Avaliação do potencial biotecnológico do óleo da semente de Euterpe oleracea Mart. In vitro e In vivo e da sua nanoemulsão
Autor(a) principal: | |
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Data de Publicação: | 2024 |
Tipo de documento: | Tese |
Idioma: | por |
Título da fonte: | Biblioteca Digital de Teses e Dissertações da UFMA |
Texto Completo: | https://tedebc.ufma.br/jspui/handle/tede/tede/5384 |
Resumo: | Açaí seed oil, Euterpe oleracea Mart., is rich in unsaturated, saturated, monounsaturated and polyunsaturated fatty acids with anti-inflammatory, antioxidant, antimicrobial, antinociceptive, anticancer, antiatherogenic and healing activities. The açaí seed is a residue produced after pulping, being a source of bioactive compounds; therefore, it is necessary to study its pharmacological properties and contribute to sustainable development. The objective of this was to evaluate the biotechnological potential of Euterpe oleracea Mart seed oil. - in vitro and in vivo - and its Nanoemulsion. The fruits used were collected in the Maracanã Ecological Park (Parque da Juçara), in São Luís, Maranhão. Samples of the plant were collected between the months of August and November, and exsicata were prepared, duly stored in the Rosa Mochel Herbarium. Subsequently, the oil was extracted with n-hexane solvent in a Soxhlet extractor and analyzed as methyl esters, weighing 100 mg of the oil for analysis of the chromatographic profile by gas chromatography combined with mass spectrometry (GC-MS) to identify the fatty acids. and its physical-chemical characterization. After understanding the physical chemistry of the oil, the açaí seed oil nanoemulsion (NE- OEO) was developed based on the calculation of the Hydrophilic Lipophilic Balance (EHL), followed by stability analysis, chemical characterization and morphology. To determine the average particle size and polydispersity index, the formulations were evaluated using the dynamic light scattering technique (Nano ZS, Malvern Instruments Ltd., UK) and Zeta Potential on the Nano ZS equipment (Malvern Instruments Ltd., UK). Both the oil and NE-OEO showed physicochemical patterns within normal limits. In other words, the oil showed acidity (0.3556± 0.0003), humidity (1.25±1.64), saponification index (189 ± 1.04), refractive index (1.4707±0.00005 ), incineration residue (ash) (0.42±0.42) and density (0.928±0.00005). And NE-OEO showed normal patterns regarding droplet size (238.37 ± 3.96), homogeneity (PDI) (0.38 ± 0.38), Zeta potential (-9.59 ± 0.11), pH (7.0 ± 0.00.) and turbidity (0.267± 0.00). Regarding phenolic compounds, NE-OEO presented the highest concentration, 146.00 ± 0.259 mg, while the oil had 127.40 ± 0.449 mg EAG g-1 of phenolic compounds, and total flavonoids values were found to be 62.62 ± 0.930 mg for the oil and 113.80 ± 0.454 mg EQ g-1 for NE-OEO. The antioxidant activity was proven based on the EC50 evaluated in the DPPH and ABTS assay, with the oil being 375.698 μg/mL in DPPH, and nanoemulsions with a significantly lower EC50, equivalent to 229.845 μg/mL. The Trolox control presented an even lower EC50. In the ABTS test, the EC50 of açaí oil was 272.0208 μg/mL, while açaí oil nanoemulsions had an even lower EC50 value, measuring 201.2895 μg/mL. Trolox also showed a significantly lower EC50 compared to oil and NE-OEO, these data confirm the antioxidant capacity of the oil and its NE- OEO. In vitro analyzes were performed on the non-tumor cell lines mouse RAW 264.7 macrophages, and human HaCat keratinocytes and cervical cancer tumor lines SiHa (HPV-16) and HeLa (HPV-18). Non-tumor cell lines were treated with concentrations (7.8 to 1000 ug/mL) of the oil and NE-OEO for 24, 48 and 72 hours, and tumor cells were treated with concentrations (0.631 to 100 ug/mL) of the oil, made analyzes of cell viability, cell death, morphology, healing and clonogenicity. After in vitro analyses, the effect of açaí seed oil and NE-OEO was analyzed in vivo in female Swiss mice. The animals were treated with different concentrations of the oil (100, 200 and 300 mg/kg) and NE-OEO (100 mg/kg) applied 0.5 mL intraperitoneally every two weeks. Açaí seed oil is rich in polyphenols, flavonoids, saturated fatty acids (49.27%) and unsaturated fatty acids (50.73%), among which unsaturated fatty acids, 29.73% are monounsaturated, 20 .85% are polyunsaturated. Lauric, myristic (C14:0), palmitic, linoleic, and oleic fatty acids were the most identified. Viability assay with MTT ((3-(4,5- Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)) demonstrated a cytotoxic effect of the oil on the SiHa (HPV-16) and HeLa (HPV- 18) and non-cytotoxic to the non-tumor cell lines HaCat, as well as the oil and NE-OEO were not cytotoxic to the non-tumor cell line RAW 264.7. Açaí seed oil induced morphological changes in HeLa and SiHa cells, inducing a reduction in cell viability, inhibiting cell migration and colony formation in cervical cancer cell lines. The annexin V-FITC assay showed cell death by early and late apoptosis at 24 and 48 hours after treatment with açaí seed oil. In in vivo experiments, the use of oil at a dose of 100, 200, 300 mg/kg and NE-OEO at a dose of 100 mg/kg were not toxic to the animals, in accordance with the normal patterns seen in the liver and kidney; Furthermore, it did not interfere with the behavior or weight of the animals, however, it induced an immunomodulatory response with a significant increase in IL10 and IL17. Our results suggest the biotechnological potential of the oil and NE-OEO in antitumor activity for cervical cancer, and in the development of colloidal systems for drug delivery. Furthermore, the use of açaí seed residue will contribute to both the local economy and sustainable development.. |
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NASCIMENTO, Maria do Desterro Soares Brandãohttp://lattes.cnpq.br/3958174822396319NASCIMENTO, Maria do Desterro Soares Brandãohttp://lattes.cnpq.br/3958174822396319SILVA, Ana Lúcia Abreuhttp://lattes.cnpq.br/8288733951324759FAUSTINO-ROCHA, Ana IsabelTELES, Amanda Marahttp://lattes.cnpq.br/3933255152524601OLIVEIRA, Rui Miguel Gil da Costahttp://lattes.cnpq.br/6785759461393904http://lattes.cnpq.br/0528263981302728BORGES, Katia Regina Assunção2024-07-08T12:39:42Z2024-04-26BORGES, Katia Regina Assunção. Avaliação do potencial biotecnológico do óleo da semente de Euterpe oleracea Mart. In vitro e In vivo e da sua nanoemulsão. 2024. 112 f. Tese (Programa de Pós-Graduação em Biotecnologia - RENORBIO/CCBS) - Universidade Federal do Maranhão, São Luís, 2024.https://tedebc.ufma.br/jspui/handle/tede/tede/5384Açaí seed oil, Euterpe oleracea Mart., is rich in unsaturated, saturated, monounsaturated and polyunsaturated fatty acids with anti-inflammatory, antioxidant, antimicrobial, antinociceptive, anticancer, antiatherogenic and healing activities. The açaí seed is a residue produced after pulping, being a source of bioactive compounds; therefore, it is necessary to study its pharmacological properties and contribute to sustainable development. The objective of this was to evaluate the biotechnological potential of Euterpe oleracea Mart seed oil. - in vitro and in vivo - and its Nanoemulsion. The fruits used were collected in the Maracanã Ecological Park (Parque da Juçara), in São Luís, Maranhão. Samples of the plant were collected between the months of August and November, and exsicata were prepared, duly stored in the Rosa Mochel Herbarium. Subsequently, the oil was extracted with n-hexane solvent in a Soxhlet extractor and analyzed as methyl esters, weighing 100 mg of the oil for analysis of the chromatographic profile by gas chromatography combined with mass spectrometry (GC-MS) to identify the fatty acids. and its physical-chemical characterization. After understanding the physical chemistry of the oil, the açaí seed oil nanoemulsion (NE- OEO) was developed based on the calculation of the Hydrophilic Lipophilic Balance (EHL), followed by stability analysis, chemical characterization and morphology. To determine the average particle size and polydispersity index, the formulations were evaluated using the dynamic light scattering technique (Nano ZS, Malvern Instruments Ltd., UK) and Zeta Potential on the Nano ZS equipment (Malvern Instruments Ltd., UK). Both the oil and NE-OEO showed physicochemical patterns within normal limits. In other words, the oil showed acidity (0.3556± 0.0003), humidity (1.25±1.64), saponification index (189 ± 1.04), refractive index (1.4707±0.00005 ), incineration residue (ash) (0.42±0.42) and density (0.928±0.00005). And NE-OEO showed normal patterns regarding droplet size (238.37 ± 3.96), homogeneity (PDI) (0.38 ± 0.38), Zeta potential (-9.59 ± 0.11), pH (7.0 ± 0.00.) and turbidity (0.267± 0.00). Regarding phenolic compounds, NE-OEO presented the highest concentration, 146.00 ± 0.259 mg, while the oil had 127.40 ± 0.449 mg EAG g-1 of phenolic compounds, and total flavonoids values were found to be 62.62 ± 0.930 mg for the oil and 113.80 ± 0.454 mg EQ g-1 for NE-OEO. The antioxidant activity was proven based on the EC50 evaluated in the DPPH and ABTS assay, with the oil being 375.698 μg/mL in DPPH, and nanoemulsions with a significantly lower EC50, equivalent to 229.845 μg/mL. The Trolox control presented an even lower EC50. In the ABTS test, the EC50 of açaí oil was 272.0208 μg/mL, while açaí oil nanoemulsions had an even lower EC50 value, measuring 201.2895 μg/mL. Trolox also showed a significantly lower EC50 compared to oil and NE-OEO, these data confirm the antioxidant capacity of the oil and its NE- OEO. In vitro analyzes were performed on the non-tumor cell lines mouse RAW 264.7 macrophages, and human HaCat keratinocytes and cervical cancer tumor lines SiHa (HPV-16) and HeLa (HPV-18). Non-tumor cell lines were treated with concentrations (7.8 to 1000 ug/mL) of the oil and NE-OEO for 24, 48 and 72 hours, and tumor cells were treated with concentrations (0.631 to 100 ug/mL) of the oil, made analyzes of cell viability, cell death, morphology, healing and clonogenicity. After in vitro analyses, the effect of açaí seed oil and NE-OEO was analyzed in vivo in female Swiss mice. The animals were treated with different concentrations of the oil (100, 200 and 300 mg/kg) and NE-OEO (100 mg/kg) applied 0.5 mL intraperitoneally every two weeks. Açaí seed oil is rich in polyphenols, flavonoids, saturated fatty acids (49.27%) and unsaturated fatty acids (50.73%), among which unsaturated fatty acids, 29.73% are monounsaturated, 20 .85% are polyunsaturated. Lauric, myristic (C14:0), palmitic, linoleic, and oleic fatty acids were the most identified. Viability assay with MTT ((3-(4,5- Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)) demonstrated a cytotoxic effect of the oil on the SiHa (HPV-16) and HeLa (HPV- 18) and non-cytotoxic to the non-tumor cell lines HaCat, as well as the oil and NE-OEO were not cytotoxic to the non-tumor cell line RAW 264.7. Açaí seed oil induced morphological changes in HeLa and SiHa cells, inducing a reduction in cell viability, inhibiting cell migration and colony formation in cervical cancer cell lines. The annexin V-FITC assay showed cell death by early and late apoptosis at 24 and 48 hours after treatment with açaí seed oil. In in vivo experiments, the use of oil at a dose of 100, 200, 300 mg/kg and NE-OEO at a dose of 100 mg/kg were not toxic to the animals, in accordance with the normal patterns seen in the liver and kidney; Furthermore, it did not interfere with the behavior or weight of the animals, however, it induced an immunomodulatory response with a significant increase in IL10 and IL17. Our results suggest the biotechnological potential of the oil and NE-OEO in antitumor activity for cervical cancer, and in the development of colloidal systems for drug delivery. Furthermore, the use of açaí seed residue will contribute to both the local economy and sustainable development..O óleo da semente de açaí, Euterpe oleracea Mart., é rico em ácidos graxos insaturados, saturados, monoinsaturados e polinsaturados com atividade anti- inflamatória, antioxidante, antimicrobiana, antinociceptiva, anticancerígena, antiaterogênica e cicatrizante. A semente do açaí é um resíduo produzido após o despolpamento, sendo fonte de compostos bioativos; assim, se faz necessário estudar suas propriedades farmacológicas e contribuir para o desenvolvimento sustentável. O objetivo deste foi avaliar o potencial biotecnológico do óleo da semente de Euterpe oleracea Mart. - in vitro e in vivo - e da sua Nanoemulsão. Os frutos utilizados foram coletados no Parque Ecológico do Maracanã (Parque da Juçara), em São Luís, Maranhão. Amostras da planta foram coletadas entre os meses agosto e novembro, e realizada confecção da exsicata, devidamente armazenada no Herbário Rosa Mochel. Posteriormente, o óleo foi extraído com solvente n-hexano em extrator Soxhlet e analisado como ésteres metílicos, pesando-se 100 mg do óleo para análise do perfil cromatográfico por cromatografia gasosa combinada com espectrometria de massa (CG-EM) para identificação dos ácidos graxos e sua caracterização físico- química. Após conhecer a físico-química do óleo, foi desenvolvida a nanoemulsão de óleo de semente de açaí (NE-OEO) a partir do cálculo do Equilíbrio Hidrofílico Lipofílico (EHL), seguido da análise de estabilidade, caracterização química e morfologia. Para determinar o tamanho médio das partículas e o índice de polidispersão, as formulações foram avaliadas através da técnica de espalhamento dinâmico de luz (Nano ZS, Malvern Instruments Ltd., UK) e do Potencial Zeta no equipamento Nano ZS (Malvern Instruments Ltd., UK). Tanto o óleo quanto a NE-OEO mostraram padrões físico-químicos dentro da normalidade. Ou seja, o óleo mostrou acidez (0,3556± 0,0003), umidade (1,25±1,64), índice de saponificação (189 ± 1,04), índice de refração (1,4707±0,00005), resíduo de incineração (cinzas) (0,42±0,42) e densidade (0,928±0,00005). E a NE-OEO mostrou os padrões normais quanto tamanho das gota (238,37 ± 3,96), homogeneidade (PDI) (0,38 ± 0,38), potencial Zeta (-9,59 ± 0,11), pH (7,0 ± 0,00.) e turbidez (0,267± 0,00). Com relação aos compostos fenólicos, NE-OEO apresentou maior concentração, 146,00 ± 0,259 mg, enquanto o óleo foi de 127,40 ± 0,449 mg EAG g-1 de compostos fenólicos, e flavonoides totais foram encontrados os valores 62,62 ± 0,930 mg para o óleo e 113,80 ± 0,454 mg EQ g-1 para a NE-OEO. A atividade antioxidante foi comprovada a partir do EC50 avaliado no ensaio de DPPH e ABTS, sendo, no DPPH, o óleo 375,698 μg/mL, e nanoemulsões com EC50 significativamente menor, equivalente a 229,845 μg/mL. O controle Trolox, apresentou um EC50 ainda mais baixo. No teste ABTS, o EC50 do óleo de açaí foi de 272,0208 μg/mL, enquanto as nanoemulsões de óleo de açaí apresentaram um valor de EC50 ainda menor, medindo 201,2895 μg/mL. O Trolox também apresentou um EC50 significativamente mais baixo em comparação com óleo e NE-OEO, esses dados confirmam a capacidade antioxidante do óleo e sua NE-OEO. Foram realizadas análises in vitro nas linhagens celulares não tumorais macrófagos RAW 264.7 de camundongo, e queratinócitos humano HaCat e linhagens tumorais de câncer cervical SiHa (HPV-16) e HeLa (HPV-18). As linhagens não tumorais foram tratadas com concentrações (7.8 a 1000 ug/mL) do óleo e NE-OEO por 24, 48 e 72 horas, e as células tumorais foram tratadas com concentrações (0,631 a 100 ug/mL) do óleo, feitas as análises de viabilidade celular, morte celular, morfologia, cicatrização e clonogenicidade. Após as análises in vitro, foi analisado o efeito do óleo e da NE- OEO da semente de açaí em in vivo em camundongo Swiss fêmeas. Os animais foram tratados com diferentes concentrações do óleo (100, 200 e 300 mg/kg) e NE-OEO (100 mg/kg) aplicados 0,5 mL intraperitonealmente em com quinzenalmente. O óleo da semente de açaí é rico em polifenois, flavonoides, ácidos graxos saturados (49,27%l) e ácidos graxos insaturados (50,73%), dentre os quais os ácidos graxos insaturados, 29,73% são monoinsaturados, 20,85% são poliinsaturados. Os ácidos graxos láurico, mirístico (C14:0), palmítico, linoleico, e oleico, foram os majoritariamente identificados. Ensaio de viabilidade com MTT ((3-(4,5- Dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide)) demonstrou efeito citotóxico do óleo na linhagem de câncer cervical SiHa (HPV-16) e HeLa (HPV-18) e não citotóxico nas linhagens não tumorais HaCat, bem como o óleo e NE-OEO não foram citotóxico para a linhagem não tumoral RAW 264.7. O óleo da semente de açaí induziu alterações morfológicas nas células HeLa e SiHa, induzindo redução de viabilidade celular, inibiu a migração celular e formação de colônia nas linhagens celulares de câncer cervical. O ensaio com anexina V-FITC evidenciou morte celular por apoptose inicial e tardia em 24 e 48 hs após tratamento com o óleo da semente de açaí. Nos experimentos in vivo, o uso óleo na dose de 100, 200, 300 mg/kg e NE-OEO na dose 100 mg/kg não foram tóxicos para os animais, de acordo os padrões normais vistos no fígado e rim; além disso, não interferiu no comportamento e peso dos animais, no entanto, induziram resposta imunomoduladora com aumento significativo de IL10 e IL17. Nossos resultados sugerem o potencial biotecnológico do óleo e da NE-OEO em atividade antitumoral para câncer cervical, e em desenvolvimento de sistemas coloidais para entrega de fármacos. Além disso, o aproveitamento do resíduo da semente de açaí contribuirá tanto para a economia local, quanto para o desenvolvimento sustentável.Submitted by Jonathan Sousa de Almeida (jonathan.sousa@ufma.br) on 2024-07-08T12:39:42Z No. of bitstreams: 1 KATIAREGINAASSUNÇÃOBORGES.pdf: 5954140 bytes, checksum: 6cf29f9cc1599a2865eb312240bb8adc (MD5)Made available in DSpace on 2024-07-08T12:39:42Z (GMT). No. of bitstreams: 1 KATIAREGINAASSUNÇÃOBORGES.pdf: 5954140 bytes, checksum: 6cf29f9cc1599a2865eb312240bb8adc (MD5) Previous issue date: 2024-04-26application/pdfporUniversidade Federal do MaranhãoPROGRAMA DE PÓS-GRADUAÇÃO EM BIOTECNOLOGIA - RENORBIO/CCBSUFMABrasilDEPARTAMENTO DE PATOLOGIA/CCBSóleo de açaí;Euterpe oleracea Mart;câncer cervical;nanoemulsão.açaí oil;Euterpe oleracea Mart;cervical cancer;nanoemulsion.MedicamentosCiências da SaúdeAvaliação do potencial biotecnológico do óleo da semente de Euterpe oleracea Mart. In vitro e In vivo e da sua nanoemulsãoAssessment of the biotechnological potential of Euterpe oleracea Mart seed oil. 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dc.title.por.fl_str_mv |
Avaliação do potencial biotecnológico do óleo da semente de Euterpe oleracea Mart. In vitro e In vivo e da sua nanoemulsão |
dc.title.alternative.eng.fl_str_mv |
Assessment of the biotechnological potential of Euterpe oleracea Mart seed oil. In vitro and in vivo and its nanoemulsion |
title |
Avaliação do potencial biotecnológico do óleo da semente de Euterpe oleracea Mart. In vitro e In vivo e da sua nanoemulsão |
spellingShingle |
Avaliação do potencial biotecnológico do óleo da semente de Euterpe oleracea Mart. In vitro e In vivo e da sua nanoemulsão BORGES, Katia Regina Assunção óleo de açaí; Euterpe oleracea Mart; câncer cervical; nanoemulsão. açaí oil; Euterpe oleracea Mart; cervical cancer; nanoemulsion. Medicamentos Ciências da Saúde |
title_short |
Avaliação do potencial biotecnológico do óleo da semente de Euterpe oleracea Mart. In vitro e In vivo e da sua nanoemulsão |
title_full |
Avaliação do potencial biotecnológico do óleo da semente de Euterpe oleracea Mart. In vitro e In vivo e da sua nanoemulsão |
title_fullStr |
Avaliação do potencial biotecnológico do óleo da semente de Euterpe oleracea Mart. In vitro e In vivo e da sua nanoemulsão |
title_full_unstemmed |
Avaliação do potencial biotecnológico do óleo da semente de Euterpe oleracea Mart. In vitro e In vivo e da sua nanoemulsão |
title_sort |
Avaliação do potencial biotecnológico do óleo da semente de Euterpe oleracea Mart. In vitro e In vivo e da sua nanoemulsão |
author |
BORGES, Katia Regina Assunção |
author_facet |
BORGES, Katia Regina Assunção |
author_role |
author |
dc.contributor.advisor1.fl_str_mv |
NASCIMENTO, Maria do Desterro Soares Brandão |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/3958174822396319 |
dc.contributor.referee1.fl_str_mv |
NASCIMENTO, Maria do Desterro Soares Brandão |
dc.contributor.referee1Lattes.fl_str_mv |
http://lattes.cnpq.br/3958174822396319 |
dc.contributor.referee2.fl_str_mv |
SILVA, Ana Lúcia Abreu |
dc.contributor.referee2Lattes.fl_str_mv |
http://lattes.cnpq.br/8288733951324759 |
dc.contributor.referee3.fl_str_mv |
FAUSTINO-ROCHA, Ana Isabel |
dc.contributor.referee4.fl_str_mv |
TELES, Amanda Mara |
dc.contributor.referee4Lattes.fl_str_mv |
http://lattes.cnpq.br/3933255152524601 |
dc.contributor.referee5.fl_str_mv |
OLIVEIRA, Rui Miguel Gil da Costa |
dc.contributor.referee5Lattes.fl_str_mv |
http://lattes.cnpq.br/6785759461393904 |
dc.contributor.authorLattes.fl_str_mv |
http://lattes.cnpq.br/0528263981302728 |
dc.contributor.author.fl_str_mv |
BORGES, Katia Regina Assunção |
contributor_str_mv |
NASCIMENTO, Maria do Desterro Soares Brandão NASCIMENTO, Maria do Desterro Soares Brandão SILVA, Ana Lúcia Abreu FAUSTINO-ROCHA, Ana Isabel TELES, Amanda Mara OLIVEIRA, Rui Miguel Gil da Costa |
dc.subject.por.fl_str_mv |
óleo de açaí; Euterpe oleracea Mart; câncer cervical; nanoemulsão. |
topic |
óleo de açaí; Euterpe oleracea Mart; câncer cervical; nanoemulsão. açaí oil; Euterpe oleracea Mart; cervical cancer; nanoemulsion. Medicamentos Ciências da Saúde |
dc.subject.eng.fl_str_mv |
açaí oil; Euterpe oleracea Mart; cervical cancer; nanoemulsion. |
dc.subject.cnpq.fl_str_mv |
Medicamentos Ciências da Saúde |
description |
Açaí seed oil, Euterpe oleracea Mart., is rich in unsaturated, saturated, monounsaturated and polyunsaturated fatty acids with anti-inflammatory, antioxidant, antimicrobial, antinociceptive, anticancer, antiatherogenic and healing activities. The açaí seed is a residue produced after pulping, being a source of bioactive compounds; therefore, it is necessary to study its pharmacological properties and contribute to sustainable development. The objective of this was to evaluate the biotechnological potential of Euterpe oleracea Mart seed oil. - in vitro and in vivo - and its Nanoemulsion. The fruits used were collected in the Maracanã Ecological Park (Parque da Juçara), in São Luís, Maranhão. Samples of the plant were collected between the months of August and November, and exsicata were prepared, duly stored in the Rosa Mochel Herbarium. Subsequently, the oil was extracted with n-hexane solvent in a Soxhlet extractor and analyzed as methyl esters, weighing 100 mg of the oil for analysis of the chromatographic profile by gas chromatography combined with mass spectrometry (GC-MS) to identify the fatty acids. and its physical-chemical characterization. After understanding the physical chemistry of the oil, the açaí seed oil nanoemulsion (NE- OEO) was developed based on the calculation of the Hydrophilic Lipophilic Balance (EHL), followed by stability analysis, chemical characterization and morphology. To determine the average particle size and polydispersity index, the formulations were evaluated using the dynamic light scattering technique (Nano ZS, Malvern Instruments Ltd., UK) and Zeta Potential on the Nano ZS equipment (Malvern Instruments Ltd., UK). Both the oil and NE-OEO showed physicochemical patterns within normal limits. In other words, the oil showed acidity (0.3556± 0.0003), humidity (1.25±1.64), saponification index (189 ± 1.04), refractive index (1.4707±0.00005 ), incineration residue (ash) (0.42±0.42) and density (0.928±0.00005). And NE-OEO showed normal patterns regarding droplet size (238.37 ± 3.96), homogeneity (PDI) (0.38 ± 0.38), Zeta potential (-9.59 ± 0.11), pH (7.0 ± 0.00.) and turbidity (0.267± 0.00). Regarding phenolic compounds, NE-OEO presented the highest concentration, 146.00 ± 0.259 mg, while the oil had 127.40 ± 0.449 mg EAG g-1 of phenolic compounds, and total flavonoids values were found to be 62.62 ± 0.930 mg for the oil and 113.80 ± 0.454 mg EQ g-1 for NE-OEO. The antioxidant activity was proven based on the EC50 evaluated in the DPPH and ABTS assay, with the oil being 375.698 μg/mL in DPPH, and nanoemulsions with a significantly lower EC50, equivalent to 229.845 μg/mL. The Trolox control presented an even lower EC50. In the ABTS test, the EC50 of açaí oil was 272.0208 μg/mL, while açaí oil nanoemulsions had an even lower EC50 value, measuring 201.2895 μg/mL. Trolox also showed a significantly lower EC50 compared to oil and NE-OEO, these data confirm the antioxidant capacity of the oil and its NE- OEO. In vitro analyzes were performed on the non-tumor cell lines mouse RAW 264.7 macrophages, and human HaCat keratinocytes and cervical cancer tumor lines SiHa (HPV-16) and HeLa (HPV-18). Non-tumor cell lines were treated with concentrations (7.8 to 1000 ug/mL) of the oil and NE-OEO for 24, 48 and 72 hours, and tumor cells were treated with concentrations (0.631 to 100 ug/mL) of the oil, made analyzes of cell viability, cell death, morphology, healing and clonogenicity. After in vitro analyses, the effect of açaí seed oil and NE-OEO was analyzed in vivo in female Swiss mice. The animals were treated with different concentrations of the oil (100, 200 and 300 mg/kg) and NE-OEO (100 mg/kg) applied 0.5 mL intraperitoneally every two weeks. Açaí seed oil is rich in polyphenols, flavonoids, saturated fatty acids (49.27%) and unsaturated fatty acids (50.73%), among which unsaturated fatty acids, 29.73% are monounsaturated, 20 .85% are polyunsaturated. Lauric, myristic (C14:0), palmitic, linoleic, and oleic fatty acids were the most identified. Viability assay with MTT ((3-(4,5- Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide)) demonstrated a cytotoxic effect of the oil on the SiHa (HPV-16) and HeLa (HPV- 18) and non-cytotoxic to the non-tumor cell lines HaCat, as well as the oil and NE-OEO were not cytotoxic to the non-tumor cell line RAW 264.7. Açaí seed oil induced morphological changes in HeLa and SiHa cells, inducing a reduction in cell viability, inhibiting cell migration and colony formation in cervical cancer cell lines. The annexin V-FITC assay showed cell death by early and late apoptosis at 24 and 48 hours after treatment with açaí seed oil. In in vivo experiments, the use of oil at a dose of 100, 200, 300 mg/kg and NE-OEO at a dose of 100 mg/kg were not toxic to the animals, in accordance with the normal patterns seen in the liver and kidney; Furthermore, it did not interfere with the behavior or weight of the animals, however, it induced an immunomodulatory response with a significant increase in IL10 and IL17. Our results suggest the biotechnological potential of the oil and NE-OEO in antitumor activity for cervical cancer, and in the development of colloidal systems for drug delivery. Furthermore, the use of açaí seed residue will contribute to both the local economy and sustainable development.. |
publishDate |
2024 |
dc.date.accessioned.fl_str_mv |
2024-07-08T12:39:42Z |
dc.date.issued.fl_str_mv |
2024-04-26 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/doctoralThesis |
format |
doctoralThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
BORGES, Katia Regina Assunção. Avaliação do potencial biotecnológico do óleo da semente de Euterpe oleracea Mart. In vitro e In vivo e da sua nanoemulsão. 2024. 112 f. Tese (Programa de Pós-Graduação em Biotecnologia - RENORBIO/CCBS) - Universidade Federal do Maranhão, São Luís, 2024. |
dc.identifier.uri.fl_str_mv |
https://tedebc.ufma.br/jspui/handle/tede/tede/5384 |
identifier_str_mv |
BORGES, Katia Regina Assunção. Avaliação do potencial biotecnológico do óleo da semente de Euterpe oleracea Mart. In vitro e In vivo e da sua nanoemulsão. 2024. 112 f. Tese (Programa de Pós-Graduação em Biotecnologia - RENORBIO/CCBS) - Universidade Federal do Maranhão, São Luís, 2024. |
url |
https://tedebc.ufma.br/jspui/handle/tede/tede/5384 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
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application/pdf |
dc.publisher.none.fl_str_mv |
Universidade Federal do Maranhão |
dc.publisher.program.fl_str_mv |
PROGRAMA DE PÓS-GRADUAÇÃO EM BIOTECNOLOGIA - RENORBIO/CCBS |
dc.publisher.initials.fl_str_mv |
UFMA |
dc.publisher.country.fl_str_mv |
Brasil |
dc.publisher.department.fl_str_mv |
DEPARTAMENTO DE PATOLOGIA/CCBS |
publisher.none.fl_str_mv |
Universidade Federal do Maranhão |
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Biblioteca Digital de Teses e Dissertações da UFMA |
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Biblioteca Digital de Teses e Dissertações da UFMA |
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