Fisetin attenuates the adverse effects of freezing and thawing procedures on the biological characteristics of human asthenoteratozoospermia samples

Reprod Biol. 2025 Dec;25(4):101060. doi: 10.1016/j.repbio.2025.101060. Epub 2025 Aug 27.

Abstract

Sperm cryopreservation is a conventional method to preserve sperm cells for a long time. This technique may cause various effects on sperm parameters. Therefore, applying beneficial antioxidants to decrease the adverse effects of freezing is noteworthy. Fisetin is a compound with antioxidant and anti-inflammatory properties. The main aim of the present study is to investigate the protective and beneficial effects of fisetin against cryo-damage on sperm functional parameters. In this experimental study, we analyzed 20 semen samples from asthenoteratozoospermic (AT) patients. Each sample was divided into three treatment groups: (1) fresh control (non-frozen), (2) cryopreservation medium (without fisetin), and (3) cryomedium supplemented with 50 μM fisetin. Freezing and thawing procedures were performed via the conventional method. Post-thaw analyses revealed that cryopreservation significantly reduced sperm motility, chromatin integrity, and mitochondrial membrane potential while increasing DNA fragmentation, malondialdehyde (MDA) levels, and apoptosis (p < 0.05). Fisetin supplementation markedly improved progressive motility (p < 0.05), reduced non-motile sperm percentage (p < 0.05), and decreased DNA fragmentation and MDA levels (p < 0.05). Additionally, it enhanced chromatin condensation and reduced apoptosis rates (p < 0.05). Fisetin attenuates cryo-damage through its antioxidant and anti-apoptotic properties, improving post-thaw sperm quality. Thus, incorporating fisetin into cryopreservation media could enhance sperm viability for assisted reproductive technologies (ART).

Keywords: Apoptosis; Cryopreservation; DNA fragmentation; Fisetin; Oxidative stress; Sperm.

MeSH terms

  • Adult
  • Antioxidants* / pharmacology
  • Apoptosis / drug effects
  • Asthenozoospermia*
  • Cryopreservation* / methods
  • Cryoprotective Agents / pharmacology
  • DNA Fragmentation / drug effects
  • Flavonoids* / pharmacology
  • Flavonols
  • Freezing / adverse effects
  • Humans
  • Male
  • Membrane Potential, Mitochondrial / drug effects
  • Semen Preservation* / methods
  • Sperm Motility / drug effects
  • Spermatozoa* / drug effects

Substances

  • Flavonols
  • fisetin
  • Flavonoids
  • Cryoprotective Agents
  • Antioxidants