Effects of Modified Cryoprotectants on the Expression of Heat Shock Protein 90 (HSP90) and Zona Pellucida II (ZPII) of Post-Thawing Sperm Quality

Authors

• Silvia Werdhy Lestari, Department of Medical Biology, Faculty of Medicine, Universitas Indonesia, Jakarta, IndonesiaFollow
• Manggiasih Dwiayu Larasati, College of Health Sciences, Bachelor of Midwifery Program, Gatot Soebroto Military Hospital, Jakarta, IndonesiaFollow
• Gita Pratama, Department of Obstetry and Gynecology, Faculty of Medicine, Universitas Indonesia, Jakarta, IndonesiaFollow
• Mulyoto Pangestu, Education Program in Reproduction and Development, Department of Obstetrics and Gynecology, School of Clinical Sciences, Monash University, Victoria, AustraliaFollow
• Muhammad Thoriq, Undergraduate study of Medicine, Faculty of Medicine, Universitas Indonesia, Depok, IndonesiaFollow
• Dantha Rafael Sitompul, Undergraduate study of Medicine, Faculty of Medicine, Universitas Indonesia, Depok, IndonesiaFollow

Abstract

Sperm cryopreservation is a basic element of assisted reproductive technology, yet it often compromises cell viability through thermal and osmotic stress. The cellular response involves HSP90, a stress marker and ZPII, which is essential for sperm-oocyte interaction and preventing polyspermy. In addition, this modified cryoprotectant combines glycerol and raffinose is expected to reduce damage and increase spermatozoa's recovery rate after cryopreservation. To address these challenges, we investigated the expression levels of HSP90 and ZPII to assess the protective capacity of modified cryoprotectant formulations. These parameters were analyzed pre- and post-cryopreservation to establish whether these novel mediums superiorly preserve sperm integrity and functional potential compared to standard protocols. Ten samples from fertile and infertile normozoospermia, infertile asthenozoospermia, infertile teratozoospermia, and infertile asthenoteratozoospermia couples were investigated before freezing (fresh) and after-thawing with modified cryoprotectants. The expression levels of HSP90 and ZPII proteins, were measured using immunocytochemistry. The neck, midpiece, and tail regions of human sperm were the primary sites of HSP90 expression, while ZPII exhibited a similar pattern, but primarily in the tail region. Furthermore, there was an increase in HSP90 expression due to cryopreservation at all positive intensities, as well as the IHC score. In contrast, there was a decrease in ZPII expression due to cryopreservation at all positive intensities, as well as the IHC score. In term of cryopreservation, HSP90 expression increased due to its function as cell stress marker, while ZPII expression decreased due to its role as protective molecule.

Keywords

Cryoprotectant modification, HSP90, Post-thawing sperm quality, Sperm Cryopreservation, ZPII

Subject Area

Biology

Article Type

Article

First Page

2485

Last Page

2495

Creative Commons License

This work is licensed under a Creative Commons Attribution 4.0 International License.

How to Cite this Article

Lestari, Silvia Werdhy; Larasati, Manggiasih Dwiayu; Pratama, Gita; Pangestu, Mulyoto; Thoriq, Muhammad; and Sitompul, Dantha Rafael (2026) "Effects of Modified Cryoprotectants on the Expression of Heat Shock Protein 90 (HSP90) and Zona Pellucida II (ZPII) of Post-Thawing Sperm Quality," Baghdad Science Journal: Vol. 23: Iss. 7, Article 10.
DOI: https://doi.org/10.21123/2411-7986.5354