ABSTRACT
The encapsulation of essential oils (EOs) in protein-based biopolymeric matrices stabilized with surfactant ensures protection and physical stability of the EO against unfavorable environmental conditions. Accordingly, this study prepared zein nanoparticles loaded with eucalyptus essential oil (Z-EEO) and Litsea cubeba essential oil (Z-LEO), stable and with antifungal activity against Colletotrichum lindemuthianum, responsible for substantial damage to bean crops. The nanoparticles were prepared by nanoprecipitation with the aid of ultrasound treatment and characterized. The nanoparticles exhibited a hydrodynamic diameter close to 200 nm and PDI < 0.3 for 120 days, demonstrating the physical stability of the carrier system. Scanning electron microscopy and Transmission electron microscopy revealed that the nanoparticles were smooth and uniformly distributed spheres. Fourier-transform infrared spectroscopy showed interaction between zein and EOs through hydrogen bonding and hydrophobic interactions. Thermogravimetric analysis demonstrated the thermal stability of the nanoparticles compared to pure bioactive compounds. The nanoparticles exhibited a dose-dependent effect in inhibiting the fungus in in vitro testing, with Z-EEO standing out by inhibiting 70.0 % of the mycelial growth of C. lindemuthianum. Therefore, the results showed that zein has great potential to encapsulate hydrophobic compounds, improving the applicability of the bioactive compound as a biofungicide, providing protection for the EO.
Subject(s)
Antifungal Agents , Eucalyptus , Litsea , Nanoparticles , Oils, Volatile , Zein , Zein/chemistry , Antifungal Agents/pharmacology , Antifungal Agents/chemistry , Oils, Volatile/chemistry , Oils, Volatile/pharmacology , Eucalyptus/chemistry , Nanoparticles/chemistry , Litsea/chemistry , Colletotrichum/drug effects , Spectroscopy, Fourier Transform Infrared , Drug StabilityABSTRACT
The purpose of this study was to determine the chemical profile of Litsea cubeba essential oil, carry out an in vitro evaluation of its antioxidant potential and its cytotoxicity, as well as its antifungal and antimicotoxigenic activities against Fusarium verticillioides. Most of the compounds observed in the EO were neral (32.75%) and geranial (37.67%). The radical scavenging capacity of 2,2-diphenyl-1-picrylhydrazyl and 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid was 104.4 and 56.4 mmol Trolox mg-1, respectively, indicating good antioxidant activity. The EO studied by us revealed cytotoxic effect against HT-29 and HeLa cancer cells. The Minimum Inhibitory and Minimum Fungicidal Concentrations against F. verticillioides were both 125 µg mL-1. Morphological investigation, performed by fluorescence microscopy and scanning electron microscopy, showed that hyphae and microconidia structures underwent changes after treatment with the EO. Analyses performed with the EO strongly reduced the mycelial development of F. verticillioides and the synthesis of fumonisins B1 and B2 in dose-dependence effect compared (P < 0.01) with the fungal control (105 conidia mL-1) and positive control (fludioxonil + metalaxyl-M). Thus, the results obtained in vitro suggest that L. cubeba EO has excellent antioxidant, fungicidal, and antimycotoxigenic effects.
Subject(s)
Antifungal Agents/pharmacology , Antioxidants/pharmacology , Fusarium/drug effects , Litsea/chemistry , Oils, Volatile/pharmacology , Acyclic Monoterpenes/analysis , Antifungal Agents/chemistry , Antineoplastic Agents, Phytogenic/chemistry , Antineoplastic Agents, Phytogenic/pharmacology , Antioxidants/chemistry , Dose-Response Relationship, Drug , Fumonisins/metabolism , Fusarium/metabolism , HT29 Cells , HeLa Cells , Humans , Microbial Sensitivity Tests , Mycelium/drug effects , Oils, Volatile/administration & dosage , Oils, Volatile/chemistryABSTRACT
BACKGROUND: The study was conducted to evaluate the in vitro thrombolytic activity, and in vivo analgesic, anti-inflammatory and antipyretic potentials of different hydrocarbon soluble extracts of Litsea glutinosa leaves for the first time widely used in the folkloric treatments in Bangladesh. This work aimed to create new insights on the fundamental mechanisms of the plant extracts involved in these activities. RESULTS: In thrombolytic activity assay, a significant clot disruption was observed at dose of 1 mg/mL for each of the extracts (volume 100 µL) when compared to the standard drug streptokinase. The n-hexane, ethyl acetate, chloroform, and crude methanolic extracts showed 32.23 ± 0.26, 37.67 ± 1.31, 43.13 ± 0.85, and 46.78 ± 0.9% clot lysis, respectively, whereas the positive control streptokinase showed 93.35 ± 0.35% disruption at the dose of 30,000 I.U. In hot plate method, the highest pain inhibitory activity was found at a dose of 500 mg/kg of crude extract (15.54 ± 0.37 sec) which differed significantly (P <0.01 and P <0.001) with that of the standard drug ketorolac (16.38 ± 0.27 sec). In acetic acid induced writhing test, the crude methanolic extract showed significant (P <0.01 and P <0.001) analgesic potential at doses 250 and 500 mg/kg body weight (45.98 and 56.32% inhibition, respectively), where ketorolac showed 64.36% inhibition. In anti-inflammatory activity test, the crude methanolic extract showed significant (P <0.001) potential at doses 250 and 500 mg/kg body weight (1.51 ± 0.04 and 1.47 ± 0.03 mm paw edema, respectively), where ketorolac showed 1.64 ± 0.05 mm edema after 3 h of carrageenan injection. In antipyretic activity assay, the crude extract showed notable reduction in body temperature (32.78 ± 0.46°C) at dose of 500 mg/kg-body weight, when the standard (at dose 150 mg/kg-body weight) exerted 33.32 ± 0.67°C temperature after 3 h of administration. CONCLUSIONS: Our results yield that the crude hydroalcoholic extract has better effects than the other in all trials. In the context, it can be said that the leaves of L. glutinosa possess remarkable pharmacological effects, and justify its traditional use as analgesic, antipyretic, anti-inflammatory, and thrombolytic agent.
Subject(s)
Analgesics/therapeutic use , Anti-Inflammatory Agents, Non-Steroidal/therapeutic use , Antipyretics/therapeutic use , Fibrinolytic Agents/therapeutic use , Litsea/chemistry , Phytotherapy , Acetic Acid , Animals , Edema/chemically induced , Edema/drug therapy , Female , Fibrinolysis/drug effects , Humans , Male , Medicine, Traditional , Methanol , Mice , Plant Extracts/pharmacology , Plant Leaves/chemistryABSTRACT
BACKGROUND: The study was conducted to evaluate the in vitro thrombolytic activity, and in vivo analgesic, anti-inflammatory and antipyretic potentials of different hydrocarbon soluble extracts of Litsea glutinosaleaves for the first time widely used in the folkloric treatments in Bangladesh. This work aimed to create new insights on the fundamental mechanisms of the plant extracts involved in these activities. RESULTS: In thrombolytic activity assay, a significant clot disruption was observed at dose of 1 mg/mL for each of the extracts (volume 100 µL) when compared to the standard drug streptokinase. The n-hexane, ethyl acetate, chloroform, and crude methanolic extracts showed 32.23 ± 0.26, 37.67 ± 1.31, 43.13 ± 0.85, and 46.78 ± 0.9% clot lysis, respectively, whereas the positive control streptokinase showed 93.35 ± 0.35% disruption at the dose of 30,000 I.U. In hot plate method, the highest pain inhibitory activity was found at a dose of 500 mg/kg of crude extract (15.54 ± 0.37 sec) which differed significantly (P <0.01 and P <0.001) with that of the standard drug ketorolac (16.38 ± 0.27 sec). In acetic acid induced writhing test, the crude methanolic extract showed significant (P <0.01 and P <0.001) analgesic potential at doses 250 and 500 mg/kg body weight (45.98 and 56.32% inhibition, respectively), where ketorolac showed 64.36% inhibition. In anti-inflammatory activity test, the crude methanolic extract showed significant (P <0.001) potential at doses 250 and 500 mg/kg body weight (1.51 ± 0.04 and 1.47 ± 0.03 mm paw edema, respectively), where ketorolac showed 1.64 ± 0.05 mm edema after 3 h of carrageenan injection. In antipyretic activity assay, the crude extract showed notable reduction in body temperature (32.78 ± 0.46°C) at dose of 500 mg/kg-body weight, when the standard (at dose 150 mg/kg-body weight) exerted 33.32 ± 0.67°C temperature after 3 h of administration. CONCLUSIONS: Our results yield that the crude hydroalcoholic extract has better effects than the other in all trials. In the context, it can be said that the leaves of L. glutinosa possess remarkable pharmacological effects, and justify its traditional use as analgesic, antipyretic, anti-inflammatory, and thrombolytic agent.