Are the Morphokinetics of Embryos Obtained from HPV-Positive Sperm Altered? A Retrospective Cohort Study

The objective of the study was to evaluate the impact of male human papillomavirus (HPV) sexually transmitted infection on sperm parameters, embryo morphokinetics, and clinical pregnancy outcomes in assisted reproductive technology (ART) procedures.

Between 2019 and 2023, 3,360 ART cycles were performed at the private IVF center MOMO' Fertilife (Bisceglie, Italy), including 1,035 first-time cycles. Of the 787 males tested for seminal HPV, 62 tested positive.

A study group of 57 HPV-positive and a matched control group of 57 HPV-negative males were selected using our clinic management software. Exclusion criteria included female age ≥42, advanced endometriosis, and HPV positivity in both partners.

Couples undergoing homologous ART with only the male partner testing HPV positive were included, while those with both partners positive were excluded. HPV-positive sperm was used without standard preparation, and all semen procedures followed WHO guidelines. All couples enrolled in the study provided written informed consent, which specified that spermatozoa from HPV-positive participants would be used without undergoing standard sperm preparation procedures.

ICSI was the only ART procedure performed in all cycles included in this study. Semen samples were collected following 3-5 days of abstinence and analyzed within 60 min post-ejaculation. Sperm concentration, motility, and morphology were assessed using phase contrast microscopy. For ICSI procedures in both groups, a standardized "horizontal swim-up" technique developed in-house was employed, an alternative method of semen preparation for ICSI developed in our center. Following liquefaction, an aliquot of the semen sample was sent to the virology laboratory, while a duplicate was stored at -80°C. In cases of inconclusive results, reanalysis was conducted using the frozen sample. IVF laboratory personnel were blinded to HPV status. Sperm quality, the morphokinetics of 210 resulting embryos, and pregnancy outcomes were analyzed. Statistical tests included the Student's t test, Shapiro-Wilk test, Mann-Whitney U test, and Chi-square or Fisher's exact tests.

The study found no statistically significant differences in embryo morphokinetics or ART outcomes between HPV-positive and HPV-negative groups. Pregnancy rates (33.3% vs. 31.6%) and first-trimester miscarriage rates (1 case each) were comparable between the two treatment groups, which also presented similar blastocyst transfer timing and transfer of high-quality embryos. Sperm parameters showed no clinically relevant variation.

The retrospective cohort design and limited sample size represent study limitations. Furthermore, the absence of data on potentially relevant confounders constrains the analysis. Specifically, abortion rate assessment was limited to the first trimester. The lack of extended follow-up data on long-term pregnancy and neonatal outcomes further restricts the conclusions.

Male HPV infection did not adversely affect embryo development or ART success rates. These findings suggest that routine HPV screening in male partners undergoing ART may not be necessary.