Effects Of Povidone Iodine Concentration On Proliferation Of The Cultured Vascular Endothelial Cells

Objective:It is not clear whether povidone iodine(PVP-I)influences the cell proliferation when being used clinically as a common disinfectants.We observed the proliferation and the activity of SOD of the vascular endothelial cell line ECV-304.Material and Methods:1.Cell culture and morphology:The vascular endothelial cell lines ECV-304 were routinely grown in RPMI 1640 medium containing 10%fetal calf serum,penicilin and streptomycin in a humidified incubator at 37℃and 5%CO₂.The cell suspension was prepared with density at 0.8～1.0×10⁵ cells per milliliter and planted in 24 wells.2.MTT analysis:The ECV-304 cells were cultured with PVP-I at various concentration of 50μg/L,100μg/L,......4000μg/L for 4,8,12,24hours.The proliferation was measured by means of MTT analysis and the morphology of the cells was observed.3.Flow Cytometry:Cells were washed with phosphate-buffered saline(PBS), fixed,suspended in PBS and then analyzed using flow cytometer.Data analysis was performed with Cell Quest software.The SPR was used to represent the activity of cell proliferation,SPR=S/(G0/G1+S+G2/M).4.Immunocytochemistry:Detect expression of PCNA and Ki-67 was detected.5.SOD activity:SOD activity was measured by kit.Results:1.Counting and proliferation activation:It has no significant differences among all the concentrations of PVP-I compared with the control group except 3500～4000μg/L at 4h.PVP-I significantly increased proliferation activities of vascular endothelial cells in a certain range of concentration 900-3000μg/L at 8h,800-2500μg/L at 12h and 100-1800μg/L at 24h.The proliferation was significant decreased compared with the control group when the concentration of PVP-I is higher than 3000μg/L after 12h. The morphology of the cells changed with the increasing of PVP-I concentration.2.SPR:The SPR of PVP-I 1000μg/L was higher than the the control group and the PVP-I 4000μg/L group was lower than the control group.3.Immunocytochemistry:Expressions of PCNA and Ki-67 was increased by the increase of the PVP-I concentration when the concentration of PVP-I was lower than 500μg/L,but it was reversed when the concentration was higher than 1000μg/L.4.SOD activity:The SOD activity was influnced by PVP-I.The SOD activity increased when the concentration of PVP-I was lower than 1000μg/L and decreased when the concentration of PVP-I was higher than 1000μg/L.Conclusions:1.The povidone iodine effect on the proliferation activity of endothelial cells is dose-and time-dependent.PVP-I has the diphasic action on the ECV-304 cell line. The proliferation was promoted when the cells were incubated for short time or when PVP-I was at low concentration.The proliferation was inhibited when cells were incubated for long time or when PVP-I was at high concentration.There is a time-dose-effect relationship.2.The effect of different concentrations of povidone iodine on proliferation activity of cultured ECV-304 ceils may be related to free radicle,cell cycle and apoptosis.