综述
ENGLISH ABSTRACT
角膜缘干细胞缺乏动物模型建立方法研究进展
郭笑霄
李新宇 [综述]
作者及单位信息
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DOI: 10.3760/cma.j.cn115989-20211130-00660
Review of methods for establishing limbal stem cell deficiency in animal model
Guo Xiaoxiao
Li Xinyu
Authors Info & Affiliations
Guo Xiaoxiao
Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
Li Xinyu
Department of Ophthalmology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China
·
DOI: 10.3760/cma.j.cn115989-20211130-00660
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摘要

角膜缘干细胞缺乏(LSCD)是眼科常见疾病,严重的LSCD是角膜病致盲的主要原因之一。LSCD新疗法的探索仍是当今研究的热点。建立LSCD动物模型有助于更深入了解LSCD的发病机制以及评估治疗方法的有效性。目前,LSCD建模的主要方式有机械法、灼烧法、化学法、化学+手术法及基因敲减法。机械法成熟可靠,应用广泛,但可能会引起严重的机械损伤。其中,使用旋转毛刺工具造模损伤均匀且创伤小,但对于工具的使用需要一定的经验积累。灼烧法造模表型稳定,但操作复杂可控性差。化学法造模快速简单,病因与临床病例的吻合度较高,但损伤的深度及范围不可控。化学+手术法角膜表面光滑平整,但易导致角膜中央与角膜缘厚度不一致,影响新生血管生长。基因敲减法复杂且动物饲养困难,应用较少。虽然造模方法众多,但各研究中的造模质量参差不齐。因此,建立可靠、成功率高的LSCD动物模型十分必要。本文就近年来常用LSCD动物模型的建立方法及优缺点进行综述,以期为后续构建合适模型提供借鉴。

角膜缘干细胞缺乏;动物模型;造模
ABSTRACT

Limbal stem cell deficiency (LSCD) is a common ocular surface disease.Severe LSCD can cause irreversible vision loss and the only effective treatment for severe LSCD is keratoplasty, which is limited by the number of donors.Exploring new effective treatments in lieu of keratoplasty remains a hot topic in current research.Since new treatments need to be critically evaluated in reliable animal models that reflect the pathological mechanisms of human LSCD, the establishment of an appropriate animal model is crucial in any experiment.Up to now, the main modeling methods for LSCD are mechanical method, thermal injury, chemical method, chemical method combined with surgery, and gene knock-down.Mechanical method is mature, reliable and widely used, but it can cause serious mechanical damage.Rotate burr tool could be the best mechanical method because of uniform damage and the small trauma, but it requires accumulation of experience.Cornea suffered from thermal injury performs stable, but the operation is complicated and uncontrollable.Chemical method is quick and simple, and the etiology is highly consistent with clinical cases, but the depth and extent of injury are uncontrollable.The chemical method combined with surgery could ensure the smoothness and flattening of the corneal surface, but the thickness of the central cornea and the limbus may be inconsistent, which affects neovascularization.The gene knockdown method is not widely used because of its complicated operations.Although there are different types of methods, the quality of modeling in different studies is not consistent.Therefore, selecting a reliable and efficient animal model of LSCD is not convenient for researchers.This review discussed current methods to establish LSCD model, including their advantages and disadvantages, therefore to provide inspiration for future model improvement.

Limbal stem cell deficiency;Animal model;Modeling methods
Li Xinyu, Email: nc.defudabe.umjt.hjtuynix
引用本文

郭笑霄,李新宇. 角膜缘干细胞缺乏动物模型建立方法研究进展[J]. 中华实验眼科杂志,2024,42(02):202-208.

DOI:10.3760/cma.j.cn115989-20211130-00660

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角膜缘干细胞缺乏(limbal stem cell deficiency,LSCD)是由于角膜缘干细胞(limbal stem cell,LSC)数量减少和/或功能异常导致的角膜上皮稳态失衡,临床上表现为角膜上皮细胞结膜化、持续或反复的角膜上皮缺损,可伴有角膜新生血管、眼表炎症反应或角膜瘢痕的形成,最终可导致角膜盲 [ 1 ]。根据病因,LSCD分为获得性LSCD和遗传性LSCD两大类,目前在我国眼表化学烧伤和热烧伤是LSCD的首要病因。根据角膜上皮结膜化的受累范围是否为全周累及,LSCD可分为部分性和完全性两大类。根据临床检查发现的角膜和角膜缘受累程度,LSCD国际共识进行了临床分期,其中病变未累及视轴或角膜中央直径5 mm区域为LSCD Ⅰ期,累及角膜中央直径5 mm区域为LSCD Ⅱ期,累及全角膜为LSCD Ⅲ期。LSCD的分类及分期对于选择治疗方式和设计手术方案至关重要 [ 1 ]。目前,LSCD的治疗主要是通过手术行角膜缘干细胞移植(limbal stem cell transplantation,LSCT),由于LSCT需要切取健康的角膜缘组织进行移植,该治疗方式本身会对供给处角膜缘造成创伤;此外,对于单次LSCT未能治愈的LSCD,需多次重复手术,而健康的角膜缘组织来源比较受限。因此,利用培养的LSC进行移植也被推荐用于LSCD的治疗 [ 2 , 3 , 4 ],但远期效果并不明确,依然面临着诸多挑战 [ 5 ]。因此,需要进行更为全面的基础研究,对LSCD的发生机制及发展演变过程进行更为深入的探索。而建立理想的动物疾病模型是对LSCD进行细胞水平和分子水平研究以及评价新的治疗措施必不可少的方法和手段。近年来,国内外许多研究对LSCD的造模方法进行了报道,涉及了不同动物种类、不同干预方式。本文综述了近年来LSCD研究常用的动物模型,总结分析动物模型的建立方法、优缺点以及用于研究LSCD的动物和类型选择,以期为后续研究人员构建合适模型提供参考。
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