信恢复到受伤的组织，即完整的表皮细胞再生，伤口网站。 <img alt="图8" src="/files/ftp_upload/3341/3341fig8.jpg" /> 图8。组织学样本显示含有皮下隧道的耳朵部分。愈合组织比正常耳薄，原有的骨骼肌是缺席，由纤维组织所取代。然而，皮肤完好。

<ol>
	<li>Ninomiya, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+vascular+bed+in+the+rabbit+ear:+microangiography+and+scanning+electron+microscopy+of+vascular+corrosion+casts.">The vascular bed in the rabbit ear: microangiography and scanning electron microscopy of vascular corrosion casts.</a> Anat. Histol. Embryol. 29, 301-305 (2000).</li><li>Sen, C. K., Gordillo, G. M., Roy, S., Kirsner, R., Lambert, L., Hunt, T. K., Gottrup, F., Gurtner, G. C., Longaker, M. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+skin+wounds:+a+major+and+snowballing+threat+to+public+health+and+the+economy.">Human skin wounds: a major and snowballing threat to public health and the economy.</a> Wound Repair. 17, 763-771 (2009).</li><li>Boulton, A. J., Vileikyte, L., Ragnarson-Tennvall, G., Apelqvist, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+global+burden+of+diabetic+foot+disease.">The global burden of diabetic foot disease.</a> Lancet. 366, 1719-1724 (2005).</li><li>Edwards, J., Stapley, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Debridement+of+diabetic+foot+ulcers.">Debridement of diabetic foot ulcers.</a> Cochrane Database Syst. Rev. 1, 1-40 (2010).</li><li>Werdin, F., Tennenhaus, M., Schaller, H. E., Rennekampff, H. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evidence-based+Management+Strategies+for+Treatment+of+Chronic+Wounds.">Evidence-based Management Strategies for Treatment of Chronic Wounds.</a> Eplasty. 9, 169-179 (2009).</li><li>Wu, Y., Chen, L., Scott, P. G., Tredget, E. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesenchymal+stem+cells+enhance+wound+healing+through+differentiation+and+angiogenesis.">Mesenchymal stem cells enhance wound healing through differentiation and angiogenesis.</a> Stem Cells. 25, 2648-2659 (2007).</li><li>Bennett, J. P., Matthews, R., Faulk, W. P. <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+chronic+ulceration+of+the+legs+with+human+amnion.">Treatment of chronic ulceration of the legs with human amnion.</a> Lancet. 1, 1153-1156 (1980).</li><li>Gottrup, F., Agren, M. S., Karlsmark, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Models+for+use+in+wound+healing+research:+a+survey+focusing+on+in+vitro+and+in+vivo+adult+soft+tissue.">Models for use in wound healing research: a survey focusing on in vitro and in vivo adult soft tissue.</a> Wound Repair. 8, 83-96 (2000).</li><li>Lindblad, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+in+wound+healing+research:+do+we+need+more.">Animal models in wound healing research: do we need more.</a> Wound Repair. 8, 81-82 (2000).</li><li>LoGerfo, F. W., Gibbons, G. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vascular+disease+of+the+lower+extremities+in+diabetes+mellitus.">Vascular disease of the lower extremities in diabetes mellitus.</a> Endocrinol. Metab. Clin. North Am. 25, 439-445 (1996).</li><li>Gottrup, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oxygen,+wound+healing+and+the+development+of+infection.+Present+status.">Oxygen, wound healing and the development of infection. Present status.</a> Eur. J. Surg. 168, 260-263 (2002).</li><li>Niinikoski, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+blood+and+oxygen+supply+on+the+biochemistry+of+repair.">The effect of blood and oxygen supply on the biochemistry of repair.</a> , 56-71 (2002).</li><li>Schaffer, M., Witte, M., Becker, H. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Models+to+study+ischemia+in+chronic+wounds.">Models to study ischemia in chronic wounds.</a> Int. J. Low Extrem. Wounds. 1, 104-111 (2002).</li><li>Niinikoski, J., Gottrup, F., Hunt, T. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+oxygen+in+wound+repair.">The role of oxygen in wound repair.</a> , 165-174 (1991).</li><li>Harding, K. G., Morris, H. L., Patel, G. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Science,+medicine+and+the+future:+healing+chronic+wounds.">Science, medicine and the future: healing chronic wounds.</a> BMJ. 324, 160-163 (2002).</li><li>Lindblad, W. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Editorial:+How+should+one+study+wound+healing.">Editorial: How should one study wound healing.</a> Wound Repair. 14, 515-515 (2007).</li><li>Chien, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ischemic+rabbit+ear+model+created+by+minimally+invasive+surgery.">Ischemic rabbit ear model created by minimally invasive surgery.</a> Wound Repair Regen. 15, 928-935 (2007).</li><li>Joseph, J., Townsend, F. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+healing+of+defects+in+immobile+skin+in+rabbits.">The healing of defects in immobile skin in rabbits.</a> Br. J. Surg. 48, 557-564 (1961).</li><li>James, G. A., Swogger, E., Wolcott, R., Pulcini, E., Secor, P., Sestrich, J., Costerton, J. W., Stewart, P. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biofilms+in+chronic+wounds.">Biofilms in chronic wounds.</a> Wound Repair Regen. 16, 37-44 (2008).</li><li>Niitsuma, J., Yano, H., Togawa, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Experimental+study+of+decubitus+ulcer+formation+in+the+rabbit+ear+lobe.">Experimental study of decubitus ulcer formation in the rabbit ear lobe.</a> J. Rehabil. Res. Dev. 40, (2003).</li><li>Salcido, R., Popescu, A., Ahn, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Animal+models+in+pressure+ulcer+research.">Animal models in pressure ulcer research.</a> J. Spinal Cord Med. 30, 107-116 (2007).</li></ol>

没有参与这项研究的利息的金融冲突。

糖尿病的影响，在美国，2,3 23.6万人，有15-20％的患者发展足部溃疡，在其一生中，4,5在每年的治疗费用超过25亿美元。3,6然而，即使是最好的治疗愈合率只有50％，并实现这往往是暂时的，复发率高达66％的糖尿病患者7。3,6当溃疡发生在糖尿病患者的主要并发症，如感染和截肢的大幅度增加。尽管近一个世纪广泛的研究和发展成千上万的敷料产品，尚没有证明是有...

在手术过程中的监测<ol><li>一个加热垫，以保持正常的温度。 </li><li>经皮血氧监视器。 </li><li>温度显示器。 </li></ol>手术<ol><li> ＃3＃15刀片与刀。 </li><li>两个双钳（对皮肤和其他软组织的另一个）。 </li><li> 2-3对小夹子。 </li><li>对一个或两个微型剪刀（用于组织切割）。 </li><li>一对小剪刀（用于切割缝合）。 </li><li>一个持针器。 </li><li>一些不锈钢拳（伤口）。 </li></ol>

a simplified technique for producing an ischemic wound model

目前糖尿病伤口研究中的一个主要障碍是缺乏缺血性损伤模型，可以安全地用于糖尿病动物。工作在非缺血性伤口的药物可能无法正常工作，在人类糖尿病患者的伤口，因为血管病变是一个主要因素，阻碍了这些伤口的愈合。我们在2007年发表的文章描述了一种微创外科技术创造一只兔子耳缺血性损伤模型。从那时起，我们已进一步简化，易于操作的过程。一只耳朵上，提出了三个小的皮肤切口上的血管蒂，从耳根1-2厘米。中央动脉结扎和切断神经。整个颅捆绑切断结扎，留下完整的只有尾分行。圆周皮下隧道，通过切口，以减少皮下组织，肌肉，神经，和小型船只。另一只耳朵被用来作为非缺血性的控制。腹侧方Ø四个伤口F各耳。这种技术产生配对比较4缺血性伤口和4个非缺血性伤口在一个动物。手术后，缺血耳朵是凉爽和紫绀，呈减少运动和耳动脉中的脉冲缺乏。缺血耳的皮肤温度为1-10°C下比正常耳，这种差异维持一个月以上。在缺血性比对照耳耳耳组织高能磷酸含量较低。伤口愈合时间长于在缺血性耳在非缺血性耳时使用了相同的待遇。该技术目前已超过80只，其中23例糖尿病（糖尿病的时间从2个星期至2年不等）。没有一个单一的兔子已开发的任何手术并发症，如出血，感染，或在皮肤切口破裂。该模型具有许多优点，如小皮中断，缺血时间较长，成功率较高，许多其他车型相比。它长约n是安全地使用在动物抵抗力降低，也可以修改，以满足不同的测试要求。

Watch this Scientific Journal Video about A Simplified Technique for Producing an Ischemic Wound Model at JoVE.com

A Simplified Technique for Producing an Ischemic Wound Model

我们已经开发出一种微创技术，创建除以中央动脉和神经，颅神经血管束兔子缺血耳朵伤口模型。皮下隧道，然后切开皮下组织。此过程会导致皮肤的破坏最小，可以安全地用于糖尿病动物。

一个生产缺血性损伤模型的简化技术

One major obstacle in current diabetic wound research is a lack of an ischemic wound model that can be safely used in diabetic animals. Drugs that work ...

a-simplified-technique-for-producing-an-ischemic-wound-model

Universidad de Cartagena UDEC

Research

JoVE Journal

Medicine

17.2K 次观看.  University of Louisville. 我们已经开发出一种微创技术，创建除以中央动脉和神经，颅神经血管束兔子缺血耳朵伤口模型。皮下隧道，然后切开皮下组织。此过程会导致皮肤的破坏最小，可以安全地用于糖尿病动物。

视频: A Simplified Technique for Producing an Ischemic Wound Model

Gene Transfer for Ischemic Heart Failure in a Preclinical Model

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine their efficacy and safety.
Gene therapy using viral vectors is one of the most promising approaches for treating cardiac diseases. Viral delivery of various different genes by changing the carrier gene has immeasurable therapeutic potential.
In this video, the full process of an animal model of heart failure creation followed by gene transfer is presented using a swine model. First, myocardial infarction is created by occluding the proximal left anterior descending coronary artery. Heart remodeling results in chronic heart failure. Unique to our model is a fairly large scar which truly reflects patients with severe heart failure who require aggressive therapy for positive outcomes. After myocardial infarct creation and development of scar tissue, an intracoronary injection of virus is demonstrated with simultaneous nitroglycerine infusion. Our injection method provides simple and efficient gene transfer with enhanced gene expression. This combination of a myocardial infarct swine model with intracoronary virus delivery has proven to be a consistent and reproducible methodology, which helps not only to test the effect of individual gene, but also compare the efficacy of many genes as therapeutic candidates.

A method of gene transfer for the treatment of ischemic heart failure is described using a swine model of myocardial infarction. Our simple and reproducible method enables us to readily evaluate the efficacy of various gene transfers with a very simple and reproducible way.

缺血性心力衰竭在临床前模型的转基因

Various emerging technologies are being developed for patients with heart failure. Well-established preclinical evaluations are necessary to determine ...

科研

医学

Murine Model of Wound Healing

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. Impaired wound healing, which occurs in diabetic patients for example, can lead to severe unfavorable outcomes such as amputation. There is, therefore, an increasing impetus to develop novel agents that promote wound repair. The testing of these has been limited to large animal models such as swine, which are often impractical. Mice represent the ideal preclinical model, as they are economical and amenable to genetic manipulation, which allows for mechanistic investigation. However, wound healing in a mouse is fundamentally different to that of humans as it primarily occurs via contraction. Our murine model overcomes this by incorporating a splint around the wound. By splinting the wound, the repair process is then dependent on epithelialization, cellular proliferation and angiogenesis, which closely mirror the biological processes of human wound healing. Whilst requiring consistency and care, this murine model does not involve complicated surgical techniques and allows for the robust testing of promising agents that may, for example, promote angiogenesis or inhibit inflammation. Furthermore, each mouse acts as its own control as two wounds are prepared, enabling the application of both the test compound and the vehicle control on the same animal. In conclusion, we demonstrate a practical, easy-to-learn, and robust model of wound healing, which is comparable to that of humans.

A murine model of cutaneous wound healing that can be used to assess therapeutic compounds in physiological and pathophysiological settings.

伤口愈合的小鼠模型

Wound healing and repair are the most complex biological processes that occur in human life. After injury, multiple biological pathways become activated. ...

Remote Limb Ischemic Preconditioning:  A Neuroprotective Technique in Rodents

Sublethal ischemia protects tissues against subsequent, more severe ischemia through the upregulation of endogenous mechanisms in the affected tissue. Sublethal ischemia has also been shown to upregulate protective mechanisms in remote tissues. A brief period of ischemia (5-10 min) in the hind limb of mammals induces self-protective responses in the brain, lung, heart and retina. The effect is known as remote ischemic preconditioning (RIP). It is a therapeutically promising way of protecting vital organs, and is already under clinical trials for heart and brain injuries. This publication demonstrates a controlled, minimally invasive method of making a limb &#8211; specifically the hind limb of a rat &#8211; ischemic. A blood pressure cuff developed for use in human neonates is connected to a manual sphygmomanometer and used to apply 160 mmHg pressure around the upper part of the hind limb. A probe designed to detect skin temperature is used to verify the ischemia, by recording the drop in skin temperature caused by pressure-induced occlusion of the leg arteries, and the rise in temperature which follows release of the cuff. This method of RIP affords protection to the rat retina against bright light-induced damage and degeneration.

Remote ischemic preconditioning (RIP) is a method of conditioning tissues against damaging stress. We have established a method of remote ischemia at the hind limb, by inflating a sphygmomanometer cuff for 5-10 min. The neuroprotective capabilities of RIP have been demonstrated in a model of retinal degeneration in rodents.

远程肢体缺血预适应:在啮齿动物的保护技术

Sublethal ischemia protects tissues against subsequent, more severe ischemia through the upregulation of endogenous mechanisms in the affected tissue. ...

A Mouse Fetal Skin Model of Scarless Wound Repair

Early in utero, but not in postnatal life, cutaneous wounds undergo regeneration and heal without formation of a scar. Scarless fetal wound healing occurs across species but is age dependent. The transition from a scarless to scarring phenotype occurs in the third trimester of pregnancy in humans and around embryonic day 18 (E18) in mice. However, this varies with the size of the wound with larger defects generating a scar at an earlier gestational age. The emergence of lineage tracing and other genetic tools in the mouse has opened promising new avenues for investigation of fetal scarless wound healing. However, given the inherently high rates of morbidity and premature uterine contraction associated with fetal surgery, investigations of fetal scarless wound healing in vivo require a precise and reproducible surgical model. Here we detail a reliable model of fetal scarless wound healing in the dorsum of E16.5 (scarless) and E18.5 (scarring) mouse embryos.

During mammalian development, early gestational skin wounds heal without a scar. Here we detail a reliable and reproducible model of fetal scarless wound healing in the cutaneous dorsum of E16.5 (scarless) and E18.5 (scarring) mouse embryos.

对无瘢痕修复鼠标胎儿皮肤模型

Early in utero, but not in postnatal life, cutaneous wounds undergo regeneration and heal without formation of a scar. Scarless fetal wound healing occurs ...

A Piglet Model of Neonatal Hypoxic-Ischemic Encephalopathy

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the world&#8217;s 2.9 million neonatal deaths. Animal models of HIE have contributed to the understanding of the pathophysiology in HIE, and have highlighted the dynamic process that occur in brain injury due to perinatal asphyxia. Thus, animal studies have suggested a time-window for post-insult treatment strategies. Hypothermia has been tested as a treatment for HIE in pdiglet models and subsequently proven effective in clinical trials. Variations of the model have been applied in the study of adjunctive neuroprotective methods and piglet studies of xenon and melatonin have led to clinical phase I and II trials1,2. The piglet HIE model is further used for neonatal resuscitation- and hemodynamic studies as well as in investigations of cerebral hypoxia on a cellular level. However, it is a technically challenging model and variations in the protocol may result in either too mild or too severe brain injury. In this article, we demonstrate the technical procedures necessary for establishing a stable piglet model of neonatal HIE. First, the newborn piglet (&#60; 24 hr old, median weight 1500 g) is anesthetized, intubated, and monitored in a setup comparable to that found in a neonatal intensive care unit. Global hypoxia-ischemia is induced by lowering the inspiratory oxygen fraction to achieve global hypoxia, ischemia through hypotension and a flat trace amplitude integrated EEG (aEEG) indicative of cerebral hypoxia. Survival is promoted by adjusting oxygenation according to the aEEG response and blood pressure. Brain injury is quantified by histopathology and magnetic resonance imaging after 72 hr.

Hypoxic-ischemic encephalopathy following perinatal asphyxia can be studied using animal models. We demonstrate the procedures necessary for establishing a piglet model of neonatal hypoxic-ischemic encephalopathy.

新生儿缺氧缺血性脑病的小猪模型

Birth asphyxia, which causes hypoxic-ischemic encephalopathy (HIE), accounts for 0.66 million deaths worldwide each year, about a quarter of the ...

Demonstration of the Rat Ischemic Skin Wound Model

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, and unrelieved pressure due to immobility. Animal models have been developed that attempt to mimic these conditions for the purpose of furthering our understanding of the complexity of chronic wounds. The model described herein is a rat ischemic skin flap model that permits a prolonged reduction of blood flow resulting in wounds that become ischemic and resemble a chronic wound phenotype (reduced vascularization, increased inflammation and delayed wound closure). It consists of a bipedicled dorsal flap with 2 ischemic wounds placed centrally and 2 non-ischemic wounds lateral to the flap as controls. A novel addition to this ischemic skin flap model is the placement of a silicone sheet beneath the flap that functions as a barrier and a splint to prevent revascularization and reduce contraction as the wounds heal. Despite the debate of using rats for wound healing studies due to their quite distinct anatomic and physiologic differences compared to humans (i.e., the presence of a panniculus carnosus muscle, short life-span, increased number of hair follicles, and their ability to heal infected wounds) the modifications employed in this model make it a valuable alternative to previously developed ischemic skin flap models.

The rat, due to its size, availability, and rather docile behavior, has been utilized as a research model for many years. The goal of this protocol is to utilize the rat as an ischemic skin wound healing model to provide valuable insight into the pathophysiology of chronic wounds.

在大鼠缺血性皮肤创伤模型的演示

The propensity for chronic wounds in humans increases with ageing, disease conditions such as diabetes and impaired cardiovascular function, and ...

Murine Left Anterior Descending (LAD) Coronary Artery Ligation: An Improved and Simplified Model for Myocardial Infarction

Ischemic heart disease (IHD), or acute coronary syndrome (ACS), is one of the leading causes of death in the United States. IHD is characterized by reduced blood supply to the heart, resulting in the loss of oxygen to and the ensuing necrosis of the heart muscle. The MI model has gained popularity for its use as a short-term ischemia-reperfusion model and a long-term permanent ligation model. Below, we describe a reliable method for the permanent ligation of the LAD. With mouse genetic engineering technology becoming more advanced, and with an increasing availability of quality murine surgical instruments, the mouse has become a popular model for MI surgeries. Our surgical model incorporates the use of an easily reversible anesthetic for the rapid recovery of the mouse; a minimally invasive endotracheal intubation without involving a tracheotomy; and a thoracentesis through the original thoracotomy site without creating an additional incision in the chest, as is done in some other methods, to effectively remove excess blood and air from the chest cavity. This method is comparatively less invasive than other methods, which dramatically reduces surgical and post-surgical complications and mortality and improves reproducibility.

Murine Left Anterior Descending LAD Coronary Artery Ligation: An Improved and Simplified Model for Myocardial Infarction

We provide a reliable method for left anterior descending artery (LAD) ligation in a mouse model. This method is comparatively less invasive than other methods, involving endotracheal intubation, a left-sided thoracotomy approach, and thoracentesis. This method can be used as a model for both acute and chronic myocardial infarction (MI).

鼠左前降支（LAD）冠状动脉结扎:改进和简化模型的心肌梗死

Ischemic heart disease (IHD), or acute coronary syndrome (ACS), is one of the leading causes of death in the United States. IHD is characterized by ...

Corneal Epithelial Abrasion with Ocular Burr As a Model for Cornea Wound Healing

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to injury. In fact, the most common type of eye injury found in clinic is a corneal abrasion. Here, we utilize an ocular burr to induce an abrasion resulting in removal of the corneal epithelium in vivo on anesthetized mice. This method allows for targeted and reproducible epithelial disruption, leaving other areas intact. In addition, we describe the visualization of the abraded epithelium with fluorescein staining and provide concrete advice on how to visualize the abraded cornea. Then, we follow the timeline of wound healing 0, 18, and 72 h after abrasion, until the wound is re-epithelialized. The epithelial abrasion model of corneal injury is ideal for studies on epithelial cell proliferation, migration and re-epithelialization of the corneal layers. However, this method is not optimal to study stromal activation during wound healing, because the ocular burr does not penetrate to the stromal cell layers. This method is also suitable for clinical applications, for example, pre-clinical test of drug effectiveness.

This protocol describes a method to inflict an abrasion to the ocular surface of the mouse, and to follow the wound healing process thereafter. The protocol takes advantage of an ocular burr to partially remove the surface epithelium of the eye in anaesthetized mice.

眼毛刺角膜上皮磨损与角膜创面愈合的模型研究

The murine cornea provides an excellent model to study wound healing. The cornea is the outermost layer of the eye, and thus is the first defense to ...

An Intact Pericardium Ischemic Rodent Model

This protocol has shown that the pericardium and its contents play an essential anti-fibrotic role in the ischemic rodent model (coronary ligation to induce myocardial injury). The majority of pre-clinical myocardial infarction models require the disruption of pericardial integrity with loss of the homeostatic cellular milieu. However, recently a methodology has been developed by us to induce myocardial infarction, which minimizes pericardial damage and retains the heart&#39;s resident immune cell population. An improved cardiac functional recovery in mice with an intact pericardial space following coronary ligation has been observed. This method provides an opportunity to study inflammatory responses in the pericardial space following myocardial infarction. Further development of the labeling techniques can be combined with this model to understand the fate and function of pericardial immune cells in regulating the inflammatory mechanisms that drive remodeling in the heart, including fibrosis.

This protocol outlines the steps for inducing myocardial infarction in mice while preserving the pericardium and its contents.

完整的心包缺血性啮齿动物模型

This protocol has shown that the pericardium and its contents play an essential anti-fibrotic role in the ischemic rodent model (coronary ligation to ...

A Simplified Model for Heterotopic Heart Valve Transplantation in Rodents

There is an urgent clinical need for heart valve replacements that can grow in children. Heart valve transplantation is proposed as a new type of transplant with the potential to deliver durable heart valves capable of somatic growth with no requirement for anticoagulation. However, the immunobiology of heart valve transplants remains unexplored, highlighting the need for animal models to study this new type of transplant. Previous rat models for heterotopic aortic valve transplantation into the abdominal aorta have been described, though they are technically challenging and costly. For addressing this challenge, a renal subcapsular transplant model was developed in rodents as a practical and more straightforward method for studying heart valve transplant immunobiology. In this model, a single aortic valve leaflet is harvested and inserted into the renal subcapsular space. The kidney is easily accessible, and the transplanted tissue is securely contained in a subcapsular space that is well vascularized and can accommodate a variety of tissue sizes. Furthermore, because a single rat can provide three donor aortic leaflets and a single kidney can provide multiple sites for transplanted tissue, fewer rats are required for a given study. Here, the transplantation technique is described, providing a significant step forward in studying the transplant immunology of heart valve transplantation.

This protocol describes a simple and efficient method for the transplantation of aortic valve leaflets under the renal capsule to allow for the study of alloreactivity of heart valves.