Search

Fibrinolytic enzymes are promising in treating cardiovascular diseases due to their capacity to dissolve blood clots. The fibrinolytic enzyme from Arthrospira platensis (FEAP) was purified by ion exchange chromatography to investigate its ability to activate plasminogen, as well as its thrombolytic and fibrinogenolytic potential. Subsequently, two different cytotoxic assays (MTT and NR) and hemolysis test were performed to evaluate FEAP's safety. Furthermore, cell migration and the genotoxic and hemolytic potential were also investigated. The purified enzyme showed thrombus degradation of 43 % and thrombolytic action directly on fibrin, which can reduce possible side effects, such as hemorrhage. MTT assay was more sensitive to determine the enzyme cytotoxicity, which decreased the viability of breast cancer tumor cells (Sarcoma-180 and MDA-MB-231) and macrophages (J774A.1). In addition, the enzyme also exhibited non-hemolytic, antimetastatic, and non-genotoxic characteristics. These findings are innovative for a fibrinolytic protease and may indicate that it is safe for people undergoing cancer treatment, reducing side effects such as hemorrhage, in addition to inhibiting tumor cells and preventing metastasis, which can help with chemotherapy treatment., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024. Published by Elsevier B.V.)

This work reports the immobilization of a fibrinolytic protease (FP) from Mucor subtilissimus UCP 1262 on Fe 3 O 4 magnetic nanoparticles (MNPs) produced by precipitation of FeCl 3 ·6H 2 O and FeCl 2 ·4H 2 O, coated with polyaniline and activated with glutaraldehyde. The FP was obtained by solid state fermentation, precipitated with 40-60% ammonium sulfate, and purified by DEAE-Sephadex A50 ion exchange chromatography. The FP immobilization procedure allowed for an enzyme retention of 52.13%. The fibrinolytic protease immobilized on magnetic nanoparticles (MNPs/FP) maintained more than 60% of activity at a temperature of 40 to 60 °C and at pH 7 to 10, when compared to the non-immobilized enzyme. MNPs and MNPs/FP did not show any cytotoxicity against HEK-293 and J774A.1 cells. MNPs/FP was not hemolytic and reduced the hemolysis induced by MNPs from 2.07% to 1.37%. Thrombus degradation by MNPs/FP demonstrated that the immobilization process guaranteed the thrombolytic activity of the enzyme. MNPs/FP showed a total degradation of the γ chain of human fibrinogen within 90 min. These results suggest that MNPs/FP may be used as an alternative strategy to treat cardiovascular diseases with a targeted release through an external magnetic field., (Copyright © 2022 Elsevier Inc. All rights reserved.)