Aneta Ostróżka-Cieślik

Comparing the effect of Biolasol® and HTK solutions on maintaining proper homeostasis, indicating the kidney storage efficiency prior to transplantation.

BACKGROUND Maintaining proper homeostasis involving normal physiological level of extra- and intracellular solutions is one of the factors that determine restoring the life functions of a transplanted organ. The aim of this study was to demonstrate the effectiveness of the newly developed Biolasol(®) solution in kidney storage and to compare its protective effect to the standard HTK solution. MATERIAL/METHODS Isolated porcine kidneys were perfused, preserved (24 and 48 h) and reperfused with Biolasol(®) and HTK solutions. The perfusate samples were used to analyze pH; the amount of released indicator enzymes aspartate aminotransferase (AST), alanine aminotransferase (ALT), and lactate dehydrogenase (LDH); and the concentration of sodium, potassium and magnesium. RESULTS Kidney storage in the HTK liquid may cause metabolic acidosis after 24 and 48 hour preservation (pH drop below 6.8). pH of perfusates sampled after perfusion and reperfusion with Biolasol(®) solution was within the range 6.8-7.7. The content of sodium ions during perfusion and reperfusion was the closest to the reference values while using the Biolasol(®) solution. Only Biolasol(®) ensured normal homeostasis of Mg2+ ions. In the presence of the HTK solution their level was significantly (more than 1000%) higher than the normal physiological value. For both solutions, ALT and AST activities were within the normal range or differed only slightly. CONCLUSIONS Biolasol(®) and HTK solutions protect kidneys against ischemic damage. Still, Biolasol(®) offers better homeostasis maintenance, which may suggest it more effectively preserves kidneys prior to transplantation.

The effect of selected antioxidants on the kinetics of changes in the stability of an HTK solution: a technical note.

Summary and ConclusionsThe effect of selected antioxidants (vitamin C, cysteine, fumaric acid) on the stability of a 0.3-mM solution of HTK has been determined. The stability of the HTK solution was determined using the changes in histidine content at elevated temperatures. The rate of the amino acid decomposition in this solution was 42.3% lower than in a solution without an antioxidant.Vitamin C is the most effective antioxidant. An HTK solution stored at +5°C is stable for 450 days with vitamin C added to it, for 265 days with cysteine, for 242 days with fumaric acid, and for 260 days with no antioxidant.

The Effect of HTK Solution Modification by Addition of Prolactin on Biochemical Indices Reflecting Ischaemic Damage to Porcine Kidney

INTRODUCTION The aim of our study was to evaluate the impact of perfusion with HTK solution, modified by the addition of prolactin (PRI), on selected biochemical parameters of porcine renal damage within 24 and 48 hours after the onset of cold ischemia time. METHODS Each study group consisted of 10 adult pigs. During harvesting the kidneys were rinsed with Ringers solution (group 1), HTK (group 2), and HTK+PRL in a dose of 0.2 mg/dL, 0.02 mg/dL, and 0.01 mg/dL in groups 3, 4 and 5, respectively. The levels of lactate dehydrogenase, asparagine (AST) and alanine aminotransferases, lactates, total protein, potassium and calcium were determined in the perfusate. After 24 and 48 hours the rinsing procedure and the abovementioned tests were repeated. RESULTS After 24 hours of storage, in 4 groups, significantly lower levels of LDH (U/L) were recorded compared with HTK solution alone, namely 235 ± 93 versus 271 ± 125 (perfusion minute, 0), and 55 ± 21 versus 125 ± 94 (30th minute). Similar behavior pattern was presented by AST (U/L) and potassium (mmol/L), and the results were 31 ± 8 versus 35 ± 12 and 16 ± 10 versus 29 ± 14, and 12 ± 3 versus 16 ± 3 and 10 ± 1 versus 13 ± 1, respectively. The changes described above were not observed in the 48th hour of reperfusion. CONCLUSION Our study results indicate the possibility of cytoprotective action of PRL after adding it to the fluid perfusing kidneys during cold ischemia. This effect, observed after 24 hours of storage, was to a considerable extent dose dependent. In our experiment the effect was pronounced only at 0.02 mg/dL supply of PRL.

Hepatoprotective Effect of Prolactin and Cysteine Contained in Perfusion and Preservation Solutions on Porcine Liver Stored in Simple Hypothermia

INTRODUCTION The aim of our study was to determine the results of histidine-tryptophan-ketoglutarate (HTK) solutions modified by the addition of the antioxidant cysteine (Cys), and of prolactin (PRL) on storage of isolated porcine livers. METHODS We measured in the media of isolated livers stored for 24 hours in HTK (control group) or modified HTK+Cys (0.3 mmol/L)+PRL (3 IU/L study group) the amounts of aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH), lactic acid as well as Ca (II), Mg (II), Na (I) and K (I) ions during a 30-minute perfusion after 24 hours of storage. RESULTS All tested markers were released more slowly into HTK+Cys+PRL with less release of K(I) and Mg(II) and greater of Na(I) and Ca(II) ions. CONCLUSIONS Addition of the Cys and PRL to HTK positively affected 24-hour storage of isolated livers.

The Influence of Condition on Permeation of Ca(II) Ions from Solutions of Selected Calcium’s Salts Through Model Membrane

The permeation of calcium’s ions from calcium solutions of fumarate, gluconate, and citrate through model membrane from the donor chamber to the acceptor chamber has been examined. Process was traced depending on the concentration of the appropriate calcium’s salts (1, 2.5, and 5 mmol/l) and pH value of acceptor environment (1.3, 6.2, and 7.4) which imitated natural conditions appearing in the digestive tract. The amount of permeating Ca(II) ions (percent) and their Ca(II) availability AUC (0–6 h) has been determined. In dependence on the conditions, penetration was as follows: 30.3–95.2% of calcium ions from fumarate solution; 73.0–90.1% of Ca(II) from citrate solution; and 19.0–95.0% of Ca from gluconate solution. The investigation indicates that the amount of permeated Ca(II) ions and their availability are connected with the concentration of the calcium salt and pH of acceptor environment. Fumarate and citrate are available at pH value of acceptor environment 1.3 and 6.2 and gluconate at the pH value of 6.2 and 7.4. These substances are practically unavailable from the acceptor environment at pH value 1.3 for gluconate and 7.4 for fumarate. Results suggest that calcium citrate can be available for organism independently from pH value of acceptor environment.

The Effect of HTK Solution Modification by Addition of Thyrotropin and Corticotropin on Biochemical Indices Reflecting Ischemic Damage to Porcine Kidney

INTRODUCTION The aim of our study was to evaluate the impact of perfusion with HTK (histidine-tryptophan-ketoglutarate, Custodiol®, Dr. Franz Kohler Chemie, Germany) solution, modified by the addition of porcine thyroid-stimulating hormone (TSH) and corticotropin (ACTH), on selected biochemical parameters of porcine renal damage within 24 and 48 hours after the onset of cold ischemia time. METHODS Each study group consisted of 10 adult pigs. During harvesting the kidneys were rinsed with Ringer solution (group 1), HTK (group 2), HTK-TSH (1 μg/dL) or HTK-ACTH (1 μg/dL) in groups 3 and 4. The solutions were cooled to 4°C-6°C. Within 30 minutes of the first perfusion, the discharged fluid was clear and the kidneys cooled to 4°C. The levels of lactate dehydrogenase, asparagine and alanine aminotransferases, lactates, total protein, potassium, calcium, and pH were determined in the perfusate. After 24 and 48 hours the rinsing procedure and the above-mentioned tests were repeated. Differences between the means of 2 independent samples were tested with a nonparametric Mann-Whitney U test. RESULTS As the result of hormone addition, in both time intervals it was possible to observe considerably lower protein concentrations (g/L) in perfusates compared with HTK solution, without an addition. At 24 hours, we measured following values: 36 ± 4, 8 ± 3 and 6 ± 1 versus 48 hours, 34 ± 1, 2 ± 1, and 4 ± 1 in groups 2, 3, and 4. A similar pattern was observed with LDH (U/L) at 48 hours: 662 ± 89, 374 ± 151, and 386 ± 111, respectively. Lactate concentrations (mmol/L) were then significantly higher: 1.4 ± 0.3 in the TSH group and 1.2 ± 0.5 in the ACTH group as opposed to 0.2 ± 0.1 in unmodified HTK group. CONCLUSION We observed the possibility of cytoprotective actions of TSH and ACTH addition to the perfusion fluid during cold ischemia, positive effects that were especially visible upon prolonged 48-hour storage.

Tips for optimizing organ preservation solutions

Organ injury during ischemia is one of the clinical problems of todays transplantation. It occurs during warm ischemia time (WIT) when the blood flow is cut off and during cold ischemia when a graft is chilled in situ until the circulation is restored to the recipient organism. Fast cooling of the organ slows down metabolism and activates intracellular enzymes, which minimizes the effects of warm ischemia. Unfortunately, hypothermia also results in inhibition of ATP synthesis, cell swelling and intracellular acidity. That is why research is continually being conducted to develop new fluids for rinsing and storing organs, as well as to optimize the composition of those that are already in use, which will allow for longer and more effective graft storage and restoration of their optimal functions after transplantation. This article provides current information on rinsing and storage fluids available on the global market. It also discusses tips for the fluid modifications with hormones and micronutrients.