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[Effects of urokinase plasminogen activator on cultured human retinal epithelial cells]
E V Boiko1, D S Mal'tsev2, V O Polyakova3
1Military Medical Academy named after S.M. Kirov, Ministry of Defense of the Russian Federation, 5 Klinicheskaya St., Saint-Petersburg, Russian Federation, 194044; Saint-Petersburg branch of the Academician S.N. Fyodorov IRTC 'Eye Microsurgery', 21 Yaroslava Gasheka St., Saint-Petersburg, Russian Federation, 192283.
Aim:
to study the effects of urokinase plasminogen activator (UPA) on the human retinal pigment epithelium (hRPE) cell culture.
Material And Methods:
The toxicity of 50 U/ml UPA was studied with the trypan blue exclusion test. Cell migration was assessed by the wound healing and modified Boyden chamber assays. Additionally, cell morphology, trypsin resistance, and Ki67 expression were investigated.
Results:
Trypan exclusion test did not reveal any cytotoxicity of 50 U/ml UPA against hRPE cells. The agent appeared able to induce cellular cluster formation and increase the number of spindle-shaped cells (6.4±2.4 cells/field and 67.3±3.2 cells/field in the controls and in the presence of 50 U/ml UPA, respectively, p<0.001). Cell migration in the Boyden chamber also showed a statistically significant increase (1.75-fold, p=0.012). Monolayer wounds were found to heal at an accelerated rate (p<0.05). This effect was dose-dependent, just like the increase in Ki67-positive cells (from 2.5 to 50 U/ml). Moreover, there was a reduction in trypsin resistance of the hRPE cells (the number of resistant cells in the control and 50 U/ml UPA cultures was 5.2±1.7 cells/field and 0.46±0.32 cells/field, respectively, p<0.001).
Conclusion:
UPA, at concentrations of 50 U/ml or less, demonstrates no cytotoxicity against the hRPE cells. The effects of UPA on hRPE include stimulation of epithelial-mesenchymal transition, migration, proliferation, and intercellular interaction. At that, changes in migratory and proliferative activity are dose-dependent.
