化学交联剂抑制基质金属蛋白酶以阻止圆锥角膜的角膜退化。

IF 3 2区 医学 Q1 OPHTHALMOLOGY Experimental eye research Pub Date : 2024-12-15 DOI:10.1016/j.exer.2024.110208
Adhithya Subramanian Gopalakrishnan, Sumaiya Sirajudeen, Nasrin Banu, Jessica Nunes, Divya T Rajendran, Seema Yadav, Namperumalsamy Venkatesh Prajna, Rachel Williams, Dharmalingam Kuppamuthu, Ramprasad Obula Giridhara Gopalan
{"title":"化学交联剂抑制基质金属蛋白酶以阻止圆锥角膜的角膜退化。","authors":"Adhithya Subramanian Gopalakrishnan, Sumaiya Sirajudeen, Nasrin Banu, Jessica Nunes, Divya T Rajendran, Seema Yadav, Namperumalsamy Venkatesh Prajna, Rachel Williams, Dharmalingam Kuppamuthu, Ramprasad Obula Giridhara Gopalan","doi":"10.1016/j.exer.2024.110208","DOIUrl":null,"url":null,"abstract":"<p><p>The need for better and simpler alternative crosslinking strategies to treat keratoconus (KC) is becoming essential as there is only a single approved way to treat it. Recently, conventional UV-A Riboflavin crosslinking is proven to have some disadvantages such as causing damage to the corneal endothelium and inducing keratocyte apoptosis. A chemical cross-linker (CXL) using carbodiimide chemistry and an octanedioic acid spacer is found effective in stiffening the cornea and has the potential to be developed as an alternative therapy to halt KC progression. In order to investigate the molecular changes induced by the cross-linker, we have analyzed the effect of the cross-linker on the activity of matrix metalloproteases (MMPs) in epithelial and stromal layers of KC corneas and in vitro cellular systems to determine its role in stiffening the KC cornea. At well-optimized concentration, KC corneal buttons were treated with the CXL and the stiffening of the cornea was measured. The collagen fibril assembly in the stroma was analyzed using transmission electron microscopy and the activity of MMPs 2 and 9 were visualized using gelatin zymography. KC corneal fibroblasts in culture and tumor necrosis factor-α (TNF-α) induced human corneal epithelial (HCE) cell line were treated with CXL and secretion of MMPs 1, 2, 3 and 9 were analyzed by enzyme-linked immunosorbent assay (ELISA). We found that the CXL stiffened the KC corneas comparable to the normal corneas, with very less cytotoxicity. The collagen fiber assembly was reorganized in an orderly fashion and fibril density and diameter increased after CXL treatment. The activity of MMPs and cathepsin G in the epithelial and stromal layers of KC tissues decreased post-treatment. Secretion and activity of MMPs from the corneal epithelial and stromal cells after CXL treatment were significantly reduced while the epithelial lysyl oxidase activity increased. The CXL, intended to stop the KC progression, modified the extracellular matrix collagen assembly in the stroma and decreased the secretion of a group of metalloproteases and their activity. We have demonstrated a set of molecular changes effected by the CXL, which might aid in the stiffening of the KC cornea.</p>","PeriodicalId":12177,"journal":{"name":"Experimental eye research","volume":" ","pages":"110208"},"PeriodicalIF":3.0000,"publicationDate":"2024-12-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Inhibition of matrix metalloproteases by a chemical cross-linker to halt the corneal degradation in keratoconus.\",\"authors\":\"Adhithya Subramanian Gopalakrishnan, Sumaiya Sirajudeen, Nasrin Banu, Jessica Nunes, Divya T Rajendran, Seema Yadav, Namperumalsamy Venkatesh Prajna, Rachel Williams, Dharmalingam Kuppamuthu, Ramprasad Obula Giridhara Gopalan\",\"doi\":\"10.1016/j.exer.2024.110208\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>The need for better and simpler alternative crosslinking strategies to treat keratoconus (KC) is becoming essential as there is only a single approved way to treat it. Recently, conventional UV-A Riboflavin crosslinking is proven to have some disadvantages such as causing damage to the corneal endothelium and inducing keratocyte apoptosis. A chemical cross-linker (CXL) using carbodiimide chemistry and an octanedioic acid spacer is found effective in stiffening the cornea and has the potential to be developed as an alternative therapy to halt KC progression. In order to investigate the molecular changes induced by the cross-linker, we have analyzed the effect of the cross-linker on the activity of matrix metalloproteases (MMPs) in epithelial and stromal layers of KC corneas and in vitro cellular systems to determine its role in stiffening the KC cornea. At well-optimized concentration, KC corneal buttons were treated with the CXL and the stiffening of the cornea was measured. The collagen fibril assembly in the stroma was analyzed using transmission electron microscopy and the activity of MMPs 2 and 9 were visualized using gelatin zymography. KC corneal fibroblasts in culture and tumor necrosis factor-α (TNF-α) induced human corneal epithelial (HCE) cell line were treated with CXL and secretion of MMPs 1, 2, 3 and 9 were analyzed by enzyme-linked immunosorbent assay (ELISA). We found that the CXL stiffened the KC corneas comparable to the normal corneas, with very less cytotoxicity. The collagen fiber assembly was reorganized in an orderly fashion and fibril density and diameter increased after CXL treatment. The activity of MMPs and cathepsin G in the epithelial and stromal layers of KC tissues decreased post-treatment. Secretion and activity of MMPs from the corneal epithelial and stromal cells after CXL treatment were significantly reduced while the epithelial lysyl oxidase activity increased. The CXL, intended to stop the KC progression, modified the extracellular matrix collagen assembly in the stroma and decreased the secretion of a group of metalloproteases and their activity. We have demonstrated a set of molecular changes effected by the CXL, which might aid in the stiffening of the KC cornea.</p>\",\"PeriodicalId\":12177,\"journal\":{\"name\":\"Experimental eye research\",\"volume\":\" \",\"pages\":\"110208\"},\"PeriodicalIF\":3.0000,\"publicationDate\":\"2024-12-15\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Experimental eye research\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1016/j.exer.2024.110208\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"OPHTHALMOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Experimental eye research","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1016/j.exer.2024.110208","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"OPHTHALMOLOGY","Score":null,"Total":0}
引用次数: 0

摘要

由于目前只有一种经批准的治疗圆锥角膜的方法,因此需要更好、更简单的交联治疗方法。近年来,传统的UV-A核黄素交联方法被证明存在损伤角膜内皮、诱导角膜细胞凋亡等缺点。一种化学交联剂(CXL)使用碳二亚胺化学和辛烷二酸间隔剂被发现可以有效地硬化角膜,并且有潜力作为一种替代疗法来阻止KC的进展。为了研究交联剂诱导的分子变化,我们分析了交联剂对KC角膜上皮和基质层以及体外细胞系统中基质金属蛋白酶(MMPs)活性的影响,以确定其在KC角膜硬化中的作用。在最佳浓度下,用CXL处理KC角膜按钮,并测量角膜的硬化。用透射电镜分析基质中胶原原纤维的组装,用明胶酶谱法观察MMPs 2和9的活性。用CXL处理培养的KC角膜成纤维细胞和肿瘤坏死因子-α (TNF-α)诱导的人角膜上皮(HCE)细胞株,酶联免疫吸附试验(ELISA)检测MMPs 1、2、3和9的分泌情况。我们发现CXL使KC角膜硬化,与正常角膜相比,细胞毒性非常小。CXL处理后,胶原纤维组装有序重组,纤维密度和直径增加。KC组织上皮和间质层的MMPs和组织蛋白酶G活性降低。CXL处理后,角膜上皮细胞和基质细胞中MMPs的分泌和活性显著降低,而上皮赖氨酸氧化酶活性升高。为了阻止KC的进展,CXL改变了基质中的细胞外基质胶原蛋白组装,降低了一组金属蛋白酶的分泌及其活性。我们已经证明了一组受CXL影响的分子变化,这可能有助于KC角膜的硬化。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
Inhibition of matrix metalloproteases by a chemical cross-linker to halt the corneal degradation in keratoconus.

The need for better and simpler alternative crosslinking strategies to treat keratoconus (KC) is becoming essential as there is only a single approved way to treat it. Recently, conventional UV-A Riboflavin crosslinking is proven to have some disadvantages such as causing damage to the corneal endothelium and inducing keratocyte apoptosis. A chemical cross-linker (CXL) using carbodiimide chemistry and an octanedioic acid spacer is found effective in stiffening the cornea and has the potential to be developed as an alternative therapy to halt KC progression. In order to investigate the molecular changes induced by the cross-linker, we have analyzed the effect of the cross-linker on the activity of matrix metalloproteases (MMPs) in epithelial and stromal layers of KC corneas and in vitro cellular systems to determine its role in stiffening the KC cornea. At well-optimized concentration, KC corneal buttons were treated with the CXL and the stiffening of the cornea was measured. The collagen fibril assembly in the stroma was analyzed using transmission electron microscopy and the activity of MMPs 2 and 9 were visualized using gelatin zymography. KC corneal fibroblasts in culture and tumor necrosis factor-α (TNF-α) induced human corneal epithelial (HCE) cell line were treated with CXL and secretion of MMPs 1, 2, 3 and 9 were analyzed by enzyme-linked immunosorbent assay (ELISA). We found that the CXL stiffened the KC corneas comparable to the normal corneas, with very less cytotoxicity. The collagen fiber assembly was reorganized in an orderly fashion and fibril density and diameter increased after CXL treatment. The activity of MMPs and cathepsin G in the epithelial and stromal layers of KC tissues decreased post-treatment. Secretion and activity of MMPs from the corneal epithelial and stromal cells after CXL treatment were significantly reduced while the epithelial lysyl oxidase activity increased. The CXL, intended to stop the KC progression, modified the extracellular matrix collagen assembly in the stroma and decreased the secretion of a group of metalloproteases and their activity. We have demonstrated a set of molecular changes effected by the CXL, which might aid in the stiffening of the KC cornea.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Experimental eye research
Experimental eye research 医学-眼科学
CiteScore
6.80
自引率
5.90%
发文量
323
审稿时长
66 days
期刊介绍: The primary goal of Experimental Eye Research is to publish original research papers on all aspects of experimental biology of the eye and ocular tissues that seek to define the mechanisms of normal function and/or disease. Studies of ocular tissues that encompass the disciplines of cell biology, developmental biology, genetics, molecular biology, physiology, biochemistry, biophysics, immunology or microbiology are most welcomed. Manuscripts that are purely clinical or in a surgical area of ophthalmology are not appropriate for submission to Experimental Eye Research and if received will be returned without review.
期刊最新文献
Establishment and evaluation of rabbit model for corneal ectasia by photorefractive keratectomy. Single-cell transcriptomic profiling of rat iridocorneal angle at perinatal stages: revisiting the development of periocular mesenchyme. Priming and release of cytokine IL-1β in microglial cells from the retina. Screening of a retinal-targeting Adeno-Associated Virus (AAV) via DNA shuffling. TAT-N24 enhances retinal ganglion cell survival by suppressing ZBP1-PANoptosome-mediated PANoptosis in an acute glaucoma mouse model.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1