Effect of glucose concentration, osmolality, and sterilization process of peritoneal dialysis fluids on cytokine production by peripheral blood mononuclear cells and polymorphonuclear cell functions in vitro

Abstract

We sought to investigate the effects of high glucose concentration, osmolality, and heat sterilization of peritoneal dialysis fluids on tumor necrosis factor-alpha (TNF-alpha) production by peripheral blood mononuclear cells (PBMC) and polymorphonuclear cell (PMN) functions. Blood samples were obtained from eight healthy volunteers. PBMCs and PMNs were harvested by centrifugation with Ficoll-Hypaque (Sigma, St Louis, MO). PBMC were incubated with an equal volume of test fluids and RPMI for 4 hours (pH equilibrated), followed by incubation for 20 hours in RPMI with or without endotoxin (10 ng/mL). Total TNF-alpha production was measured by radioimmunoassay. PMNs were incubated with pH-adjusted test fluids for 30 minutes. After incubation, phagocytosis was determined by the uptake of 14C-labeled Staphylococcus aureus, oxidative burst by reduction of ferricytochrome C to ferrouscytochrome C on stimulation with phorbol myristate acetate, and enzyme release by measurement of endotoxin-stimulated bactericidal/permeability increasing factor. To study the effects of increasing glucose concentration and osmolality on PBMC and PMN functions, we compared conventional 1.5% Dianeal (1.5%D), (Baxter Healthcare Corp, Deerfield, IL) 2.5% Dianeal (2.5%D), 4.25% Dianeal (4.25%D), and control (RPMI for PBMCs and Hank's balanced salt solution for PMNs). PMNs exposed to 4.25%D exhibited an inhibition of phagocytosis, phorbol myristate acetate (PMA)-stimulated oxidative burst, and bactericidal/permeability increasing factor release compared with control, 1.5%D, or 2.5%D. To study the effects of increased osmolality when controlled for glucose concentration, we compared 1.5%D with 1.5%D in which osmolality was increased to that of 4.25%D with the addition of either sodium chloride (1.5%D+NaCl) or mannitol (1.5%D+M). High osmolality induced higher TNF-alpha production by unstimulated PBMCs and decreased TNF-alpha production by endotoxin-stimulated PBMCs. PMN functions were also inhibited by high osmolality. To study the effects of increased glucose concentration when controlled for osmolality, we compared 4.25%D with 1.5%D+NaCl and 1.5%D+M. High glucose concentration induced an increase in TNF-alpha production by unstimulated PBMCs, a decrease in TNF-alpha production by endotoxin-stimulated PBMCs, and an inhibition of PMN functions. Finally, to investigate the effects of heat sterilization, we compared 4.25%D (heat sterilized) to a filter-sterilized 4.25%D (4.25%D-F). The filter-sterilized fluid induced less changes in PBMC and PMN functions compared with the heat-sterilized fluid. These data suggest that the high glucose concentration, high osmolality, and heat sterilization of peritoneal dialysis fluids adversely affect PBMC and PMN functions. These effects could predispose continuous ambulatory peritoneal dialysis patients to peritonitis, compromise host defense during infection, and jeopardize long-term survival of the peritoneal membrane.