Abstract This study investigated the effect of Folliculinum 6 cH on the oocyte meiosis resumption and viability rates, progesterone production and mitochondrial activity after in vitro maturation of cumulus-oocyte complexes (COCs) in sheep. Sheep ovaries were collected at a local slaughterhouse and COCs were recovered by slicing technique. The selected COCs were maturated in TCM199 (Control treatment), or control medium supplemented with 0.05% ethanol (v/v) (the vehicle of the homeopathic preparation – Ethanol treatment) or with Folliculinum 6 cH. After 24 h of in vitro maturation (IVM), oocytes were mechanically denuded and incubated with Hoechst 33342 and MitoTracker (0.5 μM) Orange CMTMRos for analysis of viability and chromatin configuration, and mitochondrial activity, respectively. The results showed that Folliculinum 6 cH addition increased oocyte degeneration and reduced meiotic resumption compared to the control (P < 0.05). Interestingly, the percentages meiotic resumption and oocyte maturation were lower in the Folliculinum 6 cH treatment compared to its vehicle (Ethanol treatment) (P < 0.05). On the other hand, when the treatments were compared, higher mitochondrial activity was observed in the Ethanol treatment (P < 0.05). In conclusion, contrary to its vehicle, the addition of Folliculinum 6 cH to the IVM medium promoted oocyte degeneration and affected negatively the mitochondrial distribution, impairing meiosis resumption.

Resumo Este trabalho teve por objetivo avaliar o efeito dos medicamentos homeopáticos (Pulsatilla nigricans e hormônio folículo estimulante homeopático - FSH) e um complexo homeopático (Bos Stress Fertilis) na foliculogênese inicial, utilizando o cultivo in vitro de folículos pré-antrais suínos como modelo in vitro. Para tanto, fragmentos ovarianos foram cultivados por um ou sete dias em α-MEM+ na ausência (controle cultivado) ou presença de FSH homeopático (6 cH), Pulsatilla (6 CH), Bos Stress Fertilis (6 CH), álcool cereal (50% - v/v) ou FSH recombinante (50 ng/ml) adicionados diariamente. Os fragmentos ovarianos não cultivados (controle fresco) ou cultivados por um e sete dias foram processados para histologia clássica. Somente o composto homeopático Bos Stress Fertilis foi eficiente em manter o percentual de sobrevivência folicular após sete dias de cultivo semelhante ao controle não cultivado e α-MEM+. Em relação ao crescimento folicular, somente a adição de FSH homeopático aumentou o diâmetro folicular quando comparado ao controle não cultivado e α-MEM+ após um dia de cultivo. Dessa forma, pode-se concluir que a adição dos medicamentos homeopáticos Bos Stress fertilis e FSH homeopático (6 CH) melhoraram, respectivamente, a sobrevivência e o crescimento in vitro de folículos pré-antrais suínos inclusos em fragmentos de tecido ovariano.

Abstract This study investigated the effect of homeopathic medicine (Pulsatilla nigricans and homeopathic follicle stimulating hormone - FSH) and the complex (Bos Stress Fertilis) on the initial folliculogenesis, using the in vitro culture of preantral follicles as in vitro model. For this, swine ovarian fragments were cultured for one or seven days in α-MEM+ in the absence (cultured control) or presence of homeopathic FSH (6 cH), Pulsatilla (6 CH), Bos Stress Fertilis (6 CH), grain alcohol (50% v / v) or recombinant FSH (50 ng / ml) added daily. Uncultured ovarian fragments (fresh control) or cultured for one and seven days were processed for classical histology. Only the homeopathic complex Bos Stress Fertilis maintained the percentage of follicular survival after seven days of culture in relation to the uncultured control and α-MEM+. Regarding follicular growth, only the addition of homeopathic FSH increased the follicular diameter when compared to the uncultured control and a-MEM+. Thus, it can be concluded that the addition of the homeopathic remedies Bos Stress fertilis and homeopathic FSH (6 CH) improved, respectively, survival and in vitro growth of swine preantral follicles included in ovarian tissue.