Stache Nadine, Sterenczak Katharina A, Sperlich Karsten, Marfurt Carl F, Allgeier Stephan, Köhler Bernd, Mikut Ralf, Bartschat Andreas, Reichert Klaus-Martin, Guthoff Rudolf F, Stachs Angrit, Stachs Oliver, Bohn Sebastian
Department of Ophthalmology, Rostock University Medical Center, Rostock, Germany.
Department of Obstetrics and Gynecology, University of Rostock, Rostock, Germany.
Quant Imaging Med Surg. 2022 Oct;12(10):4734-4746. doi: 10.21037/qims-22-15.
The purpose of the present proof-of-concept study was to use large-area confocal laser scanning microscopy (CLSM) mosaics to determine the migration rates of nerve branching points in the human corneal subbasal nerve plexus (SNP).
Three healthy individuals were examined roughly weekly over a total period of six weeks by large-area confocal microscopy of the central cornea. An in-house developed prototype system for guided eye movement with an acquisition time of 40 s was used to image and generate large-area mosaics of the SNP. Kobayashi-structures and nerve entry points (EPs) were used as fixed structures to enable precise mosaic registration over time. The migration rate of 10 prominent nerve fiber branching points per participant was tracked and quantified over the longitudinal period.
Total investigation times of 10 minutes maximum per participant were used to generate mosaic images with an average size of 3.61 mm (range: 3.18-4.42 mm). Overall mean branching point migration rates of (46.4±14.3), (48.8±15.5), and (50.9±13.9) µm/week were found for the three participants with no statistically significant difference. Longitudinal analyses of nerve branching point migration over time revealed significant time-dependent changes in migration rate only in participant 3 between the last two measurements [(63.7±12.3) and (43.0±12.5) µm/week, P<0.01]. Considering individual branching point dynamics, significant differences in nerve migration rate from the mean were only found in a few exceptions.
The results of this proof-of-concept study have demonstrated the feasibility of using confocal microscopy to study the migration rates of corneal subbasal nerves within large areas of the central human cornea (>1 mm). The ability to monitor dynamic changes in the SNP opens a window to future studies of corneal nerve health and regenerative capacity in a number of systemic and ocular diseases. Since corneal nerves are considered part of the peripheral nervous system, this technique could also offer an objective diagnostic tool and biomarker for disease- or treatment-induced neuropathic changes.
本概念验证研究的目的是使用大面积共聚焦激光扫描显微镜（CLSM）拼接图像来确定人角膜基底神经丛（SNP）中神经分支点的迁移速率。
对三名健康个体进行了为期六周的检查，大致每周检查一次，通过对中央角膜进行大面积共聚焦显微镜检查。使用内部开发的用于引导眼球运动的原型系统，采集时间为40秒，对SNP进行成像并生成大面积拼接图像。使用小林结构和神经入口点（EP）作为固定结构，以便随着时间的推移进行精确的拼接图像配准。在纵向研究期间，对每位参与者10个突出的神经纤维分支点的迁移速率进行跟踪和量化。
每位参与者的总检查时间最长为10分钟，用于生成平均大小为3.61毫米（范围：3.18 - 4.42毫米）的拼接图像。三名参与者的总体平均分支点迁移速率分别为（46.4±14.3）、（48.8±15.5）和（50.9±13.9）微米/周，无统计学显著差异。对神经分支点随时间迁移的纵向分析显示，仅在参与者3的最后两次测量之间迁移速率有显著的时间依赖性变化[（63.7±12.3）和（43.0±12.5）微米/周，P<0.01]。考虑到单个分支点的动态变化，仅在少数例外情况下发现神经迁移速率与平均值有显著差异。
本概念验证研究的结果证明了使用共聚焦显微镜研究人中央角膜大面积区域（>1毫米）内角膜基底神经迁移速率的可行性。监测SNP动态变化的能力为未来研究多种全身性和眼部疾病中的角膜神经健康和再生能力打开了一扇窗口。由于角膜神经被认为是周围神经系统的一部分，该技术还可为疾病或治疗引起的神经病变变化提供一种客观的诊断工具和生物标志物。
