ABSTRACT
Antifibrotic activity of intranasally administered conjugates of pluronics L31 and F68 with hyaluronate-endo-ß-N-acetylhexosaminidase was studied in C57Bl/6 mice under conditions of single and repeated bleomycin-induced injury to the alveolar epithelium. Conjugates were prepared using the technique of protein immobilization with ionizing radiation. We demonstrate that in cases of single and repeated injuries to the alveolar epithelium, the conjugates administered during phases of inflammation or deposition of fibrotic masses prevent the development of pulmonary fibrosis. The conjugates demonstrated more pronounced antifibrotic activity than hyaluronate-endo-ß-N-acetylhexosaminidase. The conjugate based on hydrophobic pluronic L31 showed higher effectiveness in comparison with the conjugate based on amphiphilic pluronic F68.
Subject(s)
Enzymes, Immobilized/pharmacology , Fibrinolytic Agents/pharmacology , Hyaluronic Acid/chemistry , Poloxamer/chemistry , Pulmonary Fibrosis/prevention & control , beta-N-Acetylhexosaminidases/pharmacology , Administration, Intranasal , Animals , Bleomycin , Enzymes, Immobilized/chemistry , Fibrinolytic Agents/chemistry , Hydrophobic and Hydrophilic Interactions , Mice , Mice, Inbred C57BL , Pulmonary Alveoli/drug effects , Pulmonary Alveoli/pathology , Pulmonary Fibrosis/chemically induced , Pulmonary Fibrosis/pathology , Radiation, Ionizing , beta-N-Acetylhexosaminidases/chemistryABSTRACT
The effect of immobilized hyaluronidase on stem and progenitor cells of the lungs was studied on the model of partially reversible toxic bleomycin-induced pulmonary fibrosis in C57Bl/6 mice. During the inflammation phase, immobilized hyaluronidase reduced infiltration of alveolar interstitium with hemopoietic stem cells Sca-1(+), c-Kit(+), CD34(-), (CD3, CD45R (B220), Ly6C, Ly6G (Gr1), CD11b (Mac1), TER-119)(-). Improvement of histological parameters of bleomycin lungs during the phase of collagen fiber deposition after the treatment was accompanied by accumulation of mesenchymal multipotent stromal cells (CD31(-), CD34(-), CD45(-), CD44(+), CD73(+), CD90(+), CD106(+)decrease in the population of pan-hemopoietic cells (CD45(+)), accelerated restoration of the content of endothelial cells, and inhibition of clonal activity of fibroblast precursors (CD45(-)).
Subject(s)
Enzymes, Immobilized/administration & dosage , Hyaluronoglucosaminidase/administration & dosage , Pulmonary Fibrosis/pathology , Stem Cells/metabolism , Animals , Antigens, CD/metabolism , Bone Marrow/immunology , Bone Marrow/pathology , Lung/pathology , Mice, Inbred C57BL , Pulmonary Fibrosis/drug therapy , Pulmonary Fibrosis/immunology , Stem Cells/drug effectsABSTRACT
We have demonstrated the possibility of stimulation of the function of various types of precursor cells with hyaluronidase modified with chondroitin sulfate. Parenteral administration of modified hyaluronidase increased the number of fibroblast, granulomonocyte, and erythroid CFU in the hemopoietic tissue. The changes in the pool of mesenchymal progenitor cells were more pronounced in comparison with those induced by native enzyme.
Subject(s)
Bone Marrow Cells/physiology , Chondroitin Sulfates/chemistry , Hyaluronoglucosaminidase/pharmacology , Stem Cells/physiology , Animals , Bone Marrow Cells/drug effects , Cell Adhesion , Cell Count , Female , Hyaluronoglucosaminidase/chemistry , Male , Mice, Inbred CBA , Regenerative Medicine , Stem Cells/drug effectsABSTRACT
The possibility of boosting antifibrotic activity of testicular hyaluronidase immobilized on polyethylene oxide with spiperone was studied on the bleomycin models of a single (partially reversible pneumofibrosis) and repeated (irreversible pneumofibrosis) injuries to the alveolar epithelium in C57Bl/6 mice. The antifibrotic effect was more pronounced after successive treatment with immobilized hyaluronidase and spiperone than after individual treatment with each of the compounds: no collagen deposition in the parenchyma of bleomycin-damaged lungs was found. The decrease in inflammatory cell (lymphocytes, macrophages, neutrophils, plasma cells) infiltration of the alveoli and alveolar tracts interstitium in mice treated by immobilized hyaluronidase and spiperone did not differ from the anti-inflammatory effect of spiperone monotherapy.
Subject(s)
Anti-Inflammatory Agents/pharmacology , Hyaluronoglucosaminidase/pharmacology , Pulmonary Fibrosis/drug therapy , Spiperone/pharmacology , Animals , Anti-Inflammatory Agents/therapeutic use , Bleomycin , Collagen/metabolism , Drug Evaluation, Preclinical , Drug Therapy, Combination , Enzymes, Immobilized/pharmacology , Enzymes, Immobilized/therapeutic use , Hyaluronoglucosaminidase/therapeutic use , Lung/drug effects , Lung/metabolism , Lung/pathology , Mice , Mice, Inbred C57BL , Pulmonary Fibrosis/chemically induced , Pulmonary Fibrosis/prevention & control , Spiperone/therapeutic useABSTRACT
Antifibrotic activity of testicular hyaluronidase, immobilized on polyethylenoxide and obtained by electron beam synthesis, was studied on the model of bleomycin injuries to the alveolar epithelium (irreversible pneumofibrosis) in C57Bl/6 mice and compared to the effect of testicular hyaluronidase. Intranasal therapy with immobilized and testicular hyaluronidases prevented the deposition of fibrotic mass in the parenchyma of "bleomycin" lungs. The effect of immobilized hyaluronidase was more pronounced than that of testicular hyaluronidase. The studied compounds were virtually inessential for infiltration of the alveolar interstitium and alveolar tracts by lymphocytes, macrophages, neutrophils, and plasma cells. Unchanged histoarchitectonics of bleomycin-damaged lungs in immobilized hyaluronidase therapy was due to suppression of the progenitor fibroblast cells (CD45(-)).
Subject(s)
Anti-Inflammatory Agents/administration & dosage , Hyaluronoglucosaminidase/administration & dosage , Idiopathic Pulmonary Fibrosis/drug therapy , Animals , Anti-Inflammatory Agents/chemistry , Bleomycin , Drug Carriers/administration & dosage , Drug Carriers/chemistry , Enzymes, Immobilized/administration & dosage , Enzymes, Immobilized/chemistry , Hyaluronoglucosaminidase/chemistry , Idiopathic Pulmonary Fibrosis/chemically induced , Idiopathic Pulmonary Fibrosis/pathology , Mice , Mice, Inbred C57BL , Polyethylenes/chemistry , Pulmonary Alveoli/pathology , Respiratory Mucosa/pathologyABSTRACT
We studied the effect of hyaluronate-endo-ß-N-acetylhexosaminidase on the secretory function of the liver and bone marrow microenvironment cells in chronic hepatitis. Enhanced production of substances stimulating parenchymal tissue-specific precursors and stem cell homing factors by liver cells was revealed. At the same time, production of SDF-1 and other chemoattractants and adhesion factors of progenitor cells by bone marrow elements was reduced.
Subject(s)
Bone Marrow Cells/metabolism , Chemokine CXCL12/biosynthesis , Hepatitis, Chronic/metabolism , Liver/metabolism , Polyethylene Glycols/pharmacology , Stem Cells/drug effects , beta-N-Acetylhexosaminidases/pharmacology , Animals , Bone Marrow Cells/drug effects , Cell Movement/drug effects , Cellular Microenvironment/drug effects , Hyaluronoglucosaminidase , Liver/cytology , Liver/drug effects , Male , Mice , Mice, Inbred C57BL , Stem Cells/metabolismABSTRACT
Pegylated hyaluronate-endo-ß-N-acetylhexosaminidase was shown to potentiate significantly the hemostimulatory effect of pegylated erythropoietin. It was found that enhanced production of hemopoietin by adherent and non-adherent cells of the hemopoiesis-inducing microenvironment and elevated serum content of endogenous erythropoietin along with increased susceptibility of erythroid precursors to pegylated erythropoietin underlay this phenomenon.
Subject(s)
Anemia/drug therapy , Erythropoietin/pharmacology , Hematinics/pharmacology , Polyethylene Glycols/pharmacology , beta-N-Acetylhexosaminidases/pharmacology , Anemia/chemically induced , Animals , Bone Marrow Cells/drug effects , Carboplatin , Cell Differentiation , Cell Proliferation , Cells, Cultured , Drug Synergism , Drug Therapy, Combination , Erythroid Cells/drug effects , Erythropoietin/administration & dosage , Hematinics/administration & dosage , Mice , Mice, Inbred CBA , Polyethylene Glycols/administration & dosage , Recombinant Proteins/administration & dosage , Recombinant Proteins/pharmacology , Stem Cells/drug effects , Stem Cells/physiology , beta-N-Acetylhexosaminidases/administration & dosageABSTRACT
Using the model of lung fibrosis induced by intratracheal administration of bleomycin we studied anti-fibrotic activity of combined treatment with neuroleptic haloperidol and hyaluronidase immobilized on polyethylene oxide using electron-beam synthesis. It was shown that successive administration of immobilized hyaluronidase and the neuroleptic drug inhibits deposition of collagen fibers in the bleomycin-treated lungs. Combined treatment with the test compounds reduced swelling of the alveolar epithelium, exudation and infiltration of the alveolar interstitium and alveolar passages by inflammatory cells, and desquamation of alveolocytes into alveolar lumen, so that the alveolar-capillary membrane function was preserved.
Subject(s)
Antipsychotic Agents/therapeutic use , Enzymes, Immobilized/therapeutic use , Haloperidol/therapeutic use , Hyaluronoglucosaminidase/therapeutic use , Pulmonary Fibrosis/drug therapy , Respiratory Mucosa/drug effects , Animals , Bleomycin , Capillaries/drug effects , Collagen/metabolism , Connective Tissue/drug effects , Drug Therapy, Combination , Haloperidol/pharmacology , Hyaluronoglucosaminidase/pharmacology , Inflammation/drug therapy , Lung/drug effects , Lung/pathology , Mice , Mice, Inbred C57BL , Polyethylene Glycols , Pulmonary Alveoli/drug effects , Pulmonary Alveoli/pathology , Pulmonary Fibrosis/chemically induced , Receptors, Dopamine/metabolism , Receptors, Serotonin/metabolismABSTRACT
Hyaluronidase immobilized on polyethylenoxide obtained by electron bean synthesis was administered intranasally and intravenously to C57Bl/6 mice after intratracheal bleomycin and the enzyme effects on the development of pneumofibrosis in animals were studied. Intranasal immobilized hyaluronidase prevented connective tissue growth in the lungs exposed to bleomycin and virtually did not modulate the infiltration of the alveolar and alveolar duct interstitium by inflammatory cells (lymphocytes, macrophages, neutrophils, plasma cells). The antifibrotic effect developed sooner after intranasal inoculation of immobilized hyaluronidase and was more pronounced than after intranasal native hyaluronidase. Intravenous injection of immobilized hyaluronidase did not modify the inflammatory process and deposition of collagen fibrils in the lung parenchyma in pneumofibrosis.
Subject(s)
Connective Tissue/drug effects , Enzymes, Immobilized/therapeutic use , Hyaluronoglucosaminidase/therapeutic use , Inflammation/drug therapy , Pulmonary Fibrosis/drug therapy , Animals , Bleomycin , Connective Tissue Cells/drug effects , Hyaluronic Acid/metabolism , Hyaluronoglucosaminidase/administration & dosage , Hyaluronoglucosaminidase/metabolism , Inflammation/chemically induced , Leukocyte Count , Lung/drug effects , Lung/pathology , Lymphocyte Count , Lymphocytes/immunology , Macrophages/immunology , Mice , Mice, Inbred C57BL , Neutrophils/immunology , Plasma Cells/immunology , Polyethylene Glycols/administration & dosage , Pulmonary Fibrosis/chemically inducedABSTRACT
Pegylated hyaluronate-endo-ß-N-acetylhexosaminidase considerably potentiates the hemostimulating effects of erythropoietin due to intensification of proliferation and differentiation of erythroid precursors against the background of enhanced secretion of hemopoietins by nonadherent hemopoiesis-inducing environment cells and elevation of serum erythropoietin concentration. The use of the enzyme allows 10-fold reduction of the maximum effective erythropoietin dose.
Subject(s)
Anemia/drug therapy , Erythropoietin/metabolism , Erythropoietin/pharmacology , Polyethylene Glycols/metabolism , beta-N-Acetylhexosaminidases/metabolism , Animals , Carboplatin , Dose-Response Relationship, Drug , Drug Synergism , Enzyme-Linked Immunosorbent Assay , Erythrocyte Count , Erythropoietin/blood , Female , Hemoglobins/metabolism , Male , Mice , Mice, Inbred CBA , Nanotechnology/methods , Recombinant Proteins , Statistics, NonparametricABSTRACT
Pharmacological characteristics of somatotropin pegylated using electron-beam synthesis nanotechnology (PEG-STH) were studied. Oral PEG-STH stimulated the intensity of protein and lipid metabolism and endochondral bone growth without modifying the processes of periosteal and endosteal bone formation. Specific activity of this substance administered orally significantly surpassed that of parenteral non-modified growth hormone.
Subject(s)
Bone Development/drug effects , Human Growth Hormone/pharmacology , Lipid Metabolism/drug effects , Nanotechnology , Polyethylene Glycols , Proteins/metabolism , Animals , Male , Rats , Rats, WistarABSTRACT
Hemopoiesis-stimulating activity of immobilized oligonucleotide preparation was studied on the model of cytostatic myelosuppression induced by injection of cyclophosphamide and 5-fluorouracil. Immobilized oligonucleotides stimulated regeneration of erythro- and granulocytopoiesis in the bone marrow under conditions of cytostatic treatment. The counts of neutrophilic granulocytes and platelets in the peripheral blood increased. The stimulatory effect of the drug was more manifest in animals with active behavior. The mechanism of immobilized oligonucleotide effect was based on stimulation of functional activity of erythroid and granulocytic macrophage precursors.
Subject(s)
Erythropoiesis/drug effects , Hematinics/administration & dosage , Myelopoiesis/drug effects , Oligonucleotides/administration & dosage , Administration, Oral , Animals , Blood Platelets/cytology , Blood Platelets/drug effects , Bone Marrow/drug effects , Bone Marrow/physiology , Cell Count , Cyclophosphamide/administration & dosage , Cytostatic Agents/administration & dosage , Erythropoiesis/physiology , Fluorouracil/administration & dosage , Granulocytes/cytology , Granulocytes/drug effects , Hematinics/chemistry , Injections, Intraperitoneal , Leukocytes, Mononuclear/cytology , Leukocytes, Mononuclear/drug effects , Mice , Mice, Inbred CBA , Myelopoiesis/physiology , Oligonucleotides/chemistryABSTRACT
Hypoglycemic effect of hyaluronate-endo-ß-N-acetylhexosaminidase immobilized by electron-beam synthesis nanotechnology (imHEA-HA) was studied in experimental insulin-dependent and insulin-independent diabetes mellitus. The drug exhibited a hypoglycemic effect of its own and potentiated the pharmacological effect of exogenous insulin injected in vivo. Studies on liver cell culture demonstrated an increase of cell sensitivity to insulin after treatment with imHEA-HA.
Subject(s)
Enzymes, Immobilized/therapeutic use , Glycosaminoglycans/chemistry , Hypoglycemic Agents/therapeutic use , Nanotechnology/methods , beta-N-Acetylhexosaminidases/chemistry , beta-N-Acetylhexosaminidases/therapeutic use , Animals , Diabetes Mellitus/drug therapy , Hyaluronoglucosaminidase/metabolism , Hypoglycemic Agents/chemistry , MiceABSTRACT
High hepatoprotective activity of granulocytic CSF and hyaluronidase immobilized using electron-beam immobilization technology was demonstrated on the model of CCl(4)-induced hepatitis: the preparations produced anticholestatic, anti-inflammatory, and antisclerotic effects. These effects developed against the background of stimulation of bone marrow multipotent precursor cells and their mobilization into circulation accompanied by an increase in the content of parenchymatous progenitor cells in the liver. The most pronounced positive effect was observed in combined treatment with the test preparations.
Subject(s)
Enzymes, Immobilized/therapeutic use , Granulocyte Colony-Stimulating Factor/therapeutic use , Hepatitis, Chronic/drug therapy , Hyaluronoglucosaminidase/therapeutic use , Animals , Male , Mice , Nanotechnology , Rats , Rats, Wistar , Regenerative MedicineABSTRACT
We studied the effect of mobilization of bone marrow multipotent stem cells induced by intragastric administration of pegylated hyaluronate-endo-ß-N-acetylhexosaminidase (Peg-HEAHA) on hemopoiesis under conditions of experimental chronic hepatitis. Peg-HEAHA increased the counts of hemopoietic precursors in the hemopoietic tissue against the background of their decelerated maturation. This was paralleled by a short-term decrease in the count of neutrophilic granulocyte in the bone marrow and a slight increase of neutrophil count in the peripheral blood.
Subject(s)
Electrons , Hematopoiesis/drug effects , Hepatitis, Chronic/drug therapy , Hyaluronic Acid/chemistry , Nanoparticles/therapeutic use , beta-N-Acetylhexosaminidases/chemistry , Animals , Hepatitis, Chronic/metabolism , Male , Mice , Mice, Inbred C57BL , Nanoparticles/chemistry , Nanotechnology/methods , Regenerative Medicine/methodsABSTRACT
Immobilized hyaluronidase (nanotechnology method of electron-beam synthesis) exhibited high hepatoprotective activity on the model of Cl4-induced hepatitis. This agent produced anticholestatic, anti-inflammatory, and antisclerotic effects. These effects were shown to accompany stimulation of multipotent bone marrow precursors, mobilization of these cells into the peripheral blood, and cell migration to the target organ increasing the number of parenchymal progenitor cells in the liver. The mechanisms for targeted migration of progenitor cells suggest a decrease in SDF-1 production by bone marrow stromal cells and increase in the synthesis of this factor by microenvironmental cells of the liver tissue.
Subject(s)
Cytoprotection , Enzymes, Immobilized/therapeutic use , Hepatitis, Animal/drug therapy , Hyaluronoglucosaminidase/therapeutic use , Liver/drug effects , Multipotent Stem Cells/drug effects , Animals , Biomarkers/metabolism , Bone Marrow Cells/cytology , Bone Marrow Cells/drug effects , Carbon Tetrachloride , Cell Movement/drug effects , Cellular Microenvironment/drug effects , Chemokine CXCL12/metabolism , Enzymes, Immobilized/administration & dosage , Enzymes, Immobilized/chemistry , Hepatitis, Animal/metabolism , Hepatitis, Animal/pathology , Hyaluronoglucosaminidase/administration & dosage , Hyaluronoglucosaminidase/chemistry , Liver/metabolism , Liver/pathology , Mice , Multipotent Stem Cells/cytology , Nanotechnology , Rats , Stromal Cells/cytology , Stromal Cells/drug effectsABSTRACT
In vitro experiments demonstrated increased colony-forming capacity of erythroid, granulomonocytic, and mesenchymal progenitors of the bone marrow and parenchymal progenitor elements of the liver after treatment with immobilized hyaluronidase. Increased sensitivity of these progenitor cells to erythropoietin, granulocyte colony-stimulating factor, fibroblast growth factor, and stem cell factor, respectively, was demonstrated. Immobilized hyaluronidase enhanced the formation of tissue-specific hepatic CFU against the background of reduced yield of stromal precursors in liver tissue culture containing insulin.
Subject(s)
Enzymes, Immobilized/pharmacology , Hematopoietic Stem Cells/drug effects , Hyaluronoglucosaminidase/pharmacology , Mesenchymal Stem Cells/drug effects , Animals , Bone Marrow Cells/cytology , Bone Marrow Cells/drug effects , Colony-Forming Units Assay , Enzymes, Immobilized/chemistry , Erythropoietin/pharmacology , Fibroblast Growth Factors/pharmacology , Granulocyte Colony-Stimulating Factor/pharmacology , Hematopoietic Stem Cells/cytology , Hyaluronoglucosaminidase/chemistry , Insulin/metabolism , Insulin/pharmacology , Liver/cytology , Liver/drug effects , Male , Mesenchymal Stem Cells/cytology , Mice , Mice, Inbred CBA , Nanotechnology , Stem Cell Factor/pharmacology , Tissue Culture TechniquesABSTRACT
We evaluated whether immobilized hyaluronidase can modify the hematotropic effect of immobilized granulocyte CSF (G-CSF). The preparation of immobilized hyaluronidase (50 arb. units per mouse) potentiated the specific effect of immobilized G-CSF on granulomonocytopoiesis. The preparation was shown to facilitate the indirect effect of immobilized G-CSF on hemopoiesis (stimulation of the erythroid and lymphoid hemopoietic stems). These changes were accompanied by an increase in functional activity of hemopoietic precursor cells, secretion of humoral factors by bone marrow myelokaryocytes, and concentration of hemopoietins in the serum.
Subject(s)
Granulocyte Colony-Stimulating Factor/pharmacology , Granulocytes/drug effects , Hematopoiesis/drug effects , Hyaluronoglucosaminidase/metabolism , Animals , Bone Marrow/drug effects , Bone Marrow Cells/drug effects , Cyclophosphamide/pharmacology , Enzymes, Immobilized , Hematopoietic Cell Growth Factors/blood , Hematopoietic Stem Cells/drug effects , MiceABSTRACT
The effects of immobilized granulocyte colony-stimulating factor (mediated by cells of the hemopoiesis-inducing microenvironment) on hemopoietic precursors of various classes were studied on the model of cytostatic-induced myelosuppression (administration of cyclophosphamide). The action of this preparation was compared with that of the standard preparation of granulocyte colony-stimulating factor. Thy 1,2(+)cells potentiated the effects of immobilized and standard granulocyte colony-stimulating factors on granulocyte-erythroid-macrophage-megakaryocyte precursors. Stromal cells were shown to potentiate the influence of these agents on granulocyte precursors. Induction of proliferation of precursor cells by the immobilized factor mediated by cells of the hemopoiesis-inducing microenvironment persisted for a longer period compared to that induced by the standard product.
Subject(s)
Granulocyte Colony-Stimulating Factor/pharmacology , Stromal Cells/metabolism , Th1 Cells/metabolism , Th2 Cells/metabolism , Animals , Cell Differentiation/drug effects , Cell Proliferation , Cyclophosphamide/pharmacology , Erythroid Cells/immunology , Granulocytes/immunology , Hematopoiesis/drug effects , Hematopoietic System/drug effects , Macrophages/immunology , Megakaryocytes/immunology , Mice , Mice, Inbred CBA , Stromal Cells/immunology , Th1 Cells/immunology , Th2 Cells/immunologyABSTRACT
The hemostimulating effect of erythropoietin immobilized by the nanotechnology method of electron-beam synthesis was studied on the model of carboplatin-induced myelosuppression. Subcutaneous injection or oral administration of immobilized erythropoietin was followed by stimulation of erythropoiesis. The effect was most pronounced after parenteral treatment with this agent. The increase in proliferative activity and maturation of erythroid precursor cells serve as a factor determining acceleration of reparative processes in the hemopoietic tissue.