Clinical and Experimental Reproductive Medicine

The Korean Society for Reproductive Medicine (대한생식의학회)
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Retinoic acid loaded with chitosan nanoparticles improves spermatogenesis in scrotal hyperthermia in mice

  • Fatemeh Mazini (Department of Anatomical Science, Kermanshah University of Medical Sciences) ;
  • Mohammad-Amin Abdollahifar (Urogenital Stem Cell Research Center, Shahid Beheshti University of Medical Sciences) ;
  • Hassan Niknejad (Department of Pharmacology, School of Medicine, Shahid Beheshti University of Medical Sciences) ;
  • Asma Manzari-Tavakoli (ATMP Department, Breast Cancer Research Center, Motamed Cancer Institute, ACECR) ;
  • Mohsen Zhaleh (Department of Anatomical Science, Kermanshah University of Medical Sciences) ;
  • Reza Asadi-Golshan (Department of Anatomy, School of Medicine, Tehran University of Medical Sciences) ;
  • Ali Ghanbari (Department of Anatomical Science, Kermanshah University of Medical Sciences)
  • Received : 2023.05.16
  • Accepted : 2023.07.03
  • Published : 2023.12.31
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Abstract

Objective: High temperatures can trigger cellular oxidative stress and disrupt spermatogenesis, potentially leading to male infertility. We investigated the effects of retinoic acid (RA), chitosan nanoparticles (CHNPs), and retinoic acid loaded with chitosan nanoparticles (RACHNPs) on spermatogenesis in mice induced by scrotal hyperthermia (Hyp). Methods: Thirty mice (weighing 25 to 30 g) were divided into five experimental groups of six mice each. The groups were as follows: control, Hyp induced by a water bath (43 ℃C for 30 minutes/day for 5 weeks), Hyp+RA (2 mg/kg/day), Hyp+CHNPs (2 mg/kg/72 hours), and Hyp+RACHNPs (4 mg/kg/72 hours). The mice were treated for 35 days. After the experimental treatments, the animals were euthanized. Sperm samples were collected for analysis of sperm parameters, and blood serum was isolated for testosterone measurement. Testis samples were also collected for histopathology assessment, reactive oxygen species (ROS) evaluation, and RNA extraction, which was done to compare the expression levels of the bax, bcl2, p53, Fas, and FasL genes among groups. Additionally, immunohistochemical staining was performed. Results: Treatment with RACHNPs significantly increased stereological parameters such as testicular volume, seminiferous tubule length, and testicular cell count. Additionally, it increased testosterone concentration and improved sperm parameters. We observed significant decreases in ROS production and caspase-3 immunostaining in the RACHNP group. Moreover, the expression levels of bax, p53, Fas, and FasL significantly decreased in the groups treated with RACHNPs and RA. Conclusion: RACHNPs can be considered a potent antioxidative and antiapoptotic agent for therapeutic strategies in reproductive and regenerative medicine.

Keywords

Acknowledgement

This article was financially supported by the Research Department of the Kermanshah University of Medical Sciences School of Medicine (Grant No: 4010057).

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