Biological and analytical studies of peritoneal dialysis solutions

Abstract

The purpose of our work was to conduct biological and analytical studies of the peritoneal dialysis (PD) solutions containing glucose and sodium lactate and establish correlations between cell viability of the Vero cell line and values of analytical indexes of the tested solutions. The results of this study confirm the cytotoxicity of the PD solutions even compared with the isotonic solution of sodium chloride, which may be due to the low pH of the solutions, presence of glucose degradation products (GDPs) and high osmolarity of the solutions, and unphysiological concentrations of glucose and sodium lactate. However, it is not yet known what factors or their combination and to what extent cause the cytotoxicity of PD solutions. In the neutral red (NR) test the weak, almost middle (r = -0.496 and 0.498, respectively) and unexpected correlations were found between reduced viability of monkey kidney cells and increased pH of the PD solutions and between increased cell viability and increased absorbance at 228 nm of the tested PD solutions. These two correlations can be explained by a strong correlation (r = -0.948) between a decrease in pH and an increase in the solution absorbance at 228 nm. The opposite effect was observed in the MTT test. The weak, but expected correlations (r = 0.32 and -0.202, respectively) were found between increased cell viability and increased pH in the PD solutions and between decreased cell viability and increased absorbance at 228 nm of the tested PD solutions. The middle and weak correlations (r = 0.56 and 0.29, respectively) were detected between increased cell viability and increased lactate concentration in the NR test and MTT test. The data of these correlations can be partially explained by the fact that a correlation with a coefficient r = -0.34 was found between decreased pH in the solutions and increased lactate concentration. The very weak correlations (0.138 and 0.196, respectively) were found between increased cell viability and increased glucose concentration in the NR test and MTT test. These experimental data indicate that pH is the dominating factor, which determines almost all of the established correlations. However, the character of the correlations is quite different: the higher the pH, the greater was the cell viability in the MTT test, and conversely, the higher the pH, the lower was the cell viability in the NR test. Secondly, the unexpected correlation coefficient was determined as -0.473 between decreased cell viability in the MTT test and increased cell viability in the NR test. Moreover, this phenomenon indicates that the mitochondrial enzyme succinate dehydrogenase is more vulnerable to the action of PD solutions than membrane permeability. Finally, we conclude that the NR test is not suitable for comparative studies of PD solutions which differ in pH, as it is pH dependent and does not enable the comparison of plausible cell viability.

Keywords

• cell viability
• kidney cells (Vero line)
• MTT
• neutral red
• solutions for peritoneal dialysis