Development of a high-resolution three-dimensional optical coherence tomography for characterization of epithelial cell products for corneal cell therapy.

IF 1.2 Q4 OPHTHALMOLOGY
Taiwan Journal of Ophthalmology Pub Date : 2025-08-19 eCollection Date: 2025-07-01 DOI:10.4103/tjo.TJO-D-25-00087
Hsu-Hang Yeh, Vladlen Klochkov, Chia-Ying Tsai, Wei-Lun Huang, Sheng-Lung Huang, Wei-Li Chen
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引用次数: 0

Abstract

Purpose: This study aimed to develop a high-resolution full-field optical coherence tomography (HR-FFOCT) system for automated three-dimensional (3D) imaging of cultivated epithelial cell sheets and biopsied tissue used in corneal regenerative therapy.

Materials and methods: A commercial HR-FFOCT system (ApolloVue® S100, AMO, Taiwan), originally for dermatological imaging, was re-engineered for <2 μm resolution imaging of rabbit limbal and oral mucosal tissues for limbal stem cell deficiency treatment. Modifications included a piezoelectric transducer for precise Z-stack acquisition, customized LED illumination for registration, and a specialized platform for culture dishes. The system enabled en face and cross-sectional imaging with 3D reconstruction. Rabbit-derived products for cultivated limbal epithelial transplantation (CLET) and cultivated oral mucosal epithelial transplantation (COMET) were imaged before cultivation. Morphology, stratification, and protein expression were analyzed and validated with immunocytochemistry (ICC).

Results: The optimized system produced high-resolution en face and Z-stack images with accurate alignment, capturing stratified epithelial layers in CLET and oral mucosal tissue. Reconstructed 3D images revealed structural detail consistent with ICC-verified expression of junctional proteins, including occludin and actin. Both two-dimensional and 3D visualization of biopsied oral mucosal tissue was achieved. The system enabled noninvasive monitoring of epithelial sheet architecture and thickness without sectioning.

Conclusion: The reconstructed HR-FFOCT system provides a noninvasive, real-time imaging platform for assessing epithelial cell sheets and biopsied tissue in corneal regenerative therapy. It offers potential for standardizing quality evaluation of cell-based products before transplantation and advancing translational applications in regenerative medicine.

Abstract Image

Abstract Image

Abstract Image

用于角膜细胞治疗的上皮细胞产物表征的高分辨率三维光学相干断层扫描的发展。
目的:本研究旨在开发一种高分辨率的全视野光学相干断层扫描(HR-FFOCT)系统,用于角膜再生治疗中培养上皮细胞片和活检组织的自动三维成像。材料和方法:最初用于皮肤成像的商用HR-FFOCT系统(ApolloVue®S100, AMO,台湾)被重新设计用于结果:优化后的系统产生高分辨率的正面和z堆叠图像,具有精确的对齐,捕获CLET和口腔粘膜组织中的分层上皮层。重建的3D图像显示了与icc验证的连接蛋白表达一致的结构细节,包括occludin和actin。实现了口腔粘膜组织活检的二维和三维可视化。该系统无需切片即可实现对上皮片结构和厚度的无创监测。结论:重建的HR-FFOCT系统为角膜再生治疗中上皮细胞片和活检组织的评估提供了一个无创的实时成像平台。它为移植前标准化细胞产品的质量评估和推进再生医学的转化应用提供了潜力。
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
CiteScore
1.80
自引率
9.10%
发文量
68
审稿时长
19 weeks
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