Volume 65, Issue 2, Pages (February 2004)

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Volume 65, Issue 2, Pages 442-451 (February 2004) The uremic solutes p-cresol and indoxyl sulfate inhibit endothelial proliferation and wound repair Laetitia Dou, Emilie Bertrand, Claire Cerini, Valerie Faure, Jose Sampol, Raymond Vanholder, Yvon Berland, Philippe Brunet Kidney International Volume 65, Issue 2, Pages 442-451 (February 2004) DOI: 10.1111/j.1523-1755.2004.00399.x Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 1 Effect of free water-soluble uremic solutes on endothelial cell proliferation. Endothelial cells were incubated in medium without and with free water-soluble solutes at mean uremic concentrations. After incubation, endothelial cell proliferation was measured by enzyme-linked immunosorbent assay (ELISA) analyzing 5-bromo-2-deoxy-uridine (BrdU) incorporation. Data are expressed as mean ± SEM. The number of experiments is between brackets. Kidney International 2004 65, 442-451DOI: (10.1111/j.1523-1755.2004.00399.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 2 Effect of protein-bound uremic solutes on endothelial cell proliferation. Endothelial cells were incubated with protein-bound solutes at mean uremic concentrations or with control medium, without (A) or with (B) of 4% human serum albumin (HSA). After incubation, endothelial cell proliferation was measured by enzyme-linked immunosorbent assay (ELISA) analyzing 5-bromo-2-deoxy-uridine (BrdU) incorporation. Data are expressed as mean ± SEM of change in proliferation versus control. The number of experiments is between brackets. Determination of significant differences versus control was performed by the Student paired t test. *P < 0.05 vs. control; ***P < 0.001 vs. control. Kidney International 2004 65, 442-451DOI: (10.1111/j.1523-1755.2004.00399.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 3 Effect of diluents of uremic solutes on endothelial cell proliferation. Endothelial cells were incubated with culture medium supplemented or not with diluents of uremic solutes. After incubation, endothelial cell proliferation was measured by enzyme-linked immunosorbent assay (ELISA) analyzing 5-bromo-2-deoxy-uridine (BrdU) incorporation. Data are expressed as mean ± SEM of six independent experiments. Kidney International 2004 65, 442-451DOI: (10.1111/j.1523-1755.2004.00399.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 4 Effect of p-cresol (pc) on endothelial cell proliferation. Endothelial cells were incubated in medium without and with different concentrations of p-cresol. Experiments were undertaken without (A) and with (B) 4% human serum albumin (HSA) added to the medium. After incubation, endothelial cell proliferation was measured by enzyme-linked immunosorbent assay (ELISA) analyzing 5-bromo-2-deoxy-uridine (BrdU) incorporation. Data are expressed as mean ± SEM of seven (A) or ten (B) independent experiments. Determination of significant differences was performed by analyis of variance (ANOVA) followed by a Bonferroni's multiple comparison test. **P < 0.01 vs. control; ***P < 0.001 vs control. Kidney International 2004 65, 442-451DOI: (10.1111/j.1523-1755.2004.00399.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 5 Effect of indoxyl sulfate (IS) on endothelial cell proliferation. Endothelial cells were incubated in medium without and with increasing concentration of indoxyl sulfate. Experiments were undertaken without (A) and with (B) 4% human serum albumin (HSA) added to the medium. After incubation, endothelial cell proliferation was measured by enzyme-linked immunosorbent assay (ELISA) analyzing 5-bromo-2-deoxy-uridine (BrdU) incorporation. Data are expressed as mean ± SEM of five (A) or seven (B) independent experiments. Determination of significant differences was performed by analysis of variance (ANOVA) followed by a Bonferroni's multiple comparison test. *P < 0.05 vs. control; **P < 0.01 vs. control; ***P < 0.001 vs. control. Kidney International 2004 65, 442-451DOI: (10.1111/j.1523-1755.2004.00399.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 6 Pictures of endothelial wound repair in presence of p-cresol. Endothelial monolayers were wounded, incubated with control medium or with p-cresol, and endothelial wound repair was analyzed under videomicroscopy. Pictures of wounds before addition of control medium (A) or p-cresol (C). Pictures of wounds after a 24-hour incubation with control medium (B) or with p-cresol at 50 μg/mL (D). In presence of control medium, the original wound was almost completely repaired after 24hours of incubation (A and B). In presence of p-cresol, wound repair was inhibited (C and D). Kidney International 2004 65, 442-451DOI: (10.1111/j.1523-1755.2004.00399.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 7 Effect of p-cresol (pc) on repair of wounded endothelial monolayers. Endothelial monolayers were wounded and incubated with increasing concentrations of p-cresol. Experiments were undertaken without (A) and with (B) 4% human serum albumin (HSA) added to the medium. Endothelial wound repair was analyzed under videomicroscopy. Data are expressed as mean ± SEM of six independent experiments. Determination of significant differences was performed by analysis of variance (ANOVA) followed by a Bonferroni's multiple comparison test. **P < 0.01 vs. control; ***P < 0.001 vs. control. Kidney International 2004 65, 442-451DOI: (10.1111/j.1523-1755.2004.00399.x) Copyright © 2004 International Society of Nephrology Terms and Conditions

Figure 8 Effect of indoxyl sulfate (IS) on repair of wounded endothelial monolayers. Endothelial monolayers were wounded and incubated with increasing concentrations of indoxyl sulfate. Experiments were undertaken without (A) and with (B) 4% human serum albumin (HSA) added to the medium. Endothelial wound repair was analyzed under videomicroscopy. Data are expressed as mean ± SEM of six independent experiments. Determination of significant differences was performed by analysis of variance (ANOVA) followed by a Bonferroni's multiple comparison test. **P < 0.01 vs. control; ***P < 0.001 vs. control. Kidney International 2004 65, 442-451DOI: (10.1111/j.1523-1755.2004.00399.x) Copyright © 2004 International Society of Nephrology Terms and Conditions