Potential pro-inflammatory impact of scleral lens midday fogging on human corneal epithelial cells: An in vitro study.

IF 4.1 3区 医学 Q1 OPHTHALMOLOGY
Contact Lens & Anterior Eye Pub Date : 2024-10-01 Epub Date: 2024-05-17 DOI:10.1016/j.clae.2024.102187
Maria K Walker, Carolina Lema, Rachel Redfern
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引用次数: 0

Abstract

Purpose: Midday fogging (MDF) occurs when particulate material accumulates in the fluid reservoir (FR) beneath scleral lenses (SL), and its impact on epithelial cells is unknown. This study examines the in vitro pro-inflammatory effect of the FR on human corneal epithelial cells in varying degrees of MDF.

Methods: Normal SL neophytes were recruited to wear SL 8 h daily for 4 days. Following 8 h on days 1 and 4, optical coherence tomography (OCT) images were acquired for MDF quantification using ImageJ, and the FR was collected. FR samples from the same eye were later pooled, diluted 2-fold and applied on human telomerase-immortalized corneal epithelial (hTCEpi) cells cultured on Terasaki microwell plates. Tumor necrosis factor (TNF)-α and culture media were used as positive and negative controls, respectively. After a 30-minute treatment, the nuclear factor-kappa B (NF-κB) pathway was measured by NF-κB-p65 immunofluorescence and images were analyzed with ImageJ. Pearson's correlation was conducted to determine the association between median nuclear fluorescence and MDF.

Results: Fourteen FR samples with a mean volume of 22 ± 16 µl were tested. Mean MDF severity following 8 h of SL wear was 25 ± 17 units (range 7 - 64). The median nuclear fluorescence (NF-κB-p65 translocation) in cultured hTCEpi cells ranged from 31.43 to 45.16 while the negative and positive controls were 44.71 ± 1.72 and 108.77 ± 68.38, respectively. Although a potential positive trend between MDF and median nuclear fluorescence was observed, Pearson's correlation analysis revealed no significant association (r = +0.48, P = 0.09).

Conclusions: The results suggest that the FR can trigger NF-κB-p65 translocation in hTCEpi cells, which may be associated with MDF severity. This study introduces the use of Terasaki microwell plates for immunofluorescence studies of the FR. The technique is simple, minimizes sample usage, and does not require expensive instrumentation.

巩膜透镜正午雾化对人类角膜上皮细胞的潜在促炎影响:体外研究。
目的:巩膜晶体(SL)下的储液器(FR)中积聚颗粒物质时,就会发生中午起雾(MDF)现象,其对角膜上皮细胞的影响尚不清楚。本研究探讨了在不同程度的 MDF 情况下,FR 对人类角膜上皮细胞的体外促炎作用:方法:招募正常的角膜塑形镜新手,每天佩戴角膜塑形镜 8 小时,连续 4 天。在第 1 天和第 4 天佩戴 8 小时后,使用 ImageJ 采集光学相干断层扫描(OCT)图像以量化 MDF,并采集 FR。随后将同一只眼睛的 FR 样品集中起来,稀释 2 倍,并应用于在 Terasaki 微孔板上培养的人端粒酶永久化角膜上皮细胞(hTCEpi)。肿瘤坏死因子(TNF)-α 和培养基分别作为阳性和阴性对照。处理 30 分钟后,用 NF-κB-p65 免疫荧光法测定核因子-kappa B(NF-κB)通路,并用 ImageJ 分析图像。通过皮尔逊相关性来确定核荧光中位数与 MDF 之间的关联:测试了 14 份平均体积为 22 ± 16 µl 的 FR 样品。佩戴 SL 8 小时后,MDF 的平均严重程度为 25 ± 17 单位(范围为 7 - 64)。在培养的 hTCEpi 细胞中,核荧光(NF-κB-p65 转位)的中位数为 31.43 至 45.16,而阴性和阳性对照组分别为 44.71 ± 1.72 和 108.77 ± 68.38。虽然观察到 MDF 与核荧光中位数之间存在潜在的正相关趋势,但皮尔逊相关分析显示两者之间没有显著关联(r = +0.48,P = 0.09):结果表明,FR 可触发 NF-κB-p65 在 hTCEpi 细胞中的转位,这可能与 MDF 的严重程度有关。本研究介绍了使用寺崎微孔板对 FR 进行免疫荧光研究的方法。该技术操作简单,样本用量最少,而且不需要昂贵的仪器。
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
7.60
自引率
18.80%
发文量
198
审稿时长
55 days
期刊介绍: Contact Lens & Anterior Eye is a research-based journal covering all aspects of contact lens theory and practice, including original articles on invention and innovations, as well as the regular features of: Case Reports; Literary Reviews; Editorials; Instrumentation and Techniques and Dates of Professional Meetings.
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