Modification of Preservative Fluids with Antioxidants in Terms of Their Efficacy in Liver Protection before Transplantation.

Transplantation is currently the only effective treatment for patients with end-stage liver failure. In recent years, many advanced studies have been conducted to improve the efficiency of organ preservation techniques. Modifying the composition of the preservation fluids currently used may improve graft function and increase the likelihood of transplantation success. The modified fluid is expected to extend the period of safe liver storage in the peri-transplantation period and to increase the pool of organs for transplantation with livers from marginal donors. This paper provides a literature review of the effects of antioxidants on the efficacy of liver preservation fluids. Medline (PubMed), Scopus, and Cochrane Library databases were searched using a combination of MeSH terms: "liver preservation", "transplantation", "preservation solution", "antioxidant", "cold storage", "mechanical perfusion", "oxidative stress", "ischemia-reperfusion injury". Studies published up to December 2023 were included in the analysis, with a focus on publications from the last 30 years. A total of 45 studies met the inclusion criteria. The chemical compounds analyzed showed mostly bioprotective effects on hepatocytes, including but not limited to multifactorial antioxidant and free radical protective effects. It should be noted that most of the information cited is from reports of studies conducted in animal models, most of them in rodents.

Hydrogel Formulations for Topical Insulin Application: Preparation, Characterization and In Vitro Permeation across the Strat-M® Membrane.

Insulin has shown efficacy in the treatment of hard-to-heal wounds, which is mainly due to its role in regulating oxidative stress and inflammatory reactions. The aim of this study was to develop an insulin-hydrogel carrier based on Sepineo™ P 600 and Sepineo™ PHD 100 for application to lesional skin. Preformulation studies of the developed formulations were performed in terms of analysis of the pharmaceutical availability of insulin from the hydrogels through the Strat-M® membrane, and rheological and texture measurements. Insulin is released in a prolonged manner; after a time of 6.5 h, 4.01 IU/cm2 (53.36%) and 3.69 IU/cm2 (47.4%) of the hormone were released from the hydrogel based on Sepineo™ P 600 and Sepineo™ PHD 100, respectively. Rheological analysis showed that the hydrogels tested belong to non-Newtonian, shear-thinning systems with yield stress. The insulin-hydrogel based on Sepineo™ P 600 and Sepineo™ PHD 100 shows optimal application properties. The results obtained provide a basis for further preclinical and clinical studies.

Effect of Copper on the Function of Isolated Porcine Kidneys Stored Using Simple Hypothermia.

Renal ischemia in the peri-transplant period causes a number of changes that adversely affect the initiation of normal vital functions in grafts after transplantation. To minimise the extent of ischemic damage, organs are stored in preservation fluid. The components of the fluid are supposed to ensure stabilisation of the cell cytoskeleton, protect against oxygen free radicals, reduce cell swelling, and ensure endothelial cell integrity. The aim of this study was to analyse the protective effect of Cu2+, as a component of Biolasol preservative fluid, in the prevention of nephron damage occurring during the graft storage period. Analyses of the effectiveness of copper in the presence of prolactin added to Biolasol fluid were also carried out. Forty isolated pig slaughter kidneys were used in the study, avoiding the use of laboratory animals. The kidneys were stored using simple hypothermia. After 2 h and 48 h of graft storage, selected biochemical indicators of renal function were determined in the collected perfusates. The addition of Cu2+ at a dose of 1 µg/L to the composition of Biolasol fluid was found to affect the generation of ischemic damage in the isolated pig kidney. The intensity of the occurrence of these processes is exacerbated by the presence of prolactin at a dose of 0.1 µg/L.

The Potential of Pharmaceutical Hydrogels in the Formulation of Topical Administration Hormone Drugs.

Hormones have attracted considerable interest in recent years due to their potential use in treatment of many diseases. Their ability to have a multidirectional effect leads to searching for new and increasingly effective drugs and therapies. Limitations in formulating drug forms containing hormones are mainly due to their low enzymatic stability, short half-life and limited bioavailability. One of the solutions may be to develop a hydrogel as a potential hormone carrier, for epidermal and transdermal application. This review discusses the main research directions in developing this drug formulation. The factors determining the action of hormones as drugs are presented. An analysis of hydrogel substrates and permeation enhancers that have the potential to enhance the efficacy of hormones applied to the skin is reviewed.

Effect of Manganese on the Efficacy of Preservative Solution in Protecting Isolated Porcine Kidneys.

BACKGROUND: Manganese is a bioelement essential for the proper functioning of the body. Mn2+ is a cofactor for many enzymes, including manganese superoxide dismutase. Together with the enzymes, it participates in cell defense against reactive oxygen species, immune response, and energy production in mitochondria. The aim of this study was to evaluate the effect of manganese added to Biolasol solution on the storage efficiency of isolated pig kidneys. METHODS: Twenty kidneys from Polish Great White pigs were used in the study. Biolasol solution was modified with the addition of Mn2+ at a concentration of 1 µg/L. Grafts were assigned to 2 groups: A/Biolasol (n = 10) and B/Biolasol + Mn2+ (n = 10). The kidneys were washed with the developed solutions, then stored using the static method for 48h under hypothermia (4°C) and washed again. Markers of normal renal function were analyzed in the collected perfusates-aspartate aminotransferase (AST), alanine aminotransferase (ALT), protein, creatinine, urea, and electrolytes. RESULTS: After 48 hours and 30 minutes of renal preservation, activities/concentrations of biochemical parameters in the Biolasol + Mn2+ group increased (ALT: by 33%; AST: by 86%; urea: by 18%; creatinine: 3.25-fold [P < .05]) vs Biolasol. K+ ion concentration was 33% lower in the Biolasol group after 48 hours and 30 minutes (P < .05) vs group B. The presence of manganese in the composition of Biolasol solution influenced the maintenance of optimal Na+ ion concentration. CONCLUSIONS: Manganese added to the Biolasol solution formulation at a dose of 1 µg/L showed low efficacy in protecting nephrons. We do not recommend adding manganese to the preservative solution formulation.

The Effect of Antioxidant Added to Preservation Solution on the Protection of Kidneys before Transplantation.

Ischemia-reperfusion injury is a key clinical problem of transplantology. Current achievements in optimizing organ rinse solutions and storage techniques have significantly influenced the degree of graft damage and its survival after transplantation. In recent years, intensive research has been carried out to maintain the viability of tissues and organs outside the integral environment of the body. Innovative solutions for improving the biochemical functions of the stored organ have been developed. The article discusses directions for modifying preservation solutions with antioxidants. Clinical and experimental studies aimed at optimizing these fluids, as well as perfusion and organ preservation techniques, are presented.

Pre-Formulation Studies: Physicochemical Characteristics and In Vitro Release Kinetics of Insulin from Selected Hydrogels.

Insulin loaded to the polymer network of hydrogels may affect the speed and the quality of wound healing in diabetic patients. The aim of our research was to develop a formulation of insulin that could be applied to the skin. We chose hydrogels commonly used for pharmaceutical compounding, which can provide a form of therapy available to every patient. We prepared different gel formulations using Carbopol® UltrezTM 10, Carbopol® UltrezTM 30, methyl cellulose, and glycerin ointment. The hormone concentration was 1 mg/g of the hydrogel. We assessed the influence of model hydrogels on the pharmaceutical availability of insulin in vitro, and we examined the rheological and the texture parameters of the prepared formulations. Based on spectroscopic methods, we evaluated the influence of model hydrogels on secondary and tertiary structures of insulin. The analysis of rheograms showed that hydrogels are typical of shear-thinning non-Newtonian thixotropic fluids. Insulin release from the formulations occurs in a prolonged manner, providing a longer duration of action of the hormone. The stability of insulin in hydrogels was confirmed. The presence of model hydrogel carriers affects the secondary and the tertiary structures of insulin. The obtained results indicate that hydrogels are promising carriers in the treatment of diabetic foot ulcers. The most effective treatment can be achieved with a methyl cellulose-based insulin preparation.

Effectiveness Assessment of a Modified Preservation Solution Containing Thyrotropin or Follitropin Based on Biochemical Analysis in Perfundates and Homogenates of Isolated Porcine Kidneys after Static Cold Storage.

In this paper, we assess the nephroprotective effects of thyrotropin and follitropin during ischaemia. The studies were performed in vitro in a model of isolated porcine kidneys stored in Biolasol (FZNP, Biochefa, Sosnowiec, Poland) and modified Biolasol (TSH: 1 µg/L; FSH 1 µg/L). We used the static cold storage method. The study was carried out based on 30 kidneys. The kidneys were placed in 500 mL of preservation solution chilled to 4 °C. The samples for biochemical tests were collected during the first kidney perfusion (after 2 h of storage) and during the second perfusion (after 48 h of storage). The results of ALT, AST, and LDH activities confirm the effectiveness of Biolasol + p-TSH in maintaining the structural integrity of renal cell membranes. Significantly reduced biochemical parameters of kidney function, i.e., creatinine and protein concentrations were also observed after 48 h storage. The protective effect of Biasol + p-TSH is most pronounced after 2 h of storage, suggesting a mild course of damage thereafter. A mild deterioration of renal function was observed after 48 h. The results of our analyses did not show any protective effect of Biolasol + p-FSH on the kidneys during ischaemia.

Biochemical Studies in Perfundates and Homogenates of Isolated Porcine Kidneys after Flushing with Zinc or Zinc-Prolactin Modified Preservation Solution Using a Static Cold Storage Technique.

Zinc is an effective anti-inflammatory and antioxidant trace element. The aim of this study was to analyse the protective effect of zinc and zinc-prolactin systems as additives of preservation solutions in the prevention of nephron damage caused during ischemia. The study used a model for storing isolated porcine kidneys in Biolasol®. The solution was modified with the addition of Zn at a dose of 1 µg/L and Zn: 1 µg/L with prolactin (PRL): 0.1 µg/L. After 2 h and 48 h of storage, the levels of alanine aminotransferase, aspartate aminotransferase, lactate dehydrogenase, sodium, potassium, creatinine and total protein were determined. Zinc added to the Biolasol® composition at a dose of 1 µg/L showed minor effectiveness in the protection of nephrons. In turn, Zn2+ added to Biolasol + PRL (PRL: 0.1 µg/L) acted as a prolactin inhibitor. We do not recommend the addition of Zn(II) (1 µg/L) and Zn(II) (1 µg/L) + PRL (0.1 µg/L) to the Biolasol solution.

Therapeutic Potential of Selenium as a Component of Preservation Solutions for Kidney Transplantation.

Selenium has strong antioxidant properties and diverse effects on the immune system. The aim of the study was to analyse the protective effect of selenium as a component of a kidney preservation solution on the prevention of ischemia-reperfusion injury of nephrons. The solution was modified by the addition of Se (1 µg/L), prolactin (0.1 µg/L) and Se with prolactin (1 µg/L Se + 0.1 µg/L PRL). The study used a model for storing isolated porcine kidneys in Biolasol® (modified Biolasol®), which minimizes ischemia-reperfusion injury of grafts. The introduction of Se4+ ions at a dose of 1 µg/L into the Biolasol® preservation solution in the form of Na2SeO3 caused an increase in the activity/concentration of the analysed biochemical parameters: aspartate transaminase, alanine transaminase, urea and protein. This suggests an adverse effect of Se4+ on nephron function during ischemia-reperfusion. The best graft protection was obtained by using Biolasol® modified with the addition of selenium (IV) at a dose of 1 µg/L and prolactin at a concentration of 0.1 µg/L. We proposed the mechanism of prolactin action in the metabolic reduction of selenite (SO32-) during ischemia/reperfusion.

The Role of Hormones and Trophic Factors as Components of Preservation Solutions in Protection of Renal Function before Transplantation: A Review of the Literature.

Transplantation is currently a routine method for treating end-stage organ failure. In recent years, there has been some progress in the development of an optimal composition of organ preservation solutions, improving the vital functions of the organ and allowing to extend its storage period until implantation into the recipient. Optimizations are mostly based on commercial solutions, routinely used to store grafts intended for transplantation. The paper reviews hormones with a potential nephroprotective effect, which were used to modify the composition of renal perfusion and preservation solutions. Their effectiveness as ingredients of preservation solutions was analysed based on a literature review. Hormones and trophic factors are innovative preservation solution supplements. They have a pleiotropic effect and affect normal renal function. The expression of receptors for melatonin, prolactin, thyrotropin, corticotropin, prostaglandin E1 and trophic factors was confirmed in the kidneys, which suggests that they are a promising therapeutic target for renal IR (ischemia-reperfusion) injury. They can have anti-inflammatory, antioxidant and anti-apoptotic effects, limiting IR injury.

The Effectiveness of Rinsing Organs With Selected Preservation Solutions Based on the Analysis of Their Physicochemical Parameters.

BACKGROUND: One of the key elements of successful transplantation is effectively rinsing off the blood and preserving organs under controlled hypothermia. The aim of the study was to analyze the physicochemical parameters of Biolasol (Biochefa, Sosnowiec, Poland) and histidine-tryptophan-ketoglutarate (HTK) (Custodiol) solutions, which are recommended for perfusion and preservation of abdominal parenchymal organs. METHODS: Biolasol and HTK solution were used for the study. The solutions were subjected to physicochemical analysis involving pH, density, osmolarity, viscosity, refractive index, zeta potential, hydrodynamic diameter, and rheological properties. Rheological parameters were associated with morphologic features of fluids. RESULTS: HTK and Biolasol are non-Newtonian systems with pseudoplastic properties and yield stress. The solutions begin to flow under shear stress greater than the ultimate stress. In addition, a nonlinear relationship of their viscosity as a function of velocity gradient (shear rate) was observed. CONCLUSIONS: The solutions reproduce blood properties, which leads to the conclusion about their effective filling of the vascular bed and high efficiency in organ rinsing.

Evaluation of the potential of nanoparticles containing active substances in selected chronic diseases.

Currently, over 80% of all deaths result from the incidence of chronic diseases. The challenge of modern medicine is to develop innovative and effective methods of diagnosis and therapy of these disorders. Different types of particles can be obtained with the use of nanotechnology, including nanoliposomes, solid lipid nanoparticles (SLN), nanospheres, dendrimers, as well as carbon nanotubes (CNT) or fullerenes. All of these nanoparticles (NPs) are suggested to have potential, both in medicine and in diagnosis of many diseases, giving a chance for recovery or longer life for the patients. The studies concerning the usage of NPs show their effective role in most cases. However, there are also concerns about their toxicity or long-term adverse effects. The aim of this literature review was to discuss the results of the latest available studies concerning the efficacy of selected drug-loaded nanocarriers in several chronic diseases, i.e., cardiac disorders, cancer, Alzheimer's disease (AD), Parkinson's disease (PD), and wound healing. We also focused our attention on the methodology of NPs preparation, materials used for their preparation as well as on positive and negative aspects of these nanocarriers.

Effect of Lutropin Concentration on the Efficiency of Isolated Porcine Kidney Storage in Modified Biolasol Solution.

BACKGROUND: Lutropin, a luteinizing hormone (LH), is a glycoprotein hormone secreted by gonadotropic cells of the anterior pituitary gland. LH receptors are a promising therapeutic target. The aim of the study was to analyze the protective effect of LH as a component of preservation solutions on the prevention of ischemia-reperfusion injury in isolated porcine kidneys. METHODS: The study used Biolasol solution (Biochefa, Sosnowiec, Poland) modified with the addition of ascorbic acid and LH at a concentration of 0.01 µg/L, 0.1 µg/L, and 1 µg/L. The study was carried out on 40 kidneys from 20 Polish large white pigs. Kidneys were divided into 4 groups, then washed out of the blood, and cooled by complete immersion in a 500-mL preservation solution at 4°C. The total storage time for grafts in the preservation solutions was 2 hours. After this time, the kidneys were perfused. Perfundates intended for biochemical analyses were collected from the renal vein after 0 minutes and after 30 minutes of perfusion. Another perfusion was performed after 24-hour renal storage. The activity of aspartate aminotransferase and alanine aminotransferase, as well as sodium, potassium, total protein, and urea concentrations was determined. RESULTS: LH as a component of preservation solutions affects the degree of ischemia-reperfusion injury of isolated porcine kidneys. CONCLUSIONS: The results obtained suggest that the efficacy of LH in improving ischemic kidney function is dependent on the dose used. The best graft protection was obtained using the lowest concentration of LH: 0.01 µg/L.

Pharmacological Benefits and Risk of Using Hormones in Organ Perfusion and Preservation Solutions in the Aspect of Minimizing Hepatic Ischemia-Reperfusion Injury during Storage.

For several years, research has been carried out on the effectiveness of solutions for perfusion and preservation of organs, including the liver. There is a search for an optimal pharmacological composition of these solutions, allowing to preserve or improve vital functions of the organ for as long as possible until it is transplanted into a recipient. Hormones due to their properties, often resulting from their pleiotropic effects, may be a valuable component for optimizing the composition of liver perfusion and preservation solutions. The paper presents the current state of knowledge on liver perfusion and preservation solutions modified with hormones. It also shows the characteristics of the hormones evaluated, taking into account their physiological functions in the body.

The Effect of Modified Biolasol Solution on the Efficacy of Storing Isolated Porcine Kidneys.

Biolasol is a newly developed solution for storing the liver, pancreas, kidneys, and heart by simple hypothermia. It exhibits high efficacy in maintaining structural and functional integrity of the graft prior to its transplantation. The solution was modified by the addition of ascorbic acid (0.088g/l) and ascorbic acid with prolactin (1 µg/l PRL + 0.088g/l vitamin C). The effectiveness of the obtained solutions in the protection of nephrons of isolated porcine kidneys was assessed based on the analysis of the activity of ALT (alanine aminotransferase), AST (aspartate aminotransferase), and LDH (lactate dehydrogenase) as well as lactate concentration determined in perfundates collected after 2 h (0' and 30' preservation) and 48 h (0' and 30' preservation) of graft storage. It has been found that the synergistic action of Biolasol components determines the integrity and stability of cell membranes, which in turn affects the proper functioning of the organ after transplantation. The addition of ascorbic acid and prolactin to Biolasol affects the maintenance of the normal cytoskeleton of the stored graft.

Tips for optimizing organ preservation solutions.

Organ injury during ischemia is one of the clinical problems of today's 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.

[Lysosomal storage diseases in selected clinical cases]. / Lizosomalne choroby spichrzeniowe w wybranych przypadkach klinicznych.

Lysosomal storage diseases (LSDs) are metabolic disorders that result from a situation when enzymes taking part in different processes exist in too small an amount or don't exist whatsoever. They exhibit great clinical diversification. The intensity of the symptoms depends on residual enzyme activity, type of stored material and storage speed. The common feature of all LSDs is their progressive course and reduced life expectancy. The symptoms can appear in the first few weeks of life and the patients are then referred for medical treatment to pediatricians. Older patients are referred for medical treatment to dermatologists, cardiologists, neurologists or orthopedists. There are few described cases in Poland which is due to the fact that these diseases are considered rare.

Influence of trace elements on stabilization of aqueous solutions of ascorbic acid.

Together with vitamin C, zinc, selenium, manganese, and magnesium play a vital role in the preservation of organs scheduled for transplantation. In the present study, it is shown that addition of 1 mg/l of these elements influences the stability of 0.3 mM ascorbic acid solutions. The solution's stability was estimated using an accelerated stability test. The concentration of vitamin C was measured using a validated spectrophotometric method, which uses the reduction of 2,6-dichlorophenoloindophenol by ascorbic acid. Elevated temperatures, the factor accelerating substances' decomposition reaction rate, were used in the tests. The research was conducted at two temperatures at intervals of 10 °C: 80 ± 0.1 and 90 ± 0.1 °C. It was stated that the studied substances' decomposition occurred in accordance with the equation for first-order reactions. The function of the logarithmic concentration (log%C) over time was revealed to be rectilinear. This dependence was used to determine the kinetics of decomposition reaction rate parameters. The stabilization of vitamin C solutions was measured as the time in which 10 % of the substance decomposed at 20 and 0 °C. Addition of Se(IV) or Mg(II) ions significantly increase the stability of ascorbic acid solution (∼34 and ∼16 %, respectively), but Zn(II) causes a significant decrease in stability by ∼23 %. Addition of Mn(II) has no significant influence on vitamin C stability.

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.