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Abstract
Purpose
To evaluate the response and cellular damage of cultured human pterygial cells according to the concentration and exposure time of topical cyclosporin.
Methods
Human pterygial cells were exposed to a cyclosporin A concentrations of 0.1 µg/ml (0.0001%), 1 µg/ml (0.0001%), 10 µg/ml (0.001%), 100 µg/ml (0.01%), or 500 µg/ml (0.05%) for 5 or 10 minutes. An MTT-based colorimetric assay was performed to assess the metabolic activity of cellular proliferation, and a lactate dehydrogenase (LDH) leakage assay was used to determine cellular damage. The extra-cellular matrix of PIP, laminin and MMP were evaluated, and the measurement of pro-inflammatory cytokine, TNF-α and IL-1b. IL-6, IL-8 was performed using ELISA kits.
Results
The pterygial cellular inhibitory effect of cyclosporin was similar to that of the control according to the concentration and exposure time (p > 0.05). Compared with the control, the level of LDH did not show a statistically significant difference between concentration and exposure time (p > 0.05). There was no significant difference of inhibitory effects by PIP, laminin, or MMP between the experimental and control groups (p > 0.05). The production of TNF-α and IL from the experimental pterygial cells due to the effect of cyclosporin was not significantly different from that of the control at a longer exposure time or stronger concentration (p > 0.05).
Conclusions
The response of pterygial cells to topical cyclosporin A at concentrations less than 0.05% for less than 10 minutes of exposure time showed no prevention of pterygial recurrence. With regard to cellular damage, little effects on inhibition of PIP, laminin, MMP, IL, and TNF-α were observed compared with that of the control.
Figures and Tables
Figure 1
Metabolic activities of cultured human pterygial cells measured with MTT assays by a scanning spectrometer (ELISA reader). There is no significant difference of cellular viabilities between the experimental and control groups with increasing concentration and longer exposure time (p > 0.05).
Figure 2
LDH titers of cultured human pterygial cells exposed in cyclosporine. There is no significant difference of LDH activities in cultured pterygial cells between the experimental and control groups according to concentration and exposure time (p > 0.05).
Figure 3
PIP level of cultured human pterygial cells exposed in cyclosporine. There is no significant difference of PIP levels in cultured pterygial cells between the experimental and control groups according to concentration and exposure time (p > 0.05).
Figure 4
Laminin level of cultured human pterygial cells exposed to cyclosporin. There is no significant difference of laminin levels in cultured pterygial cells between the experimental and control groups according to concentration and exposure time (p > 0.05).
Figure 5
MMP level of cultured human pterygial cells exposed to cyclosporine. There is no significant difference of MMP-1 (upper), MMP-2 (middle), MMP-9 (lower) levels in cultured pterygial cells between the experimental and control groups according to concentration and exposure time (p > 0.05).
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