Name: intravitreal injections in the ovine eye
Uploaded: 2022-07-05T14:00:00
Description: Watch this Scientific Journal Video about Intravitreal Injections in the Ovine Eye at JoVE.com

作者要感谢Steve Heap博士（BVSc，CertVOphthal）在建立该协议和执行Murray等人描述的注射方面的帮助15。作者还感谢来自CureKids New Zealand，Canterbury医学研究基金会，Neurogene Inc和Batten Disease Support and Research Association的资助。

所有实验方案均由林肯大学动物伦理委员会批准，并符合美国国立卫生研究院关于在研究中照顾和使用动物的指南和新西兰动物福利法案（1999）。边境代尔绵羊在14 岁出生时被诊断出来，并在林肯大学的研究农场饲养。三只3个月大的纯合子（CLN5-/-）母羊接受了左眼单次IVT注射，未经治疗的右眼作为内部对照。将视网膜电图和病理学数据与历史健康和受影响的对照数据?...

<ol>
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L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Longitudinal+in+vivo+monitoring+of+the+CNS+demonstrates+the+efficacy+of+gene+therapy+in+a+sheep+model+of+CLN5+Batten+disease.">Longitudinal in vivo monitoring of the CNS demonstrates the efficacy of gene therapy in a sheep model of CLN5 Batten disease.</a> Molecular Therapy. 26 (10), 2366-2378 (2018).</li><li>Murray, S. J., Mitchell, N. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Natural+history+of+retinal+degeneration+in+ovine+models+of+CLN5+and+CLN6+neuronal+ceroid+lipofuscinoses.">Natural history of retinal degeneration in ovine models of CLN5 and CLN6 neuronal ceroid lipofuscinoses.</a> Scientific Reports. 12 (1), 3670 (2022).</li><li>Russell, K. N., Mitchell, N. L., Wellby, M. P., Barrell, G. K., Palmer, D. 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J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intravitreal+gene+therapy+protects+against+retinal+dysfunction+and+degeneration+in+sheep+with+CLN5+Batten+disease.">Intravitreal gene therapy protects against retinal dysfunction and degeneration in sheep with CLN5 Batten disease.</a> Experimental Eye Research. 207, 108600 (2021).</li><li>Ross, 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=Outer+retinal+transduction+by+AAV2-7m8+following+intravitreal+injection+in+a+sheep+model+of+CNGA3+achromatopsia.">Outer retinal transduction by AAV2-7m8 following intravitreal injection in a sheep model of CNGA3 achromatopsia.</a> Gene Therapy. , (2021).</li><li>Boyd, R. 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=Photoreceptor-targeted+gene+delivery+using+intravitreally+administered+AAV+vectors+in+dogs.">Photoreceptor-targeted gene delivery using intravitreally administered AAV vectors in dogs.</a> Gene Therapy. 23 (2), 223-230 (2016).</li><li>Dalkara, 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=In+vivo-directed+evolution+of+a+new+adeno-associated+virus+for+therapeutic+outer+retinal+gene+delivery+from+the+vitreous.">In vivo-directed evolution of a new adeno-associated virus for therapeutic outer retinal gene delivery from the vitreous.</a> Science Translational Medicine. 5 (189), (2013).</li><li>Gearhart, P. M., Gearhart, C., Thompson, D. A., Petersen-Jones, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Improvement+of+visual+performance+with+intravitreal+administration+of+9-cis-retinal+in+Rpe65-mutant+dogs.">Improvement of visual performance with intravitreal administration of 9-cis-retinal in Rpe65-mutant dogs.</a> Archives of Ophthalmology. 128 (11), 1442-1448 (2010).</li><li>Ross, 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=Evaluation+of+photoreceptor+transduction+efficacy+of+capsid-modified+adeno-associated+viral+vectors+following+intravitreal+and+subretinal+delivery+in+sheep.">Evaluation of photoreceptor transduction efficacy of capsid-modified adeno-associated viral vectors following intravitreal and subretinal delivery in sheep.</a> Human Gene Therapy. 31 (13-14), 719-729 (2020).</li><li>Kotterman, M. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antibody+neutralization+poses+a+barrier+to+intravitreal+adeno-associated+viral+vector+gene+delivery+to+non-human+primates.">Antibody neutralization poses a barrier to intravitreal adeno-associated viral vector gene delivery to non-human primates.</a> Gene Therapy. 22 (2), 116-126 (2015).</li><li>Whitehead, M., Osborne, A., Yu-Wai-Man, P., Martin, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Humoral+immune+responses+to+AAV+gene+therapy+in+the+ocular+compartment.">Humoral immune responses to AAV gene therapy in the ocular compartment.</a> Biological Reviews. 96 (4), 1616-1644 (2021).</li><li>Yun, C., Oh, J., Hwang, S. -. Y., Kim, S. -. W., Huh, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Subconjunctival+hemorrhage+after+intravitreal+injection+of+anti-vascular+endothelial+growth+factor.">Subconjunctival hemorrhage after intravitreal injection of anti-vascular endothelial growth factor.</a> Graefe's Archive for Clinical and Experimental Ophthalmology. 253 (9), 1465-1470 (2015).</li><li>Christensen, L., Cerda, A., Olson, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Real-time+measurement+of+needle+forces+and+acute+pressure+changes+during+intravitreal+injections.">Real-time measurement of needle forces and acute pressure changes during intravitreal injections.</a> Clinical &amp; Experimental Ophthalmology. 45 (8), 820-827 (2017).</li><li>Allmendinger, A., Butt, Y. L., Mueller, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraocular+pressure+and+injection+forces+during+intravitreal+injection+into+enucleated+porcine+eyes.">Intraocular pressure and injection forces during intravitreal injection into enucleated porcine eyes.</a> European Journal of Pharmaceutics and Biopharmaceutics. 166, 87-93 (2021).</li><li>Ross, M., Ofri, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+future+of+retinal+gene+therapy:+Evolving+from+subretinal+to+intravitreal+vector+delivery.">The future of retinal gene therapy: Evolving from subretinal to intravitreal vector delivery.</a> Neural Regeneration Research. 16 (9), 1751-1759 (2021).</li><li>Henein, C., 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=Hydrodynamics+of+intravitreal+injections+into+liquid+vitreous+substitutes.">Hydrodynamics of intravitreal injections into liquid vitreous substitutes.</a> Pharmaceutics. 11 (8), 371 (2019).</li><li>Park, I., Park, H. S., Kim, H. K., Chung, W. K., Kim, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Real-time+measurement+of+intraocular+pressure+variation+during+automatic+intravitreal+injections:+An+ex-vivo+experimental+study+using+porcine+eyes.">Real-time measurement of intraocular pressure variation during automatic intravitreal injections: An ex-vivo experimental study using porcine eyes.</a> PloS One. 16 (8), 0256344 (2021).</li><li>Willekens, K., 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=Intravitreally+injected+fluid+dispersion:+Importance+of+injection+technique.">Intravitreally injected fluid dispersion: Importance of injection technique.</a> Investigative Ophthalmology &amp; Visual Science. 58 (3), 1434-1441 (2017).</li><li>Peynshaert, K., Devoldere, J., De Smedt, S. C., Remaut, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+and+ex+vivo+models+to+study+drug+delivery+barriers+in+the+posterior+segment+of+the+eye.">In vitro and ex vivo models to study drug delivery barriers in the posterior segment of the eye.</a> Advanced Drug Delivery Reviews. 126, 44-57 (2018).</li><li>Kiss, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Vector+Considerations+for+Ocular+Gene+Therapy.+Adeno-associated+virus+vectors+offer+a+safe+and+effective+tool+for+gene+delivery.">Vector Considerations for Ocular Gene Therapy. Adeno-associated virus vectors offer a safe and effective tool for gene delivery.</a> Retinal Physician. 17, 40-45 (2020).</li><li>Kleine Holthaus, S. -. 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玻璃体内注射是人类眼科中最常见的外科手术之一，已被证明可有效将AAV介导的基因疗法输送到绵羊的视网膜。我们之前已经证明了AAV9的功效。CLN5基因治疗在玻璃体内使用CLN5 NCL15减轻绵羊视网膜功能障碍和变性。希望将这种给药途径转化为人类NCL患者也将证明是有益的。
小容量IVT注射到羊眼的方案相对简单和非侵入性，易于重现，并且易于非专家学习。?...

有几种方法可用于将治疗剂输送到视网膜，包括玻璃体内 （IVT）、视网膜下、脉络膜上、眼周或局部给药。每种给药途径都涉及克服诸如血液 - 视网膜屏障或内部和外部限制膜之类的屏障，并且根据所输送的药物和特定的视网膜靶标1，2具有不同的疗效率。
IVT药物输送涉及将注射到眼睛的玻璃体中，玻璃体是一种占据眼睛后房的凝胶状物质。玻璃体的主要功能是维持眼球的形状并保持眼组织（如晶状体和视网膜）就位。玻璃体液主要由水组成，少量胶原蛋白、透明质酸和其他非胶原蛋白3。IVT注射是一种简单而常见的程序，常规用于治疗各种眼部疾病，包括年龄相关性黄斑变性，糖尿病黄斑水肿，糖尿病视网膜病变，视网膜静脉阻塞和几种遗传性视网膜营养不良4，5。
神经元蜡样脂褐素糖（NCL;巴顿病）是一组致命的溶酶体贮积病，会导致大脑和视网膜严重退化。目前有13种已知的NCL变异，由不同基因（CLN1-8，CLN10-14）的突变引起，主要影响儿童，但发病年龄和疾病严重程度为6。NCL具有共同的进行性症状，包括认知和运动衰退，癫痫发作和视力丧失。NCL无法治愈;然而，脑定向酶替代疗法目前正在针对CLN2疾病7，8进行临床试验，AAV介导的基因疗法在临床前研究中显示出巨大的前景，CLN5基因治疗的临床试验预计将于2022年开始9，10。
许多其他物种发展出天然存在的NCL形式，包括猫，狗，羊和牛。NCL的两种绵羊模型目前正在新西兰积极研究中：Borderdale绵羊的CLN5疾病模型和南汉普郡绵羊的CLN6疾病模型。受影响的绵羊表现出人类疾病的许多临床和病理特征，包括视网膜萎缩和视力丧失10，11。尽管在患有CLN5疾病的绵羊中进行脑定向CLN5基因治疗可以预防或阻止脑萎缩和临床衰退，但接受治疗的绵羊仍然会失去视力9。这突出了治疗视网膜以保护视力和保持更好生活质量的必要性，从而建立了绵羊眼部基因治疗方案。
羊眼代表了人眼的良好模型，因为它在眼球尺寸、玻璃体体积和视网膜结构10、12、13 方面相似。本文详细介绍了IVT将小体积（≤100μL）治疗性病毒载体输送到羊眼的手术方案。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>1 mL low dead-space safety syringe with permanently attached 0.5 inch needle</td>
			<td>Fisher Scientific, Auckland, New Zealand</td>
			<td>05-561-28</td>
			<td>Covidien Monoject Tuberculin Safety syringe or similar</td>
		</tr>
		<tr>
			<td>1.5 mL microcentrifuge tube</td>
			<td>Sigma Aldrich</td>
			<td>HS4323</td>
			<td>Autoclave tubes to sterilise prior to use</td>
		</tr>
		<tr>
			<td>Anesthesia machine with gas bench and monitor&#160;</td>
			<td>Hyvet Anesthesia, Christchurch, New Zealand</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Antibiotic eye drops&#160;</td>
			<td>Teva Pharma Ltd, Auckland, New Zealand</td>
			<td></td>
			<td>Commercial name: Chlorafast (0.5% chloramphenicol)</td>
		</tr>
		<tr>
			<td>BrightMount plus anti-fade mounting medium</td>
			<td>Abcam, Cambridge, United Kingdom</td>
			<td>ab103748</td>
			<td></td>
		</tr>
		<tr>
			<td>DAPI (4&#8242; ,6-diamidino-2-phenylindole dihydrochloride)</td>
			<td>Sigma Aldrich, St. Louis, Missouri, United States</td>
			<td>10236276001</td>
			<td></td>
		</tr>
		<tr>
			<td>Diazepam sedative</td>
			<td>Ilium, Troy Laboratories Pty Ltd, Tauranga, New Zealand</td>
			<td></td>
			<td>5 mg/mL</td>
		</tr>
		<tr>
			<td>Endotracheal tubes</td>
			<td>Flexicare Medical Ltd, Mountain Ash, United Kingdom</td>
			<td></td>
			<td>Standard, cuffed. Sizes 7, 7.5, or 8 depending on sheep size</td>
		</tr>
		<tr>
			<td>Eye speculum</td>
			<td>Capes Medical Ltd, Tauranga, New Zealand</td>
			<td>KP151/14</td>
			<td>Nopa Barraquer-Colibri (10 mm)</td>
		</tr>
		<tr>
			<td>Fenestrated surgical drape</td>
			<td>Amtech Medical Ltd, Whanganui, New Zealand</td>
			<td>DI583</td>
			<td>Or similar&#160;</td>
		</tr>
		<tr>
			<td>Filter Tips</td>
			<td>Interlab, Auckland, New Zealand</td>
			<td></td>
			<td>10, 200, and 1,000 &#181;L&#160;</td>
		</tr>
		<tr>
			<td>Formaldehyde solution (37%)</td>
			<td>Fisher Scientific, Auckland, New Zealand</td>
			<td>AJA809-2.5PL</td>
			<td>Make up to 10% in distilled water with 0.9% NaCl</td>
		</tr>
		<tr>
			<td>Goat anti-rabbit Alexa Fluor 594</td>
			<td>Invitrogen Carlsbad, CA, USA</td>
			<td>&#160;A-11012</td>
			<td>Use at a dilution of 1:500</td>
		</tr>
		<tr>
			<td>Isoflurane anesthetic</td>
			<td>Attane, Bayer Animal Health, Auckland, New Zealand</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Ketamine HCl anesthetic/analgesic</td>
			<td>PhoenixPharm Distributors Ltd, Auckland, New Zealand</td>
			<td></td>
			<td>100 mg/mL</td>
		</tr>
		<tr>
			<td>Laryngoscope (veterinary)</td>
			<td>KaWe Medical, Denmark</td>
			<td></td>
			<td>Miller C blade, size 2</td>
		</tr>
		<tr>
			<td>Needles&#160;</td>
			<td>Capes Medical Ltd, Tauranga, New Zealand</td>
			<td>302025</td>
			<td>BD Hypodermic Needles, or similar</td>
		</tr>
		<tr>
			<td>Non-steroidal anti-inflammatory</td>
			<td>Boehringer Ingelheim (NZ) Ltd, Auckland, New Zealand</td>
			<td>49402/008</td>
			<td>Commercial name: Metacam 20 (20 mg/mL meloxicam)</td>
		</tr>
		<tr>
			<td>Non-toothed forceps</td>
			<td>Capes Medical Ltd, Tauranga, New Zealand</td>
			<td>AB864/16</td>
			<td>Or similar&#160;</td>
		</tr>
		<tr>
			<td>Non-toothed hemostat</td>
			<td>Capes Medical Ltd, Tauranga, New Zealand</td>
			<td>AA150/12</td>
			<td>Or similar&#160;</td>
		</tr>
		<tr>
			<td>Normal goat serum</td>
			<td>Thermo Fisher Scientific, Christchurch, New Zealand</td>
			<td>16210072</td>
			<td></td>
		</tr>
		<tr>
			<td>Oxygen (medical)</td>
			<td>BOC Gas, Christchurch, New Zealand</td>
			<td></td>
			<td>D2 cylinder, gas code 180</td>
		</tr>
		<tr>
			<td>Phosphate buffered saline&#160;</td>
			<td>Thermo Fisher Scientific, Christchurch, New Zealand</td>
			<td>10010023</td>
			<td>Sterile, filtered</td>
		</tr>
		<tr>
			<td>Povidone-Iodine solution</td>
			<td>Capes Medical Ltd, Tauranga, New Zealand</td>
			<td>005835</td>
			<td>Commercial name: Betadine (10% povidone-iodine)</td>
		</tr>
		<tr>
			<td>Rabbit anti-cow glial fibrillary acidic protein (GFAP)</td>
			<td>Dako, Glostrup, Denmark</td>
			<td>Z0334</td>
			<td>Use at a dilution of 1:2,500</td>
		</tr>
		<tr>
			<td>Self-complementary adeno-associated virus serotype 9, containing the chicken beta action (CBh) promoter and codon-optimized ovine CLN5</td>
			<td>University of North Carolina Vector Core, NC, USA.</td>
			<td>scAAV9/CBh-oCLN5opt</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Chloride 0.9% IV Solution</td>
			<td>Capes Medical Ltd, Tauranga, New Zealand</td>
			<td>AHB1322</td>
			<td>Commercial name: Saline solution&#160;</td>
		</tr>
		<tr>
			<td>Subcutaneous antibiotics</td>
			<td>Intervet Schering Plough Animal Health Ltd, Wellington, New Zealand</td>
			<td></td>
			<td>Commercial name: Duplocillin LA (150,000 IU/mL procaine penicillin and 115,000 IU/mL benzathine penicillin)</td>
		</tr>
		<tr>
			<td>Surgical sharp blunt curved scissors&#160;</td>
			<td>Capes Medical Ltd, Tauranga, New Zealand</td>
			<td>SSSHBLC130</td>
			<td></td>
		</tr>
		<tr>
			<td>Terumo Syringe Luer Lock</td>
			<td>Amtech Medical Ltd, Whanganui, New Zealand</td>
			<td>SH159/SH160</td>
			<td>Sterile syringes; 10 mL for drawing up induction drugs, 20 mL for drawing up saline</td>
		</tr>
		<tr>
			<td>Virkon Disinfectant Powder</td>
			<td>EBOS Group Ltd, Christchurch, NZ</td>
			<td>28461115</td>
			<td></td>
		</tr>
	</tbody>
</table>

intravitreal injections in the ovine eye

有几种方法可以将治疗剂输送到视网膜，包括玻璃体内 （IVT）、视网膜下、脉络膜上、眼周或局部给药。IVT药物输送涉及注射到眼睛的玻璃体中，玻璃体是一种充满眼睛后房并保持眼球形状的凝胶状物质。虽然IVT途径的特异性目标性不如视网膜下递送，但它的侵入性要小得多，并且在临床环境中广泛用于一系列眼部疾病。 
我们之前证明了玻璃体内递送腺相关病毒（AAV）介导的基因治疗产物（AAV9.CLN5）在绵羊中具有天然存在的CLN5形式的神经元蜡样脂褐质沉着症（NCL）。受影响的绵羊一只眼睛接受IVT基因治疗，另一只未经治疗的眼睛作为内部对照。治疗后15个月内，接受治疗的眼睛保持视网膜结构和功能，而未经治疗的眼睛在尸检时表现出进行性下降和严重萎缩。根据绵羊研究，CLN5基因治疗产品于2021年9月被美国食品药品监督管理局批准为候选研究新药（IND）。本文详细介绍了IVT将治疗性病毒载体递送到绵羊眼的手术方案。

该研究小组先前已证明 IVT 递送 CLN5 基因治疗载体在减轻 CLN5 NCL 绵羊视网膜功能障碍和变性的功效15.受影响的绵羊接受单次 100 μL IVT 注射包装在 AAV 血清型 9 （AAV9） 载体 （AAV9.CLN5）进入一只眼睛，对侧眼睛作为未经治疗的内部对照。从注射年龄（3个月）到终末期疾病（18个月）每月评估视力。对治疗和未治疗的眼睛以及年龄匹配的健康和受CLN5影响的对照组进行了视网膜组织学...

Watch this Scientific Journal Video about Intravitreal Injections in the Ovine Eye at JoVE.com

Intravitreal Injections in the Ovine Eye

在羊眼中进行玻璃体内注射，目的是将病毒介导的基因治疗传递到视网膜。

绵羊眼玻璃体内注射

There are several methods for the delivery of therapeutic agents to the retina, including intravitreal (IVT), subretinal, suprachoroidal, periocular, or ...

玻璃体内注射可用于将治疗剂输送到视网膜。这里介绍了玻璃体内注射到羊眼的方案，作为人眼的模型。玻璃体内注射比其他眼部注射的侵入性要小得多。玻璃体内注射可以成功地用于递送基因治疗产品，并治疗具有天然存在的神经元蜡样脂褐素糖的绵羊视网膜变性。一旦绵羊被麻醉，通过使用20毫升注射器用1至5%聚维酮碘溶液冲洗眼睛，开始对要注射的眼睛进行无菌消毒。然后，将无菌的开窗手术窗帘放在眼睛上。将眼窥器贴合眼睑以保持眼睛睁开。用镊子抓住眼睛背侧的延髓结膜，并向腹内侧旋转眼球。将针插入眼睛外侧巩膜后方约 7 毫米处，并向后倾斜以避开晶状体。单次注射100微升作为推注，尽可能靠近视网膜，而不干扰视网膜表面。在取出窥器和悬垂物之前，用大约 10 至 15 毫升的 1% 聚维酮碘溶液冲洗眼睛，然后加入 10 毫升盐水。视网膜电图分析显示，治疗的眼睛的视网膜功能保留，而未治疗的眼睛以与CLN5影响的动物类似的方式下降。在接受治疗的眼睛中，视网膜组织学几乎正常化，总视网膜厚度与中央视网膜中的健康对照动物相当。相比之下，未经处理的视网膜厚度与受CLN5影响的动物相当。神经胶质纤维酸性蛋白（GFAP）是视网膜应激和星形胶质细胞的标志物，在治疗的眼睛中的表达低于未经治疗的眼睛，表明治疗后疾病相关的炎症减弱。随着玻璃体内注射到羊眼的方案的建立，绵羊现在可用于测试许多玻璃体内递送治疗剂的安全性和有效性。

Research

JoVE Journal

Medicine

A Mouse Model of the Cornea Pocket Assay for Angiogenesis Study

一个小鼠模型角膜血管生成研究掌上含量

A normal cornea is clear of vascular tissues. However, blood vessels can be induced to grow and survive in the cornea when potent angiogenic factors are ...

科研

医学

Development of an Audio-based Virtual Gaming Environment to Assist with Navigation Skills in the Blind

基于音频的虚拟游戏环境的发展，以帮助盲人的导航技巧

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio ...

Heterotopic Mucosal Engrafting Procedure for Direct Drug Delivery to the Brain in Mice

异位黏膜移植程序直接给药到小鼠的大脑

Delivery of therapeutics into the brain is impeded by the presence of the blood-brain barrier (BBB) which restricts the passage of polar and high ...

Performing Subretinal Injections in Rodents to Deliver Retinal Pigment Epithelium Cells in Suspension

视网膜下进行注射的鼠害提供视网膜色素上皮细胞悬浮

The conversion of light into electrical impulses occurs in the outer retina and is accomplished largely by rod and cone photoreceptors and retinal pigment ...

Trabecular Meshwork Response to Pressure Elevation in the Living Human Eye

小梁响应压升高在活的人类眼睛

The mechanical characteristics of the trabecular meshwork (TM) are linked to outflow resistance and intraocular pressure (IOP) regulation. The rationale ...

Fluorescent Dye Labeling of Erythrocytes and Leukocytes for Studying the Flow Dynamics in Mouse Retinal Circulation

用于研究小鼠视网膜循环中流动动力学的红细胞和白细胞的荧光染料标记

The retinal and choroidal blood flow dynamics may provide insight into the pathophysiology and sequelae of various ocular diseases, such as glaucoma, ...

Transvaginal Mesh Insertion in the Ovine Model

经阴道网格插入在牛模型中

This protocol describes mesh insertion into the rectovaginal septum in sheep using a single vaginal incision technique, with and without the trocar-guided ...

Ovine Lumbar Intervertebral Disc Degeneration Model Utilizing a Lateral Retroperitoneal Drill Bit Injury

Ovine Lumbar椎间盘退变模型利用横向腹膜后位钻伤

Intervertebral disc degeneration is a significant contributor to the development of back pain and the leading cause of disability worldwide. Numerous ...

Ultrahigh Resolution Mouse Optical Coherence Tomography to Aid Intraocular Injection in Retinal Gene Therapy Research

超高分辨率小鼠光学相干层析成像辅助眼内注射在视网膜基因治疗中的应用

HR-SD-OCT is utilized to monitor the progression of photoreceptor degeneration in live mouse models, assess the delivery of therapeutic agents into the ...