In vitro effect of autologous platelet-rich plasma on H2 O2 -induced oxidative stress in human spermatozoa.

BACKGROUND: Among new therapies emerging in the medical field, the use of platelet-rich plasma (PRP) in human reproduction has not yet been explored. OBJECTIVES: This study aimed at investigating the effect of autologous PRP on sperm parameters in the presence and absence of H2 O2 . MATERIALS AND METHODS: Semen samples were collected from 30 healthy men in their fourth decade attending Azoury IVF clinic. Spermatozoa, cultured in the presence or absence of 10 µM H2 O2 , were left untreated or treated with increasing concentrations of PRP (2, 5, and 10%). After 24-h incubation, ROS levels were assessed and sperm parameters were evaluated. RESULTS: Our results highlight the harmful effect of H2 O2 on sperm parameters, showing an increase in the percentage of reactive oxygen species (ROS)-positive cells, vacuolization, and sperm DNA fragmentation, and a decrease in progressive and total motility in the H2 O2 -treated group compared to non-stressed spermatozoa. When samples were treated with PRP, an improvement of the studied parameters was noted mainly with 2% PRP, thus regarded as the best concentration to achieve a positive effect on sperm parameters. Indeed, non-stressed and stressed spermatozoa treated with 2% PRP showed a significant increase in progressive and total motility, coupled with a decrease in ROS-positive cells, DNA fragmentation, vacuolization, and dead cells compared to the untreated group. In contrast, no significant difference in cell morphology was found between the two groups. Moreover, 2% PRP treatment enhanced sperm parameters and prevented cell death in H2 O2 -exposed spermatozoa as compared to freshly collected semen. DISCUSSION: We suggest that PRP because of its wide arrays of growth factors included in his alpha granules contributes to the inhibition of ROS through the antioxidant, anti-apoptotic activity. CONCLUSION: Autologous PRP improves the quality of the sperm, more so in the presence of an H2 O2 -induced OS.

Cryopreservation media differentially affect sperm motility, morphology and DNA integrity.

INTRODUCTION: Human sperm freezing is very widely used for male fertility preservation. This procedure consists in adding cryoprotectants to the spermatozoa followed by cooling and storing the spermatozoa at a subzero temperature. Many standardized cryopreservation media are available on the market. However, these media differ in their chemical composition and there are no sufficient data to optimize their classification. Therefore, the aim of this study was to compare five commercially available sperm cryopreservation media, which have not been compared together, in terms of motility, morphology and DNA integrity. MATERIALS AND METHODS: One-hundred semen samples were obtained from 10 fertile participants and 90 infertile men. Each sample was evaluated before freezing for motility, morphology and DNA fragmentation index (DFI). Then, it was equally divided into five aliquots. Each aliquot was cryopreserved using one of the five media (A, B, C, D, and E). The same parameters were re-evaluated after the addition of the cryopreservation media in the fertile group, and after sperm thawing in fertile and infertile groups. RESULTS: The results showed that the five selected cryopreservation media had negative effects on sperm motility and morphology per se. In the infertile group, the cryosurvival factor was significantly lower in cryomedium A when compared to the four other media (p < 0.001). In addition, a significantly higher percentage of sperm with coiled tail was detected in cryomedium E compared to cryomedium A (p < 0.05) and to cryomedium B (p < 0.001) after thawing, in the infertile group. Furthermore, the sperm DFI was significantly higher in cryomedia A (p < 0.001), B (p < 0.001), C (p < 0.01), D (p < 0.01) and E (p < 0.05) compared to that of the fresh semen derived from infertile participants. CONCLUSION: This study indicates that the recovery rate of competent spermatozoa, after cryopreservation, is still critical in infertile men. Therefore, frozen semen sample should be used only when necessary.