Effects of Two Types of Melatonin-Loaded Nanocapsules with Distinct Supramolecular Structures: Polymeric (NC) and Lipid-Core Nanocapsules (LNC) on Bovine Embryo Culture Model.
Melatonin has been used as a supplement in culture medium to improve the efficiency of in vitro produced mammalian embryos. Through its ability to scavenge toxic oxygen derivatives and regulate cellular mRNA levels for antioxidant enzymes, this molecule has been shown to play a protective role against damage by free radicals, to which in vitro cultured embryos are exposed during early development. In vivo and in vitro studies have been performed showing that the use of nanocapsules as active substances carriers increases stability, bioavailability and biodistribution of drugs, such as melatonin, to the cells and tissues, improving their antioxidant properties. These properties can be modulated through the manipulation of formula composition, especially in relation to the supramolecular structures of the nanocapsule core and the surface area that greatly influences drug release mechanisms in biological environments. This study aimed to evaluate the effects of two types of melatonin-loaded nanocapsules with distinct supramolecular structures, polymeric (NC) and lipid-core (LNC) nanocapsules, on in vitro cultured bovine embryos. Embryonic development, apoptosis, reactive oxygen species (ROS) production, and mRNA levels of genes involved in cell apoptosis, ROS and cell pluripotency were evaluated after supplementation of culture medium with non-encapsulated melatonin (Mel), melatonin-loaded polymeric nanocapsules (Mel-NC) and melatonin-loaded lipid-core nanocapsules (Mel-LNC) at 10-6, 10-9, and 10-12 M drug concentrations. The highest hatching rate was observed in embryos treated with 10-9 M Mel-LNC. When compared to Mel and Mel-NC treatments at the same concentration (10-9 M), Mel-LNC increased embryo cell number, decreased cell apoptosis and ROS levels, down-regulated mRNA levels of BAX, CASP3, and SHC1 genes, and up-regulated mRNA levels of CAT and SOD2 genes. These findings indicate that nanoencapsulation with LNC increases the protective effects of melatonin against oxidative stress and cell apoptosis during in vitro embryo culture in bovine species.
Programa de Pós-Graduação em Biotecnologia (PPGB), Grupo de Pesquisa em Oncologia Celular e Molecular, Biotecnologia/Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, 96010-900, RS, Brazil.
Eliza Rossi
ER
Komninou
Programa de Pós-Graduação em Biotecnologia (PPGB), Grupo de Pesquisa em Oncologia Celular e Molecular, Biotecnologia/Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, 96010-900, RS, Brazil.
Mariana Härter
MH
Remião
Programa de Pós-Graduação em Biotecnologia (PPGB), Grupo de Pesquisa em Oncologia Celular e Molecular, Biotecnologia/Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, 96010-900, RS, Brazil.
Caroline Gomes
CG
Lucas
Programa de Pós-Graduação em Biotecnologia (PPGB), Grupo de Pesquisa em Oncologia Celular e Molecular, Biotecnologia/Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, 96010-900, RS, Brazil.
William Borges
WB
Domingues
In Vitro Brasil S/A, Mogi Mirim, 13800-970, SP, Brazil.
Andrea Cristina
AC
Basso
Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Av. Ipiranga, 2752, Porto Alegre, 90610-000, RS, Brazil.
Denise Soledade
DS
Jornada
Programa de Pós-Graduação em Biotecnologia (PPGB), Grupo de Pesquisa em Oncologia Celular e Molecular, Biotecnologia/Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, 96010-900, RS, Brazil.
João Carlos
JC
Deschamps
Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Av. Ipiranga, 2752, Porto Alegre, 90610-000, RS, Brazil.
Ruy Carlos Ruver
RC
Beck
Departamento de Química Orgânica, Instituto de Química, Universidade Federal do Rio Grande do Sul, Porto Alegre, 91501-970, RS, Brazil.
Adriana Raffin
AR
Pohlmann
Department of Animal Science, McGill University, Sainte Anne de Bellevue, H9X 3V9, QC, Canada.
Vilceu
V
Bordignon
Programa de Pós-Graduação em Biotecnologia (PPGB), Grupo de Pesquisa em Oncologia Celular e Molecular, Biotecnologia/Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, 96010-900, RS, Brazil.
Fabiana Kömmling
FK
Seixas
Programa de Pós-Graduação em Biotecnologia (PPGB), Grupo de Pesquisa em Oncologia Celular e Molecular, Biotecnologia/Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, 96010-900, RS, Brazil.
Vinicius Farias
VF
Campos
Faculdade de Farmácia, Universidade Federal do Rio Grande do Sul, Av. Ipiranga, 2752, Porto Alegre, 90610-000, RS, Brazil.
Silvia Stanisçuaski
SS
Guterres
Programa de Pós-Graduação em Biotecnologia (PPGB), Grupo de Pesquisa em Oncologia Celular e Molecular, Biotecnologia/Centro de Desenvolvimento Tecnológico, Universidade Federal de Pelotas, Pelotas, 96010-900, RS, Brazil.
Tiago
T
Collares
D000818:Animals; D000975:Antioxidants / Q000737:chemistry / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D002374:Catalase / Q000235:genetics / Q000378:metabolism; D002417:Cattle; D003470:Culture Media / Q000737:chemistry; D004337:Drug Carriers / Q000737:chemistry / Q000494:pharmacology*; D004339:Drug Compounding; D004622:Embryo, Mammalian / Q000187:drug effects* / Q000502:physiology; D047108:Embryonic Development / Q000187:drug effects; D005260:Female; D005307:Fertilization in Vitro; D018507:Gene Expression Regulation, Developmental; D008297:Male; D008550:Melatonin / Q000737:chemistry / Q000494:pharmacology*; D053769:Nanocapsules / Q000737:chemistry; D011091:Polyesters / Q000737:chemistry*; D011109:Polymethacrylic Acids / Q000737:chemistry*; D011247:Pregnancy; D017382:Reactive Oxygen Species / Q000037:antagonists & inhibitors / Q000378:metabolism; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1 / Q000235:genetics / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000235:genetics / Q000378:metabolism; D000099156:Superoxide Dismutase 2
D000975:Antioxidants; D053148:Caspase 3; D002374:Catalase; D003470:Culture Media; D004337:Drug Carriers; D008550:Melatonin; D053769:Nanocapsules; D011091:Polyesters; D011109:Polymethacrylic Acids; D017382:Reactive Oxygen Species; D000071425:Src Homology 2 Domain-Containing, Transforming Protein 1; D013482:Superoxide Dismutase; D051028:bcl-2-Associated X Protein; D000099156:Superoxide Dismutase 2; C016240:polycaprolactone; C038300:methylmethacrylate-methacrylic acid copolymer