ABSTRACT
The aim of this work was to investigate the in vitro cytotoxic effect of previously developed nanocapsules, nanoemulsion, and microemulsion based on bullfrog oil (BFO) against human melanoma cells (A2058). The nanosystems were produced as described in previous studies and characterized according to droplet/particle distribution and zeta potential. The biocompatibility was evaluated by the determination of the hemolytic potential against human erythrocytes. The cytotoxicity assessment was based on MTT and cell death assays, determination of Reactive Oxygen Species (ROS) levels, and cell uptake. The nanosystems were successfully reproduced and showed hemolytic potential smaller than 10% at all oil concentrations (50 and 100 µg.mL-1) (p < 0.05). The MTT assay revealed that the nanosystems decreased the mitochondrial activity up to 92 ± 2% (p < 0.05). The study showed that the free BFO induced cell apoptosis, while all the nanostructured systems caused cell death by necrosis associated with a ROS overproduction. This can be related to the increased ability of the nanostructured systems to deliver the BFO across all cellular compartments (membrane, cytoplasm, and nucleus). Finally, these results elucidate the in vitro BFO nanosystems cytotoxic effect against human melanoma cells (A2058), revealing the emulsified ones as the most cytotoxic systems. Overall, the findings suggest that the safety and antineoplastic activity of these systems can be further investigated by in vivo studies.
Subject(s)
Antineoplastic Agents/pharmacology , Melanoma/drug therapy , Nanostructures , Skin Neoplasms/drug therapy , Animals , Antineoplastic Agents/isolation & purification , Antineoplastic Agents/toxicity , Apoptosis/drug effects , Cell Line, Tumor , Emulsions , Erythrocytes/drug effects , Erythrocytes/metabolism , Hemolysis/drug effects , Humans , Melanoma/pathology , Mitochondria/metabolism , Nanocapsules , Oils/isolation & purification , Oils/pharmacology , Oils/toxicity , Particle Size , Rana catesbeiana/metabolism , Reactive Oxygen Species/metabolism , Skin Neoplasms/pathologyABSTRACT
Bullfrog oil is a natural product extracted from the Rana catesbeiana Shaw adipose tissue and used in folk medicine for the treatment of several diseases. The aim of this study was to evaluate the extraction process of bullfrog oil, to develop a suitable topical nanoemulsion and to evaluate its efficacy against melanoma cells. The oil samples were obtained by hot and organic solvent extraction processes and were characterized by titration techniques and gas chromatography mass spectrometry (GC-MS). The required hydrophile-lipophile balance and the pseudo-ternary phase diagram (PTPD) were assessed to determine the emulsification ability of the bullfrog oil. The anti-tumoral activity of the samples was assessed by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay for normal fibroblast (3T3) and melanoma (B16F10) cell lines. Both extraction methods produced yielded around 60% and the oil was mainly composed of unsaturated compounds (around 60%). The bullfrog oil nanoemulsion obtained from PTPD presented a droplet size of about 390 nm and polydispersity = 0.05 and a zeta potential of about -25 mV. Both the bullfrog oil itself and its topical nanoemulsion did not show cytotoxicity in 3T3 linage. However, these systems showed growth inhibition in B16F10 cells. Finally, the bullfrog oil presented itself as a candidate for the development of pharmaceutical products free from cytotoxicity and effective for antineoplastic therapy.
Subject(s)
Antineoplastic Agents/isolation & purification , Biological Products/therapeutic use , Melanoma, Experimental/drug therapy , Oils/therapeutic use , Rana catesbeiana , 3T3 Cells , Animals , Antineoplastic Agents/therapeutic use , Antineoplastic Agents/toxicity , Biomedical Research/trends , Cell Line, Tumor , Drug Screening Assays, Antitumor , Emulsions , HeLa Cells , Humans , Mice , Oils/chemistry , Oils/isolation & purification , Oils/toxicityABSTRACT
Microemulsion-based animal oils, alone or associated with polymers have been extensively used in pharmacy, medicine and cosmetics, since the major lipid constituents of the oils show several biological activities. Despite showing antimicrobial activity, there are no reports in the literature regarding the effects of bullfrog oil on cytotoxic activity against tumor cells. The aim of the present study was to synthesize, characterise and evaluate the in vitro effects on melanoma cell line (B16F10) of bullfrog oil microemulsions associated or not with chitosan, surfactant and bullfrog oil (CSBO) and surfactant and bullfrog oil (SBO), respectively. The microemulsions were developed and their physical-chemical characteristics were evaluated by light microscopy, dynamic light scattering, atomic force microscopy and zeta potential. The microemulsions showed regular spherical shapes, high polydispersity and excellent (+82.2 ± 1.0 mV) to low (-16.0 ± 0.5 mV), colloidal stability. The systems significantly decreased the in vitro cell viability of melanoma skin cancer by up to 90.2% (CSBO) and 91.8% (SBO); while free bullfrog oil showed no effects. The results obtained from microemulsions of bullfrog oil indicate the potential of the microemulsions developed, alone or in combination with other chemotherapeutic agents, for future use in biomedical approaches aiming towards cancer therapy.
Subject(s)
Antineoplastic Agents/toxicity , Cell Survival/drug effects , Chitosan/chemistry , Emulsions/toxicity , Oils/toxicity , Rana catesbeiana/metabolism , Animals , Antineoplastic Agents/chemistry , Biological Products , Cell Line, Tumor , Emulsions/chemistry , Melanoma , Oils/chemistry , Particle SizeABSTRACT
The sucupira extract is used in folk medicine as an alternative treatment for various diseases, being known to the essential oil extracted from the leaves of actions, trunk bark, tubers and roots, but the use of seed is little discussed. Aiming to determine the toxicity was held this experiment, with six groups of mice receiving the organic fraction of the seed extract of the white sucupira (FOESSB) at different doses every other day. Bioassays of spontaneous activity revealed that FOESSB provided a list of characteristic behavioral responses, signaling stimulating effects and depressant on the central nervous system, effects on the SNA among others, though , the effect dose of relationships found in responses associated with significant effects on the CNS, it is recommended that, for future studies lower doses are employed to 385.71mg / kg.
O extrato da sucupira é usado pela medicina popular como alternativa de tratamento para diversas enfermidades, sendo conhecidas às ações do óleo essencial extraído das folhas, casca do tronco, tubérculos e raiz, porém o uso da semente é pouco discutida. Objetivando determinar a toxicidade, realizou-se este experimento, com seis grupos de camundongos que receberam a fração orgânica do extrato da semente da sucupira branca (FOESSB) em doses diferentes em dias alternados. Os bioensaios da atividade espontânea revelaram que a FOESSB proporcionou um elenco de respostas comportamentais característico, sinalizando efeitos estimulantes e depressores sobre o SNC, efeitos sobre o SNA entre outros, muito embora, pelas relações de dose efeito encontradas em respostas associadas a importantes efeitos sobre o SNC, recomenda-se que, para futuros trabalhos sejam empregadas doses inferiores a 385,71mg/Kg. Não foram encontradas diferenças significativas na avaliação macroscópica e pesagens efetuadas sobre órgãos dos camundongos.
Subject(s)
Animals , Mice , Phytotherapy , Plants, Medicinal/toxicity , Oils/toxicity , Autonomic Nervous System , Central Nervous SystemABSTRACT
The sucupira extract is used in folk medicine as an alternative treatment for various diseases, being known to the essential oil extracted from the leaves of actions, trunk bark, tubers and roots, but the use of seed is little discussed. Aiming to determine the toxicity was held this experiment, with six groups of mice receiving the organic fraction of the seed extract of the white sucupira (FOESSB) at different doses every other day. Bioassays of spontaneous activity revealed that FOESSB provided a list of characteristic behavioral responses, signaling stimulating effects and depressant on the central nervous system, effects on the SNA among others, though , the effect dose of relationships found in responses associated with significant effects on the CNS, it is recommended that, for future studies lower doses are employed to 385.71mg / kg. (AU)
O extrato da sucupira é usado pela medicina popular como alternativa de tratamento para diversas enfermidades, sendo conhecidas às ações do óleo essencial extraído das folhas, casca do tronco, tubérculos e raiz, porém o uso da semente é pouco discutida. Objetivando determinar a toxicidade, realizou-se este experimento, com seis grupos de camundongos que receberam a fração orgânica do extrato da semente da sucupira branca (FOESSB) em doses diferentes em dias alternados. Os bioensaios da atividade espontânea revelaram que a FOESSB proporcionou um elenco de respostas comportamentais característico, sinalizando efeitos estimulantes e depressores sobre o SNC, efeitos sobre o SNA entre outros, muito embora, pelas relações de dose efeito encontradas em respostas associadas a importantes efeitos sobre o SNC, recomenda-se que, para futuros trabalhos sejam empregadas doses inferiores a 385,71mg/Kg. Não foram encontradas diferenças significativas na avaliação macroscópica e pesagens efetuadas sobre órgãos dos camundongos. (AU)
Subject(s)
Animals , Mice , Plants, Medicinal/toxicity , Phytotherapy , Oils/toxicity , Autonomic Nervous System , Central Nervous SystemABSTRACT
The objective of this study was to evaluate the environmental behavior of different types of automotive lubricant oils. Based on respirometry assays the biodegradability was monitored, and toxicological tests were executed to assess the lubricants toxicity before and after microbial activity. Used oil was the most biodegradable, however, it was the most toxic. Also, all lubricants presented toxicity even after biodegradation due to 40% Eruca sativa germination inhibition and a low LC50 to Eisenia foetida (0.50-0.25 mL). Moreover, used automotive lubricants have a high toxicity because of polycyclic aromatic hydrocarbons concentration that establishes them as a potential carcinogen.