Name: urethral stricture induction followed by buccal mucosa graft urethroplasty in a rat model
Uploaded: 2023-04-28T13:00:00
Description: Watch this Scientific Journal Video about Urethral Stricture Induction Followed by Buccal Mucosa Graft Urethroplasty in a Rat Model at JoVE.com

我们感谢北里斯本大学医院中心放射科主任João Leitão和北里斯本大学医院中心放射科技术员Catia Fernandes在实现尿道造影方面的合作。

所有动物手术均根据指令 2010/63/EU 进行。这些程序得到了机构动物福利机构的批准，该机构获得了葡萄牙动物保护主管当局DGAV的许可（许可证号为0421/000/000/2021）。本研究使用12-14周龄的雄性Wistar Han IGS（Crl：WI（Han）大鼠（400-500g）。这些动物是从商业来源获得的（见 材料表）。
1.溶液的制备
<ol><li>麻醉液<ol><li>用 2.3 mL 美托咪定（1 mg/mL;0.715 mg/kg 体...

<ol>
	<li>Souza, G. F., Calado, A. A., Delcelo, R., Ortiz, V., Macedo, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Histopathological+evaluation+of+urethroplasty+with+dorsal+buccal+mucosa:+an+experimental+study+in+rabbits.">Histopathological evaluation of urethroplasty with dorsal buccal mucosa: an experimental study in rabbits.</a> International Brazilian Journal of Urology. 34 (3), 345-354 (2008).</li><li>Andrich, D. E., Mundy, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Urethral+strictures+and+their+surgical+treatment.">Urethral strictures and their surgical treatment.</a> BJU International. 86 (5), 571-580 (2000).</li><li>Bhargava, S., Chapple, C. R., Bullock, A. J., Layton, C., MacNeil, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tissue-engineered+buccal+mucosa+for+substitution+urethroplasty.">Tissue-engineered buccal mucosa for substitution urethroplasty.</a> BJU International. 93 (6), 807-811 (2004).</li><li>Fu, Q., 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=Substitution+urethroplasty+for+anterior+urethral+stricture+repair:+comparison+between+lingual+mucosa+graft+and+pedicled+skin+flap.">Substitution urethroplasty for anterior urethral stricture repair: comparison between lingual mucosa graft and pedicled skin flap.</a> Scandinavian Journal of Urology. 51 (6), 479-483 (2017).</li><li>Blain, B., 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=Vascular+grafts+in+the+rat+model:+an+anatomic+study.">Vascular grafts in the rat model: an anatomic study.</a> Microsurgery. 21 (3), 80-83 (2001).</li><li>Schmauss, D., Weinzierl, A., Schmauss, V., Harder, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Common+rodent+flap+models+in+experimental+surgery.">Common rodent flap models in experimental surgery.</a> European Surgical Research. 59 (3-4), 255-264 (2018).</li><li>You, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+rat+orthotopic+renal+transplantation+model+for+renal+allograft+rejection.">A rat orthotopic renal transplantation model for renal allograft rejection.</a> Journal of Visualized Experiments. (180), e63464 (2022).</li><li>Park, J. H., 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=Balloon-expandable+biodegradable+stents+versus+self-expandable+metallic+stents:+a+comparison+study+of+stent-induced+tissue+hyperplasia+in+the+rat+urethra.">Balloon-expandable biodegradable stents versus self-expandable metallic stents: a comparison study of stent-induced tissue hyperplasia in the rat urethra.</a> Cardiovascular and Interventional Radiology. 42 (9), 1343-1351 (2019).</li><li>Mart&#237;n-Cano, F., 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=Histological+and+immunohistochemical+changes+in+the+rat+oral+mucosa+used+as+an+autologous+urethral+graft.">Histological and immunohistochemical changes in the rat oral mucosa used as an autologous urethral graft.</a> Journal of Pediatric Surgery. 48 (7), 1557-1564 (2013).</li><li>Hofer, M. D., 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=Analysis+of+primary+urethral+wound+healing+in+the+rat.">Analysis of primary urethral wound healing in the rat.</a> Urology. 84 (1), e1-7 (2014).</li><li>Tavukcu, H. H., 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=Protective+effect+of+platelet-rich+plasma+on+urethral+injury+model+of+male+rats.">Protective effect of platelet-rich plasma on urethral injury model of male rats.</a> Neurourology and Urodynamics. 37 (4), 1286-1293 (2018).</li></ol>

颊黏膜移植物的尿道成形术是尿道重建的主要基石。然而，应开发创新程序来优化已经描述的程序，并建立新的程序，例如组织工程材料和生物移植物，以减少并发症和发病率。已经发表了几种程序来建立临床前模型和定义手术技术。Souza 等人 1 进行了一项包括 12 只新西兰兔子的研究。进行腹侧纵向皮肤切开，将尿道从白膜中动员出来，然后切除尿道背段，形成缺损。同时，从...

尿道重建是泌尿外科医生在狭窄、创伤或先天性缺损的情况下处理尿道损伤的主要挑战1.出于治愈目的，尿道成形术是大多数患者的首选治疗方法，长尿道缺损（>2 cm）和前尿道缺损需要某种形式的替代尿道成形术2。许多组织已被用作尿道替代品，包括生殖器或生殖器外区域、膀胱壁粘膜或广泛的颊粘膜2 的全层或裂层皮肤移植物。颊粘膜移植物有几个优点，例如来自潮湿和无毛的环境、易于收获、抗感染、厚上皮、降低假性憩室形成的可能性以及薄层，允许早期吸收和接种3.与皮瓣相比，移植物没有血液供应，取决于受体的血管床才能存活4.
移植物或皮瓣的动物模型已被广泛用于开发或改进手术技术，研究和了解组织生理学、潜在机制和失败原因，以及评估创新治疗策略5,6。虽然较大的动物有助于技术执行，但啮齿动物，即大鼠和小鼠，更容易处理和维护，对疾病有抵抗力，更具成本效益，而且重要的是，具有研究分子机制的工具，这对于测试创新疗法至关重要5,6。在大鼠中使用不同的组织，即皮肤、骨骼、肌肉6、血管5，甚至实体器官7，已经描述了几种皮瓣和移植物模型。然而，在小鼠模型中，对用于尿道重建或组织工程的移植物的研究很少。
然而，转化科学的进步依赖于模仿疾病的动物模型。到目前为止，由于尿道重建在其狭窄后立即进行，因此尚未解决局部病理生理学环境。在此，本研究旨在使用颊粘膜移植物在局部病理生理学环境中进行尿道重建。为此，在重建前 1 周诱导尿道狭窄。该实验模型在大鼠身上进行，可以测试创新疗法，并在未来研究其分子机制和临床优势。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Atipamezole</td>
			<td>OrionPharma</td>
			<td></td>
			<td>ANTISEDAN (atipamezole) is indicated for the reversal of the sedative and analgesic effects.</td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>RichterPharma</td>
			<td></td>
			<td>Buprenorphine is a derivative of the opioid alkaloid thebaine that is a more potent (25-40 times) and longer lasting analgesic than morphine.</td>
		</tr>
		<tr>
			<td>Carl Zeiss Opmi-1 FC Surgical Stereo Microscope</td>
			<td>Carl Zeiss Microscopy, Germany</td>
			<td></td>
			<td>OPMI 1 FC from ZEISS symbolizes quality, precision and reliability. The manual system is easy to use and delivers high fidelity images with the legendary ZEISS optics.</td>
		</tr>
		<tr>
			<td>Carprofen</td>
			<td>Zoetis</td>
			<td></td>
			<td>Carprofen is a non-steroidal anti-inflammatory drug (NSAID) of the propionic acid class.</td>
		</tr>
		<tr>
			<td>Catheter 22 G. x 1&#39;&#39;</td>
			<td>B Braun</td>
			<td>Introcan Safety</td>
			<td>IV Catheter of fluorinated ethylene propylene (FEP) with firmer construction for 
			arterial access.</td>
		</tr>
		<tr>
			<td>Enrofloxacin</td>
			<td>Bayer</td>
			<td></td>
			<td>Enrofloxacin is a fluoroquinolone antibiotic.</td>
		</tr>
		<tr>
			<td>Fentanyl</td>
			<td>Braun</td>
			<td>3644960</td>
			<td>Fentanyl is a powerful synthetic opioid analgesic.</td>
		</tr>
		<tr>
			<td>Flumazenil</td>
			<td>Fresenius Kabi</td>
			<td></td>
			<td>Benzodiazepine antagonist is used for the complete or partial reversal of the sedative effects caused by benzodiazepines.</td>
		</tr>
		<tr>
			<td>High temperature cautery</td>
			<td>Fiab</td>
			<td>F7244</td>
			<td>Disposable cautery, sterile, high temperature (1200 &#176;C), 28 mm fine tip.</td>
		</tr>
		<tr>
			<td>Instillagel gel</td>
			<td>Farco-pharma</td>
			<td></td>
			<td>Cellulose-based lubricant with local anaesthetic and disinfecting properties.</td>
		</tr>
		<tr>
			<td>Iopromide</td>
			<td>Bayer</td>
			<td></td>
			<td>Non-ionic injectable contrast medium, with iodine.</td>
		</tr>
		<tr>
			<td>Laser Doppler imaging system (perfusion imager moorLDI2-HIR and dedicated software)</td>
			<td>MoorLDI-V6.0, Moor Instruments Ltd, Axminster, UK</td>
			<td>5710</td>
			<td>The angiogenesis models uses Laser Doppler imaging to assess blood perfusion in the hind limbs, one of which is ligated surgically. Dedicated measurement and comparison software allows the definition of regions of interest for blood flow assessment on the ischaemic versus non-ischaemic limb to establish a &#34;reperfusion ratio&#34; which can be assessed as often as needed and over a number of days on the same subject.</td>
		</tr>
		<tr>
			<td>Lubrithal Eye Gel</td>
			<td>Dechra</td>
			<td></td>
			<td>Eye gel used in animals for prevention of dry eyes during anaesthesia. Lubricating and moisturising action on cornea and conjunctiva.</td>
		</tr>
		<tr>
			<td>Male Wistar Han IGS (Crl:WI(Han) rats</td>
			<td>Charles River Laboratories, Spain</td>
			<td></td>
			<td>Twelve to fourteen-week-old</td>
		</tr>
		<tr>
			<td>Metedomidin</td>
			<td>Virbac</td>
			<td>037/01/07RFVPT</td>
			<td>Medetomidine is a synthetic drug used as surgical anesthetic.</td>
		</tr>
		<tr>
			<td>Midazolam</td>
			<td>Labesfal</td>
			<td></td>
			<td>Benzodiazepine medication is used for anesthesia and procedural sedation.</td>
		</tr>
		<tr>
			<td>Monoplan Angiography System</td>
			<td>Philips Medical Systems</td>
			<td>Azurion 7 M20</td>
			<td>A stationary diagnostic fluoroscopic x-ray system specifically designed to optimize the capability of users to visually and quantitatively evaluate the anatomy and function of blood vessels of the heart, brain and other organs, as well as the lymphatic system.</td>
		</tr>
		<tr>
			<td>Mosquito forceps</td>
			<td>Henry Schein</td>
			<td>102-4346</td>
			<td>Hartman-Mosquito Hemostatic Forcep Curved 3-1/2&#34; Stainless Steel</td>
		</tr>
		<tr>
			<td>Needle Holder</td>
			<td>Henry Schein</td>
			<td>100-2570</td>
			<td>Needle holder Mayo-Hegar, stainless steel, 14 cm</td>
		</tr>
		<tr>
			<td>Ophtalmic Needle Holder</td>
			<td>Asico</td>
			<td>AE-6143</td>
			<td>Needle holder barranquer most delicate without lock</td>
		</tr>
		<tr>
			<td>Pentobarbital Sodium</td>
			<td>Ecuphar</td>
			<td></td>
			<td>Pentobarbital Sodium is the sodium salt of pentobarbital used for euthanasia.</td>
		</tr>
		<tr>
			<td>Pointed Forceps</td>
			<td>Aesculap</td>
			<td>BD335R</td>
			<td>Microforceps, 0.30 mm tip</td>
		</tr>
		<tr>
			<td>Polysorb 6.0</td>
			<td>Medtronic (Covidien)</td>
			<td>UL-101</td>
			<td>Coated Synthetic Absorbable Suture aimed to reduce the inflammatory reaction in tissues, followed by gradual encapsulation of the suture by fibrous connective tissue.</td>
		</tr>
		<tr>
			<td>Providone-Iodine</td>
			<td>Mylan</td>
			<td></td>
			<td>Povidone-iodine 10% is an antiseptic drug, used as a disinfectant before and after surgery.</td>
		</tr>
		<tr>
			<td>Scalpel Blade n&#186;11</td>
			<td>B Braun</td>
			<td>BB511</td>
			<td>Carbon steel, sterile</td>
		</tr>
		<tr>
			<td>Spring Scissor</td>
			<td>Henry Schein</td>
			<td>600-4826</td>
			<td>Surgical scissors 31 castroviejo</td>
		</tr>
		<tr>
			<td>Surgical Forceps</td>
			<td>Aesculap</td>
			<td>BD33R</td>
			<td>Microforceps, 0.20mm tip</td>
		</tr>
		<tr>
			<td>Surgical Scissor</td>
			<td>Aesculap</td>
			<td>MA873R</td>
			<td>Micro Iris Scissor, curved shrap tips</td>
		</tr>
		<tr>
			<td>SurgiPro 7.0</td>
			<td>Medtronic (Coviden)</td>
			<td>VP-702-X</td>
			<td>Non-Absorbable Monofilament Polypropylene Suture indicated for use in general soft tissue ligation.</td>
		</tr>
	</tbody>
</table>

urethral stricture induction followed by buccal mucosa graft urethroplasty in a rat model

尿道重建是泌尿科医生的一个重要专业领域。当需要尿道移植时，颊粘膜被认为是最佳选择，尽管在某些情况下，它不合适或需要优化以修复给定的狭窄。因此，开发创新程序并评估其在实验模型中的假定成功对于满足临床需求至关重要。为此，本研究描述了一种方案，其中通过电烙术诱导Wistar大鼠尿道狭窄。1 周后，使用从下唇采集的颊粘膜移植物进行尿道重建，并以腹侧高嵌方式放置。逆行尿道造影显示，与狭窄诱导后的相应值相比，尿道成形术后的尿道直径显着改善。此外，使用激光多普勒通过血液灌注分析评估移植物的位置。正如预期的那样，深蓝色区域对应于非血管化的颊粘膜移植物。该手术可以成功地模拟尿道损伤和组织调节的正常病理生理过程，以及使用颊粘膜移植物以可重复的方式进行尿道重建，并可作为未来基于组织工程或尿道移植物的研究基础。

共12只雄性Wistar大鼠，体重400-500 g，12-14周龄，用于尿道狭窄诱导。1 周后进行了逆行尿道造影 （RUG1）8，证实了该技术的成功。在狭窄诱导水平上以毫米为单位测量尿道直径。在此之后，在大鼠尿道的腹面进行颊粘膜移植物的尿道成形术。在尿道成形术后14天，将相同的大鼠提交到第二次逆行尿道造影（RUG2），并在移植物放置水平上以毫米为单位测量尿道直径。平均 RUG1 和 RUG2 ?...

Watch this Scientific Journal Video about Urethral Stricture Induction Followed by Buccal Mucosa Graft Urethroplasty in a Rat Model at JoVE.com

Urethral Stricture Induction Followed by Buccal Mucosa Graft Urethroplasty in a Rat Model

在本方案中，在Wistar大鼠中开发了尿道狭窄诱导，然后用颊粘膜移植物进行尿道重建。进行逆行尿道造影和激光多普勒评估，验证尿道重建（狭窄形成后）和移植物放置。

大鼠模型中的尿道狭窄诱导后进行颊粘膜移植尿道成形术

Urethral reconstruction is an important area of expertise for urologists. The buccal mucosa is considered the best option when urethral grafting is ...

在该方案中，使用颊粘膜移植物的尿道重建首次在局部病理生理环境中开发，这对于模拟疾病至关重要。该技术使我们能够开发创新疗法，并以转化方式研究其分子机制和临床优势。首先，将保护性眼凝胶涂在麻醉大鼠的双眼上。将动物置于热垫上的背侧褥疮位置，并将解剖显微镜放置在大鼠上方进行外科手术。缩回阴茎包皮后，在阴茎腺体背侧放置浅表七零缝合线。将针架留在原位以保持阴茎缩回。将一根 22 号静脉导管放入尿道，使用润滑凝胶进行导尿。用手术刀刀片在皮肤上做一个一厘米长的腹侧切口。用镊子和弹簧剪刀解剖组织层，直到尿道暴露于中轴水平。用电烙装置在尿道的一侧和阴茎中轴的腹侧施加电流。用六零可吸收缝合线闭合切口。取出导尿管和牵引缝合线。将大鼠置于腹侧褥疮位置，并皮下注射抗镇静溶液。用聚维酮碘清洁麻醉大鼠的下唇、颊粘膜、阴茎和腹部皮肤。在下唇的两侧和中间放置三条七零缝合线。在每个里面放一只蚊子，使嘴唇缩回，露出内部粘膜。使用弹簧剪刀和尖镊子，从下唇的颊内粘膜中采集四毫米的移植物。将移植物放入装有无菌 0.9% 盐水的小受体中。用海绵压缩供体区域。取下先前放置的下唇缝合线，露出阴茎。将一根 22 号静脉导管放入尿道，使用润滑凝胶进行烧灼。使用弹簧剪刀，做一个环形的冠状下切口，并将阴茎脱到根部。解剖剩余层以暴露尿道。在冠状沟远端约3毫米处做一个4毫米的纵向腹侧切口，在狭窄处刮出尿道。在刮刀的每一侧放置两条七零保持缝合线。在每个里面留下一只蚊子以缩回尿道。在刮刀的两端放置两条不可吸收的七哦缝合线。将颊粘膜移植物以腹侧高嵌方式放置，粘膜侧朝向尿道腔。将其中一条缝合线穿过移植物，并用运行缝合线进行半椭圆。拔下导尿管并重新定位皮肤。用六零可吸收间断缝合线闭合环形冠下切口。取下牵引缝合线，然后如前所述施用抗镇静溶液。狭窄诱导大鼠输尿管直径为0.92 mm，移植尿道成形术两周后改善至1.59 mm。与尿道成形术前动物相比，尿道成形术后的平均血流量显着减少。电流过大会导致尿道闭合的风险。移植物必须使粘膜朝向尿道腔，并采用水密缝合线，以避免瘘管。可以进行任何类型的移植物尿道成形术，可以评估不同类型的移植物。这种实验模型使我们能够优化移植物的使用并探索新的治疗策略，例如对转化研究产生影响的组织工程材料，以满足临床需求。

Research

JoVE Journal

Medicine

A Model of Chronic Nutrient Infusion in the Rat

在大鼠模型慢性营养输液

Chronic exposure to excessive levels of nutrients is postulated to  affect the function of several organs and tissues and to contribute to the  ...

科研

医学

Ultrasound Imaging-guided Intracardiac Injection to Develop a Mouse Model of Breast Cancer Brain Metastases Followed by Longitudinal MRI

超声成像引导下腔内注射到患乳腺癌脑转移其次是纵向的MRI的小鼠模型

Breast cancer brain metastasis, occurring in 30% of breast cancer patients at stage IV, is associated with high mortality. The median survival is only 6 ...

Acute Brain Trauma in Mice Followed By Longitudinal Two-photon Imaging

急性脑外伤小鼠其次是纵向双光子成像

Although acute brain trauma often results from head damage in different accidents and affects a substantial fraction of the population, there is no ...

Candida albicans Biofilm Development on Medically-relevant Foreign Bodies in a Mouse Subcutaneous Model Followed by Bioluminescence Imaging

Candida albicans Biofilm Development on Medically-relevant Foreign Bodies in a Mouse Subcutaneous Model Followed by Bioluminescence Imaging

白色念珠菌生物膜发展在医学上，国外相关机构在小鼠皮下型号其次是生物发光成像

Candida albicans biofilm development on biotic and/or abiotic surfaces represents a specific threat for hospitalized patients. So far, C. albicans ...

A Rat Model of Ventricular Fibrillation and Resuscitation by Conventional Closed-chest Technique

室颤由传统的封闭胸技术大鼠模型和复苏

A rat model of electrically-induced ventricular fibrillation followed by cardiac resuscitation using a closed chest technique that incorporates the basic ...

Induction of Accelerated Atherosclerosis in Mice: The "Wire-Injury" Model

小鼠加速动脉硬化的诱导："电线损伤"模型

Atherosclerosis is a proliferative fibro-inflammatory disease developing in the arterial wall, inducing a deficient blood flow or a lack of blood flow. ...

Vein Interposition Model: A Suitable Model to Study Bypass Graft Patency

静脉介入型号：研究旁路移植通畅一个合适的模型

Bypass grafting is an established treatment method for coronary artery disease. Graft patency continues to be the Achilles heel of saphenous vein grafts. ...

Using a Chemical Biopsy for Graft Quality Assessment

使用化学活检进行移植质量评估

Kidney transplantation is a life-saving treatment for a large number of people with end-stage renal dysfunction worldwide. The procedure is associated ...

Acupuncture in a Rat Model of Asthma

哮喘鼠模型中的针灸

A high platform can fix rats without restriction and completely expose the acupoints on the back during acupuncture manipulation. This article describes ...

Organ Ischemia-Reperfusion Injury by Simulating Hemodynamic Changes in Rat Liver Transplant Model

器官缺血-输液损伤模拟大鼠肝脏移植模型的血动力学变化

Orthotopic liver transplantation (OLT) in rats is a tried and proven animal model used for preoperative, intraoperative, and postoperative studies, ...