Name: an anesthesia, surgery, and harvest method for the evaluation of transpedicular screws using an in vivo porcine lumbar spine model
Uploaded: 2017-05-31T12:30:00.000+00:00
Duration: 9 min 7 s
Description: Watch this Scientific Journal Video about An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model at JoVE.com

本研究得到了韩国春武国立大学医院生物医学研究所（CNUH-BRI）资助的赠款（CNUH-BRI-2012-02-005）的支持。

春武国立大学体制动物护理与使用委员会批准了本研究。动物的治疗，使用和处理遵循所有指导方针和政策。保持手术室在24°C。 麻醉<ol><li>适应12个月的小型猪在实验单位至少一周。进行临床检查，测量呼吸频率，心率和体温。在麻醉程序前不要喂养每只小型猪12小时。 </li><li>将阿托品（0.05mg / kg）和氯胺酮（20mg / kg）/西拉星（2mg / kg）注射入耳后侧颈部肌肉区域进行预处理。 </li><li>镇静后，将橡皮筋紧贴耳朵底部，用局部酒精清洁耳朵。 </li><li>将一根针头上的塑料导管置于耳静脉中并取出橡皮筋。确认导管正确放置。流感用肝素化生理盐水导管，用胶带固定。 </li><li> 对于气管内气管插管，将微型猪置于胸骨放置位置。在助手的帮助下，用适当的吊索握住猪的下巴，张开嘴巴。 <ol><li>将喉镜尖端插入咽腔，以从软腭中取代会厌。使用喉镜刀片的尖端使声带可见，并在呼气期间将气管内导管推入气管。 </li><li>感觉自由通过空气进行适当的插管，并检查小型猪左右两侧呼吸声的胸部听诊。 </li></ol></li><li>使用10 mL注射器，用空气将气管插管的适当体积的空气填充袖带，并使用胶带将管固定到鼻孔。 </li><li>通过内皮提供2.0％浓度的异氟烷，一种吸入麻醉剂用于长时间麻醉acheal插管。测试角膜和睑板反射以确认麻醉并使用眼睛上的软膏以防止干燥。 </li><li>在麻醉期间至少每5分钟监测心血管系统，呼吸系统和体温，直到微型猪恢复。 </li><li>在脊柱手术前30分钟，缓慢应用30 mg / kg头孢唑啉，第一代头孢菌素抗生素。 </li><li>使用IV管线施用5-10mL / kg / h的温（37℃）盐水以维持体内平衡，并提供50μg/ kg / min的芬太尼以控制疼痛。 </li><li>脊柱手术后，当强烈的吞咽反射明显时，进行拔管。 </li><li>将小型猪带入房间并进行监测，直到从麻醉中恢复。当微型猪充分意识到时，提供食物和水。 </li><li>每天服用3 mg / kg的恩氟沙星抗生素，每天服用4.4 mg / kg卡洛芬，用于控制头痛。 </li><li>莫尼每天去猪，直到缝合线去除。 </li></ol>脊柱手术<ol><li>按照制造商的指导方针，将经椎弓根螺钉和后固定器系统高压灭菌。 </li><li>当猪处于仰卧位时，使用剃须刀将小型猪的背部从中心向左或向右约10厘米处剃去。用聚维酮碘溶液和70％酒精清洁皮肤。 </li><li>使用手术刀从腰椎的第二棘突进行纵向中线切口至第一中位神经嵴。通过皮下组织和筋膜解剖到棘突的尖端。 </li><li>使用Cobb电梯从底层薄片骨膜下降下椎旁肌。解剖沿着棘突和椎板限于小关节。 </li><li>打开入口点的浅层皮层（其略低于L3的哺乳动物进程）到两边的L5），用毛刺或羊毛。 </li><li>将导销插入开放位置，平行于上端板并与棘突成20度角。使用C型臂或便携式的前后/横向X射线定义理想的起点。 </li><li>根据X光片将椎弓根探针插入25 mm。使用椎弓根探测装置通过椎弓根和身体触诊确认完整的骨内轨迹。 </li><li>将六个椎弓根螺钉插入准备的L3至L5的椎弓根，直到螺钉头固定好。将植入物头的侧面开口指向所需的方向，并将水平位置与杆的轨迹对齐。 </li><li>将两根杆分别插入椎弓根螺钉头的两侧。使用通用手柄将套筒和螺母放入椎弓根螺钉头。 </li><li>使用直套扳手松开螺母，并使用反扭矩扳手牢固地拧紧螺母。 </li><li>确认职位of椎弓根螺钉使用便携式的前后/横向X射线。 注意:等待小型猪醒来，检查后肢的步态和运动功能，以确定螺丝是否被严重植入。 </li><li>使用带有抽吸的球泡灌注注射器，用3L生理盐水冲洗手术部位。 </li><li>在手术部位放置硅胶排水管，取出硅胶头。使用1.0公尺可吸收的缝合线关闭椎旁肌和皮下。用2.0公尺不吸收尼龙缝合线缝合皮肤。 </li><li>用聚维酮碘消毒缝合部位，并使用灭菌的纱布和胶带进行敷料。 </li></ol>收获程序<ol><li>术后12周，将耳塞后侧颈部肌肉区域注射赛拉嗪（2mg / kg）和氯胺酮（10mg / kg）进行预处理。 </li><li>镇静后，将15mg / kg KCl直接给予耳静脉导管用于安乐死。 </li><li>在以前的手术瘢痕病变中进行纵向中线切口。解剖软组织和椎旁肌肉。 </li><li>暴露腰椎，椎板，棒，螺母和横向过程的棘突从L3到L5。 </li><li>使用反扭矩扳手和杆拆下螺母。使用振荡锯切割L2-3光盘空间和L5-S1光盘空间。 </li><li>用Cobb电梯和塔镊解剖L3-5脊柱的中部和前部两侧。收获后，如果不能立即测试脊椎，请将其包裹在用盐水浸泡的纱布中并储存在-20°C。 </li></ol>

<ol>
	<li>Greenfield, R. T., Grant, R. E., Bryant, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pedicle+screw+fixation+in+the+management+of+unstable+thoracolumbar+spine+injuries.">Pedicle screw fixation in the management of unstable thoracolumbar spine injuries.</a> Orthop Rev. 21 (6), 701-706 (1992).</li><li>Upasani, V. V., 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=Pedicle+screw+surface+coatings+improve+fixation+in+nonfusion+spinal+constructs.">Pedicle screw surface coatings improve fixation in nonfusion spinal constructs.</a> Spine. 34 (4), 335-343 (2009).</li><li>Halvorson, T. L., Kelley, L. A., Thomas, K. A., Whitecloud, T. S., Cook, S. 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N., 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=Pedicular+fixation+in+the+osteoporotic+spine:+a+pilot+in+vivo+study+on+long-term+ovariectomized+sheep.">Pedicular fixation in the osteoporotic spine: a pilot in vivo study on long-term ovariectomized sheep.</a> J Orthop Res. 20 (6), 1217-1224 (2002).</li><li>Fini, M., 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=Biological+assessment+of+the+bone-screw+interface+after+insertion+of+uncoated+and+hydroxyapatite-coated+pedicular+screws+in+the+osteopenic+sheep.">Biological assessment of the bone-screw interface after insertion of uncoated and hydroxyapatite-coated pedicular screws in the osteopenic sheep.</a> J Biomed Mater Res A. 66 (1), 176-183 (2003).</li><li>Branemark, R., Ohrnell, L. O., Skalak, R., Carlsson, L., Branemark, P. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Biomechanical+characterization+of+osseointegration:+an+experimental+in+vivo+investigation+in+the+beagle+dog.">Biomechanical characterization of osseointegration: an experimental in vivo investigation in the beagle dog.</a> J Orthop Res. 16 (1), 61-69 (1998).</li><li>McLain, R. F., Yerby, S. A., Moseley, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+morphometry+of+L4+vertebrae:+comparison+of+large+animal+models+for+the+human+lumbar+spine.">Comparative morphometry of L4 vertebrae: comparison of large animal models for the human lumbar spine.</a> Spine. 27 (8), E200-E206 (2002).</li><li>Giavaresi, G., 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=In+vivo+preclinical+evaluation+of+the+influence+of+osteoporosis+on+the+anchorage+of+different+pedicle+screw+designs.">In vivo preclinical evaluation of the influence of osteoporosis on the anchorage of different pedicle screw designs.</a> Eur Spine J. 20 (8), 1289-1296 (2011).</li><li>Hasegawa, T., 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=Hydroxyapatite-coating+of+pedicle+screws+improves+resistance+against+pull-out+force+in+the+osteoporotic+canine+lumbar+spine+model:+a+pilot+study.">Hydroxyapatite-coating of pedicle screws improves resistance against pull-out force in the osteoporotic canine lumbar spine model: a pilot study.</a> Spine J. 5 (3), 239-243 (2005).</li><li>Smorgick, Y., 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=Single-+versus+multilevel+fusion+for+single-level+degenerative+spondylolisthesis+and+multilevel+lumbar+stenosis:+four-year+results+of+the+spine+patient+outcomes+research+trial.">Single- versus multilevel fusion for single-level degenerative spondylolisthesis and multilevel lumbar stenosis: four-year results of the spine patient outcomes research trial.</a> Spine. 38 (10), 797-805 (2013).</li><li>Busscher, I., Ploegmakers, J. 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经脊椎螺钉在猪脊柱中的评估需要很多时间和精力。首先，小型猪是大型动物。对于动物护理和麻醉，研究人员需要一个专门的协议。第二，手术应保持与人体手术相似的环境。评估脊椎椎弓根螺钉的目的是开发一种可应用于人类的高效螺钉。第三，评估经椎弓根螺钉的长期稳定性需要在脊柱手术后大约三个月。因此，椎弓根螺钉固定领域的研究人员需要通过准确的规划来规范协议。尽管已经对?...

Transpedicular screw fixation is a gold-standard treatment for degenerative lumbar spine and bursting fracture because it involves three columns of the spine and achieves stabilization1,2. However, most patients who undergo such surgery also have osteoporosis3,4. Many studies have evaluated the fixation strength and the osseointegration status of transpedicular screws, because the loosening of pedicle screws currently in use has been reported in patients with osteoporosis5,6.
The porcine spine is similar to the human spine in terms of size. It is less expensive compared to a primate model7. Furthermore, an in vivo mechanical study has demonstrated that the quadruped porcine spine is essentially loaded in the same way as that of the human spine8, which is why many researchers use porcine spines for studies on the prevention of pedicle screw loosening. However, it takes several months to study pedicle screws in the porcine spine because identifying the long-term stability of pedicle screws takes times. In order to compare different types of screws in the vertebral body, it is necessary to insert the screws in similar positions. Therefore, researchers should be well-acquainted with proper anesthesia techniques, standardized surgical protocols, and harvest procedures before performing any experiments. Here, we describe a detailed method for anesthesia, surgery, and harvest for the evaluation of pedicle screw fixation using a porcine spine model, including ex vivo imaging, histology, and strength testing.

<table><tbody><tr><td>Miniature pig</td><td>OrientBio</td><td></td><td></td></tr><tr><td>Atropine</td><td>Jeil pharmaceutical</td><td>A04900241</td><td>Anesthesia</td></tr><tr><td>Over-the needle plastic catheter</td><td>BD</td><td>REF382412</td><td>Maintenance of IV line</td></tr><tr><td>Ketamine</td><td>Yuhan</td><td>A04502441</td><td>Anesthesia</td></tr><tr><td>Xylazine</td><td>Bayer Korea</td><td>A00800071</td><td>Anesthesia</td></tr><tr><td>Laryngoscope</td><td>Karl storz</td><td></td><td>Intubation</td></tr><tr><td>Endotracheal tube</td><td>Covidien</td><td></td><td>Intubation</td></tr><tr><td>Isoflurane</td><td>JW pharmaceutical Co</td><td>A02104781</td><td>Anesthesia</td></tr><tr><td>Eye ointment</td><td>Hanlim pharma</td><td>A37851721</td><td>Protection of pig&#039;s eye</td></tr><tr><td>Cefazolin</td><td>Donga pharma</td><td>A01503951</td><td>Antibiotics</td></tr><tr><td>Saline</td><td>JW pharmaceutical Co</td><td>A02151392</td><td>Maintenance of fluid homeostasis</td></tr><tr><td>Fentanyl</td><td>Hana pharm</td><td>C03200032</td><td>Pain control</td></tr><tr><td>Enrofloxacin</td><td>Bayer</td><td>93106-60-6&amp;nbsp;</td><td>Antibiotics</td></tr><tr><td>Morphine</td><td>Myungmoon pharma</td><td>C03700091</td><td>Pain control</td></tr><tr><td>Meloxicam</td><td>Boehringer Ingelheim</td><td>A07600711</td><td>Antibiotics</td></tr><tr><td>Povidone-iodine</td><td>Hyundai pharma</td><td></td><td>Wound dressing</td></tr><tr><td>Scalpel blade size 15</td><td>Braun</td><td>&amp;nbsp;I1 BB515</td><td>Skin incision</td></tr><tr><td>Cobb elevator</td><td>Codman</td><td>65-2546</td><td>Dissection of muscle</td></tr><tr><td>Burr</td><td>Medtronic</td><td></td><td>Making of starting point of screw</td></tr><tr><td>Rongeur</td><td>Aesculap</td><td>FO515R</td><td>Making of starting point of screw</td></tr><tr><td>Guide pin (K-wire)</td><td>CE</td><td>01067803</td><td>Guidance of screw trajectory</td></tr><tr><td>C-arm</td><td>GE</td><td>OEC 9800 plus</td><td>Guidance of screw trajectory</td></tr><tr><td>Portable X-ray</td><td>Siemens</td><td>Mobile XP hybrid</td><td>Guidance of screw trajectory</td></tr><tr><td>Pedicle probe</td><td>OtisBiotech</td><td>SPI-02-01</td><td>Guidance of screw trajectory</td></tr><tr><td>Pedicle sounding device</td><td>OtisBiotech</td><td>SPI-03-01</td><td>Guidance of screw trajectory</td></tr><tr><td>Pedicle screw</td><td>OtisBiotech</td><td>MS-40025</td><td></td></tr><tr><td>Posterior fixator systems</td><td>OtisBiotech</td><td></td><td></td></tr><tr><td>Rod&amp;nbsp;</td><td>OtisBiotech</td><td>ROD-60140</td><td>Rigid fixation between screws</td></tr><tr><td>Universal handle</td><td>OtisBiotech</td><td>SPI-08-01</td><td>To fix the screws to the rod</td></tr><tr><td>Straight socket wrench</td><td>OtisBiotech</td><td>SPI-06-01</td><td>To fix the screws to the rod</td></tr><tr><td>counter torque wrench</td><td>OtisBiotech</td><td>SPI-07-01</td><td>To fix the screws to the rod</td></tr><tr><td>Bulb irrigation syringe</td><td>Hyupsug medical</td><td>HS-IR-140</td><td>Irrigation</td></tr><tr><td>Silicone drain</td><td>Sewon medical</td><td>2205-006</td><td>To drain the fluid at the surgical site</td></tr><tr><td>3.0 metric absorbable suture</td><td>Ethicon</td><td>BA1673H</td><td>Muscle suture</td></tr><tr><td>2.0 metric nonabsorbable nylon suture</td><td>Ethicon</td><td>W1626T</td><td>Skin suture</td></tr><tr><td>Gauze</td><td>Kingphar Korea</td><td>KP120-06</td><td></td></tr><tr><td>Pentobarbital</td><td>Hanlim pharma</td><td>645301221</td><td>Euthanasia</td></tr><tr><td>Oscillating saw</td><td>Zimmer</td><td></td><td>Harvest spine</td></tr><tr><td>Tower forceps</td><td>Aesculap</td><td>BF461R</td><td>Harvest spine</td></tr></tbody></table>

an anesthesia, surgery, and harvest method for the evaluation of transpedicular screws using an in vivo porcine lumbar spine model

椎弓根螺钉固定术是治疗脊柱疾病的黄金标准。然而，许多研究报道了脊柱手术后松动椎弓根螺钉的问题，这是一个严重的问题。为了解决这个问题，已经对多种类型的椎弓根螺钉进行了检查，以确定脊柱骨中具有良好的固定强度和骨整合的那些。由于解剖尺寸，机械特性和成本，猪脊柱是评估椎弓根螺钉的人类脊柱的良好替代品。尽管有几项研究报道，在猪模型中椎弓根螺钉是有效的，但没有研究已经描述了使用猪模型评估椎弓根螺钉的详细方案。在这里，我们描述了使用体内猪腰椎模型评估经椎弓根螺钉的详细方法。这里提供的麻醉，脊柱手术和收获的技术细节将有助于评估t睾丸螺钉固定模型。

这里描述了使用体内猪腰椎模型来评估经椎弓根螺钉的麻醉，手术和收获的详细方案。该协议适用于许多下游分析，包括机械测试（ 图1 ），定量微CT评估（ 图2 ）和组织学（ 图3 ）。代表性机械测试（ 图1 ）显示平均提取扭转峰值扭矩。它代表了使用机械测试仪的椎弓根螺钉和骨骼之间的结合强度。评估了三种类...

Watch this Scientific Journal Video about An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model at JoVE.com

An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model

在这里，我们介绍一种通过使用体内猪腰椎模型来评估椎弓根螺钉的方法。

用于评估椎弓根螺钉的麻醉，手术和收获方法<em&gt;体内</em&gt;猪腰椎模型

Pedicle screw fixation is the gold standard for the treatment of spinal diseases. However, many studies have reported the issue of loosening pedicle ...

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Unit 1

An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model

Local Anesthetics

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7.6K 次观看.  Chonbuk National University School of Medicine, Chonbuk National University Hospital. 在这里，我们介绍一种通过使用体内猪腰椎模型来评估椎弓根螺钉的方法。 

视频: An Anesthesia, Surgery, and Harvest Method for the Evaluation of Transpedicular Screws Using an In Vivo Porcine Lumbar Spine Model

In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

In vivo imaging using two-photon microscopy 1 in mice that have been genetically engineered to express fluorescent proteins in specific cell types 2-3 has significantly broadened our knowledge of physiological and pathological processes in numerous tissues in vivo 4-7. In studies of the central nervous system (CNS), there has been a broad application of in vivo imaging in the brain, which has produced a plethora of novel and often unexpected findings about the behavior of cells such as neurons, astrocytes, microglia, under physiological or pathological conditions 8-17. However, mostly technical complications have limited the implementation of in vivo imaging in studies of the living mouse spinal cord. In particular, the anatomical proximity of the spinal cord to the lungs and heart generates significant movement artifact that makes imaging the living spinal cord a challenging task.
We developed a novel method that overcomes the inherent limitations of spinal cord imaging by stabilizing the spinal column, reducing respiratory-induced movements and thereby facilitating the use of two-photon microscopy to image the mouse spinal cord in vivo. This is achieved by combining a customized spinal stabilization device with a method of deep anesthesia, resulting in a significant reduction of respiratory-induced movements. This video protocol shows how to expose a small area of the living spinal cord that can be maintained under stable physiological conditions over extended periods of time by keeping tissue injury and bleeding to a minimum. Representative raw images acquired in vivo detail in high resolution the close relationship between microglia and the vasculature. A timelapse sequence shows the dynamic behavior of microglial processes in the living mouse spinal cord. Moreover, a continuous scan of the same z-frame demonstrates the outstanding stability that this method can achieve to generate stacks of images and/or timelapse movies that do not require image alignment post-acquisition. Finally, we show how this method can be used to revisit and reimage the same area of the spinal cord at later timepoints, allowing for longitudinal studies of ongoing physiological or pathological processes in vivo.

In vivo Imaging of the Mouse Spinal Cord Using Two-photon Microscopy

A minimally invasive protocol to stabilize the mouse spinal column and perform repetitive in vivo spinal cord imaging using two-photon microscopy is described. This method combines a spinal stabilization device and an anesthetic regimen to minimize respiratory-induced movements and produce raw imaging data that require no alignment or other post-processing.

在体内脊髓的鼠标线使用双光子显微镜成像

In vivo imaging using two-photon microscopy 1 in mice that have been genetically engineered to express fluorescent proteins in specific cell types 2-3 has ...

科研

神经科学

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by repeated, time-lapse in vivo imaging in live rodents. These studies were made possible by straightforward surgical procedures, which enabled optical access for a prolonged period of time without repeat surgical procedures. In contrast, analogous studies of the spinal cord have been previously limited to only a few imaging sessions, each of which required an invasive surgery. As previously described, we have developed a spinal chamber that enables continuous optical access for upwards of 8 weeks, preserves mechanical stability of the spinal column, is easily stabilized externally during imaging, and requires only a single surgery. Here, the design of the spinal chamber with its associated surgical implements is reviewed and the surgical procedure is demonstrated in detail. Briefly, this video will demonstrate the preparation of the surgical area and mouse for surgery, exposure of the spinal vertebra and appropriate tissue debridement, the delivery of the implant and vertebral clamping, the completion of the chamber, the removal of the delivery system, sealing of the skin, and finally, post-operative care. The procedure for chronic in vivo imaging using nonlinear microscopy will also be demonstrated. Finally, outcomes, limitations, typical variability, and a guide for troubleshooting are discussed.

A Procedure for Implanting a Spinal Chamber for Longitudinal In Vivo Imaging of the Mouse Spinal Cord

In this video, we describe a procedure for implanting a chronic optical imaging chamber over the dorsal spinal cord of a live mouse. The chamber, surgical procedure, and chronic imaging are reviewed and demonstrated.

一个程序，植入脊柱商会纵<em&gt;在体内</em&gt;鼠标脊髓成像

Studies in the mammalian neocortex have enabled unprecedented resolution of cortical structure, activity, and response to neurodegenerative insults by ...

Laminotomy for Lumbar Dorsal Root Ganglion Access and Injection in Swine

Dorsal root ganglia (DRG) are anatomically well defined structures that contain all primary sensory neurons below the head. This fact makes DRG attractive targets for injection of novel therapeutics aimed at treating chronic pain. In small animal models, laminectomy has been used to facilitate DRG injection because it involves surgical removal of the vertebral bone surrounding each DRG. We demonstrate a technique for intraganglionic injection of lumbar DRG in a large animal species, namely, swine. Laminotomy is performed to allow direct access to DRG using standard neurosurgical techniques, instruments, and materials. Compared with more extensive bone removal via laminectomy, we implement laminotomy to conserve spinal anatomy while achieving sufficient DRG access. Intraoperative progress of DRG injection is monitored using a non-toxic dye. Following euthanasia on post-operative day 21, the success of injection is determined by histology for intraganglionic distribution of 4&#39;,6-diamidino-2-phenylindole (DAPI). We inject a biologically inactive solution to demonstrate the protocol. This method could be applied in future preclinical studies to target therapeutic solutions to DRG. Our methodology should facilitate testing the translatability of intraganglionic small animal paradigms in a large animal species. Additionally, this protocol may serve as a key resource for those planning preclinical studies of DRG injection in swine.

We describe a method for laminotomy in swine that provides access to lumbar dorsal root ganglia (DRG) for intraganglionic injection. Injection progress is monitored intraoperatively and histologically confirmed up to 21 days after surgery. This protocol could be used for future preclinical studies involving DRG injection.

Laminotomy 对猪腰背根神经节通路和注射的治疗

Dorsal root ganglia (DRG) are anatomically well defined structures that contain all primary sensory neurons below the head. This fact makes DRG attractive ...

Laminectomy and Spinal Cord Window Implantation in the Mouse

This protocol describes a method for spinal cord laminectomy and glass window implantation for in vivo imaging of the mouse spinal cord. An integrated digital vaporizer is utilized to achieve a stable plane of anesthesia at a low-flow rate of isoflurane. A single vertebral spine is removed, and a commercially available cover-glass is overlaid on a thin agarose bed. A 3D-printed plastic backplate is then affixed to the adjacent vertebral spines using tissue adhesive and dental cement. A stabilization platform is used to reduce motion artifact from respiration and heartbeat. This rapid and clamp-free method is well-suited for acute multi-photon fluorescence microscopy. Representative data are included for an application of this technique to two-photon microscopy of the spinal cord vasculature in transgenic mice expressing eGFP:Claudin-5 &#8212; a tight junction protein.

This protocol describes implantation of a glass window onto the spinal cord of a mouse to facilitate visualization by intravital microscopy.

拉米内切手术和脊髓窗口植入小鼠

This protocol describes a method for spinal cord laminectomy and glass window implantation for in vivo imaging of the mouse spinal cord. An integrated ...

In Vivo Mouse Model of Spinal Implant Infection

Spine implant infections portend poor outcomes as diagnosis is challenging and surgical eradication is at odds with mechanical spinal stability. The purpose of this method is to describe a novel mouse model of spinal implant infection (SII) that was created to provide an inexpensive, rapid, and accurate in vivo tool to test potential therapeutics and treatment strategies for spinal implant infections.
In this method, we present a model of posterior-approach spinal surgery in which a stainless-steel k-wire is transfixed into the L4 spinous process of 12-week old C57BL/6J wild-type mice and inoculated with 1 x 103 CFU of a bioluminescent strain of Staphylococcus aureus Xen36 bacteria. Mice are then longitudinally imaged for bioluminescence in vivo on post-operative days 0, 1, 3, 5, 7, 10, 14, 18, 21, 25, 28, and 35. Bioluminescence imaging (BLI) signals from a standardized field of view are quantified to measure in vivo bacterial burden.
To quantify bacteria adhering to implants and peri-implant tissue, mice are euthanized and the implant and surrounding soft tissue are harvested. Bacteria are detached from the implant by sonication, cultured overnight and then colony forming units (CFUs) are counted. The results acquired from this method include longitudinal bacterial counts as measured by in vivo S. aureus bioluminescence (mean maximum flux) and CFU counts following euthanasia.
While prior animal models of instrumented spine infection have involved invasive, ex vivo tissue analysis, the mouse model of SII presented in this paper leverages noninvasive, real time in vivo optical imaging of bioluminescent bacteria to replace static tissue study. Applications of the model are broad and may include utilizing alternative bioluminescent bacterial strains, incorporating other types of genetically engineered mice to contemporaneously study host immune response, and evaluating current or investigating new diagnostic and therapeutic modalities such as antibiotics or implant coatings.

The protocol describes a novel in vivo mouse model of spinal implant infection where a stainless-steel k-wire implant is infected with bioluminescent Staphylococcus aureus Xen36. Bacterial burden is monitored longitudinally with bioluminescent imaging and confirmed with colony forming unit counts after euthanasia.

脊柱植入物感染的体内小鼠模型

Spine implant infections portend poor outcomes as diagnosis is challenging and surgical eradication is at odds with mechanical spinal stability. The ...

医学

Real-Time Assessment of Spinal Cord Microperfusion in a Porcine Model of Ischemia/Reperfusion

Spinal cord injury is a devastating complication of aortic repair. Despite developments for the prevention and treatment of spinal cord injury, its incidence is still considerably high and therefore, influences patient outcome. Microcirculation plays a key role in tissue perfusion and oxygen supply and is often dissociated from macrohemodynamics. Thus, direct evaluation of spinal cord microcirculation is essential for the development of microcirculation-targeted therapies and the evaluation of existing approaches in regard to spinal cord microcirculation. However, most of the methods do not provide real-time assessment of spinal cord microcirculation. The aim of this study is to describe a standardized protocol for real-time spinal cord microcirculatory evaluation using laser-Doppler needle probes directly inserted in the spinal cord. We used a porcine model of ischemia/reperfusion to induce deterioration of the spinal cord microcirculation. In addition, a fluorescent microsphere injection technique was used. Initially, animals were anesthetized and mechanically ventilated. Thereafter, laser-Doppler needle probe insertion was performed, followed by the placement of cerebrospinal fluid drainage. A median sternotomy was performed for exposure of the descending aorta to perform aortic cross-clamping. Ischemia/reperfusion was induced by supra-celiac aortic cross-clamping for a total of 48 min, followed by reperfusion and hemodynamic stabilization. Laser-Doppler Flux was performed in parallel with macrohemodynamic evaluation. In addition, automated cerebrospinal fluid drainage was used to maintain a stable cerebrospinal pressure. After completion of the protocol, animals were sacrificed, and the spinal cord was harvested for histopathological and microsphere analysis. The protocol reveals the feasibility of spinal cord microperfusion measurements using laser-Doppler probes and shows a marked decrease during ischemia as well as recovery after reperfusion. Results showed comparable behavior to fluorescent microsphere evaluation. In conclusion, this new protocol might provide a useful large animal model for future studies using real-time spinal cord microperfusion assessment in ischemia/reperfusion conditions.

Spinal cord microcirculation plays a pivotal role in spinal cord injury. Most methods do not allow real-time assessment of spinal cord microcirculation, which is essential for the development of microcirculation-targeted therapies. Here, we propose a protocol using Laser-Doppler-Flow Needle probes in a large animal model of ischemia/reperfusion.

猪缺血/再灌注模型中脊髓微灌注的实时评估

Spinal cord injury is a devastating complication of aortic repair. Despite developments for the prevention and treatment of spinal cord injury, its ...

Surgical Technique for Lumbar Spinal Catheter Insertion in Pigs Enabling Continuous Access to the Thecal Sac in a Terminal Setup

Pigs are increasingly used as a large animal model for pharmacologic CNS research due to the anatomical and physiological similarities between the porcine and human central nervous systems (CNS). However, accessing the cerebrospinal fluid (CSF) in larger pig breeds by conventional lumbar puncture techniques can be challenging due to an oblique orientation of the spinal spinous processes and a limited interlaminar space. Accordingly, an open surgical procedure for inserting a lumbar spinal catheter for continuous CSF sampling at the L4/L5 level in pigs is thoroughly described in this work. After positioning the pig and identifying the anatomical landmarks, a dorsal midline surgical incision is made to expose the spinous processes. By advancing the introducer needle, the spinal catheter is inserted inside the thecal sac of the spinal canal while leaving the bone structures of the spine intact. This method allows continuous infusion into or sampling from the porcine thecal sac with minimal bleeding or CSF leakage. The procedure is simple, time-efficient, and reproducible across different experimental setups, offering significant potential for various pre-clinical studies, including pharmacokinetic research, surgical training, and spinal cord injury models.

We present a technique for inserting a lumbar spinal catheter at the L4-L5 level in a 3-month-old Danish Landrace pig as part of a terminal research protocol, enabling continuous infusion or CSF sampling from the thecal sac.

猪腰椎导管插入手术技术，可在终端设置中连续进入鞘囊

Pigs are increasingly used as a large animal model for pharmacologic CNS research due to the anatomical and physiological similarities between the porcine ...

A Mouse Model of Lumbar Spine Instability

Intervertebral disc degeneration (IDD) is a common pathological change leading to low back pain. Appropriate animal models are desired for understanding the pathological processes and evaluating new drugs. Here, we introduced a surgically induced lumbar spine instability (LSI) mouse model that develops IDD starting from 1 week post operation. In detail, the mouse under anesthesia was operated by low back skin incision, L3&#8211;L5 spinous processes exposure, detachment of paraspinous muscles, resection of processes and ligaments, and skin closure. L4&#8211;L5 IVDs were chosen for the observation. The LSI model develops lumbar IDD by porosity and hypertrophy in endplates at an early stage, decrease in intervertebral disc volume, shrinkage in nucleus pulposus at an intermediate stage, and bone loss in lumbar vertebrae (L5) at a later stage. The LSI mouse model has the advantages of strong operability, no requirement of special equipment, reproducibility, inexpensive, and relatively short period of IDD development. However, LSI operation is still a trauma that causes inflammation within the first week post operation. Thus, this animal model is suitable for study of lumbar IDD.

We developed a lumbar intervertebral disc degeneration mouse model by resection of L3&#8211;L5 spinous processes along with supra- and inter-spinous ligaments and detachment of paraspinous muscles.

腰椎不稳定的鼠标模型

Intervertebral disc degeneration (IDD) is a common pathological change leading to low back pain. Appropriate animal models are desired for understanding ...

生物学

Pedicle Screw Placement Using an Augmented Reality Head-Mounted Display in a Porcine Model

This protocol helps assess the accuracy and workflow of an augmented reality (AR) hybrid navigation system using the Magic Leap head-mounted display (HMD) for minimally invasive pedicle screw placement. The cadaveric porcine specimens were placed on a surgical table and draped with sterile covers. The levels of interest were identified using fluoroscopy, and a dynamic reference frame was attached to the spinous process of a vertebra in the region of interest. Cone beam computerized tomography (CBCT) was performed, and a 3D rendering was automatically generated, which was used for the subsequent planning of the pedicle screw placements. Each surgeon was fitted with an HMD that was individually eye-calibrated and connected to the spinal navigation system.
Navigated instruments, tracked by the navigation system and displayed in 2D and 3D in the HMD, were used for 33 pedicle cannulations, each with a diameter of 4.5 mm. Postprocedural CBCT scans were assessed by an independent reviewer to measure the technical (deviation from the planned path) and clinical (Gertzbein grade) accuracy of each cannulation. The navigation time for each cannulation was measured. The technical accuracy was 1.0 mm &#177; 0.5 mm at the entry point and 0.8 mm &#177; 0.1 mm at the target. The angular deviation was 1.5&#176; &#177; 0.6&#176;, and the mean insertion time per cannulation was 141 s &#177; 71 s. The clinical accuracy was 100% according to the Gertzbein grading scale (32 grade 0; 1 grade 1). When used for minimally invasive pedicle cannulations in a porcine model, submillimeter technical accuracy and 100% clinical accuracy could be achieved with this protocol.

The augmented reality head-mounted display, Magic Leap, was used in combination with a conventional navigation system to place pedicle screws in a porcine model by adhering to a novel workflow. With a median insertion time of &lt;2.5 min, submillimeter technical accuracy and 100% clinical accuracy were achieved according to Gertzbein.

在猪模型中使用增强现实头戴式显示器放置椎弓根螺钉

This protocol helps assess the accuracy and workflow of an augmented reality (AR) hybrid navigation system using the Magic Leap head-mounted display (HMD) ...

<meta charset="utf8"></head><body>胸腰椎爆裂性骨折矢状面调整螺钉系统技术

Minimally Invasive Treatment for Thoracolumbar Burst Fracture Using Sagittal Alignment Screws and A Trauma Reduction Device

Thoracolumbar (TL) burst fracture is one of the most common indications for minimally invasive percutaneous pedicle screw fixation. Although the indication for surgical treatment of neurologically intact TL fractures remains under debate, studies have demonstrated that posttraumatic malalignment may lead to a deterioration in the patient&#39;s quality of life. For burst fractures with malalignment or fragments in the spinal canal, a reduction technique using ligamentotaxis is commonly used to improve long-term outcomes.
The sagittal adjusting screw (SAS) system is a monoaxial screw system with a fixed head and concave sliding saddle that allows lordotic sliding of the rod in the sagittal plane after screw insertion. SAS also has a percutaneous option and has been used for TL spine fractures. Notably, the SAS only allows motion on the sagittal plane, allowing both secure fixation and angular reduction. The SAS has certain advantages over the conventional Schanz screw system or normal mono-/multiaxial pedicle screws for TL spine fracture treatment. In addition, specialized trauma reduction devices are available for the SAS system. In this video protocol, we discuss the indication for the SAS system in TL burst fracture and describe a technique of TL burst fracture reduction and fixation using the SAS system. Additionally, we describe our recent case series with radiological evaluation, including regional kyphotic angle and percent loss of anterior vertebral body height, to evaluate the newly introduced trauma reduction device.

<meta charset="utf8"></head><body>使用矢状对位螺钉和创伤复位装置对胸腰椎爆裂性骨折进行微创治疗

The sagittal adjusting screw (SAS) system has been widely used for thoracolumbar (TL) spinal trauma. Percutaneous options and specialized trauma reduction devices are also available for the SAS system. We describe a technique for reducing TL burst fractures using an SAS system and a newly introduced trauma reduction device.

用于评估椎弓根螺钉的麻醉，手术和收获方法

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