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. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+bone+mineral+density+on+pedicle+screw+fixation.">Effects of bone mineral density on pedicle screw fixation.</a> Spine. 19 (21), 2415-2420 (1994).</li><li>Weinstein, J. N., Spratt, K. F., Spengler, D., Brick, C., Reid, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spinal+pedicle+fixation:+reliability+and+validity+of+roentgenogram-based+assessment+and+surgical+factors+on+successful+screw+placement.">Spinal pedicle fixation: reliability and validity of roentgenogram-based assessment and surgical factors on successful screw placement.</a> Spine. 13 (9), 1012-1018 (1988).</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>Kim, D. 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=Evaluation+of+Titanium-Coated+Pedicle+Screws:+In+Vivo+Porcine+Lumbar+Spine+Model.">Evaluation of Titanium-Coated Pedicle Screws: In Vivo Porcine Lumbar Spine Model.</a> World Neurosurg. 91, 163-171 (2016).</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>Smit, T. 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+use+of+a+quadrupted+as+an+in+vivo+model+for+the+study+of+the+spine-biomechanical+considrations.">The use of a quadrupted as an in vivo model for the study of the spine-biomechanical considrations.</a> Eur Spine J. 11 (2), 137-144 (2002).</li><li>Aldini, N. 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. J., Verkerke, G. J., Veldhuizen, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+anatomical+dimensions+of+the+complete+human+and+porcine+spine.">Comparative anatomical dimensions of the complete human and porcine spine.</a> Eur Spine J. 19 (7), 1104-1114 (2010).</li></ol>

作者与本文无关。

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 border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<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&#39;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&#160;</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>&#160;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&#160;</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 ...

an-anesthesia-surgery-harvest-method-for-evaluation-transpedicular

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

Local Anesthetics

SCIENTISTS IN ACTION

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

Using Quantitative Real-time PCR to Determine Donor Cell Engraftment in a Competitive Murine Bone Marrow Transplantation Model

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules that regulate HSCs. In these transplant model systems, the function of HSCs is determined by the ability of these cells to engraft and reconstitute lethally irradiated recipient mice. Commonly, the donor cell contribution/engraftment is measured by antibodies to donor- specific cell surface proteins using flow cytometry. However, this method heavily depends on the specificity and the ability of the cell surface marker to differentiate donor-derived cells from recipient-originated cells, which may not be available for all mouse strains. Considering the various backgrounds of genetically modified mouse strains in the market, this cell surface/ flow cytometry-based method has significant limitations especially in mouse strains that lack well-defined surface markers to separate donor cells from congenic recipient cells. Here, we reported a PCR-based technique to determine donor cell engraftment/contribution in transplant recipient mice. We transplanted male donor bone marrow HSCs to lethally irradiated congenic female mice. Peripheral blood samples were collected at different time points post transplantation. Bone marrow samples were obtained at the end of the experiments. Genomic DNA was isolated and the Y chromosome specific gene, Zfy1, was amplified using quantitative Real time PCR. The engraftment of male donor-derived cells in the female recipient mice was calculated against standard curve with known percentage of male vs. female DNAs. Bcl2 was used as a reference gene to normalize the total DNA amount. Our data suggested that this approach reliably determines donor cell engraftment and provides a useful, yet simple method in measuring hematopoietic cell reconstitution in murine bone marrow transplantation models. Our method can be routinely performed in most laboratories because no costly equipment such as flow cytometry is required.

Determining donor cell engraftment presents a challenge in mouse bone marrow transplant models that lack well-defined phenotypical markers. We described a methodology to quantify male donor cell engraftment in female transplant recipient mice. This method can be used in all mouse strains for the study of HSC functions.

采用实时定量PCR检测供体细胞植入一个具有竞争力的小鼠骨髓移植模型

Murine bone marrow transplantation models provide an important tool in measuring hematopoietic stem cell (HSC) functions and determining genes/molecules ...

科研

医学

An Improved Method for Rapid Intubation of the Trachea in Mice

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity is a commonly used model to study both acute lung injury and fibrosis, and multiple methods have been developed for administering bleomycin (and other toxic agents) into the lungs. However, many of these approaches, such as transtracheal instillation, have inherent drawbacks, including the need for strong anesthetics and survival surgery. This paper reports a quick, reproducible method of intratracheal intubation that involves mild inhaled anesthesia, visualization of the trachea, and the use of a surrogate spirometer to confirm exposure. As a proof of concept, 8-12 week old C57BL/6 mice were administered either 2.0 U/kg of bleomycin or an equivalent volume of PBS, and both damage and fibrotic endpoints were measured post-exposure. This procedure allows researchers to treat a large cohort of mice in a relatively short period with little expense and minimal post-procedure care.

This article presents a rapid and simple method for administering bleomycin directly into the mouse trachea via intubation. Key advantages of this method are that it is highly reproducible, easy to master, and does not require specialized equipment or lengthy recovery times.

在小鼠气管插管快速的改进方法

Despite some anatomical and physiological differences, mouse models continue to be an essential tool for studying human lung disease. Bleomycin toxicity ...

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

The aim of this protocol presents an optimized procedure for the purification and cultivation of pBECs and to establish in vitro blood-brain barrier (BBB) models based on pBECs in mono-culture (MC), MC with astrocyte-conditioned medium (ACM), and non-contact co-culture (NCC) with astrocytes of porcine or rat origin. pBECs were isolated and cultured from fragments of capillaries from the brain cortices of domestic pigs 5-6 months old. These fragments were purified by careful removal of meninges, isolation and homogenization of grey matter, filtration, enzymatic digestion, and centrifugation. To further eliminate contaminating cells, the capillary fragments were cultured with puromycin-containing medium. When 60-95% confluent, pBECs growing from the capillary fragments were passaged to permeable membrane filter inserts and established in the models. To increase barrier tightness and BBB characteristic phenotype of pBECs, the cells were treated with the following differentiation factors: membrane permeant 8-CPT-cAMP (here abbreviated cAMP), hydrocortisone, and a phosphodiesterase inhibitor, RO-20-1724 (RO). The procedure was carried out over a period of 9-11 days, and when establishing the NCC model, the astrocytes were cultured 2-8 weeks in advance. Adherence to the described procedures in the protocol has allowed the establishment of endothelial layers with highly restricted paracellular permeability, with the NCC model showing an average transendothelial electrical resistance (TEER) of 1249 &#177; 80 &#937; cm2, and paracellular permeability (Papp) for Lucifer Yellow of 0.90 10-6 &#177; 0.13 10-6 cm sec-1 (mean &#177; SEM, n=55). Further evaluation of this pBEC phenotype showed good expression of the tight junctional proteins claudin 5, ZO-1, occludin and adherens junction protein p120 catenin. The model presented can be used for a range of studies of the BBB in health and disease and, with the highly restrictive paracellular permeability, this model is suitable for studies of transport and intracellular trafficking.

Improved Method for the Establishment of an In Vitro Blood-Brain Barrier Model Based on Porcine Brain Endothelial Cells

The aim of the protocol is to present an optimized procedure for the establishment of an in vitro blood-brain barrier (BBB) model based on primary porcine brain endothelial cells (pBECs). The model shows high reproducibility, high tightness, and is suitable for studies of transport and intracellular trafficking in drug discovery.

一种基于猪脑内皮细胞的体外血脑屏障模型的改进方法

The aim of this protocol presents an optimized procedure for the purification and cultivation of pBECs and to establish in vitro blood-brain barrier (BBB) ...

In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The amount of the different tissues in the callus provides important information on the fracture healing progress. Available in vivo techniques to longitudinally monitor the callus tissue development in preclinical fracture-healing studies using small animals include digital radiography and &#181;CT imaging. However, both techniques are only able to distinguish between mineralized and non-mineralized tissue. Consequently, it is impossible to discriminate cartilage from fibrous tissue. In contrast, magnetic resonance imaging (MRI) visualizes anatomical structures based on their water content and might therefore be able to noninvasively identify soft tissue and cartilage in the fracture callus. Here, we report the use of an MRI-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice. The experiments demonstrated that the fixator and a custom-made mounting device allow repetitive MRI scans, thus enabling longitudinal analysis of fracture-callus tissue development.

In Vivo Evaluation of Fracture Callus Development During Bone Healing in Mice Using an MRI-compatible Osteosynthesis Device for the Mouse Femur

The evaluation of tissue development in the fracture callus during endochondral bone healing is essential to monitor the healing process. Here, we report the use of a magnetic resonance imaging (MRI)-compatible external fixator for the mouse femur to allow MRI scans during bone regeneration in mice.

在体内MRI 对小鼠股骨骨愈合过程中骨折愈伤组织发育的评价

Endochondral fracture healing is a complex process involving the development of fibrous, cartilaginous, and osseous tissue in the fracture callus. The ...

A Pre-Clinical Porcine Model of Orthotopic Heart Transplantation

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart failure. Multiple small-animal models of heart transplantation have been used to study the acute and long-term effects of novel therapies. However, few are tested and demonstrated success in clinical trials. It is of critical importance to evaluate new therapies in a clinically relevant large-animal model for efficient and reliable translation of basic studies&#39;&#160;findings. Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and previously used to investigate novel cardioprotective strategies. This procedure focuses on acute ischemia-reperfusion injury and is a reliable method to investigate novel interventions which have been tested and validated in smaller experimental models, such as the murine model. We demonstrate its usefulness in assessing cardiac performance during the early post-transplantation period and other potential possibilities enabled by the model.

Here, we describe a pre-clinical large-animal (porcine) model of orthotopic heart transplantation that has been firmly established and utilized to investigate novel cardioprotective strategies.

正交心脏移植的临床前猪模型

Fifty-years following the first successful report, cardiac transplantation remains the gold-standard treatment for eligible patients with advanced heart ...

An In Vivo Mouse Model of Total Intravenous Anesthesia During Cancer Resection Surgery

Anesthesia is a routine component of cancer care that is used for diagnostic and therapeutic procedures. The anesthetic technique has recently been implicated in impacting long-term cancer outcomes, possibly through modulation of adrenergic-inflammatory responses that impact cancer cell behavior and immune cell function. Emerging evidence suggests that propofol-based total intravenous anesthesia (TIVA) may be beneficial for long-term cancer outcomes when compared to inhaled volatile anesthesia. However, the available clinical findings are inconsistent. Preclinical studies that identify the underlying mechanisms involved are critically needed to guide the design of clinical studies that will expedite insight. Most preclinical models of anesthesia have been extrapolated from the use of anesthesia in in vivo research and are not optimally designed to study the impact of anesthesia itself as the primary endpoint. This paper describes a method for delivering propofol-TIVA anesthesia in a mouse model of breast cancer resection that replicates key aspects of clinical delivery in cancer patients. The model can be used to study mechanisms of action of anesthesia on cancer outcomes in diverse cancer types and can be extrapolated to other non-cancer areas of preclinical anesthesia research.

An In Vivo Mouse Model of Total Intravenous Anesthesia During Cancer Resection Surgery

This paper describes a method for modeling total intravenous anesthesia (TIVA) during cancer resection surgery in mice. The goal is to replicate key features of anesthesia delivery to patients with cancer. The method allows investigation of how anesthetic technique affects cancer recurrence after resection surgery.

癌症切除手术中全静脉麻醉的 体内 小鼠模型

Anesthesia is a routine component of cancer care that is used for diagnostic and therapeutic procedures. The anesthetic technique has recently been ...

Establishment and Evaluation of a Porcine Vein Graft Disease Model

Venous graft disease (VGD) is the leading cause of coronary artery bypass graft (CABG) failure. Large animal models of CABG-VGD are needed for the investigation of disease mechanisms and the development of therapeutic strategies. 
To perform the surgery, we enter the cardiac chamber through the third intercostal space and carefully dissect the internal mammary vein and immerse it in normal saline. The right main coronary artery is then treated for ischemia. The target vessel is incised, a shunt plug is placed, and the distal end of the graft vein is anastomosed. The ascending aorta is partially blocked, and the proximal end of the graft vein is anastomosed after perforation. The graft vein is checked for patency, and the proximal right coronary artery is ligated. 
CABG surgery is performed in minipigs to harvest the left internal mammary vein for its use as a vascular graft. Serum biochemical tests are used to evaluate the physiological status of the animals after surgery. Ultrasound examination shows that the proximal, middle, and distal end of the graft vessel are unobstructed. In the surgical model, turbulent blood flow in the graft is observed upon histological examination after the CABG surgery, and venous graft stenosis associated with intimal hyperplasia is observed in the graft. The study here provides detailed surgical procedures for the establishment of a repeatable CABG-induced VGD model.

In this protocol, novel pig vein bypass grafting was performed through a small incision in the left chest wall without cardiopulmonary bypass. A postoperative pathology study was done, which showed intimal thickening.

猪静脉移植病模型的建立与评价

Venous graft disease (VGD) is the leading cause of coronary artery bypass graft (CABG) failure. Large animal models of CABG-VGD are needed for the ...

3D Printing Model of a Patient's Specific Lumbar Vertebra

Selective dorsal rhizotomy (SDR) is a difficult, risky, and sophisticated operation, in which a laminectomy should not only expose an adequate surgical field of view but also protect the patient&#39;s spinal nerves from injury. Digital models play an important role in the pre-and intra-operation of SDR, because they can not only make doctors more familiar with the anatomical structure of the surgical site, but also provide precise surgical navigation coordinates for the manipulator. This study aims to create a 3D digital model of a patient-specific lumbar vertebra that can be used for planning, surgical navigation, and training of the SDR operation. The 3D printing model is also manufactured for more effective work during these processes.
Traditional orthopedic digital models rely almost entirely on computed tomography (CT) data, which is less sensitive to soft tissues. Fusion of the bone structure from CT and the neural structure from magnetic resonance imaging (MRI) is the key element for the model reconstruction in this study. The patient&#39;s specific 3D digital model is reconstructed for the real appearance of the surgical area and shows the accurate measurement of inter-structural distances and regional segmentation, which can effectively help in the preoperative planning and training of SDR. The transparent bone structure material of the 3D-printed model allows surgeons to clearly distinguish the relative relationship between the spinal nerve and the vertebral plate of the operated segment, enhancing their anatomical understanding and spatial sense of the structure. The advantages of the individualized 3D digital model and its accurate relationship between spinal nerve and bone structures make this method a good choice for preoperative planning of SDR surgery.

This study aims to create a 3D-printed model of a patient-specific lumbar vertebra, which contains both the vertebra and spinal nerve models fused from high-resolution computed tomography (HRCT) and MRI-Dixon data.

患者特定腰椎的3D打印模型

Selective dorsal rhizotomy (SDR) is a difficult, risky, and sophisticated operation, in which a laminectomy should not only expose an adequate surgical ...

用于肺结节诊断和治疗的 3D 重建

A 3D Digital Model for the Diagnosis and Treatment of Pulmonary Nodules

The three-dimensional (3D) reconstruction of pulmonary nodules using medical images has introduced new technical approaches for diagnosing and treating pulmonary nodules, and these approaches are progressively being acknowledged and adopted by physicians and patients. Nonetheless, constructing a relatively universal 3D digital model of pulmonary nodules for diagnosis and treatment is challenging due to device differences, shooting times, and nodule types. The objective of this study is to propose a new 3D digital model of pulmonary nodules that serves as a bridge between physicians and patients and is also a cutting-edge tool for pre-diagnosis and prognostic evaluation. Many AI-driven pulmonary nodule detection and recognition methods employ deep learning techniques to capture the radiological features of pulmonary nodules, and these methods can achieve a good area under-the-curve (AUC) performance. However, false positives and false negatives remain a challenge for radiologists and clinicians. The interpretation and expression of features from the perspective of pulmonary nodule classification and examination are still unsatisfactory. In this study, a method of continuous 3D reconstruction of the whole lung in horizontal and coronal positions is proposed by combining existing medical image processing technologies. Compared with other applicable methods, this method allows users to rapidly locate pulmonary nodules and identify their fundamental properties while also observing pulmonary nodules from multiple perspectives, thereby providing a more effective clinical tool for diagnosing and treating pulmonary nodules.

作者聚焦:用于肺结节诊断和治疗的 3D 数字模型  

The objective of this study is to develop a novel 3D digital model of pulmonary nodules that serves as a communication bridge between physicians and patients and is also a cutting-edge tool for pre-diagnosis and prognostic evaluation.

用于肺结节诊断和治疗的三维数字模型

The three-dimensional (3D) reconstruction of pulmonary nodules using medical images has introduced new technical approaches for diagnosing and treating ...