Acrosome reaction and chromatin integrity as additional parameters of semen analysis to predict fertilization and blastocyst rates.

BACKGROUND: Traditional semen parameters have shown little to none predictive value for fertilization and blastocyst viability for a successful pregnancy. Therefore, the purpose of this study was to explore the usefulness of incorporating the acrosome reaction (AR) and chromatin integrity to conventional semen analysis to individually predict the fertile potential of sperm samples. METHODS: A cross-sectional study was conducted in 69 participants undergoing IVF using oocyte donation. Semen samples were collected and evaluated for: AR [spontaneous (sAR) and induced (iAR)] by flow cytometry using anti-CD46-FITC, Acrosome Response to an Ionophore Challenge (ARIC), chromatin integrity by Sperm Chromatin Structure Assay (DNA Fragmentation Index-%DFI and High DNA Stainability-%HDS), WHO semen analysis, fertilization and blastocyst rates. RESULTS: The participant age was 40.0 ± 6.1 years (66% were normozoospermic). Sperm morphology, sAR, iAR, and ARIC were associated with the fertilization (ß = 3.56, R2 = 0.054; ß = - 5.92, R2 = 0.276; ß = 1.83, R2 = 0.150; and ß = 2.10, R2 = 0.270, respectively, p < 0.05). A logit model was developed to calculate the probability of fertilization (≥ 60%) for each participant, using the sperm morphology and ARIC as independent variables, followed by ROC analysis to determine a cutoff probability of 0.65 (specificity = 80.6%, sensitivity = 63.2%). %DFI was inversely associated with the viable blastocyst rate (ß = - 1.77, R2 = 0.057, p = 0.003), by the logit model and ROC analysis, a cutoff probability of 0.70 (specificity = 80.6%, sensitivity = 72.3%) was obtained to predict blastocyst viability (≥ 40%). There was no difference in the results with normozoospermic samples (n = 46). CONCLUSIONS: The incorporation of ARIC and %DFI allowed to obtain predictive models for high fertilization and blastocyst rates in an individualized way, being promising tools to improve the diagnosis of male fertility potential for research or assisted reproduction, even in men with unknown infertility.

Assessment of Prostaglandin-Endoperoxide Synthase 2 and Versican gene expression profile from the cumulus cells: association with better in vitro fertilization outcomes.

BACKGROUND: Current methods for determining superior embryo quality (morphological assessment) are unable to compensate for poor pregnancy outcomes. Due to the importance of the cumulus-oocyte complex and the value of cumulus cells (CCs) as markers of embryo health, we determined the association between the CCs gene expression of the Prostaglandin-Endoperoxide Synthase 2 (PTGS2) and Versican (VCAN) with pregnancy. METHODS: One hundred forty-nine women, suffering from infertility and undergoing IVF, were included in this study (age: 29-46 years; BMI = 25.5 ± 5.0 kg/m2). Patients underwent a standard IVF protocol. CCs were isolated during oocyte retrieval, and their RNA was isolated using Trizol. The mRNA expression of PTGS2, VCAN, and L19 was measured by qPCR. The PVL index, (PTGS2 + VCAN)*L19normalized, was determined for each oocyte. Clinical pregnancy was confirmed by ß-hCG and the presence of a fetal heartbeat. Associations were determined by ROC curves or logistic regression. RESULTS: There was no correlation between the PVL index and morphological scores. Using only single embryo transfers (SETs), we determined that the PVL index was associated with pregnancy (ß-hCG: AUC = 0.87, 95%CI: 0.74-1.00) with an optimal cutoff value of 58.2. Using the complete cohort (consisting of SETs, and patients with 2, 3, or 4 embryos transferred), the presence of at least one embryo with a PVL index score ≥ 58.2 was associated with a greater probability of achieving pregnancy (ß-hCG: odds ratio = 17.15, 95%CI: 6.82-43.18, p < 0.001). CONCLUSION: Transferring at least one embryo with a PVL index score ≥ 58.2, generates a higher chance of achieving pregnancy.