Objectives: To observe the effect of viper (Ahylysantipfarciasi) venom on attachment to collagen, migration and proliferation of rabbit Tenon's capsular fibroblast (TFs) in tissue culture.
Methods: Anti-adhesion experiment: The 3 -- 5 passage TFs suspended in DMEM medium were pre-incubated for 30 minutes in the presence of various concentrations of viper venom (0, 2.5 x 10(-4), 5.0 x 10(-4), 1.0 x 10(-3) and 5.0 x 10(-3) U/ml) at 37C, 5% CO(2), then inoculated in 24-well plate coated with rat-tail collagen. After incubation for 90 minutes, the floating cells were removed. The attached cells in each well were enumerated microscopically and determined by the value of absorption (A) of 3-(4, 5-dimethylthiazolzyl)-2, 5-diphenyl tetrazodium bromide (MTT). Anti-migration experiment: A denuded area was made when the 3rd passage of TFs was confluent into a monolayer. Cells were then exposed to viper venom (0 -- 5.0 x 10(-3) U/ml) at 37C, 5% CO(2), and the cells having migrated into the denuded area were enumerated every 6 hours. Anti-proliferative experiment: Two hours after the 3rd passage of TFs was incubated in the 24-well plate at 37C, 5% CO(2), they were exposed to different concentrations of viper venom drug (0 -- 5.0 x 10(-3) U/ml). Twenty-four and 48 hours later, the number of cells was determined by MTT method.
Results: Viper venom inhibited TFs from attaching to collagen in a dose-dependent manner and the ID(50) was 1.0 x 10(-3) U/ml. Only did 5.0 x 10(-3) U/ml of viper venom show significant difference from the control during 6 -- 48 hours. The difference of A value was not significant among all groups at 24 and 48 hr.
Conclusions: In vitro, viper venom (> 1.0 x 10(-3) U/ml) can significantly inhibit the attachment of TFs to collagen, and 5.0 x 10(-3) U/ml can inhibit the migration, but can not affect their proliferation.