Αlpha-lipoic acid and l-arginine on in vitro production of ovine embryos

Detalhes bibliográficos
Autor(a) principal: Freitas, Beatriz Cavalcanti de
Data de Publicação: 2023
Outros Autores: Silva, Jossimara de Melo, Santos, Damaris Raquel Pires dos, Aureliano, Érika Karoline de Oliveira, Lima, Andreza Mayara Carneiro, Silva, Paulo Ricardo Vieira da, Oliveira, Luana Kealy Pimentel de, Cordeiro, Mabel Freitas, Lopes Júnior, Edilson Soares
Tipo de documento: preprint
Idioma: por
Título da fonte: SciELO Preprints
Texto Completo: https://preprints.scielo.org/index.php/scielo/preprint/view/6001
Resumo: The objective of this study was to evaluate the effect of α-lipoic acid on in vitro maturation and of l-arginine on in vitro fertilization of oocytes for in vitro production of sheep embryos. Thus, the ovaries were collected at a local slaughterhouse and the oocytes collected by the method of follicular aspiration with a vacuum pump were divided equally into five maturation groups: CON, without addition of antioxidant; CIS group, with cysteamine as an antioxidant source; and the groups ALA5, ALA10 and ALA20, containing concentrations of 5.0, 10.0 and 20.0 µM of ALA. After 24h of maturation, the oocytes were evaluated concerning occurrence and degrees of expansion of the cumulus cells. The oocytes of the best IVM group were destinated for in vitro fertilization (IVF). The selected and capacitated sperm were included in five groups: CON, without addition of capacitator agent; HEP group, with heparin as a capacitator source; and ARG5, ARG10 and ARG20 groups, where IVF occurred in the same HEP medium, replacing heparin with 5 mM, 10 mM and 20 mM L-arginine, respectively. The results were expressed as a percentage and the variables of cumulus cell expansion and number of cleaved structures were compared using the chi-square test in Epi Info software (Epi Info 7.2.5, Atlanta, GA, USA, 2021). Results were considered significant when P < 0.05. Regarding the expansion of cumulus cells all groups showed 100% expansion, on the total expansion category of COCs it was observed that ALA5 and ALA20 groups did not present the same proportion of COCs with total expansion as CIS group. After in vitro fertilization (IVF), there were no statistical differences in relation to the cleaved structures between CON, HEP, ARG5 and ARG10 groups. However, it was also observed that the use of 20.0 mM of L-arginine reduced the percentage of cleaved structures when compared to the ARG10 group (P<0.05).
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spelling Αlpha-lipoic acid and l-arginine on in vitro production of ovine embryosÁcido α-lipoico y l-arginina en la producción in vitro de embriones ovinosÁcido α-lipóico e l-arginina na produção in vitro de embriões ovinosantioxidantePIVoócitoovelhazigotoantioxidantIVPoocytesheepzygoteantioxidantecigotoPIVovocitoovejaThe objective of this study was to evaluate the effect of α-lipoic acid on in vitro maturation and of l-arginine on in vitro fertilization of oocytes for in vitro production of sheep embryos. Thus, the ovaries were collected at a local slaughterhouse and the oocytes collected by the method of follicular aspiration with a vacuum pump were divided equally into five maturation groups: CON, without addition of antioxidant; CIS group, with cysteamine as an antioxidant source; and the groups ALA5, ALA10 and ALA20, containing concentrations of 5.0, 10.0 and 20.0 µM of ALA. After 24h of maturation, the oocytes were evaluated concerning occurrence and degrees of expansion of the cumulus cells. The oocytes of the best IVM group were destinated for in vitro fertilization (IVF). The selected and capacitated sperm were included in five groups: CON, without addition of capacitator agent; HEP group, with heparin as a capacitator source; and ARG5, ARG10 and ARG20 groups, where IVF occurred in the same HEP medium, replacing heparin with 5 mM, 10 mM and 20 mM L-arginine, respectively. The results were expressed as a percentage and the variables of cumulus cell expansion and number of cleaved structures were compared using the chi-square test in Epi Info software (Epi Info 7.2.5, Atlanta, GA, USA, 2021). Results were considered significant when P < 0.05. Regarding the expansion of cumulus cells all groups showed 100% expansion, on the total expansion category of COCs it was observed that ALA5 and ALA20 groups did not present the same proportion of COCs with total expansion as CIS group. After in vitro fertilization (IVF), there were no statistical differences in relation to the cleaved structures between CON, HEP, ARG5 and ARG10 groups. However, it was also observed that the use of 20.0 mM of L-arginine reduced the percentage of cleaved structures when compared to the ARG10 group (P<0.05).El objetivo de este trabajo fue evaluar el efecto del ácido α-lipoico en la maduración in vitro y la l-arginina en la fecundación in vitro de ovocitos para la producción in vitro de embriones ovinos. Así, se recolectaron ovocitos de ovarios obtenidos de mataderos mediante el método de aspiración folicular con bomba de vacío, los cuales se dividieron en cinco grupos de maduración: CON, sin adición de antioxidante, grupo CIS, con cisteamina como fuente antioxidante y los grupos ALA5, ALA10 y ALA20, que contienen concentraciones de 5, 10 y 20 µM de ácido α-lipoico. Después de 24 h de maduración, se evaluó la aparición y el grado de expansión de las células del cúmulo en los ovocitos. Los ovocitos del mejor grupo MIV fueron destinados a fecundación in vitro (FIV), siendo por tanto incluidos en gotas con medio de fecundación in vitro, junto con los espermatozoides seleccionados y capacitados, resultando en los siguientes grupos: CON, sin adición de agente capacitador; grupo HEP, con heparina como fuente de condensadores; y los grupos ARG5, ARG10 y ARG20, donde se produjo la FIV en el mismo medio HEP, reemplazando la heparina por L-arginina 5, 10 y 20 mM, respectivamente. Los resultados se expresaron en porcentajes y las variables expansión celular del cúmulo y número de estructuras escindidas se compararon mediante la prueba chicuadrado del software Epi Info (Epi Info 7.2.5, Atlanta, GA, USA, 2021). Los resultados se consideraron significativos cuando P < 0,05. En cuanto a la expansión de las células del cúmulo, todos los grupos mostraron 100% de expansión. Evaluando los grados de expansión de las células del cúmulo, en la categoría de expansión total, se observó que los grupos ALA5 y ALA20 presentaron la misma proporción de CCO con expansión total que el grupo CIS (P<0.05), sin embargo se observa que hay una tendencia a la baja de las estructuras con plena expansión cuando se utilizan antioxidantes. En cuanto a las estructuras escindidas, fue posible identificar que no hubo diferencias estadísticas en relación a las estructuras escindidas, entre los grupos CON, HEP, ARG5 y ARG10. Sin embargo, también se observó que el uso de Larginina 20 mM redujo el porcentaje de estructuras escindidas en comparación con el grupo ARG10 (P<0.05).O objetivo deste trabalho foi avaliar o efeito do ácido α-lipóico na maturação in vitro e da l-arginina na fertilização in vitro de oócitos para produção in vitro de embriões ovinos. Desta forma, foram realizadas colheitas de oócitos oriundos de ovários obtidos em abatedouro através do método de aspiração folicular com bomba de vácuo, que foram divididos em cinco grupos de maturação: CON, sem adição de antioxidante, grupo CIS, com cisteamina como fonte antioxidante e os grupos ALA5, ALA10 e ALA20, contendo as concentrações de 5, 10, e 20 µM de ácido α-lipóico. Após 24h de maturação, os oócitos foram avaliados quanto à ocorrência e grau de expansão das células do cumulus. Os oócitos do melhor grupo MIV foram destinados à fertilização in vitro (FIV), sendo, portanto, incluídos em gotas com meio de fertilização in vitro, juntamente com os espermatozoides selecionados e capacitados, resultando nos seguintes grupos: CON, sem adição de capacitador agente; grupo HEP, com heparina como fonte de capacitor; e grupos ARG5, ARG10 e ARG20, onde a FIV ocorreu no mesmo meio HEP, substituindo a heparina por L-arginina 5, 10 e 20 mM, respectivamente. Os resultados foram expressos em porcentagem e as variáveis de expansão das células do cumulus e número de estruturas clivadas foram comparadas por meio do teste qui-quadrado do software Epi Info (Epi Info 7.2.5, Atlanta, GA, EUA, 2021). Os resultados foram considerados significativos quando P < 0,05. Em relação à expansão das células do cumulus, todos os grupos apresentaram 100% de expansão. Avaliando os graus de expansão das células do cumulus, na categoria de expansão total observou-se que os grupos ALA5 e ALA20 apresentaram a mesma proporção de CCO com expansão total que o grupo CIS (P<0,05), porém se observa que há uma tendência de queda de estruturas com expansão total quando os antioxidantes foram utilizados. Em relação às estruturas clivadas, foi possível identificar que não houve diferenças estatísticas em relação às estruturas clivadas, entre os grupos CON, HEP, ARG5 e ARG10. Contudo, também foi observado que o uso de 20 mM de L-arginina reduziu o percentual de estruturas clivadas quando comparado ao grupo ARG10 (P<0,05).SciELO PreprintsSciELO PreprintsSciELO Preprints2023-05-05info:eu-repo/semantics/preprintinfo:eu-repo/semantics/publishedVersionapplication/pdfhttps://preprints.scielo.org/index.php/scielo/preprint/view/600110.1590/SciELOPreprints.6001porhttps://preprints.scielo.org/index.php/scielo/article/view/6001/11523Copyright (c) 2023 Beatriz Cavalcanti de Freitas, Jossimara de Melo Silva, Damaris Raquel Pires dos Santos, Érika Karoline de Oliveira Aureliano, Andreza Mayara Carneiro Lima, Paulo Ricardo Vieira da Silva, Luana Kealy Pimentel de Oliveira, Mabel Freitas Cordeiro, Edilson Soares Lopes Júniorhttps://creativecommons.org/licenses/by/4.0info:eu-repo/semantics/openAccessFreitas, Beatriz Cavalcanti deSilva, Jossimara de MeloSantos, Damaris Raquel Pires dosAureliano, Érika Karoline de OliveiraLima, Andreza Mayara CarneiroSilva, Paulo Ricardo Vieira daOliveira, Luana Kealy Pimentel deCordeiro, Mabel FreitasLopes Júnior, Edilson Soaresreponame:SciELO Preprintsinstname:Scientific Electronic Library Online (SCIELO)instacron:SCI2023-04-29T22:15:58Zoai:ops.preprints.scielo.org:preprint/6001Servidor de preprintshttps://preprints.scielo.org/index.php/scieloONGhttps://preprints.scielo.org/index.php/scielo/oaiscielo.submission@scielo.orgopendoar:2023-04-29T22:15:58SciELO Preprints - Scientific Electronic Library Online (SCIELO)false
dc.title.none.fl_str_mv Αlpha-lipoic acid and l-arginine on in vitro production of ovine embryos
Ácido α-lipoico y l-arginina en la producción in vitro de embriones ovinos
Ácido α-lipóico e l-arginina na produção in vitro de embriões ovinos
title Αlpha-lipoic acid and l-arginine on in vitro production of ovine embryos
spellingShingle Αlpha-lipoic acid and l-arginine on in vitro production of ovine embryos
Freitas, Beatriz Cavalcanti de
antioxidante
PIV
oócito
ovelha
zigoto
antioxidant
IVP
oocyte
sheep
zygote
antioxidante
cigoto
PIV
ovocito
oveja
title_short Αlpha-lipoic acid and l-arginine on in vitro production of ovine embryos
title_full Αlpha-lipoic acid and l-arginine on in vitro production of ovine embryos
title_fullStr Αlpha-lipoic acid and l-arginine on in vitro production of ovine embryos
title_full_unstemmed Αlpha-lipoic acid and l-arginine on in vitro production of ovine embryos
title_sort Αlpha-lipoic acid and l-arginine on in vitro production of ovine embryos
author Freitas, Beatriz Cavalcanti de
author_facet Freitas, Beatriz Cavalcanti de
Silva, Jossimara de Melo
Santos, Damaris Raquel Pires dos
Aureliano, Érika Karoline de Oliveira
Lima, Andreza Mayara Carneiro
Silva, Paulo Ricardo Vieira da
Oliveira, Luana Kealy Pimentel de
Cordeiro, Mabel Freitas
Lopes Júnior, Edilson Soares
author_role author
author2 Silva, Jossimara de Melo
Santos, Damaris Raquel Pires dos
Aureliano, Érika Karoline de Oliveira
Lima, Andreza Mayara Carneiro
Silva, Paulo Ricardo Vieira da
Oliveira, Luana Kealy Pimentel de
Cordeiro, Mabel Freitas
Lopes Júnior, Edilson Soares
author2_role author
author
author
author
author
author
author
author
dc.contributor.author.fl_str_mv Freitas, Beatriz Cavalcanti de
Silva, Jossimara de Melo
Santos, Damaris Raquel Pires dos
Aureliano, Érika Karoline de Oliveira
Lima, Andreza Mayara Carneiro
Silva, Paulo Ricardo Vieira da
Oliveira, Luana Kealy Pimentel de
Cordeiro, Mabel Freitas
Lopes Júnior, Edilson Soares
dc.subject.por.fl_str_mv antioxidante
PIV
oócito
ovelha
zigoto
antioxidant
IVP
oocyte
sheep
zygote
antioxidante
cigoto
PIV
ovocito
oveja
topic antioxidante
PIV
oócito
ovelha
zigoto
antioxidant
IVP
oocyte
sheep
zygote
antioxidante
cigoto
PIV
ovocito
oveja
description The objective of this study was to evaluate the effect of α-lipoic acid on in vitro maturation and of l-arginine on in vitro fertilization of oocytes for in vitro production of sheep embryos. Thus, the ovaries were collected at a local slaughterhouse and the oocytes collected by the method of follicular aspiration with a vacuum pump were divided equally into five maturation groups: CON, without addition of antioxidant; CIS group, with cysteamine as an antioxidant source; and the groups ALA5, ALA10 and ALA20, containing concentrations of 5.0, 10.0 and 20.0 µM of ALA. After 24h of maturation, the oocytes were evaluated concerning occurrence and degrees of expansion of the cumulus cells. The oocytes of the best IVM group were destinated for in vitro fertilization (IVF). The selected and capacitated sperm were included in five groups: CON, without addition of capacitator agent; HEP group, with heparin as a capacitator source; and ARG5, ARG10 and ARG20 groups, where IVF occurred in the same HEP medium, replacing heparin with 5 mM, 10 mM and 20 mM L-arginine, respectively. The results were expressed as a percentage and the variables of cumulus cell expansion and number of cleaved structures were compared using the chi-square test in Epi Info software (Epi Info 7.2.5, Atlanta, GA, USA, 2021). Results were considered significant when P < 0.05. Regarding the expansion of cumulus cells all groups showed 100% expansion, on the total expansion category of COCs it was observed that ALA5 and ALA20 groups did not present the same proportion of COCs with total expansion as CIS group. After in vitro fertilization (IVF), there were no statistical differences in relation to the cleaved structures between CON, HEP, ARG5 and ARG10 groups. However, it was also observed that the use of 20.0 mM of L-arginine reduced the percentage of cleaved structures when compared to the ARG10 group (P<0.05).
publishDate 2023
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10.1590/SciELOPreprints.6001
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