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In a primary culture of orbital fibroblasts from GO patients and control subjects treated with H 2 O 2 to induce OS, the antioxidants vitamin C, N-acetyl-L-cysteine, melatonin, retinol, -carotene and vitamin E were found to reduce the manifestations OS (proliferation of fibroblasts, the level of hyaluronic acid, the level of tumor necrosis factor- (TNF-), IFN- and IL1-, which provided the basis for the possible clinical use of these substances in patients with GO (87)

Afterwards, the medium was replaced with fresh DMEM, and the cells were incubated for an additional 48 h