Einfluss der Selektiven Retinatherapie (SRT) auf inflammatorische Zellmediatoren des subretinalen Raums

Zusammenfassung

Hintergrund Ziel der Studie war es den Einfluss der Selektiven Retinatherapie (SRT) auf die Ausschüttung inflammatorischer Zellmediatoren, wie dem Komplementfaktor-3 (CC3), Tumor Growth Factor-beta2 (TGF-β2), Tumor Necrosis Factor-alpha (TNF-α) und Interferon-gamma (IFN-γ) in einem porcinen Organkulturmodell zu untersuchen.

Material und Methoden Porcine Organkulturexplantate aus retinalem Pigmentepithel (RPE), Bruch-Membran und Choroidea wurden mit 2 gepulsten Lasersystemen (SRT_YLF und SRT_YAG) behandelt (Nd : YLF, λ = 527 nm, Pulsdauer 1,7 µs und Nd : YAG, Wellenlänge 532 nm, Pulsdauer 2,4 – 3 µs). Es wurden 30 Pulse bei einer Repetitionsrate von 100 Hz und einer Spotgröße von 200 × 200 µm appliziert. Es wurde mit einer Energiedichte von 140 mJ/cm² pro Puls (auf der RPE-Zelltodschwelle) und 180 mJ/cm² pro Puls (über der RPE-Zelltodschwelle) behandelt. Die Explantate wurden in modifizierten Ussing-Kammern kultiviert und die Zellvitalität mittels Calcein-AM-Färbung untersucht. Die Sekretion und Expression der Zellmediatoren wurde mittels ELISA bzw. im Western Blot analysiert.

Ergebnisse Vier Tage nach SRT wurde die Regeneration der RPE-Zellen im Bereich der Läsion beobachtet. Ein Tag nach SRT mit 140 mJ/cm² pro Puls zeigte sich eine Reduktion der basolateralen CC3-Sekretion. Nach der Behandlung mit 180 mJ/cm² pro Puls wurde nach 4 Tagen eine verminderte Sekretion von IFN-γ beobachtet.

Schlussfolgerung Die SRT führt zu keiner Induktion der untersuchten proinflammatorischen Zytokine in vitro.

Abstract

Purpose To investigate the effect of Selective Retina Therapy (SRT) on inflammatory key factors such as complement factor-C3 (CC3), tumor growth factor-beta2 (TGF-β2), tumor necrosis factor-alpha (TNF-α) and interferon-gamma (IFN-γ).

Materials and Methods Porcine RPE-Bruchʼs membrane-choroid explants were irradiated with two SRT laser systems, SRT_YLF and SRT_YAG (Nd : YLF laser, wave length 527 nm, pulse duration 1.7 µs and Nd : YAG laser, wave length 532 nm, pulse duration 2.4 – 3 µs). Laser irradiation was performed on a spot size of 200 × 200 µm, 30 pulses, with a repetition rate of 100 Hz, and a radiant exposure of 140 (threshold RPE death) and 180 mJ/cm² per pulse (above threshold RPE death). Explants were cultivated in modified Ussing chambers and culture viability was assessed by calcein-AM cell staining. Secretion of inflammatory factors was analyzed by ELISA. Protein expression of tissue explants was assessed by Western blot.

Results Regeneration of RPE was observed after 4 days. One day after SRT with 140 mJ/cm² per pulse the secretion of basal CC3 decreased in ELISA. Following 180 mJ/cm² radiant exposure, the level of IFN-γ decreased at day 4.

Conclusion SRT does not induce the release of the pro-inflammatory factors analyzed in this in-vitro study.

Schlüsselwörter

altersabhängige Makuladegeneration - diabetisches Makulaödem - intraokulare Entzündungen - retinales Pigmentepithel - Selektive Retinatherapie

Key words

age-related macular degeneration - diabetic macular edema - intraocular inflammation - retinal pigment epithelium - selective retina therapy

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