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Jerry Huang, Ling Yeung, Nan-Kai Wang, Yen-Po Chen, Yih-Shiou Hwang, Kuan-Jen Chen, Wei-Chi Wu, Tun-Lu Chen, Chi-Chun Lai; Diluted Preserved Triamcinolone Acetonide for Internal Limiting Membrane Staining: Staining Morphology and Cytotoxicity Evaluations. Invest. Ophthalmol. Vis. Sci. 2013;54(15):5786.
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To determine the proper concentration of preserved triamcinolone acetonide (TA) suitable for internal limiting membrane (ILM) staining and peeling, and to evaluate the toxicity of various concentrations of preserved TA on in vitro retinal pigment epithelium (RPE) and retinal ganglion cells.
In the clinical study, a retrospective review of 20 consecutive patients with idiopathic macular hole and/or macular pucker who underwent surgery with pars plana vitrectomy and TA-assisted internal limiting membrane peeling was performed. Non-diluted and balanced salt solution (BSS) diluted (1:4 and 1:9) TA-assisted ILM stainings were performed and their staining morphology were assessed. The distribution patterns of the TA crystals of various concentrations were also assessed microscopically. In the in vitro study, cytotoxicities of various concentrations of preserved TA diluted in BSS on cultured human RPE cells (ARPE-19) and retinal ganglion cells (RGC5) were evaluated. Cells were treated with BSS, 4, 8, and 40mg/ml concentration of triamcinolone acetonide for 20 minutes. Toxicity was determined by WST-1 mitochondrial dehydrogenase assays.
Clinically, a 1:4 dilution of commercially-available preserved TA demonstrated the best consistency and particle aggregate size for ILM viewing in ILM peeling surgery. Cultured human RPE cells and retinal ganglion cells did not show decrease of mitochondrial dehydrogenase activity in 1:4 (8mg/ml) and 1:9 (4mg/ml) dilutions of preserved TA for 20 minutes.
Diluted preserved TA (≦8mg/ml) demonstrated no toxicity in cultured RPE cells and retinal ganglion cells. A 1:4 dilution (8mg/ml) of preserved TA appears to be a safe and cost-effective alternative for staining the ILM in ILM peeling surgery.
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