Stiffness Measurement of Retinal Capillaries and Subendothelial Matrix using Atomic Force Microscopy.
Stiffness Measurement of Retinal Capillaries and Subendothelial Matrix using Atomic Force Microscopy.
复制标题
使用原子力显微镜测量视网膜毛细血管和内皮下基质的硬度。
DOI：
10.1101/2024.02.28.582372
复制
发表时间：
2024
期刊：
影响因子：
--
通讯作者：
Ghosh,Kaustabh
中科院分区：
文献类型：
--
作者：
Tierno,IreneSantiago;Agarwal,Mahesh;Matisioudis,Nikolaos;Chandrakumar,Sathishkumar;Ghosh,Kaustabh
视网膜毛细血管变性是糖尿病视网膜病变（DR）早期的临床标志。我们最近的研究表明，糖尿病引起的视网膜毛细血管硬度增加在炎症介导的视网膜毛细血管变性中起着至关重要的作用，这是以前未被认识到的。视网膜毛细血管硬化是赖氨酸氧化酶过度表达的结果，赖氨酸氧化酶是一种交联和硬化内皮下基质的酶。由于早期治疗DR有望预防或减缓DR进展和相关的视力丧失，内皮下基质和毛细血管僵硬是早期DR管理的相关和新的治疗靶点。此外，直接测量视网膜毛细血管硬度可以作为一个关键的临床前验证步骤，用于开发新的成像技术，以无创评估动物和人类受试者的视网膜毛细血管硬度。考虑到这一观点，我们在这里提供了一个使用原子力显微镜对小鼠视网膜毛细血管和视网膜内皮下基质进行分离和刚度测量的详细方案。
Retinal capillary degeneration is a clinical hallmark of the early stages of diabetic retinopathy (DR). Our recent studies have revealed that diabetes-induced increase in retinal capillary stiffness plays a crucial and previously unrecognized causal role in inflammation-mediated degeneration of retinal capillaries. Retinal capillary stiffening results from overexpression of lysyl oxidase, an enzyme that crosslinks and stiffens the subendothelial matrix. Since tackling DR at the early stage is expected to prevent or slow down DR progression and associated vision loss, subendothelial matrix and capillary stiffness represent relevant and novel therapeutic targets for early DR management. Further, direct measurement of retinal capillary stiffness can serve as a crucial preclinical validation step for the development of new imaging techniques for non-invasive assessment of retinal capillary stiffness in animal and human subjects. With this view in mind, we here provide a detailed protocol for the isolation and stiffness measurement of mouse retinal capillaries and retinal subendothelial matrix using atomic force microscopy.