Purpose
To compare the efficacy of swim-up and DGC in improving sperm deformity and DNA fragmentation and to determine which method is better in teratozoospermic patients requiring artificial reproduction.

/pdf/efficacy-of-swim-up-versus-density-gradient-centrifugation-4md7xtonc7.pdf

Efficacy of swim-up versus density gradient centrifugation in improving sperm deformity rate and DNA fragmentation index in semen samples from teratozoospermic patients

https://www.fertstert.org/article/S0015-0282(11)01106-X/pdf

Clinical outcomes and development of children born after intracytoplasmic sperm injection (ICSI) using extracted testicular sperm or ejaculated extreme severe oligo-astheno-teratozoospermia sperm: a comparative study.

Human sperm vitrification is a new cryopreservation method. This study compared the effects of rapid freezing and vitrification on various sperm parameters, hyaluronan-binding assay and DNA fragmentation and assessed the impact of cryoprotectant agents (CPA) with vitrification. A total of 30 normo-ejaculates were prepared by swim up and the motile sperm fraction was divided into four: fresh (control), rapid freezing, and two vitrification groups (a, lacking CPA; b, with CPA). For rapid freezing, a cryovial of sperm suspension was held just above the liquid nitrogen surface, and for vitrification, 30μl suspension was dropped directly into liquid nitrogen. Sperm parameters, including motility, viability and morphology, declined after cryopreservation in both groups. DNA fragmentation was not significantly higher in the vitrification (15.7±4.4%) or rapid freezing (16.6±5.6%) groups when compared with controls (11.6±4.5%). The rates of hyaluronan binding were similar between the control and cryopreserved groups. Moreover, addition of CPA for vitrification had a neutral effect on rates of sperm recovery. In conclusion, vitrification has great potential for human sperm cryopreservation and does not require CPA, with its possible toxicity. However, it is not superior to rapid cryopreservation regarding sperm recovery rate in normozoospermia. Human sperm vitrification is a new cryopreservation method that has been introduced recently. This study compared the effects of rapid freezing with vitrification on rates of sperm parameters, hyaluronan-binding assay and DNA fragmentation after thawing/warming and assessed the impact of cryoprotectant agent (CPA) on vitrification. The study was performed on 30 ejaculates prepared using the swim-up technique. Each motile sperm suspension was divided into four: control (fresh); rapid freezing; and two vitrification groups (a, lacking CPA; b, with CPA). For rapid freezing, a cryovial of sperm suspension was held above the surface of liquid nitrogen. For vitrification, 30μl sperm suspension was dropped into liquid nitrogen directly. The rates of progressive motility (86.6±5.9%) and viability (95.8±3.9%) in controls declined significantly, to 40.0±13.0% and 63.2±7.7% for rapid freezing and 41.9±10.3% and 64.4±10.0% for vitrification, respectively. Normal sperm morphology was also significantly decreased after cryopreservation in all groups. DNA fragmentation was higher with rapid freezing compared with fresh controls (16.6±5.6% vs. 11.6±4.5%, P=0.01), but DNA fragmentation did not increase significantly in vitrified samples (15.7±4.4%). The rates of hyaluronan binding were similar between the control and cryopreserved groups. Moreover, addition of CPA for vitrification had a neutral effect on rates of sperm recovery. In conclusion, vitrification has great potential for human sperm cryopreservation and does not require CPA, with its possible toxicity. However, it is not superior to rapid cryopreservation regarding sperm recovery rate in normozoospermia.

http://ssu.ac.ir/cms/fileadmin/user_upload/ivfen/IVf_Persian/executive_comiitee/Dr_khalili/articles/Vitrification_is_not_superior_to_rapid_freezing.pdf

Vitrification is not superior to rapid freezing of normozoospermic spermatozoa: effects on sperm parameters, DNA fragmentation and hyaluronan binding.

Background
Poor sperm quality can negatively affect embryonic development and IVF outcome. This study is aimed at investigating the influence of various lifestyle factors on semen quality according to MSOME (motile sperm organelle morphology examination) criteria.

/pdf/the-combination-matters-distinct-impact-of-lifestyle-factors-4rv6tozf9z.pdf

The combination matters--distinct impact of lifestyle factors on sperm quality: a study on semen analysis of 1683 patients according to MSOME criteria.

With the motile sperm organelle morphology examination (MSOME), spermatozoa morphology may be assessed directly on motile spermatozoa at high magnification (up to 6600×). This procedure describes more precisely spermatozoa abnormalities, especially head vacuoles. However, no consensus has been established concerning normal or abnormal MSOME criteria. The aim of our study was to define MSOME vacuole criteria assessed objectively with a digital imaging system software to establish a potential relationship between conventional semen parameters. A total of 440 semen samples were obtained from males consulting in Rouen University Hospital Reproductive Biology Laboratory. Conventional semen analysis (volume, sperm concentration, progressive motility, vitality and morphology) and MSOME assessment {sperm head length, width and area as well as vacuole number, vacuole area and relative vacuole area to sperm head [RVA (%) = [vacuole area (μm(2))/head area (μm(2))] × 100)]} were performed for each semen sample. Among our 440 males, 109 presented normal conventional semen parameters and 331 abnormal ones. Sperm head vacuoles were significantly larger in abnormal semen samples (p < 0.0001). RVA was the most discriminative MSOME criterion between normal and abnormal semen samples according to ROC curves analysis, and was negatively correlated with poor sperm morphology (r = -0.53, p < 0.0001). We concluded to (i) the normal occurrence of vacuoles in sperm head whatever the normality or abnormality of semen parameters, (ii) the discriminative function of the RVA to distinguish semen samples with normal and abnormal parameters, and (iii) the strong correlation between high RVA and poor sperm morphology.

https://onlinelibrary.wiley.com/doi/pdf/10.1111/j.1365-2605.2012.01249.x

Relationship between conventional sperm parameters and motile sperm organelle morphology examination (MSOME).

Use of high-magnification microscopy for the assessment of sperm recovered after two different sperm processing methods.

Sperm vacuole improvement after antioxidant therapy

Resumen En los ultimos anos, varios autores han comenzado a estudiar los espermatozoides con microscopia de alta magnificacion. Todos ellos encuentran una clara relacion entre la presencia de vacuolas en la cabeza de los espermatozoides y las tasas de embarazo y aborto, asi como con un aumento en el porcentaje de fragmentacion del ADN. De hecho, la morfologia espermatica y la presencia o no de vacuolas nucleares parece ser un reflejo del estado fisiologico de los espermatozoides. Recientemente, el Centro de Infertilidad y Reproduccion Humana de Barcelona ha adquirido un microscopio para alta magnificacion, con la intencion de utilizarlo de manera rutinaria para diagnostico morfologico espermatico y en la denominada inyeccion intracitoplasmatica de espermatozoides morfologicamente seleccionados (IMSI). Se han disenado diferentes estudios: el efecto de los antioxidantes sobre la disminucion de las vacuolas espermaticas, que tecnica de preparacion del semen selecciona un mayor porcentaje de espermatozoides no vacuolados, etc. Nuestro objetivo ha sido incorporar una nueva herramienta para profundizar en el conocimiento del espermatozoide humano y mejorar tanto en el diagnostico del factor masculino como en los resultados de las diferentes tecnicas de reproduccion asistida.

Magnificación de espermatozoides: una nueva visión del factor masculino en reproducción asistida

Resumen Objetivo La aparicion de nuevas herramientas de estudio en el campo de la andrologia nos ha permitido evaluar nuevos marcadores de calidad seminal, como la presencia de vacuolas en el nucleo del espermatozoide, mediante el uso de microscopia de alta magnificacion, y el grado de fragmentacion del acido desoxirribonucleico (ADN) espermatico. Ambos marcadores de calidad pueden verse afectados por la presencia de radicales libres de oxigeno, que favorecen la oxidacion celular. En el presente estudio piloto se plantea valorar la utilidad de un tratamiento con antioxidantes para reducir tanto el indice de vacuolizacion espermatica, como el grado de fragmentacion del ADN de una forma significativa. Material y metodos Estudio prospectivo para el que se selecciono a un total de 65 pacientes, de los cuales 33 finalizaron el estudio. Cada paciente entrego dos muestras de semen y realizo un tratamiento con antioxidantes mediante Androferti®. Se evaluo el indice de vacuolizacion en el nucleo del espermatozoide y la fragmentacion del ADN espermatico en cada una de las muestras entregadas. Resultados Se observa un aumento estadisticamente significativo de espermatozoides con grados de vacuolizacion I y II despues de un mes de tratamiento, asi como tambien una disminucion del grado de fragmentacion del ADN espermatico. Conclusiones A pesar de que se trata de un estudio clinico no aleatorizado, parece concluyente la importancia del uso de un tratamiento antioxidante para aumentar el exito de un ciclo de reproduccion asistida, con el objetivo de disminuir la influencia negativa de malformaciones en el nucleo espermatico y de la fragmentacion del ADN.

Efecto del tratamiento con vitaminas, L-carnitina y coenzima Q10 en el índice de vacuolización y la fragmentación espermática en pacientes de fecundación in vitro

Resumen Las diferentes tecnicas de reproduccion asistida han ido evolucionando sensiblemente desde sus inicios. En su momento, la inyeccion intracitoplasmatica de espermatozoides (ICSI) permitio solucionar muchos casos de infertilidad masculina. La observacion morfologica de los ovocitos y embriones, y una adecuada clasificacion y seleccion hicieron aumentar las tasas de embarazo. Hoy se dispone de una nueva tecnica para observar a grandes aumentos los espermatozoides, permitiendo una mejor seleccion de los que se van a microinyectar, es la denominada inyeccion intracitoplasmica de espermatozoides morfologicamente seleccionados (IMSI). Esto se esta traduciendo en un aumento de las tasas de embarazo y en una importante reduccion de abortos.

Ana Monqaut

Papers

Use of high-magnification microscopy for the assessment of sperm recovered after two different sperm processing methods.

Sperm vacuole improvement after antioxidant therapy

Magnificación de espermatozoides: una nueva visión del factor masculino en reproducción asistida

Efecto del tratamiento con vitaminas, L-carnitina y coenzima Q10 en el índice de vacuolización y la fragmentación espermática en pacientes de fecundación in vitro

Es la inyección intracitoplásmica de espermatozoides morfológicamente seleccionados una técnica con futuro