Serum-derived extracellular vesicles for the treatment of severe ocular surface disease

IF 5.9 1区 医学 Q1 OPHTHALMOLOGY Ocular Surface Pub Date : 2024-08-17 DOI:10.1016/j.jtos.2024.08.009
Namita Saraf , Rajalakshmy Ayilam Ramachandran , Mou Cao , Andrew Lemoff , Hamid Baniasadi , Danielle M. Robertson
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Abstract

Purpose

Autologous serum is widely used for the treatment of severe ocular surface disease with mixed efficacy. Extracellular vesicles (EVs) are small membrane bound structures present in all body fluids, including serum. This study compared the proteomic, metabolomic, and inflammatory cytokine composition of serum-derived EVs (SDEVs) to that of the soluble free protein fraction and the subsequent capacity of SDEVs to induce corneal epithelial cell migration and inflammation.

Methods

SDEVs were isolated from human serum using size exclusion chromatography. SDEVs were analyzed using nanoparticle tracking analysis, transmission electron microscopy, and western blotting. The effects of SDEVs on corneal epithelial cell migration were tested using a standard scratch assay. Inflammatory cytokines in SDEVs and the free protein fraction were quantified using a microarray. A mutli-omics approach was further used to define SDEV cargo. The ability of SDEVs to modulate inflammation in corneal epithelial cells was quantified using ELISAs.

Results

Western blot and TEM confirmed the presence of SDEVs. Proinflammatory cytokines, along with complement proteins and TGF-β, were decreased in SDEVs compared to serum. Metabolites present in SDEVs were mostly involved in amino acid biosynthesis, the TCA cycle and oxidative phosphorylation. SDEVs exhibited pro-migratory effects similar to serum however, SDEVs did not induce secretion of IL-6 or IL-8.

Conclusions

SDEVs exhibit reduced levels of pro-inflammatory cytokines while retaining the beneficial wound healing properties of serum. Unlike serum, SDEVs do not induce inflammation. SDEVs may represent an alternative option for patients with severe ocular surface disease where traditional autologous serum has failed.

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用于治疗严重眼表疾病的血清衍生细胞外囊泡
目的:自体血清被广泛用于治疗严重的眼表疾病,但疗效参差不齐。细胞外囊泡(EVs)是存在于包括血清在内的所有体液中的小型膜结合结构。本研究比较了血清衍生EVs(SDEVs)与可溶性游离蛋白部分的蛋白质组、代谢组和炎症细胞因子组成,以及SDEVs随后诱导角膜上皮细胞迁移和炎症的能力:方法:使用尺寸排阻色谱法从人血清中分离出 SDEVs。方法:采用尺寸排阻色谱法从人类血清中分离出 SDEV,并利用纳米粒子跟踪分析、透射电子显微镜和 Western 印迹法对 SDEV 进行分析。使用标准划痕试验测试了 SDEV 对角膜上皮细胞迁移的影响。使用芯片对 SDEV 和游离蛋白部分中的炎性细胞因子进行了量化。此外,还采用了一种突变组学方法来确定 SDEV 的载体。使用 ELISAs 对 SDEVs 调节角膜上皮细胞炎症的能力进行了量化:Western印迹和TEM证实了SDEV的存在。与血清相比,SDEVs 中的促炎细胞因子、补体蛋白和 TGF-β 均有所减少。SDEVs 中的代谢物主要参与氨基酸的生物合成、TCA 循环和氧化磷酸化。SDEVs 表现出与血清相似的促迁移作用,但 SDEVs 不会诱导 IL-6 或 IL-8 的分泌:结论:SDEV 降低了促炎细胞因子的水平,同时保留了血清有益的伤口愈合特性。与血清不同,SDEVs 不会诱发炎症。对于传统自体血清治疗失败的严重眼表疾病患者来说,SDEV 可能是一种替代选择。
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来源期刊
Ocular Surface
Ocular Surface 医学-眼科学
CiteScore
11.60
自引率
14.10%
发文量
97
审稿时长
39 days
期刊介绍: The Ocular Surface, a quarterly, a peer-reviewed journal, is an authoritative resource that integrates and interprets major findings in diverse fields related to the ocular surface, including ophthalmology, optometry, genetics, molecular biology, pharmacology, immunology, infectious disease, and epidemiology. Its critical review articles cover the most current knowledge on medical and surgical management of ocular surface pathology, new understandings of ocular surface physiology, the meaning of recent discoveries on how the ocular surface responds to injury and disease, and updates on drug and device development. The journal also publishes select original research reports and articles describing cutting-edge techniques and technology in the field. Benefits to authors We also provide many author benefits, such as free PDFs, a liberal copyright policy, special discounts on Elsevier publications and much more. Please click here for more information on our author services. Please see our Guide for Authors for information on article submission. If you require any further information or help, please visit our Support Center
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