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  • Wiley  (2)
  • Lucius, Ralph  (2)
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  • Wiley  (2)
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  • 1
    In: Acta Ophthalmologica, Wiley, Vol. 92, No. 8 ( 2014-12)
    Abstract: Microglia are the local cells of the innate immunity in the retina. Toll‐like receptor ( TLR ) 3 is a receptor of the innate immune system, recognizing viral double‐stranded RNA . Retinal pigment epithelium (RPE) cells express TLR ‐3 and react to TLR ‐3 stimulation. In this study, we investigated the effect of TLR ‐3‐activated RPE on microglia. Methods Primary porcine RPE cells were prepared from freshly prepared pigs' eyes. Retinal microglia were prepared from porcine retina. Expression of the microglia marker Iba1 was evaluated using immunocytochemistry. RPE cells were treated with polyinosinic/polycytidylic acid (Poly I:C; 100 ng/ml, 1  μ g/ml, 10  μ g/ml, 100  μ g/ml) for 24 hr. Either the supernatant was applied to microglia for 6 or 24 hr or microglia cells were directly treated with Poly I:C for 6 and 24 hr. Expression of interleukin (IL)‐1ß, IL‐6, tumour necrosis factor (TNF) α , IL‐10, Cox2 and iNOS was evaluated in quantitative PCR. Phagocytosis was evaluated with a microscope‐based and a fluoroscan‐based phagocytosis assay. Results Retinal pigment epithelium (RPE) cells induce the expression of IL‐6, IL‐1ß and IL‐10 in microglia cells. Microglia cells respond to Poly I:C stimulation in a concentration‐dependent manner with the induction of IL‐1ß, IL‐6, TNF α , Cox2, iNOS and, to a lesser degree, IL‐10. Stimulation of microglia cells with supernatant of Poly I:C‐treated RPE cells further elevated IL‐6, IL‐1ß and Cox2 expression, while it reduced the expression of iNOS . No changes in phagocytosis could be detected. Conclusions TLR ‐3‐activated RPE exacerbates inflammatory response of microglia in a differentiated manner. This indicates that viral infections in the RPE may have a proinflammatory influence on retinal microglia.
    Type of Medium: Online Resource
    ISSN: 1755-375X , 1755-3768
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2014
    detail.hit.zdb_id: 2466981-7
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  • 2
    In: Lasers in Surgery and Medicine, Wiley, Vol. 53, No. 3 ( 2021-03), p. 359-369
    Abstract: The thermal stimulation therapy of the retinal pigment epithelium (TSR) is a sublethal laser technique for thermal stimulation of the retinal pigment epithelium (RPE)‐Bruch's membrane (BrM)‐complex. The aim of this study was to investigate the influence of TSR on the release of age‐related macular degeneration (AMD)‐relevant cell mediators. Study Design/Materials and Methods Porcine RPE‐BrM‐choroid explants were irradiated with a 532 nm continuous wave laser using different spot sizes (100–300 µm, duration 100 milliseconds, 15–100 mW). Cell death was investigated by calcein staining. Explants were treated with grids of sublethal spots and cultivated in modified Ussing chambers. The effect on matrix metalloproteinase‐2 (MMP‐2) and ‐9 was investigated by zymography and quantitative reverse transcription polymerase chain reaction. Secretion of vascular endothelial growth factor (VEGF), pigment epithelium derived factor (PEDF), and transforming growth factor‐β (TGF‐β) was analyzed by enzyme‐linked immunosorbent assay and expression of HSP70 was examined by western blot. Integrity of the RPE/BrM‐complex was analyzed by scanning electron microscopy. Results Laser powers of 15 mW (100 µm) and 45 mW (300 µm) did not induce RPE cell death. The integrity of the RPE/BrM‐complex was not impaired after TSR. After TSR with 300 µm spot size, we observed a significant increase of active MMP‐2 in the basal compartments. The content of PEDF significantly increased in treated explants in both compartments with 100 and 300 µm spot sizes. VEGF and TGF‐β secretion was not triggered by TSR. Conclusions TSR represents a possible RPE stimulating treatment for dry AMD. TSR increases the basal release of active MMP‐2, which might reverse age‐related thickening of BrM. VEGF secretion was not triggered by TSR while anti‐angiogenic PEDF was increased, indicating an induction of an anti‐angiogenic and neuroprotective environment. Lasers Surg. Med. © 2020 Wiley Periodicals LLC
    Type of Medium: Online Resource
    ISSN: 0196-8092 , 1096-9101
    URL: Issue
    Language: English
    Publisher: Wiley
    Publication Date: 2021
    detail.hit.zdb_id: 1475539-7
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