摘要
PURPOSE: To evaluate the effects of trypan blue (TB) on the survival of cultured human trabecular meshwork cells (HTMCs). METHODS: Primarily cultured HTMCs were exposed to 0.05, 0.10 or 0.50% TB for 1, 5 or 30 min. Cellular survival was assessed using the MTT assay and degree of apoptosis was analyzed with flow cytometry using annexin-V/propidium iodide double staining. RESULTS: Long-term exposure or high concentration of TB decreased the survival of HTMCs (p < 0.05). In flow cytometric analysis, exposure to 0.50% TB for 30 min increased the degree of apoptosis (p < 0.05). Commercial TB decreased cell survival after exposure for 5 min and increased the degree of apoptosis after exposure for 30 min (p < 0.05). CONCLUSIONS: TB may cause cellular damage of cultured HTMCs and apoptosis could be the underlying mechanism. In TB-assisted cataract surgery, TB should be used for the shortest time possible and removed completely.
Subject(s)
Humans , Apoptosis , Cataract , Cell Survival , Flow Cytometry , Trabecular Meshwork , Trypan Blue摘要
PURPOSE: To compare the effects of anti-inflammatory agents, specifically bromfenac, loteprednol, and prednisolone, on the permeability of cultured human trabecular meshwork cell (HTMC) monolayers. METHODS: HTMCs were cultured until confluency in the inner chamber of Transwell, then exposed to 1/1,000 or 1/500 diluted commercial 0.1% bromfenac, 0.5% loteprednol, and 1% prednisolone for 24 hours. The permeabilities of carboxyfluorescein through the HTMC monolayer were measured with a spectrofluorometer after 2 hours in the outer chamber. Cellular viabilities were assessed with an 3-[4,5–dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. RESULTS: Bromfenac and loteprednol diluted at 1/1,000 or 1/500 did not significantly affect the cellular survival (p > 0.05). Bromfenac did not affect the permeability via the HTMC monolayer (p > 0.05) and loteprednol decreased the permeability (p < 0.05). In addition, 1/2,000 prednisolone also decreased the permeability (p < 0.05). CONCLUSIONS: Among the anti-inflammatory agents, the non-steroidal anti-inflammatory agent bromfenac did not affect the permeability, while loteprednol and prednisolone decreased the permeability through the HTMC monolayer. Thus, loteprednol and prednisolone may decrease the trabecular outflow.
Subject(s)
Humans , Anti-Inflammatory Agents , Loteprednol Etabonate , Permeability , Prednisolone , Trabecular Meshwork摘要
PURPOSE: To compare the effects of anti-inflammatory agents, specifically bromfenac, loteprednol, and prednisolone, on the permeability of cultured human trabecular meshwork cell (HTMC) monolayers. METHODS: HTMCs were cultured until confluency in the inner chamber of Transwell, then exposed to 1/1,000 or 1/500 diluted commercial 0.1% bromfenac, 0.5% loteprednol, and 1% prednisolone for 24 hours. The permeabilities of carboxyfluorescein through the HTMC monolayer were measured with a spectrofluorometer after 2 hours in the outer chamber. Cellular viabilities were assessed with an 3-[4,5–dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay. RESULTS: Bromfenac and loteprednol diluted at 1/1,000 or 1/500 did not significantly affect the cellular survival (p > 0.05). Bromfenac did not affect the permeability via the HTMC monolayer (p > 0.05) and loteprednol decreased the permeability (p < 0.05). In addition, 1/2,000 prednisolone also decreased the permeability (p < 0.05). CONCLUSIONS: Among the anti-inflammatory agents, the non-steroidal anti-inflammatory agent bromfenac did not affect the permeability, while loteprednol and prednisolone decreased the permeability through the HTMC monolayer. Thus, loteprednol and prednisolone may decrease the trabecular outflow.