Objective: The purpose of our work was to conduct technological, analytical, and biological investigations and stability studies of peritoneal dialysis (PD) solutions containing glucose and sodium lactate in single-chamber containers.Methods: Different formulations of PD solutions were prepared and sterilized at a temperature of 121 °C during 15 m. UV-spectrophotometric determination was performed using purified water as a blank. The spectra of the solutions were run in the range of 220 to 400 nm for the identification of an absorption maximum (λmax) and measuring the absorbance at 228-230 nm and λmaxbefore and after heat sterilization. λmax of the most PD solutions after sterilization was found in the range of 273 to 281 nm. The potentiometric determination was done for pH measuring PD solutions before and after sterilization. Alternative analytical procedure of direct argent metric method was employed for fast measuring content of chloride ions. Viability of Vero cells was evaluated using 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) test, neutral red (NR) uptake assay, and sulforhodamine B (SRB) test.Results: The results showed that the absorbance of the most laboratory batches of PD solutions augments significantly with increasing an autoclave heating time to the sterilization temperature and time of its cooling after sterilization. The longer these parameters, the higher is the absorbance at 228-230 and 273-281 nm indicating 3,4-dideoxyglucoson-3-en (3,4-DGE) and 5-hydroxymethylfurfural (5-HMF) contents, respectively. Glucose degradation is practically absent at short terms of heating and cooling an autoclave while sterilized samples are preserved sterile. Stability studies showed a significant decrease in the absorbance at 228-230 nm during storage with the achievement of a nadir and the following weak increase; an elevation or decrease in the absorption maximum; a substantial decline in the pH of the solution after sterilization and an insignificant pH decrease during storage. The viability of kidney cells was the highest in the SRB test and the lowest one was in the MTT test.Conclusion: The influence of sterilization regimes on the quality of conventional PD solutions and their stability during storage were studied. The highest cytotoxicity was detected in the MTT test, and the lowest one was done in the SRB test, indicating the largest vulnerability of mitochondria under the influence of PD solutions compared to the membranes permeability, functioning of kidney cell lysosomes and the ability of cells to synthesize proteins. Our studies could be useful in the context of planning development of PD solutions with the purpose of authorization and domestic manufacture of these solutions in low-and-middle-income countries.
Water is the most common compound of hydrogen and oxygen in the nature. It is a universal solvent of many substances, and therefore chemically pure water does not exist in the nature. The water contained in the body is qualitatively different from ordinary water as it is structured water. Such crystalline structures of water are the matrix of life. Their presence gives possibility of the occurrence of important biophysical processes and biochemical reactions. Insufficient intake of water into the body or its excessive loss leads to dehydration, which is accompanied by thickening of the blood and impairing hemodynamics. Excessive intake of water into the body causes water intoxication. Purpose – to conduct a bibliosemantic analysis of the sources of the literature on the medical and biological functions of water. Research methods – bibliosemantic, analytical, logical methods and generalizion method. Water is the structural basis of cells necessary to maintain their optimal volume. It determines the spatial structure and function of biomolecules. Insufficient intake of water into the body or its excessive loss leads to an impaired hemodynamics. Excessive intake of water into the body causes water intoxication. All disoders of water-salt balance in the body can be divided into two groups: dehydration and hyperhydration. In each group, there are disorders with a decrease, increase, and no change in osmotic pressure (hypotonic, hypertonic, and isotonic disorders, respectively). Water is used in medical and pharmaceutical practice as an excipient, and for the manufacture of allopathic, homeopathic and anthroposophic medicines. The State Pharmacopoeia of Ukraine includes several articles on the use of water depending on the purpose and regulates water quality: 1) highly purified water, water for injections «in bulk» water and sterilised water for injections; 2) purified water: water «in bulk» and water in containers. Cosmetics are presented on the Ukrainian market, the main biologically active compound of which is water, in particular, natural, thermal and micellar. They are widely used in cosmetology. The biomedical function of water in the body is to preserve cell volume, provide turgor to the cells and save the body from temperature fluctuations. Disruption of water-salt balance leads to dehydration or hyperhydration. There are changes with a decrease, increase, and no change in osmotic pressure (hypotonic, hypertonic, and isotonic disorders, respectively). They cause disruption of the life of the whole organism. In pharmaceutical practice, water is widely used for the manufacture of allopathic, homeopathic and anthroposophic medicines. It can be obtained in various ways, but its quality is regulated by the relevant government regulations. In cosmetic practice, water is used not only as a basic solvent, but in the form of natural, micellar and thermal water, where it is a biologically active compound.
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. K e y w o r d s: solutions for peritoneal dialysis, cell viability, kidney cells (Vero line), neutral red, mtt.
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