Comparative study of physicochemical properties on corneal stromal lenticules following four decellularization methods.

IF 3 2区 医学 Q1 OPHTHALMOLOGY
Bingqing Sun, Zhe Zhang, Yanze Yu, Fei Xia, Yong Ma, Xuan Ding, Xiaosong Han, Ti Wang, Xingtao Zhou, Jing Zhao
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引用次数: 0

Abstract

This study compares the physicochemical properties of corneal stromal lenticules following decellularization via four methods. Human corneal stromal lenticules, derived from small incision lenticule extraction surgery, underwent decellularization with sodium dodecyl sulfate (SDS), Triton X-100 (Tx) combined with SDS, trypsin-ethylenediaminetetraacetic acid (TE), or NaCl combined with deoxyribonuclease (DNase), respectively. Lenticule DNA and glycosaminoglycan (GAG) content, immunofluorescence staining of cell nuclei and collagen, transparency, biomechanics, histological structure, and immunogenicity were examined in each group and compared with fresh lenticules. All decellularized groups exhibited effective cell removal, with no significant decrease in GAG content (all P > 0.05). DNA content decreased in all decellularization groups (all P < 0.01), most notably in the SDS and Tx + SDS groups. Additionally, collagen I and IV fluorescence intensity was reduced in the TE group only (P < 0.0001). Histological staining revealed close similarity in collagen arrangement between the Tx + SDS group and fresh lenticules. Collagen fiber density increased while spacing and diameter decreased in all decellularized groups (all P < 0.05), with partial collagen degradation detected in the TE group. Light transmittance remained above 60% in the visible light spectrum in all groups. The Young's modulus or elastic modulus did not decrease significantly among decellularized lenticules (all P > 0.05). Human leukocyte antigen (HLA)-DR, HLA-ABC, and CD45 expression decreased in the Tx + SDS and NaCl + DNase groups (all P < 0.001). Although all four decellularization methods showed varying decellularization efficacy, Tx + SDS effectively removed cells without damaging corneal morphology, extracellular matrix, or biomechanics, indicating its potential for lenticule storage, transplantation, and bio-scaffold fabrication.

四种脱细胞方法角膜基质皮孔理化性质的比较研究
本研究比较了角膜基质透镜通过四种方法脱细胞后的理化性质。人类角膜基质皮孔来自小切口皮孔提取手术,分别用十二烷基硫酸钠(SDS)、结合 SDS 的 Triton X-100 (Tx)、胰蛋白酶-乙二胺四乙酸(TE)或含脱氧核糖核酸酶(DNase)的氯化钠进行脱细胞处理。对各组皮孔的 DNA 和糖胺聚糖(GAG)含量、细胞核和胶原的免疫荧光染色强度、透明度、生物力学、组织学结构和免疫原性进行了检测,并与新鲜皮孔进行了比较。所有脱细胞组都能有效去除细胞,且 GAG 含量无明显下降(均 P > 0.05)。所有脱细胞组的 DNA 含量均有下降(均 P < 0.01),其中以 SDS 组和 Tx+SDS 组最为明显。此外,只有 TE 组的胶原 I 和 IV 荧光强度降低(P < 0.0001)。组织学染色显示,Tx+SDS 组与新鲜皮孔的胶原排列非常相似。所有脱细胞组的胶原纤维密度都有所增加,而间距和直径都有所减少(P < 0.05),TE 组出现部分胶原降解。在可见光光谱下,所有组的透光率都保持在 60% 以上。脱细胞皮孔的杨氏模量或弹性模量没有明显降低(均为 P > 0.05)。人类白细胞抗原 (HLA)-DR、HLA-ABC 和 CD45 的表达在 Tx+SDS 组和 NaCl+DNase 组均有所下降(均 P <0.001)。虽然四种脱细胞方法的脱细胞效果各不相同,但 Tx+SDS 能有效去除细胞,且不会破坏角膜形态、细胞外基质或生物力学,这表明它具有用于角膜透镜储存、移植和生物杖制造的潜力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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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.
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