Perfluoro-octanoic acid impairs sperm motility through the alteration of plasma membrane

  • I. Šabović
  • I. Cosci
  • L. De Toni
  • A. Ferramosca
  • M. Stornaiuolo
  • A. Di Nisio
  • S. Dall’Acqua
  • A. Garolla
  • C. ForestaEmail author
Original Article



Perfluoroalkyl-substances (PFAS) are chemical additives considered harmful for humans. We recently showed that accumulation of perfluoro-octanoic acid (PFOA) in human semen of exposed subjects was associated with altered motility parameters of sperm cells, suggesting direct toxicity.


To determine whether direct exposure of human spermatozoa to PFOA was associated to impairment of cell function.

Patients and methods

Spermatozoa isolated from semen samples of ten normozoospermic healthy donors were exposed up to 2 h to PFOA, at concentrations from 0.1 to 10 ng/mL. Viability and motility parameters were evaluated by Sperm Class Analyser. Cell respiratory function was assessed by both mitochondrial probe JC-1 and respiratory control ratio (RCR) determination. Sperm accumulation of PFOA was quantified by liquid chromatography–mass spectrometry. Expression of organic ion-transporters OATP1 and SLCO1B2 was assessed by immunofluorescence and respective role in PFOA accumulation was evaluated by either blockade with probenecid or membrane scavenging through β-cyclodextrin (β-CD). Plasma membrane fluidity and electrochemical potential (ΔΨp) were evaluated, respectively, with Merocyanine-540 and Di-3-ANEPPDHQ fluorescent probes.


Compared to untreated controls, a threefold increase of the percentage of non-motile sperms was observed after 2 h of exposure to PFOA regardless of the concentration of PFOA, whilst RCR was significantly reduced. Only scavenging with β-CD was effective in reducing PFOA accumulation, suggesting membrane involvement. Altered membrane fluidity, reduced ΔΨp and sperm motility loss associated with exposure to PFOA were reverted by β-CD treatment.


PFOA alters human sperm motility through plasma-membrane disruption, an effect recovered by incubation with β-CD.


Sperm motility Liquid chromatography–mass spectrometry Probenecid Membrane fluidity 



This research did not receive any specific grant from funding agencies in the public, commercial, or not-for-profit sectors.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

This study was approved by the Institutional Ethics Committee of the Hospital of Padova, Italy, (protocol number 2208P and successive amendments). The investigation was performed according to the principles of the Declaration of Helsinki.

Informed consent

All patients provided signed informed consent at enrollment.

Supplementary material

40618_2019_1152_MOESM1_ESM.docx (14 kb)
Supplementary material 1 (DOCX 14 kb)


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Copyright information

© Italian Society of Endocrinology (SIE) 2019

Authors and Affiliations

  1. 1.Department of Medicine and Unit of Andrology and Reproductive MedicineUniversity of PadovaPaduaItaly
  2. 2.Familial Cancer ClinicVeneto Institute of Oncology (IOV-IRCCS)PaduaItaly
  3. 3.Department of Biological and Environmental Sciences and TechnologiesUniversity of SalentoLecceItaly
  4. 4.Department of Pharmaceutical and Pharmacological SciencesUniversity of PadovaPaduaItaly

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