Hyperthermia versus busulfan: Finding the effective method in animal model of azoospermia induction

Abstract

Animal models of azoospermia are very applicable when evaluating new treatment methods for research purposes. The present study aimed to compare azoospermia induction in mice using busulfan or hyperthermia. To do this, about 36 adult male mice (28–30 g) were included into three experimental groups randomly (n = 12): control, busulfan (injected by a single dose of 40 mg/kg busulfan intraperitoneally) and hyperthermia (exposure to a temperature of 43°C every other day for 5 weeks). Animals were preserved for 35 and 70 days following interventions and then were sacrificed for further evaluations. After 35 days, busulfan and hyperthermia groups revealed a significant decrease in the sperm count and weight of testis compared to the control group (p < .0001). In addition, after 70 days, sperm count and weight of testis in group busulfan showed a significant increase compared to group hyperthermia (p < .01). No significant difference was observed regarding the mortality of mice between busulfan and hyperthermia groups. In group busulfan, degenerative changes in the germinal epithelium were detected in some tubules, although in group hyperthermia, degenerative changes and complete depletion of all tubules were observed. Continuous hyperthermia is a more effective method in the induction of as animal model of azoospermia compared to the busulfan.

Non‐apoptotic cell death such as pyroptosis, autophagy, necroptosis and ferroptosis acts as partners to induce testicular cell death after scrotal hyperthermia in mice

“…Our previous studies showed abnormalities in sperm parameters including sperm count, motility, viability and morphology, as well as histological changes in the testes including a decrease in the number of testicular cells and a decrease in testicular tissue volume after exposure to high temperatures that can result in reduced germ cells via apoptotic pathways (Ilkhani et al, 2020;Ziaeipour et al, 2019).…”

“…All experimental groups were anaesthetized by ketamine (100 mg/ kg) and xylazine (5 mg/kg). Then, the lower parts of the body including the scrotum were submerged in a 43°C water bath for 20 min (Ziaeipour et al, 2019).…”

“…Increased testicular temperature induces germ cell depletion and reduces spermatogenesis and fertility. Also, testicular hyperthermia leads to oligospermia and azoospermia (Hasani et al, 2020;Ilkhani et al, 2020;Khosravi et al, 2021;Ziaeipour et al, 2019). Raised testicular temperature causes the atrophy of germinal epithelium and spermatogenic arrest (Ziaeipour et al, 2019), leading to lower sperm counts; Sertoli (Cai et al, 2011) and Leydig (Kanter & Aktas, 2009) cells are also impacted by heat stress.…”

“…Also, testicular hyperthermia leads to oligospermia and azoospermia (Hasani et al, 2020;Ilkhani et al, 2020;Khosravi et al, 2021;Ziaeipour et al, 2019). Raised testicular temperature causes the atrophy of germinal epithelium and spermatogenic arrest (Ziaeipour et al, 2019), leading to lower sperm counts; Sertoli (Cai et al, 2011) and Leydig (Kanter & Aktas, 2009) cells are also impacted by heat stress. The fundamental mechanism by which germ cell destruction occurs in response to heat stress is not yet fully understood.…”

Therapeutic Effects of Edaravone on Azoospermia: Free Radical Scavenging and Autophagy Modulation in Testicular Tissue of Mice

“…Obstructive (OA) or non-obstructive (NOA) azoospermia is the medical condition defined as the complete absence of spermatozoa in semen due to obstruction, inadequate hormonal stimulation, or impaired spermatogenesis. Azoospermia and oligozoospermia also occur due to exposure to toxic substances, genetic factors, injuries, immunosuppressants, and anticancer drugs such as cyclophosphamide and busulfan ( 1 , 2 ).…”