ABSTRACT
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 BlueABSTRACT
PURPOSE: To investigate the effects of tetrahydrozoline (THZ) on the survival of cultured human trabecular meshwork cells (HTMC) and the permeability of HTMC monolayer. METHODS: Primary cultured HTMC were exposed to an adrenergic agonist (0.01, 0.1, 1.0 or 10 µM THZ) for 1 day and 3 days. Carboxyfluorescein permeability through the HTMC monolayer was measured using Transwell. Cellular viability and nitric oxide (NO) production were assessed using MTT and Griess assays, respectively. RESULTS: THZ did not affect the cellular survival (p > 0.05) or NO production (p > 0.05). THZ significantly increased the carboxyfluorescein permeability through the HTMC monolayer in a dose-dependent manner compared with non-exposed control (p < 0.05) after exposure for 1 and 3 days. CONCLUSIONS: THZ does not affect the survival of HTMC but decreases the permeability of HTMC monolayer in a dose-dependent manner. Thus, THZ may possibly decrease trabecular outflow.
Subject(s)
Humans , Adrenergic Agonists , Nitric Oxide , Permeability , Trabecular MeshworkABSTRACT
PURPOSE: To investigate the effects of Rho kinase (ROCK) inhibitor on the production of nitric oxide (NO) and expression of endothelial nitric oxide synthase (eNOS) in cultured human trabecular meshwork cells (HTMC). METHODS: Primarily cultured HTMC were exposed to 0 µM, 10 µM or 100 µM Y-27632 for 3 days and NO production was assessed using Griess assay. After 24 hours, the effect of Y-27632 on the contraction of collagen matrix and the permeability of the HTMC monolayer was determined. The expression of eNOS mRNA was assessed using reverse transcription-polymerase chain reaction (RT-PCR) and cellular survival with the 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. RESULTS: In HTMC, 10 µM and 100 µM Y-27632 significantly increased NO production after 1 day and 3 days (p = 0.020 and 0.001, respectively). At 1 day after exposure, Y-276320 significantly relaxed the collagen matrix and increased the permeability of the HTMC monolayer (all p = 0.001) and the eNOS mRNA expression (p = 0.039). CONCLUSIONS: Increased NO production may play a role in the mechanism of increased trabecular outflow associated with ROCK inhibitor.
Subject(s)
Humans , Collagen , Nitric Oxide Synthase Type III , Nitric Oxide , Permeability , rho-Associated Kinases , RNA, Messenger , Trabecular MeshworkABSTRACT
PURPOSE: To investigate the effects of amniotic membrane extract (AME) on the survival of cultured human nasal mucosa fibroblasts. METHODS: Primary cultured human nasal mucosa fibroblasts were exposed to 0, 10, 20, or 30 microg/mL AME for 3 days. The survival of the human nasal mucosa fibroblasts was measured using the MTT assay and apoptosis was evaluated with flow cytometry using annexin-V/propidium iodide double staining. RESULTS: AME decreased significantly in fibroblast proliferation after exposure to 10 microg/mL (p = 0.000), and caused significant apoptosis of the fibroblasts after exposure to 10 microg/mL (p = 0.024). CONCLUSIONS: AME decreased fibroblast proliferation in vitro at least through induction of apoptosis. Therefore, adjuvant use of AME during endonasal dacryocystorhinostomy may improve clinical outcomes.