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通过基质内注射II型胶原酶建立兔角膜扩张动物模型。

Animal model of corneal ectasia in rabbits by intrastromal injection of type II collagenase.

作者信息

Cano-Gómez L E, Casillas-Casillas E, Andrade-Lozano P, Ventura-Juárez J, Barba-Gallardo L F

机构信息

Maestría en Investigación Biomédica, Centro de Ciencias de la Salud, Universidad Autónoma de Aguascalientes, Aguascalientes, Mexico.

Departamento de Optometría, Centro de Ciencias de la Salud, Universidad Autónoma de Aguascalientes, Aguascalientes, Mexico.

出版信息

Arch Soc Esp Oftalmol (Engl Ed). 2023 Apr;98(4):206-212. doi: 10.1016/j.oftale.2023.03.005. Epub 2023 Mar 4.

Abstract

INTRODUCTION

Collagenase II has been used to induce experimental keratoconus in animal models. However, its effect when administered by intrastromal injection has not been studied, so the purpose of this study was to study the effects of intrastromal injection of collagenase II on corneal surface and corneal morphology.

METHODS

Six New Zealand rabbits were used, collagenase II was administered by intrastromal injection (5μL of 2.5mg/mL) in the right eyes and balanced salt solution in the left eyes. Keratometry was performed to evaluate curvature alteration, also at day 7 corneas were obtained and Hematoxylin-Eosin staining was performed to examine morphologic changes. Likewise, changes in type I collagen expression were investigated by Sirius Red staining and semiquantitative PCR.

RESULTS

K1, K2 and Km presented differences in the means with statistically significant changes. The morphological changes that were demonstrated were degradation and irregular arrangement of the corneal stroma, increase in the cellular density of keratocytes and slight cellular infiltration. Finally, it was demonstrated that there is greater expression of type I collagen fibers in the experimental group as opposed to the controls and the thickness of the fibers also increased due to the action of collagenase II, however, in terms of genetics there were no changes in the expression of type I collagen at molecular level between the control and experimental groups.

CONCLUSIONS

Collagenase II administered by intrastromal injection is able to induce changes in the corneal surface and stroma, being able to simulate a model of keratoconus.

摘要

引言

胶原酶II已被用于在动物模型中诱导实验性圆锥角膜。然而,其通过基质内注射给药时的效果尚未得到研究,因此本研究的目的是探讨基质内注射胶原酶II对角膜表面和角膜形态的影响。

方法

使用6只新西兰兔,右眼通过基质内注射(2.5mg/mL的5μL)给予胶原酶II,左眼给予平衡盐溶液。进行角膜曲率测量以评估曲率变化,同样在第7天获取角膜并进行苏木精-伊红染色以检查形态学变化。此外,通过天狼星红染色和半定量PCR研究I型胶原表达的变化。

结果

K1、K2和Km的平均值存在差异,且变化具有统计学意义。所显示的形态学变化为角膜基质的降解和排列不规则、角膜细胞的细胞密度增加以及轻微的细胞浸润。最后,结果表明与对照组相比,实验组中I型胶原纤维的表达更高,并且由于胶原酶II的作用,纤维厚度也增加,然而,在基因方面,对照组和实验组之间I型胶原在分子水平的表达没有变化。

结论

通过基质内注射给予胶原酶II能够诱导角膜表面和基质的变化,能够模拟圆锥角膜模型。

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