Cat epididymal semen cryopreserved with and without vitamin E: effect on sperm parameters and lipid peroxidation.

The aims of this study were to investigate: 1) if the addition of α-tocopherol (vitamin E) in three concentrations (0.3, 0.6 and 0.9 mM) is able to preserve spermatozoa integrity after thawing and 2) the effect of α-tocopherol supplementation on lipid peroxidation. Fifty four domestic cats were used in this study constituting 18 pools (3 cats per pool). Each pool was submitted at four experimental groups: group 0 (control) - epididymal sperm were frozen with a commercial Botucrio® extender; group 0.3, group 0.6 and group 0.9 - the extender was supplemented with 0.3, 0.6 and 0.9 mM of α-tocopherol, respectively. Each semen sample was evaluated for motility, progressive forward motility, morphology, sperm viability (plasma membrane integrity-PMI), hypo-osmotic swelling test (HOST), before and after thawing. The evaluation of lipid peroxidation reaction by Thiobarbituric Acid Reactive Substances (TBARS) test was performed on thawed semen only. Results demonstrated that there was no significant difference between control and the three α-tocopherol groups with regards to motility and progressive motility after thawing (P > 0.05). As expected, in fresh samples viability was significantly higher than in all the cryopreserved groups in which there was no positive influence of any of the α-tocopherol concentration used. Lipid peroxidation was higher in the supplemented groups 0.6 and 0.9 mM of α-tocopherol than in control and in 0.3 mM group. In conclusion, the addition of α-tocopherol to the commercial extender had no positive influence on reduction of lipid peroxidation. This topic deserves further investigations to better understand the effect of cryopreservation procedures on epididymal spermatozoa and to establish adequate strategies to counteract sperm cryodamages.

Cat epididymal semen cryopreserved with and without vitamin E: effect on sperm parameters and lipid peroxidation

The aims of this study were to investigate: 1) if the addition of α-tocopherol (vitamin E) in three concentrations (0.3, 0.6 and 0.9 mM) is able to preserve spermatozoa integrity after thawing and 2) the effect of α-tocopherol supplementation on lipid peroxidation. Fifty four domestic cats were used in this study constituting 18 pools (3 cats per pool). Each pool was submitted at four experimental groups: group 0 (control) – epididymal sperm were frozen with a commercial Botucrio® extender; group 0.3, group 0.6 and group 0.9 – the extender was supplemented with 0.3, 0.6 and 0.9 mM of α-tocopherol, respectively. Each semen sample was evaluated for motility, progressive forward motility, morphology, sperm viability (plasma membrane integrity-PMI), hypo-osmotic swelling test (HOST), before and after thawing. The evaluation of lipid peroxidation reaction by Thiobarbituric Acid Reactive Substances (TBARS) test was performed on thawed semen only. Results demonstrated that there was no significant difference between control and the three α-tocopherol groups with regards to motility and progressive motility after thawing (P > 0.05). As expected, in fresh samples viability was significantly higher than in all the cryopreserved groups in which there was no positive influence of any of the αtocopherol concentration used. Lipid peroxidation was higher in the supplemented groups 0.6 and 0.9 mM of α-tocopherol than in control and in 0.3 mM group. In conclusion, the addition of α-tocopherol to the commercial extender had no positive influence on reduction of lipid peroxidation. This topic deserves further investigations to better understand the effect of cryopreservation procedures on epididymal spermatozoa and to establish adequate strategies to counteract sperm cryodamages.

Cat epididymal semen cryopreserved with and without vitamin E: effect on sperm parameters and lipid peroxidation

The aims of this study were to investigate: 1) if the addition of α-tocopherol (vitamin E) in three concentrations (0.3, 0.6 and 0.9 mM) is able to preserve spermatozoa integrity after thawing and 2) the effect of α-tocopherol supplementation on lipid peroxidation. Fifty four domestic cats were used in this study constituting 18 pools (3 cats per pool). Each pool was submitted at four experimental groups: group 0 (control) epididymal sperm were frozen with a commercial Botucrio® extender; group 0.3, group 0.6 and group 0.9 the extender was supplemented with 0.3, 0.6 and 0.9 mM of α-tocopherol, respectively. Each semen sample was evaluated for motility, progressive forward motility, morphology, sperm viability (plasma membrane integrity-PMI), hypo-osmotic swelling test (HOST), before and after thawing. The evaluation of lipid peroxidation reaction by Thiobarbituric Acid Reactive Substances (TBARS) test was performed on thawed semen only. Results demonstrated that there was no significant difference between control and the three α-tocopherol groups with regards to motility and progressive motility after thawing (P > 0.05). As expected, in fresh samples viability was significantly higher than in all the cryopreserved groups in which there was no positive influence of any of the αtocopherol concentration used. Lipid peroxidation was higher in the supplemented groups 0.6 and 0.9 mM of α-tocopherol than in control and in 0.3 mM group. In conclusion, the addition of α-tocopherol to the commercial extender had no positive influence on reduction of lipid peroxidation. This topic deserves further investigations to better understand the effect of cryopreservation procedures on epididymal spermatozoa and to establish adequate strategies to counteract sperm cryodamages.(AU)

Quantificação atrial de células mononucleares de medula óssea (CMMO) infundidas via intrapericárdica em suínos submetidos a infarto agudo do miocárdio / Atrial quantification of bone marrow mononuclear cells (BMMC) infusioned by intrapericardic route in swine submitted to myocardial infarction

Nas últimas décadas a medicina regenerativa tem se destacado em todo o mundo, graças ao surgimento da terapia com células-tronco, as quais tem grande capacidade de diferenciação em diversos tipos de linhagens celulares, reconstruindo tecidos lesados. A medula óssea, fonte das células utilizadas nesse estudo, contém células-tronco adultas, hematopoiéticas e mesenquimais, pertencentes ao grupo de células mononucleares, que dentre outras funções, são capazes de levar ao remodelamento cardíaco pós-processo isquêmico, e para tanto, podem ser introduzidas no coração sob algumas vias de infusão, sendo que a mais ser recentemente estudada foi a intrapericárdica, a qual foi eleita para o desenvolvimento deste estudo, sendo a qual utilizados 6 suínos, fêmeas, com média de peso de 25Kg, divididos em 2 grupos: 3 animais induzidos ao infarto agudo do mi do miocárdio e tratados com células mononucleares de medula óssea, e 3 animais que apenas receberam as mesmas células, porém não foram induzidos ao infarto (animais controle). As células mononucleares da medula porém óssea foram coletadas e separadas por densidade Ficoll, marcadas com fluorocromo HoechstⓇ e infundidas via intrapericárdica. Após analisarmos os átrios dos animais dos 2 grupos, percebemos que houve uma distribuição homogênea, independente da presença de fator quimiotático no grupo dos animais infartados e que s células foram capazes de sair do espaço pericárdico e transmuralmente ocupar toda a estratigrafia cardíaca. Não houve diferença significativa na quantidade de células encontradas, p>0,01, quando comparamos átrios de animais infartados versus átrios de animais não infartados.

In the last decades, the regenerative medicine have been applied and investigated worldwide due to the stem cell therapy, which has shown high capacity of differentiation into a diverse cell lineage, restoring damaged tissues. In this study we applied bone marrow, which contains adult stem cells, hematopoietic and mesenchimal lineages. Mesenchimal lineage belongs to the mononuclear cell group that among its function, have demonstrated capacity for cardiac remodeling after ischemic processes and, can be introduced into the heart by infusion techniques like intrapericardic technique which has been investigated lately. In this study we investigated the application of intrapericardic technique for mononuclear cell infusion in swine induced to myocardial infarction. We used 6 female swine, averaging 25 kg distributed into control group (3 animals) and experimental group (3 animals), which animal were induced to infarction and received mononuclear cell infusion by intrapericardic technique. Control group received only mononuclear cell infusion by the same technique in normal heart. Mononuclear cells were obtained by Ficoll density and were stained by Hoescht fluorocrome. Atria analysis in both groups revealed a homogeneous distribution of the infusioned cells in the atria with no significant difference, p>0.01 between control and experimental group.