Department of Ophthalmology, Saarland University Medical Center, Kirrberger Str. 100, 66424, Homburg, Saarland, Germany,
Graefes Arch Clin Exp Ophthalmol. 2013 Nov;251(11):2585-90. doi: 10.1007/s00417-013-2465-6. Epub 2013 Sep 27.
With increasing resistance of microorganisms to antibiotics, photodynamic inactivation (PDI) may also be a potential therapeutic option in infectious keratitis. As part of the inflammatory response in infectious keratitis, keratocytes produce various interleukins. The purpose of this study was to evaluate the potential anti-inflammatory effect of PDI, analyzing interleukin-1 alpha (IL-1α), interleukine-1 beta (IL-1β), interleukin-6 (IL-6), and interleukin-8 (IL-8) secretion of human keratocytes following PDI, in vitro.
Primary human keratocytes were isolated by digestion in collagenase A (1.0 mg/ml) from human corneal buttons, and cultured in DMEM/Ham's F12 medium supplemented with 10% FCS. Keratocyte cell cultures underwent illumination using red (670 nm) light for 13 min following exposure to 100 nM concentration of the photosensitizer chlorin e6 (Ce6) in the culture medium. Five and 24 hours after PDI, secretion of IL-1α, IL-1β, IL-6, and IL-8 was measured by enzyme-linked immunoabsorbent assay (ELISA).
The secretion of IL-1α was under the measurement limit in treated and untreated cell cultures 5 and 24 h after PDI. Compared to untreated controls, IL-6 and IL-8 secretion of keratocytes decreased (p < 0.05 and 0.0001) significantly 5 hours after PDI, whereas IL-1β secretion remained unchanged. Twenty-four hours after PDI, secretion of IL-1β, IL-6, and IL-8 did not differ significantly from untreated controls.
In the short term, PDI does not have an impact on IL-1α and IL-1β secretion of keratocytes, in vitro. Photodynamic inactivation inhibits IL-6 and IL-8 secretion of keratocytes transiently (5 h), which normalizes 24 h following treatment. Through the short-term impact of chlorine e6-PDI on IL-6 and IL-8 secretion, PDI may inhibit the inflammatory cascade in at least keratocyte cultures.
随着微生物对抗生素的耐药性不断增加，光动力灭活（PDI）也可能成为感染性角膜炎的潜在治疗选择。在感染性角膜炎的炎症反应中，角膜细胞会产生各种白细胞介素。本研究旨在评估 PDI 的潜在抗炎作用，分析体外 PDI 后人类角膜细胞产生的白细胞介素 1 阿尔法（IL-1α）、白细胞介素 1 贝塔（IL-1β）、白细胞介素 6（IL-6）和白细胞介素 8（IL-8）的分泌情况。
通过胶原酶 A（1.0 mg/ml）消化从人角膜纽扣中分离原代人角膜细胞，并在补充有 10% FCS 的 DMEM/Ham's F12 培养基中培养。在用培养基中的光敏剂氯己定（Ce6）暴露 100 nM 浓度后，用红色（670nm）光对角膜细胞培养物进行 13 分钟的照射。PDI 后 5 小时和 24 小时，通过酶联免疫吸附测定（ELISA）测量 IL-1α、IL-1β、IL-6 和 IL-8 的分泌。
PDI 后 5 小时和 24 小时，处理和未处理细胞培养物中 IL-1α 的分泌均低于测量下限。与未处理对照相比，PDI 后 5 小时，角膜细胞的 IL-6 和 IL-8 分泌显著降低（p<0.05 和 0.0001），而 IL-1β 分泌保持不变。PDI 后 24 小时，IL-1β、IL-6 和 IL-8 的分泌与未处理对照无显著差异。
在短期内，PDI 对体外角膜细胞的 IL-1α 和 IL-1β 分泌没有影响。光动力灭活会短暂抑制（5 小时）角膜细胞的 IL-6 和 IL-8 分泌，在治疗后 24 小时恢复正常。通过氯己定-PDI 对 IL-6 和 IL-8 分泌的短期影响，PDI 可能会抑制至少在角膜细胞培养物中的炎症级联反应。
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