Biopharmaceutical characteristics of autologous red blood cells ghosts containing cytokines and antibiotics

Introduction: Transport systems based on autologous red blood cells for targeted drug delivery can be considered as a promising approach in the treatment of surgical infections. Experimental studies have revealed the feasibility of targeted drug delivery by encapsulation of cytokines and antibiotics into autologous erythrocyte ghosts.Purpose: To study biopharmaceutical characteristics of autologous erythrocyte ghosts containing cytokines and antibiotics (pharmacocytes). Material and methods: The erythrocyte pharmacocytes were prepared by the hypotonic hemolysis method, or  the use of human red blood cells. The association and dissociation indicators of rifampicin and cytokine substances with the erythrocyte ghosts were conducted using standard methods.Results: We have defined the following extracellular concentrations to be optimal for deposition of drug substances into pharmacocytes: for rifampicin – 10 000 ?/ml,  erythropoietin - 1000 IU / ml, TNF-a - 5000 IU / ml, IL-1-? - 5000 U / ml, IFN-? - 10 000ME / ml, IL-2 - 50 000 IU / ml, angiogenin - 0.04 mg / ml. Two types of correlations of cytokines and pharmacocytes were identified. In this study, we found that 40-60 % of the erythropoietin, IFN- ? and angiogenin were bound to red blood cells ghosts, more than 10% of which were bound irreversibly.   For TNF-a,  IL-1-? and IL-2, the red blood cells ghosts were capable of binding and depositing within 10-20 % of the input extracellular concentration, and these bindings were almost completely reversible. The rifampicin was bound by red blood cells ghosts with 5 % efficiency and also completely reversibly.Conclusion: The study has shown the effectiveness of inclusion of the studied components, such as erythropoietin, IFN-? and angiogenin into the red blood cells ghosts, with significant efficiency (40-60 %). It presents the potential of using this system in targeted delivery of cytokines and antibiotics for treatment of surgical infections, thus facilitating the reduction in toxicity and adverse systemic effects of drugs and improving the treatment results

Pharmacokinetic Properties of Cytokines in Their Targeted Delivery Based on Autologous Erythrocyte Pharmacocytes

Introduction. Using autologous erythrocytes as drug carriers for targeted delivery of cytokines to the sites of inflammation could potentially provide new opportunities for treatment of patients with purulent diseases. The targeted characteristic of erythrocytes is associated with the nature of purulent inflammation, where a large amount of erythrocytes is phagocytized and drugs encapsulated into the erythrocytes could be easily released. On the other hand, autologous erythrocytes meet all the criteria for the ideal drug carrier. They are nontoxic, not immunogenic, and able to bear a large number of drug molecules while preserving an original conformation of the drugs. Thus, in this study, we aimed to analyze pharmacokinetic profiles of IL-1? encapsulated into erythrocytes’ ghosts (pharmacocytes) in comparison to intravenously injected free IL-1?.Material and methods. Albino rats were randomly divided into two groups, each group receiving a different kind of IV injection via the tail vein. Group A (control) received 500 µg of free IL-1?, and group B received an injection of 1 ml of pharmacocytes loaded with 500 µg of test substance. At fixed time points after injection (15, 30, 60, 180, 540, 720, and 1,440 minutes) serum samples were collected. Homogenates of liver, spleen, lung, heart, kidney, and adipose tissue were obtained 24 hours after injections. Concentration of the tested substance in the collected organs and blood plasma were measured by ELISA. Results. We have observed an increased half-life period (T1/2) for encapsulated IL-1? compared to the control. T1/2 for free IL-1? was one hour, while administration of loaded pharmacocytes allowed the half-life period to increase by more than 15 fold (1,043.40 ± 137.92 min) preserving high level of IL-1? activity in the blood samples up to 24 hours. The increased time of IL-1? presence in the body when administered in the form of pharmacocytes could be explained by reduction of elimination constant (Cel) by 1.6 fold, and clearance (CLel) by more than 100 fold. We also observed an increased concentration of IL-1? in liver, spleen, and lung over at least 24 hours. When administered in free form, IL-1? disappeared from these organs within 6 hours.Conclusions. Pharmacocytes have shown to improve pharmacokinetic profiles of IL-1? by increasing the half-life period of the cytokine, reducing its clearance and elimination as well as increasing the deposition of the drug in liver, spleen and lungs. These data suggest that pharmacocytes be effective drug carriers for targeted delivery of cytokines to the sites of inflammation and have a potential for improving the treatment outcomes of purulent diseases

Biopharmaceutical characteristics of autologous red blood cells ghosts containing cytokines and antibiotics

Introduction: Transport systems based on autologous red blood cells for targeted drug delivery can be considered as a promising approach in the treatment of surgical infections. Experimental studies have revealed the feasibility of targeted drug delivery by encapsulation of cytokines and antibiotics into autologous erythrocyte ghosts. Purpose: To study biopharmaceutical characteristics of autologous erythrocyte ghosts containing cytokines and antibiotics (pharmacocytes). Material and methods: The erythrocyte pharmacocytes were prepared by the hypotonic hemolysis method, or  the use of human red blood cells. The association and dissociation indicators of rifampicin and cytokine substances with the erythrocyte ghosts were conducted using standard methods. Results: We have defined the following extracellular concentrations to be optimal for deposition of drug substances into pharmacocytes: for rifampicin – 10 000 μ/ml,  erythropoietin - 1000 IU / ml, TNF-a - 5000 IU / ml, IL-1-β - 5000 U / ml, IFN-γ - 10 000ME / ml, IL-2 - 50 000 IU / ml, angiogenin - 0.04 mg / ml. Two types of correlations of cytokines and pharmacocytes were identified. In this study, we found that 40-60 % of the erythropoietin, IFN- γ and angiogenin were bound to red blood cells ghosts, more than 10% of which were bound irreversibly.   For TNF-a,  IL-1-β and IL-2, the red blood cells ghosts were capable of binding and depositing within 10-20 % of the input extracellular concentration, and these bindings were almost completely reversible. The rifampicin was bound by red blood cells ghosts with 5 % efficiency and also completely reversibly. Conclusion: The study has shown the effectiveness of inclusion of the studied components, such as erythropoietin, IFN-γ and angiogenin into the red blood cells ghosts, with significant efficiency (40-60 %). It presents the potential of using this system in targeted delivery of cytokines and antibiotics for treatment of surgical infections, thus facilitating the reduction in toxicity and adverse systemic effects of drugs and improving the treatment results

Blueberry and cranberry extracts mitigate CCL4-induced liver damage, suppressing liver fibrosis, inflammation and oxidative stress

The current study aims to evaluate potential hepatoprotective effect of lingonberry, cranberry and blueberry polyphenols on carbon tetrachloride (CCL-4)-induced acute and subacute liver injury in rats. A total of 55 male Wistar rats, divided into six experimental and control groups. With the exception of the negative control group, all groups received an intraperitoneal injection of CCl-4, twice a week for 14 days. An extract of lingonberry, cranberry, blueberry polyphenols and the positive control, silymarin were administered daily via intragastric route, for 14 consecutive days. The untreated control group showed characteristic of classical oxidative stress-mediated liver damage with vacuolization of the hepatocyte cytoplasm, infiltration by immune cells and proliferation of collagen fibers, decrease in body weight and increase in liver weight; increased levels of AST and ALT in serum, an increased lipid peroxidation in the liver. However, the use of cranberry and blueberry polyphenols significantly suppressed liver damage, exerting an effect comparable to the hepatoprotective effect of the positive control. The extracts prevented and reduced inflammatory liver damage by reducing IL-6, TNF-α and IFN-γ levels. In conclusion, blueberry and cranberry extracts have a protective effect against acute and subacute CCl4-induced hepatotoxicity in rats

Pharmacokinetic Properties of Cytokines in Their Targeted Delivery Based on Autologous Erythrocyte Pharmacocytes

Introduction. Using autologous erythrocytes as drug carriers for targeted delivery of cytokines to the sites of inflammation could potentially provide new opportunities for treatment of patients with purulent diseases. The targeted characteristic of erythrocytes is associated with the nature of purulent inflammation, where a large amount of erythrocytes is phagocytized and drugs encapsulated into the erythrocytes could be easily released. On the other hand, autologous erythrocytes meet all the criteria for the ideal drug carrier. They are nontoxic, not immunogenic, and able to bear a large number of drug molecules while preserving an original conformation of the drugs. Thus, in this study, we aimed to analyze pharmacokinetic profiles of IL-1β encapsulated into erythrocytes’ ghosts (pharmacocytes) in comparison to intravenously injected free IL-1β. Material and methods. Albino rats were randomly divided into two groups, each group receiving a different kind of IV injection via the tail vein. Group A (control) received 500 µg of free IL-1β, and group B received an injection of 1 ml of pharmacocytes loaded with 500 µg of test substance. At fixed time points after injection (15, 30, 60, 180, 540, 720, and 1,440 minutes) serum samples were collected. Homogenates of liver, spleen, lung, heart, kidney, and adipose tissue were obtained 24 hours after injections. Concentration of the tested substance in the collected organs and blood plasma were measured by ELISA.  Results. We have observed an increased half-life period (T1/2) for encapsulated IL-1β compared to the control. T1/2 for free IL-1β was one hour, while administration of loaded pharmacocytes allowed the half-life period to increase by more than 15 fold (1,043.40 ± 137.92 min) preserving high level of IL-1β activity in the blood samples up to 24 hours. The increased time of IL-1β presence in the body when administered in the form of pharmacocytes could be explained by reduction of elimination constant (Cel) by 1.6 fold, and clearance (CLel) by more than 100 fold. We also observed an increased concentration of IL-1β in liver, spleen, and lung over at least 24 hours. When administered in free form, IL-1β disappeared from these organs within 6 hours. Conclusions. Pharmacocytes have shown to improve pharmacokinetic profiles of IL-1β by increasing the half-life period of the cytokine, reducing its clearance and elimination as well as increasing the deposition of the drug in liver, spleen and lungs. These data suggest that pharmacocytes be effective drug carriers for targeted delivery of cytokines to the sites of inflammation and have a potential for improving the treatment outcomes of purulent diseases

POLYPHENOLS AS CALORIC-RESTRICTION MIMETICS AND AUTOPHAGY INDUCERS IN AGING RESEARCH

It has been thought that caloric restriction favors longevity and healthy aging where autophagy plays a vital role. However, autophagy decreases during aging and that can lead to the development of aging-associated diseases such as cancer, diabetes, neurodegeneration, etc. It was shown that autophagy can be induced by mechanical or chemical stress. In this regard, various pharmacological compounds were proposed, including natural polyphenols. Apart from the ability to induce autophagy, polyphenols, such as resveratrol, are capable of modulating the expression of pro- and anti-apoptotic factors, neutralizing free radical species, affecting mitochondrial functions, chelating redox-active transition metal ions, and preventing protein aggregation. Moreover, polyphenols have advantages compared to chemical inducers of autophagy due to their intrinsic natural bio-compatibility and safety. In this context, polyphenols can be considered as a potential therapeutic tool for healthy aging either as a part of a diet or as separate compounds (supplements). This review discusses the epigenetic aspect and the underlying molecular mechanism of polyphenols as an anti-aging remedy. In addition, the recent advances of studies on NAD-dependent deacetylase sirtuin-1 (SIRT1) regulation of autophagy, the role of senescence-associated secretory phenotype (SASP) in cells senescence and their regulation by polyphenols have been highlighted as well. Apart from that, the review also revised the latest information on how polyphenols can help to improve mitochondrial function and modulate apoptosis (programmed cell death)

The Effect of Different Glucose Concentrations on the Antiproliferative Activity of Metformin in MCF-7 Breast Cancer Cells

The glucose-lowering drug metformin has been reported to have anticancer properties through unknown mechanisms. Other unknown factors that may influence its anticancer potential include the glycemic status of the patient. Therefore, the objective of this study is to determine the effect of different glucose environments on the antiproliferative potency and the cellular mechanism of action of metformin. Human breast cancer cells, MCF-7, were incubated in low, normal, elevated, and high glucose environments and treated with metformin. The antiproliferative potential of metformin and its effect on protein expression as well as its ability to induce cellular apoptosis and autophagy under different glucose environments, were determined using different molecular techniques. Metformin significantly inhibited cellular proliferation in a time- and glucose-concentration-dependent manner. In comparison to elevated glucose, low normal glucose alone induced a significant level of autophagy that was further increased in the presence of metformin. While glucose concentration did not appear to have an effect on the antiproliferative potency of metformin, the cellular basis of action was shown to be glucose-dependent. The antiproliferative mechanism of action of metformin in elevated and low normal glucose environments is mTOR-dependent, whereas, in the high glucose environment, the antiproliferative mechanism is independent of mTOR. This is the first study to report that both the antiproliferative potency and the cellular mechanism of action aredependent on the concentration of glucose

ANTIRADICAL AND CYTOPROTECTIVE PROPERTIES OF ALLIUM NUTANS L. HONEY AGAINST CCL4-INDUCED LIVER DAMAGE IN RATS

The aim of this study is determine the in vitro and in vivo antiradical properties and the cytoprotective activity of Allium nutans L. honey extract. The antiradical properties of the extracts were investigated in rabbit alveolar macrophages and human foreskin fibroblast (hFFs) cells in the presence of doxorubicin, a cytotoxic substance using DPPH and ABTS assays. The cytoprotective activities were determined using 18 Wistar rats divided into three different groups, a negative control, and two other groups with experimentally induced hepatotoxicity by a single intraperitoneal injection of 50% carbon tetrachloride (CCl4) oil solution. A positive control group, received drinking water only and an experimental group that was treated with Allium nutans L. honey extracts for 7 days. In vitro treatment with Allium nutans L. honey extracts resulted in 78% reduction in radical activity in DPPH and 91.6% inhibition using the ABTS. Also, honey extracts were able to preserve 100% of cell viability in the presence of the cytotoxic, doxorubicin. Furthermore, the treatment with honey extracts resulted in a significant reduction in damage to the structure of liver tissue, as well significant reduction in the levels of ALT and AST in the experimental group compared to the control group

GUT MODULATION OF DYSBIOSIS INDUCED BY DEXTRAN SULFATE SODIUM

Inflammatory bowel disease is one of the serious burdens of clinical medicine and healthcare. This study investigated the potential of a biological product based on mare's milk and metabolites of symbiotic microflora for modulation of intestinal microflora affected by dextran sulfate sodium (DSS)-induced dysbiosis. Symbiotic microflora was isolated from the stool of healthy volunteers. Lysates for the production of short-chain fatty acids of screened microorganisms were mixed with mare's milk. The activity of the biological product was evaluated on the DSS model of induced colitis. Histological changes in the intestinal epithelium were determined. The structure of the microbiome was evaluated based on the analysis of 16S rRNA microbial sequences. Histological examination of rat intestinal tissues after application of the biological product showed reduced infiltration of granulocytes, macrophages, and lymphocytes. The results of sequencing demonstrated a decrease in the biological diversity of microbiota affected by colitis. The full recovery was observed after 21 days of the application of the biological product. The product induced the structural changes of the microbiome damaged by DSS. Likewise, the number of the pathogenic intestinal microflora was decreased Representatives of SCFA producing bacteria increased concentrations of genus Lactobacillus