Effects of low molecular weight heparins and unfractionated heparin on viability of human umbilical vein endothelial cells

Abstract

The aim of this study was to investigate the effects of bemiparin, nadroparin, enoxaparin, and heparin on viability of human umbilical vein endothelial cells (HUVEC). Cultivation of HUVEC was performed in an incubator having 5 % CO2 at 37 A degrees C and with predefined supplemented medium, until cell monolayers attained confluence which occurred after 7 days. The assays were performed in the exponential growth phase of the cells. The cell viability was assessed using the cleavage of tetrazolium salts added to the culture medium. Heparin sodium, enoxaparin sodium, bemiparin sodium, and nadroparin calcium with concentrations of 100, 10, and 1 IU/100 mu L were used for the proliferation assay in which cells were incubated for 24, 48, and 72 h with these drugs. The experiments were conducted in four replicates. Among the study drugs with maximal concentration used in the experiments (100 IU/100 mu L), heparin was found to be associated with the lowest viability score in 24 and 48 h, while bemiparin showed the lowest at 72 h. Bemiparin 100 IU/100 mu L was significantly associated with lower viability score than that of bemiparin 10 IU/100 mu L and bemiparin 1 IU/100 mu L at every time interval. Among gradual concentrations of enoxaparin, however, concentration of 1 IU/100 mu L was associated with the lowest viability scores at every time point. Heparin 1 IU/100 mu L, nadroparin 100 IU/100 mu L, and enoxaparin 100 IU/100 mu L groups had the highest viability score after 72 h of incubation. Among low molecular weight heparins (LMWHs), 100 IU/100 mu L concentration of bemiparin was associated with a more pronounced effect on reducing viability of HUVEC after 72 h of incubation, while nadroparin 100 IU/100 mu L and enoxaparin 100 IU/100 mu L showed the least effects. LMWHs differ both from each other and heparin with respect to their effects on cellular viability of HUVEC in dose- and time-dependent manner.