Open AccessAdverse effects of fullerenes on endothelial cells: Fullerenol C60(OH)24 induced tissue factor and ICAM-1 membrane expression and apoptosis in vitro
Author(s) -
Monique P. Gelderman,
Olga Šimáková,
Jeffrey D. Clogston,
Anil K. Patri,
Sheena F. Siddiqui,
Alexander C. Vostal,
Jan Šimák
Publication year - 2008
Publication title -
international journal of nanomedicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.245
H-Index - 128
eISSN - 1178-2013
pISSN - 1176-9114
DOI - 10.2147/ijn.s1680
Subject(s) - apoptosis , tunel assay , dna fragmentation , umbilical vein , egta , chemistry , microbiology and biotechnology , phosphatidylserine , extracellular , calcium , intracellular , fragmentation (computing) , andrology , in vitro , biology , biochemistry , programmed cell death , membrane , medicine , phospholipid , organic chemistry , ecology
We studied the effects of a C60 water suspension at 4 microg/mL (nC60) and the water soluble fullerenol C60(OH)24 at final concentrations of 1-100 microg/mL on human umbilical vein endothelial cells (HUVECs) in culture. We found that a 24 hr treatment of HUVECs with C60(OH)24 at 100 microg/mL significantly increased cell surface expression of ICAM-1(CD54) (67 +/- 4% CD54+ cells vs. 19 +/- 2 % CD540 cells in control; p < 0.001). In addition, this treatment induced the expression of tissue factor (CD142) on HUVECs (54 +/- 20% CD142+ cells vs 4 +/- 2% CD142+ cells in control; p = 0.008) and increased exposure of phosphatidylserine (PS) (29 +/- 2% PS+ cells vs. 12 +/- 5% PS+ cells in control; p < 0.001). Analysis of cell cycle and DNA fragmentation (TUNEL) showed that both nC60 and C60(OH)24 caused G1 arrest of HUVECs and C60(OH)24 induced significant apoptosis (21 +/- 2% TUNEL+ cells at 100 microg/mL of C60(OH)24 vs. 4 +/- 2% TUNEL+ cells in control; p < 0.001). We also demonstrated that both nC60 and C60(OH)24 induced a rapid concentration dependent elevation of intracellular calcium [Ca2+]i. This could be inhibited by EGTA, suggesting that the source of [Ca2+]i in fullerene stimulated calcium flux is predominantly from the extracellular environment. In conclusion, fullerenol C60(OH)24 had both pro-inflammatory and pro-apoptotic effects on HUVECs, indicating possible adverse effects of fullerenes on the endothelium.