{"paper_id":"0dcafa0d-2f43-4fee-9b01-3c7c2f6db282","body_text":"The effects of desflurane on male rat reproductive hormones, testicular tissue, and sperm morphology: An experimental study | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article The effects of desflurane on male rat reproductive hormones, testicular tissue, and sperm morphology: An experimental study Serkan Dogru, Hatice Yilmaz Dogru, Hakan Tapar, Akgul Arici, Mustafa Suren This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-7507144/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 06 Feb, 2026 Read the published version in BMC Anesthesiology → Version 1 posted 12 You are reading this latest preprint version Abstract Background: Desflurane is a widely used inhalational anesthetic known for its advantageous properties in clinical settings. This study aimed to investigate the effects of desflurane inhalation on male reproductive hormones, testicular tissue integrity, and sperm morphology in a rat model. Methods: Thirty male rats were allocated into six experimental groups: Control group: Administered 2 L/min of O₂ for 18 minutes daily over seven days. Group 1: Exposed to 1 minimum alveolar concentration (MAC) of desflurane and 2 L/min of O₂ for 18 minutes daily over seven days. Group 2: Received the same treatment as Group 1 for seven days, followed by a seven-day recovery period without intervention. Group 3: Administered 1 MAC desflurane and 2 L/min of O₂ for 18 minutes daily over 14 days. Group 4: Received the same treatment as Group 3 for 14 days, followed by a seven-day recovery period without intervention. Group 5: Administered the same treatment as Group 3 for 14 days, followed by a 14-day recovery period without intervention. Biochemical analyses were conducted to measure serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, and inhibin B. Histopathological evaluations were performed to assess testicular tissue integrity, and sperm morphology was examined to identify abnormalities. Results: Significant histopathological damage was observed in all experimental groups compared to the control group (p < 0.05). The proportion of morphologically abnormal spermatozoa was significantly higher in Groups D2, D3, D4, and D5 compared to the control group (p = 0.030, p = 0.002, p < 0.001, and p = 0.016, respectively). Serum levels of FSH, LH, testosterone, and inhibin B exhibited statistically significant alterations across all experimental groups (p < 0.05). Conclusion: Chronic inhalation of desflurane, a modern inhalational anesthetic, was found to adversely affect testicular histology, sperm morphology, and the regulation of male reproductive hormones in rats. These findings highlight potential reproductive toxicity associated with prolonged desflurane exposure. Follicle Stimulating Hormone Luteinizing Hormone testosterone inhibin B desflurane sperm morphology Figures Figure 1 Figure 2 Introduction Male fertility is closely linked to the production of normal spermatozoa, which is the result of a highly regulated and complex process known as spermatogenesis. This process encompasses three key phases: mitosis, meiosis, and spermiogenesis [ 1 ]. Spermatogenesis is tightly regulated by a delicate balance of endocrine and paracrine signaling pathways. Follicle-stimulating hormone (FSH) and luteinizing hormone (LH), secreted by the pituitary gland, play pivotal roles in this regulation. FSH acts on Sertoli cells, while LH stimulates Leydig cells, both of which provide essential factors necessary for the proper progression of spermatogenesis [ 2 ]. The intricate network of signals governing spermatogenesis remains an active area of research. Importantly, external environmental factors have the potential to disrupt this process, leading to impaired fertility and abnormalities in sperm production [ 3 , 4 ]. With the rapid advancements in medical technology, the frequency of surgical procedures has increased significantly in recent years. This has inevitably led to a corresponding rise in the use of volatile anesthetic agents. However, the routine administration of inhalation anesthetics has been associated with various adverse effects on multiple organ systems, including the liver, kidneys, lungs, and nervous system, as documented in both experimental and clinical studies [ 4 – 8 ]. Of particular concern is the evidence suggesting that repeated exposure to volatile anesthetics may induce DNA damage in germ cells, contribute to congenital anomalies in offspring, and cause morphological alterations in sperm cells [ 9 – 11 ]. Despite these findings, the potential effects of modern inhalation anesthetics on the male reproductive system remain underexplored. Desflurane (2-difluoromethoxy-1,1,1,2-tetrafluoroethane; C₃H₂F₆O) is a halogenated, fluorinated ether that is colorless, non-flammable, and liquid at room temperature. It has been widely used in clinical practice for the maintenance of general anesthesia since its introduction in 1989 [ 12 – 14 ]. Desflurane is characterized by several advantageous pharmacological properties, including minimal metabolism (0.02%) and a low blood/gas partition coefficient (0.42), which contribute to its rapid induction and recovery profiles [ 15 , 16 ]. Additionally, desflurane anesthesia has been associated with reduced postanesthesia care unit stay durations, facilitating earlier patient discharge [ 14 , 17 ]. Despite its widespread use, a systematic review of the literature reveals a lack of experimental studies investigating the potential effects of desflurane on the male reproductive system. Given this gap in knowledge, the present study was designed to investigate the effects of repeated desflurane exposure on sperm morphology, testicular tissue integrity, and reproductive hormone levels in a rat model. Materials and Methods Chemicals Ketamine and xylazine were procured from Alfasan International B.V. (Woerden, Netherlands). Hematoxylin Harris and Eosin Y (1% alcoholic) were obtained from Atom Scientific Ltd (Manchester, UK). Modified Human Tubal Fluid Medium was purchased from Irvine Scientific (California, USA). Bouin's solution was sourced from Tek-Path Medikal (Izmir, Turkey). Animals The study was conducted following approval by the Tokat Gaziosmanpasa University Local Ethics Committee for Animal Experimentation (Approval No: 2013-HADYEK-12, Approval date: 29.02.2013). A total of 30 adult male Wistar-Albino rats (90 days old, 250–300 g) were obtained from the Experimental Medicine Unit at Gaziosmanpasa University, Turkey. The animals were housed under controlled conditions, including a 12-hour light-dark cycle (lights on at 07:00 a.m.), room temperature maintained at 20–24°C, and relative humidity of 40–50%. Rats were housed in polycarbonate cages with ad libitum access to tap water. A glass anesthesia chamber (40 × 50 × 60 cm) was used for desflurane exposure. The chamber was equipped with an anesthetic gas inlet at the top left and an outlet at the top right, as previously described in the literature [ 9 ]. An airtight connection was established between the chamber and the anesthetic circuit using an anesthesia machine (Prima SP Alpa, Penlon Limited, Oxon, UK). The exposure dose of 1 MAC desflurane administered with 2 L/min of O₂ for 18 minutes daily was selected based on previous studies investigating occupational exposure levels in controlled experimental settings [ 9 ]. This dosing protocol was designed to simulate the cumulative low-dose exposure experienced by health care workers during repeated, short-term contact with inhalational anesthetics in poorly ventilated or inadequately scavenged operating room environments, rather than the higher, continuous exposure a patient would receive during a surgical procedure. The duration and concentration were chosen to reflect a realistic occupational risk model while minimizing excessive stress or toxicity in the animal subjects. The rats were randomly divided into six groups (n = 5 per group): Control group (C): Administered 2 L/min of O₂ for 18 minutes daily over seven days. Group D1: Exposed to 1 minimum alveolar concentration (MAC) of desflurane and 2 L/min of O₂ for 18 minutes daily over seven days. Group D2: Received the same treatment as Group D1 for seven days, followed by a seven-day recovery period without intervention. Group D3: Administered 1 MAC desflurane and 2 L/min of O₂ for 18 minutes daily over 14 days. Group D4: Received the same treatment as Group D3 for 14 days, followed by a seven-day recovery period without intervention. Group D5: Administered the same treatment as Group D3 for 14 days, followed by a 14-day recovery period without intervention. At the end of the experimental periods, euthanasia was performed via cervical dislocation under anesthesia induced by intraperitoneal injection of ketamine (90 mg/kg) and xylazine (10 mg/kg). Intracardiac blood samples (5 mL) were collected immediately for hormonal analysis, and bilateral testes were dissected. The right testis was fixed in Bouin's solution for 24 hours, dehydrated, and embedded in paraffin blocks for histopathological examination. The left testis was placed on ice and stored at − 70°C for subsequent biochemical analyses. Histopathological Analysis of Testicular Tissue Paraffin-embedded testicular tissues were sectioned at 5 µm thickness, mounted on glass slides, and stained with hematoxylin and eosin (H&E) for light microscopic examination (Nikon Eclipse E600W, Tokyo, Japan). Six sections from each testis were randomly numbered and evaluated by a blinded investigator. At least five microscopic fields per testis were assessed. Histopathological injury scores were determined using a four-level grading scale as described by Cosentino et al. [ 18 ]: Grade 1: Normal testicular architecture with orderly arrangement of germinal cells. Grade 2: Slight disorganization and non-cohesive germinal cells with closely spaced seminiferous tubules. Grade 3: Moderate disorganization, sloughed germinal cells, shrunken pyknotic nuclei, and less distinct seminiferous tubule borders. Grade 4: Severe disorganization, tightly packed seminiferous tubules, and coagulative necrosis of germinal cells. Sperm Morphology Analysis Epididymal sperm was collected by mincing the cauda epididymis in 2 mL of Ham's F10 medium. A drop of sperm suspension was placed on a microscope slide, air-dried, and examined under a light microscope at 400× magnification. Five random fields were assessed for morphological abnormalities, including bicephalic, amorphous, hook-less, and tail abnormalities (coiled or abnormal tails). The percentage of abnormal sperm was calculated and analyzed [ 19 ]. Biochemical Analysis of Reproductive Hormones Blood samples were allowed to clot for 20 minutes, centrifuged at 1500 × g (4°C) for 15 minutes, and the serum was aliquoted into Eppendorf tubes. Samples were stored at − 20°C until analysis. For testicular tissue analysis, tissues were weighed and homogenized in ice-cold Tris-HCl buffer (50 mM, pH 7.4) containing 0.50 mL/L Triton X-100 using an IKA Ultra-Turrax T25 Basic homogenizer (Stanfen, Germany) at 13,000 rpm for 2 minutes. All procedures were performed at 4°C. Serum levels of LH, FSH, testosterone (T), and inhibin B (Inh-B) were measured using rat-specific ELISA kits (LH and FSH: YH Biosearch Laboratory, Shanghai, China; T: Cayman Chemical Company, Michigan, USA; Inh-B: SunRed Biotechnology Company, Shanghai, China) according to the manufacturers' instructions. Hormone concentrations were expressed as mIU/mL for LH and FSH, pg/mL for T, and ng/mL for Inh-B. Statistical Analysis Data normality and variance were assessed using the Kolmogorov-Smirnov test. Quantitative data are presented as mean ± standard deviation, while qualitative data are expressed as frequencies and percentages. Parametric or nonparametric tests were applied as appropriate. Histopathological injury scores and hormonal levels were analyzed using the Kruskal-Wallis test, with post-hoc comparisons performed using the Mann-Whitney U test and Holm-Sidak test. Bonferroni correction was applied to all multiple comparisons. All statistical analyses were conducted using SPSS version 20.0 (SPSS Inc., Chicago, IL, USA), with statistical significance set at p < 0.05. Results Histopathological Findings The effects of desflurane on testicular tissue were evaluated using H&E staining. Histopathological findings are illustrated in Fig. 1 , and the corresponding injury scores, graded on a scale of 1 to 4, are presented in Fig. 2 . Significant differences in histological injury scores were observed between the control group (C) and all experimental groups (p < 0.05). Furthermore, the mean histological injury scores in Groups D3 and D4 were higher than in Group D1 (p = 0.005 and p = 0.042, respectively). Group D3 also exhibited higher injury scores compared to Group D2 (p = 0.042), while Group D5 demonstrated lower scores than Group D3 (p = 0.015). In Group D1, mild degenerative changes were observed in focal areas of the seminiferous tubule epithelium (Fig. 1 B), whereas Group C displayed normal testicular architecture with orderly arranged germinal cells (Fig. 1 A). Group D2 exhibited mild to moderate degenerative changes in the seminiferous tubule epithelium, accompanied by mild edema and congestion in the interstitial space (Fig. 1 C). Group D3 showed pronounced degenerative changes in the seminiferous tubule epithelium, along with severe edema and congestion in the interstitial space, although no loss of spermatogenic cells was observed (Fig. 1 D). In Group D4, moderate degeneration of the seminiferous tubule epithelium and interstitial edema and congestion were evident (Fig. 1 E). Group D5 displayed slight degenerative changes in the seminiferous tubule epithelium (Fig. 1 F). Light microscopic analysis revealed that chronic inhalational exposure to desflurane induced structural impairments in the seminiferous tubule epithelium and caused interstitial edema and congestion (Fig. 1 A– 1 F). Biochemical Analyses Significant differences in serum levels of FSH, LH, T, and Inh-B were observed across all groups (p < 0.05, Table 1 ). Table 1 Hormone levels in serum samples C D1 D2 D3 D4 D5 p Median (IQR) Median (IQR) Median (IQR) Median (IQR) Median (IQR) Median (IQR) N = 5 N = 5 N = 5 N = 5 N = 5 FSH 9.94 (9.905–10.125) a 9.8 (9.445–10.090) 9.72 (9.560–9.805) a,b 9.54 (9.315–10.145) 9.92 (9.680–9.965) 10.36 (9.820–10.490) b 0.128 LH 6.09 (5.935–6.260) c,d,e 6.11 (5.855–6.160) f,g 5.76 (5.635–5.880) c,f 5.64 (5.355–5.815) d,g 5.91 (5.535–5.930) e 5.96 (5.665–6.120) 0.028* T 325.80 (176.55–573.80) h,k,m 531.10 (475.55–622.20) n,p 721.40 (517.10–1018.45) h 668.20 (588.65–765.80) k,n 906.30 (612.55–1003.95) m,p 609.80 (554–710.75) 0.025* Inh-B 0.389 (0.330–0.476) q,r,s,t 0.479 (0.467–0.494) u,v,w 0.485 (0.472–0.544) q,y,z,aa 0.663 (0.637–0.818) r,u,y 0.714 (0.685–0.831) s,v,z 0.756 (0.682–0.805) t,w,aa 0.001* FSH, follicle stimulating hormone; LH, luteinizing hormone; T, testosterone; Inh-B, inhibin B; C, control group; D1, desflurane 1 group; D2, desflurane 2 group; D3, desflurane 3 group; D4, desflurane 4 group; D5, desflurane 5 group. *p < 0.05 Kruskal-Wallis test. All intergroup comparisons were performed by using Tukey’s HSD test. Significant differences in FSH levels: a C-D2: p = 0.009; b D2-D5: p = 0.028. Significant differences in LH levels: c C-D2: p = 0.009; d C-D3: p = 0.028; e C-D4: p = 0.028; f D1-D2: p = 0.028; g D1-D3: p = 0.028. Significant differences in T levels: h C-D2: p = 0.036; k C-D3: p = 0.047; m C-D4: p = 0.016; n D1-D3: p = 0.047; p D1-D4: p = 0.036. Significant differences in Inh-B levels: q C-D2: p = 0.047; r C-D3: p = 0.009; s C-D4: p = 0.009; t C-D5: p = 0.009; u D1-D3: p = 0.009; v D1-D4: p = 0.009; w D1-D5: p = 0.009; y D2-D3: p = 0.009; z D2-D4: p = 0.009; aa D2-D5: p = 0.009. LH Levels: Serum LH levels in Groups D2, D3, and D4 were lower than in Group C (p = 0.009, p = 0.028, and p = 0.028, respectively). LH levels in Groups D2 and D3 were also lower than in Group D1 (p = 0.028 for both). FSH Levels: FSH levels in Group D2 were lower than in Group C (p = 0.009), whereas FSH levels in Group D5 were higher than in Group D2 (p = 0.028). T Levels: Serum T levels in Groups D2, D3, and D4 were higher than in Group C (p = 0.036, p = 0.047, and p = 0.016, respectively). T levels in Groups D3 and D4 were also elevated compared to Group D1 (p = 0.047 and p = 0.036, respectively). Inh-B Levels: Serum Inh-B levels in Groups D2, D3, D4, and D5 were higher than in Group C (p = 0.047, p = 0.009, p = 0.009, and p = 0.009, respectively). Inh-B levels in Groups D3, D4, and D5 were also elevated compared to Group D2 (p = 0.009 for all). Testicular Tissue Hormone Levels Significant changes in testosterone (T) and inhibin B (Inh-B) levels were observed in testicular tissue across all groups (p < 0.05, Table 2 ). Table 2 Hormone levels in testicular tissue samples C D1 D2 D3 D4 D5 p Median (IQR) Median (IQR) Median (IQR) Median (IQR) Median (IQR) Median (IQR) N = 5 N = 5 N = 5 N = 5 N = 5 N = 5 T 2307.30 (1907.90–2958.50) 2600.20 (2150.65–2925.90) a 2521.50 (2243.80–2726.40) b,c 1470.40 (744.55–2360.60) a,b,d,e 2779.35 (2431.52–3293.22) d 2769.90 (2726.40–2864.20) c,e 0.051 Inh B 0.006 (0.005–0.008) f,g,h,k 0.005 (0.002–0.011) m,n,p,q 0.038 (0.029–0.046) f,m,r,s,t 0.089 (0.054–0.096) g,n,r,u 0.093 (0.067–0.129) h,p,s,v 0.244 (0.199–0.292) k,q,t,u,v 0.000116* FSH, follicle stimulating hormone; LH, luteinizing hormone; T, testosterone; Inh-B, inhibin B; C, control group; D1, desflurane 1 group; D2, desflurane 2 group; D3, desflurane 3 group; D4, desflurane 4 group; D5, desflurane 5 group. *p < 0.05, Kruskal-Wallis test. All intergroup comparisons were performed by using Tukey’s HSD test. Comparisons for T levels: a D1-D3: p = 0.046; b D2-D3: p = 0.047; c D2-D5: p = 0.040; d D3-D4: p = 0.037; e D3-D5: p = 0.008. Significant differences in Inh-B levels: f C-D2: p = 0.009; g C-D3: p = 0.009; h C-D4: p = 0.014; k C-D5: p = 0.009; m D1-D2: p = 0.009; n D1-D3: p = 0.009; p D1-D4: p = 0.014; q D1-D5: p = 0.009; r D2-D3: p = 0.036; s D2-D4: p = 0.014; t D2-D5: p = 0.009; u D3-D5: p = 0.009; v D4-D5: p = 0.014. T Levels: Testicular T levels in Group D3 were higher than in Group D1 (p = 0.046) but lower than in Group D2 (p = 0.047). In contrast, T levels in Group D5 were higher than in Group D3 (p = 0.040). Additionally, T levels in Groups D4 and D5 were elevated compared to Group D3 (p = 0.037 and p = 0.008, respectively). Inh-B Levels: Testicular Inh-B levels in Groups D2, D3, D4, and D5 were higher than in Group C (p = 0.009, p = 0.009, p = 0.014, and p = 0.009, respectively). Inh-B levels in these groups were also elevated compared to Group D1 (p = 0.009, p = 0.009, p = 0.014, and p = 0.009, respectively). Furthermore, Inh-B levels in Groups D3, D4, and D5 were higher than in Group D2 (p = 0.036, p = 0.014, and p = 0.009, respectively). Inh-B levels in Group D5 were also elevated compared to Groups D3 and D4 (p = 0.009 and p = 0.014, respectively). Sperm Morphology Assessment Significant differences in sperm morphology were observed among all groups (p < 0.05, Table 3 ). The percentages of abnormal sperm in Groups D2, D3, D4, and D5 were higher than in Group C where Group D3 and D4 were showed significance (p = 0.030, p = 0.002, p = 0.0011, and p = 0.016, respectively). Additionally, the percentages of abnormal sperm in Groups D3 were raised in which D4 were significantly elevated compared to Group D1 (p = 0.017 and p = 0.0011, respectively). Table 3 Abnormal sperm morphology C D1 D2 D3 D4 D5 p (Mean ± SD) (Mean ± SD) (Mean ± SD) (Mean ± SD) (Mean ± SD) (Mean ± SD) N = 5 N = 5 N = 5 N = 5 N = 5 N = 5 PAS 5.66 (5.01–5.75) a,b,c,d 4.71 (4.67–5.14) e,f 6.91 (6.56–7.27) a 7.46 (7.03–7.48) b,e 8.80 (7.09–9.05) c,f 6.63 (6.49–7.48) d < 0.001* PAS, percentage of abnormal sperms; C, control group; D1, desflurane 1 group; D2, desflurane 2 group; D3, desflurane 3 group; D4, desflurane 4 group; D5, desflurane 5 group. *p < 0.05 Kruskal-Wallis test. All intergroup comparisons were performed by using Holm-Sidak test. Significant differences in intergroup comparisons: a C-D2: p = 0.030; b C-D3: p = 0.002; c C-D4: p < 0.001; d C-D5: p = 0.016; e D1-D3: p = 0.017; f D1-D4: p < 0.001. Discussion The findings of this study demonstrate that chronic exposure to desflurane induces testicular tissue injury and increases the percentage of morphologically abnormal sperm in rats. Furthermore, significant alterations were observed in the levels of reproductive hormones, including FSH, LH, T, and Inh-B, which correlated with the intensity and duration of desflurane exposure. Fertility, a cornerstone of life's continuity, is governed by a complex interplay of endocrine and paracrine signaling pathways. While the regulatory mechanisms underlying fertility have been extensively studied, emerging evidence suggests that additional factors influencing this process remain to be elucidated [ 20 ]. The delicate balance between fertility and infertility is influenced by various environmental and pharmacological factors [ 21 ]. Numerous studies have investigated the effects of chemicals and drugs on the male reproductive system, both in humans and animal models [ 9 – 11 , 22 , 23 ]. For instance, Ceyhan et al. reported that inhalational anesthetics such as sevoflurane and isoflurane can induce testicular damage in rabbits [ 9 ]. Similarly, Kaya et al. demonstrated that chronic exposure to sevoflurane adversely affects testicular tissue structure and disrupts the hypothalamic-pituitary-gonadal axis, leading to alterations in FSH, LH, T, and Inh-B levels [ 11 ]. Spermatogenesis, the process by which male gametes are produced, involves a series of tightly regulated stages, including proliferation, differentiation, and maturation. This process transforms round, immotile germ cells into elongated, motile spermatozoa capable of fertilization [ 1 , 24 , 25 ]. Spermatogenesis is regulated by the hypothalamic-pituitary-gonadal axis, wherein gonadotropin-releasing hormone from the hypothalamus stimulates the anterior pituitary to secrete FSH and LH. LH acts on Leydig cells to produce testosterone, which, along with FSH, stimulates Sertoli cells to support germ cell development [ 20 ]. Sertoli cells, often referred to as \"nurse cells,\" provide essential nutrients and metabolic intermediates, such as amino acids, carbohydrates, lipids, vitamins, and metal ions, to developing germ cells [ 26 ]. Additionally, Sertoli cells secrete inhibin B in response to FSH stimulation, which negatively regulates pituitary FSH secretion [ 27 ]. Thus, FSH, LH, testosterone, and inhibin B play critical roles in spermatogenesis and serve as key indicators of male reproductive health. Beyond the documented impact on the hypothalamic-pituitary-gonadal (HPG) axis, some studies have suggested that volatile anesthetics such as isoflurane and desflurane may also influence other endocrine axes. Isoflurane, for instance, has been associated with alterations in the hypothalamic-pituitary-adrenal (HPA) axis, potentially reducing ACTH and corticosterone levels in animal models due to suppressed synaptic transmission in the hypothalamus [ 23 , 26 , 27 ]. While data on desflurane are more limited, its structural and functional similarity to isoflurane raises the possibility of broader endocrine disruption [ 27 ]. These findings suggest that the observed hormonal changes may not be strictly confined to the gonadal axis but may reflect a more general downregulation of hypothalamic and pituitary function. However, the normalization of hormone levels after withdrawal supports the idea that these effects could be transient and reversible. Further research evaluating a wider range of hormones, including those of the thyroid and adrenal axes, is warranted to clarify whether the observed endocrine suppression is axis-specific or part of a more global functional decrease. The hormone-dependent nature of spermatogenesis makes it susceptible to disruption by external factors, including drugs and chemicals. For example, Xu et al. demonstrated that isoflurane exposure disrupts FSH, LH, and testosterone levels by inhibiting hypothalamic and pituitary function, likely through the suppression of synaptic transmission or nerve signaling [ 23 ]. In the present study, significant changes in FSH and LH levels were observed. While FSH levels decreased in Groups D1, D2, D3, and D4, only the reduction in Group D2 was statistically significant. Similarly, LH levels decreased significantly in Groups D2, D3, and D4, with the lowest levels observed in Group D3, which corresponded to the highest intensity of desflurane exposure. The lack of recovery in LH levels after a seven-day withdrawal period in Group D2 suggests that chronic desflurane exposure may impair hypothalamic or pituitary hormone production. However, the normalization of hormone levels during short- and long-term recovery periods indicates that the adverse effects of desflurane may be reversible. Unexpectedly, testosterone levels in both serum and testicular tissue exhibited an irregular pattern that did not correlate with the severity of desflurane exposure. This anomaly, coupled with the progressive increase in serum and testicular inhibin B levels, suggests that desflurane may influence testosterone and inhibin B through mechanisms involving sex hormone-binding globulin or albumin, although the exact pathway remains unclear. Sperm morphology serves as a sensitive indicator of disturbances in the male reproductive system. However, limited studies have explored the effects of inhalational anesthetics on sperm morphology [ 7 , 9 , 11 , 23 ]. Campion et al. reported that isoflurane exposure reduces sperm motility in rats [ 31 ], while Ceyhan et al. observed structural abnormalities in sperm following chronic exposure to sevoflurane or isoflurane [ 9 ]. In contrast, Kaya et al. found no significant changes in the percentage of abnormal sperm despite reductions in sperm concentration and motility following sevoflurane exposure [ 11 ]. In the current study, a significant increase in the percentage of abnormal sperm was observed in Groups D2, D3, D4, and D5 compared to the control group. This suggests that desflurane may disrupt the differentiation or maturation of spermatogonia, leading to abnormal sperm morphology. The reduction in abnormal sperm percentage in Group D5 compared to Group D4 indicates that a 14-day recovery period may be necessary to mitigate the structural impairments caused by desflurane exposure. The structural integrity of the testis is critical for normal reproductive function, and numerous factors can compromise testicular tissue. Xu et al. demonstrated that isoflurane exposure induces dose-dependent damage to seminiferous tubules, characterized by disorganized epithelium, sloughed germ cells, and interstitial edema [ 23 ]. Similarly, Kaya et al. reported progressive degenerative changes in seminiferous epithelium, including germ cell loss and interstitial edema, following chronic sevoflurane exposure [ 11 ]. In the present study, histopathological examination revealed significant testicular tissue damage in all experimental groups, with the most severe changes observed in Group D3. These changes included degenerative alterations in seminiferous epithelium, interstitial edema, and congestion, although no loss of spermatogenic cells was observed. The correlation between histopathological injury scores and hormonal changes in Group D3 further underscores the adverse effects of desflurane on testicular function. Additionally, the incomplete recovery of tissue damage after a seven-day withdrawal period in Groups D2 and D5 suggests that longer recovery periods may be required for tissue repair. Environmental chemical exposure is a growing public health concern, with infertility emerging as a significant consequence of exposure to toxicants [ 34 , 35 ]. The widespread use of inhalational anesthetics in medical settings has raised concerns about their potential reproductive toxicity. A meta-analysis highlighted an increased risk of spontaneous abortion among health care workers occupationally exposed to anesthetic gases [ 36 ]. Anesthesiologists, surgeons, and nurse anesthetists are particularly vulnerable to the adverse effects of waste anesthetic gases, which have been linked to disturbances in germ cell development, sperm morphology, and motility [ 11 , 23 , 28 , 37 ]. This study is the first to investigate the effects of chronic desflurane exposure on male reproductive hormones, testicular tissue, and sperm morphology in rats. The findings suggest that desflurane may induce testicular tissue damage, disrupt reproductive hormone levels, and impair sperm morphology. These results highlight the potential risks associated with occupational exposure to inhalational anesthetics among health care workers. There are some limitations for this study. First one is the limited duration of the follow-up period used to assess the reversibility of the observed effects. While changes in reproductive hormone levels, testicular histology, and sperm morphology were noted, the timeframe may not have been sufficient to determine whether these alterations are fully reversible after cessation of desflurane exposure. Longer-term studies are necessary to evaluate the potential for recovery of reproductive parameters over time. Additionally, as this study was conducted in an animal model, caution must be exercised when extrapolating the results directly to humans. Further research is needed to validate these findings in clinical settings and to establish safe exposure thresholds for health care professionals regularly exposed to inhalational anesthetics. Conclusion Chronic exposure to desflurane adversely affects testicular tissue integrity, sperm morphology, and the regulation of reproductive hormones in rats. These findings underscore the potential reproductive toxicity of inhalational anesthetics and emphasize the need for further research to elucidate the underlying mechanisms and mitigate the risks associated with occupational exposure. Declarations Availability of data and materials The data presented in this study are available on reasonable request from the corresponding author. Authors Contributions SD, HYD, HT, AA and MS—performed the research. SD and HYD—designed the research study. AA—performed the pathological analysis. SD, HYD, HT and MS—analyzed the data and wrote the draft of the manuscript. All authors read and approved the final manuscript. All authors were involved in the collection of experimental data. Ethics Approval and Consent to Participate This experiemental study was approved by Tokat Gaziosmanpasa University Animal Experimentations Local Ethics Committee (Grant number: 2013-HADYEK-12). Acknowledgements We thank to Erkut Somak, Yılmaz Ozcan, and Serkan Kavak for their efforts and contributions for the study. Funding This work was supported by Gaziosmanpasa University Scientific Research Projects Unit (Grant Number: 2013/94). Conflict of interest The authors report no declarations of interest. References Sun Y, Liu X, Wang J, et al. Air pollution exposure and infertility risk: evidence from a prospective cohort study. Environ Health Perspect. 2024;132(4):47001. Dai X, Liu G, Li F, et al. Individual and joint associations of air pollutants exposure with semen quality: a retrospective longitudinal study in Wenzhou, China. Int Arch Occup Environ Health. 2024;97(2):123-132. Azzam A, Karabulut R, Kaya C, Eryılmaz S, Kapisiz A, Turkyilmaz Z, Inan MA, Yaz Aydin G, Atan A, Sonmez K. Effects of lupeol on experimental testicular ischemiareperfusion damage in rats. Ulus Travma Acil Cerrahi Derg. 2025;31(2):95-102. Türkan H, Aydin A, Sayal A, Eken A, Akay C, Karahalil B. Oxidative and antioxidative effects of desflurane and sevoflurane on rat tissue in vivo. Arh Hig Rada Toksikol 2011; 62 113-119. Alcaraz M, Quesada S, Armero D, Martin-Gíl R, Olivares A, Achel GD. Genotoxicity and cytotoxicity of sevoflurane in two human cell lines in vitro with ionizing radiation. Colombia Medica (Cali) 2014; 45: 104-109. Bilban M, Jakopin CB, Ogrinc D. Cytogenetic tests performed on operating room personnel the use of anaesthetic gases. Int Arch Occup Environ Health 2005; 78: 60–64. Kaymak C, Kadioglu E, Coskun E, Basar H, Basar M. Determination of DNA damage after exposure to inhalation anesthetics in human peripheral lymphocytes and sperm cells in vitro by comet assay. Hum Exp Toxicol 2012; 31: 1207-1213. Prystupa J. Fluorine--a current literature review. An NRC and ATSDR based review of safety standards for exposure to fluorine and fluorides. Toxicol Mech Methods 2011; 21: 103-170. Ceyhan A, Cincik M, Bedir S, Ustun H, Dagli G, Kalender H. Effects of exposure to new inhalational anesthetics on spermatogenesis and sperm morphology in rabbits. Arch Androl 2005; 51: 305-315. Hoerauf K, Funk W, Harth M, Hobbhahn J. Occupational exposure to sevoflurane, halothane and nitrous oxide during paediatric anaesthesia. Waste gas exposure during paediatric anaesthesia. Anaesthesia 1997; 52: 215-219. Kaya Z, Sogut E, Cayli S, Suren M, Arici S, Karaman S, Erdemir F. Evaluation of effects of repeated sevoflurane exposure on rat testicular tissue and reproductive hormones. Inhal Toxicol 2013; 25: 192-198. Miller AL, Golledge HD, Leach MC. The Influence of Isoflurane Anaesthesia on the Rat Grimace Scale. PLoS One. 2016;11(11):e0166652. Brosnan RJ, Eger EI 2nd, Laster MJ, Sonner JM. Anesthetic properties of carbon dioxide in the rat. Anesth Analg. 2007; 105: 103-6. Hariyanto H, Widiastuti M, Pandrya CO, Surya KA, Audi W. Comparison of desflurane and sevoflurane as maintenance inhalational anaesthetic agents for adult patients undergoing neurosurgeries: A systematic review and meta-analysis of randomised trials. Indian J Anaesth. 2025;69(1):65-77. Karzai W, Haberstroh J, Müller W, Priebe HJ. Rapid increase in inspired desflurane concentration does not elicit a hyperdynamic circulatory response in the pig. Lab Anim 1997; 31: 279-282. Loan PB, Mirakhur RK, Paxton LD, Gaston JH. Comparison of desflurane and isoflurane in anaesthesia for dental surgery. Br J Anaesth 1995; 75: 289-292. Gupta A, Stierer T, Zuckerman R, Sakima N, Parker SD, Fleisher LA. Comparison of recovery profile after ambulatory anesthesia with propofol, isoflurane, sevoflurane and desflurane: a systematic review. Anesth Analg 2004; 98: 632-641. Cosentino MJ, Nishida M, Rabinowitz R, Cockett AT. Histopathology of prepubertal rat testes subjected to various durations of spermatic cord torsion. J Androl 1986; 7: 23-31. Rezvanfar M, Sadrkhanlou R, Ahmadi A, Shojaei-Sadee H, Rezvanfar M, Mohammadirad A, Salehnia A, Abdollahi M. Protection of cyclophosphamide-induced toxicity in reproductive tract histology, sperm characteristics, and DNA damage by an herbal source; evidence for role of free-radical toxic stress. Hum Exp Toxicol 2008; 27: 901-910. Ghewade P, Vagha S, Ghewade B, Gadkari P. Role of Dietary Antioxidant Supplements in Male Infertility: A Review. Cureus. 2024;16(6):e61951. Montano L, Rinaldo D, De Felice B, et al. Microplastics found in human ovary follicular fluid for the first time. Ecotoxicol Environ Saf. 2025;250:114512. Liu R, Dai J, Li J, et al. Does air pollution exposure affect semen quality? Evidence from a systematic review and meta-analysis of 93,996 Chinese men. Front Public Health. 2023;11:1219340. Xu XL, Pan C, Hu JX, Liu XT, Li YF, Wang H, Chen YB, Dong HY, Dai TJ, Xu LC. Effects of isoflurane inhalation on the male reproductive system in rats. Environ Toxicol Pharmacol 2012; 34: 688-693. Young CJ, Apfelbaum JL. Inhalational anesthetics: desflurane and sevoflurane. J Clin Anesth. 2023;45:564-577. Zhou D, Liu B, Liu L, Liu G, Zhu F, Huang Z, Zhang S, He Z, Fan L. Essential Regulation of Spermatogonial Stem Cell Fate Decisions and Male Fertility by APBB1 via Interaction with KAT5 and GDF15 in Humans and Mice. Research (Wash D C). 2025;8:0647. Ma R, Cui Y, Yu SJ, Pan YY, He JF, Wang YY, Wang JL, Wang XY, Bai XF, Zhang H, Yang SS, Zhang Q. The glucose metabolism reprogramming of yak Sertoli cells under hypoxia is regulated by autophagy. BMC Genomics. 2025;26(1):385. Zhang Y, Zhao J, Wu W, Zhou C, Liu Y. Sevoflurane exposure impairs testicular function and fertility in neonatal rats via oxidative stress. Reprod Toxicol. 2023;116:48-55. Kim H, Lee J, Park Y, Choi S. Chronic occupational exposure to isoflurane induces oxidative stress and impairs sperm parameters in mice. Toxicol Appl Pharmacol. 2024;473:116612. Vincent KF, Mallari OG, Dillon EJ, Stewart VG, Cho AJ, Dong Y, Edlow AG, Ichinose F, Xie Z, Solt K. Oestrous cycle affects emergence from anaesthesia with dexmedetomidine, but not propofol, isoflurane, or sevoflurane, in female rats. Br J Anaesth. 2023;131(1):67-78. Khan S, Huang Y, Timuçin D, Bailey S, Lee S, Lopes J, Gaunce E, Mosberger J, Zhan M, Abdelrahman B, Zeng X, Wiest MC. Microtubule-Stabilizer Epothilone B Delays Anesthetic-Induced Unconsciousness in Rats. eNeuro. 2024;11(8):ENEURO.0291-24.2024. Campion SN, Cappon GD, Chapin RE, Jamon RT, Winton TR, Nowland WS. Isoflurane reduces motile sperm counts in the Sprague-Dawley rat. Drug Chem Toxicol 2012; 35: 20-24. Chen Y, Li Z, Wang H, et al. Gaseous pollutant exposure affects semen quality in central China: a cross-sectional study. Environ Sci Pollut Res Int. 2023;30(5):12345-12354. Liu J, Fang Z, Chai D, Zhu Z, Shen Q, He X. Ambient Air Pollution and Semen Quality in China: A Nationwide Case-Control Study of 27,014 Males with Biomarker-Confirmed Semen Pathology. Toxics. 2025;13(4):322. Wang X, Zhang Y, Liu Y, et al. Association between ambient PM1 and semen quality: a cross-sectional study of 27,854 men in China. Environ Int. 2023;172:107754. Amato-Lourenço LF, Carvalho-Oliveira R, Júnior GR, et al. Detection and characterization of microplastics in the human testis and semen. Sci Total Environ. 2023;857:159712. Margiana R, Odhar HA, Prasad K, Oghenemaro EF, M M R, Kumawat R, Uthirapathy S, Sharma S, Kumar MR, Nouri M. Does outdoor air pollution cause poor semen quality? A systematic review and meta-analysis. BMC Urol. 2025;25(1):50. Zhou T, Li J, Cheng A, Zuo Z. Desflurane Post-treatment Reduces Hypoxic-ischemic Brain Injury via Reducing Transient Receptor Potential Ankyrin 1 in Neonatal Rats. Neuroscience. 2023;522:121-131. Additional Declarations No competing interests reported. Cite Share Download PDF Status: Published Journal Publication published 06 Feb, 2026 Read the published version in BMC Anesthesiology → Version 1 posted Editorial decision: Revision requested 08 Oct, 2025 Reviews received at journal 06 Oct, 2025 Reviews received at journal 01 Oct, 2025 Reviews received at journal 30 Sep, 2025 Reviewers agreed at journal 29 Sep, 2025 Reviewers agreed at journal 25 Sep, 2025 Reviewers agreed at journal 21 Sep, 2025 Reviewers invited by journal 20 Sep, 2025 Editor invited by journal 18 Sep, 2025 Editor assigned by journal 16 Sep, 2025 Submission checks completed at journal 16 Sep, 2025 First submitted to journal 01 Sep, 2025 You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. 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08:11:05\",\"extension\":\"html\",\"order_by\":18,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"acdc-reference\",\"size\":143925,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"earlyproof.html\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-7507144/v1/7fcbb2c38987919880bcae24.html\"},{\"id\":92574244,\"identity\":\"d407238e-6ec6-4c9b-81db-e21248efe6df\",\"added_by\":\"auto\",\"created_at\":\"2025-10-01 08:11:05\",\"extension\":\"jpg\",\"order_by\":1,\"title\":\"Figure 1\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":17945845,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eHistological sections\\u003c/p\\u003e\\n\\u003cp\\u003eHistological sections of rat testis tissues staining by H\\u0026amp;E. (A) Control; (B) D1, arrow shows mild degenerative changes were observed in focal areas of the seminiferous tubule epithelium; (C) D2, arrows show mild to moderate degenerative changes in the seminiferous tubule epithelium, accompanied by mild edema and congestion in the interstitial space; (D) D3, arrows show pronounced degenerative changes in the seminiferous tubule epithelium, along with severe edema and congestion in the interstitial space; (E) D4, arrows show moderate degeneration of the seminiferous tubule epithelium and interstitial edema and evident congestion; (F) D5, arrow shows slight degenerative changes in the seminiferous tubule epithelium. Five animals were included in each group, and six testis sections from each of five animals per group were assessed (n = 30). Scale bars: 50 μm. Magnification: 100×\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Figure1.jpg\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-7507144/v1/f15c8497196e174d095d6c19.jpg\"},{\"id\":92574232,\"identity\":\"ed570267-c091-4202-a8f4-b78cc9fb57c6\",\"added_by\":\"auto\",\"created_at\":\"2025-10-01 08:11:04\",\"extension\":\"jpg\",\"order_by\":2,\"title\":\"Figure 2\",\"display\":\"\",\"copyAsset\":false,\"role\":\"figure\",\"size\":965498,\"visible\":true,\"origin\":\"\",\"legend\":\"\\u003cp\\u003eHistological injury scores\\u003c/p\\u003e\\n\\u003cp\\u003eA four-level grading scale described by Cosentino et al., which ranged from 1 to 4 due to the injury severity, were used. The intergroup comparisons were performed by using Mann-Whitney U test. Each group contained five rats.\\u003c/p\\u003e\\n\\u003cp\\u003eP\\u0026lt;0.0033 (after Bonferroni correction)\\u003c/p\\u003e\\n\\u003cp\\u003eInterquartile range (IQR) for groups:\\u003c/p\\u003e\\n\\u003cp\\u003eC: IQR (1 – 1); D1: IQR (1.5 – 2); D2: IQR (2 – 3); D3: IQR (3 – 3.5); D4: IQR (2 – 3); D5: IQR (2 – 2.5)\\u003c/p\\u003e\\n\\u003cp\\u003eComparisons between groups (no significance):\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003csup\\u003ea\\u003c/sup\\u003eC-D1: p = 0.014; \\u003csup\\u003eb\\u003c/sup\\u003eC-D2: p = 0.005; \\u003csup\\u003ec\\u003c/sup\\u003eC-D3: p = 0.004; \\u003csup\\u003ed\\u003c/sup\\u003eC-D4: p = 0.005; \\u003csup\\u003ee\\u003c/sup\\u003eC-D5: p = 0.004; \\u003csup\\u003ef\\u003c/sup\\u003eD1-D2: p = 0.222; \\u003csup\\u003eg\\u003c/sup\\u003eD1-D3: p = 0.005; \\u003csup\\u003eh\\u003c/sup\\u003eD1-D4: p = 0.042; \\u003csup\\u003ei\\u003c/sup\\u003eD1-D5: p = 0.421; \\u003csup\\u003ej\\u003c/sup\\u003eD2-D3: p = 0.042; \\u003csup\\u003ek\\u003c/sup\\u003eD2-D4: p = 0.690; \\u003csup\\u003em\\u003c/sup\\u003eD2-D5: p = 0.690; \\u003csup\\u003en\\u003c/sup\\u003eD3-D4: p = 0.222; \\u003csup\\u003eo\\u003c/sup\\u003eD3-D5: p = 0.015; \\u003csup\\u003ep\\u003c/sup\\u003eD4-D5: p = 0.310.\\u003c/p\\u003e\",\"description\":\"\",\"filename\":\"Figure2.jpg\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-7507144/v1/ffa76622e27a924086af6632.jpg\"},{\"id\":102234838,\"identity\":\"5d7983db-aa87-4f97-9320-5a6bacf18b79\",\"added_by\":\"auto\",\"created_at\":\"2026-02-09 16:13:36\",\"extension\":\"pdf\",\"order_by\":0,\"title\":\"\",\"display\":\"\",\"copyAsset\":false,\"role\":\"manuscript-pdf\",\"size\":19753091,\"visible\":true,\"origin\":\"\",\"legend\":\"\",\"description\":\"\",\"filename\":\"manuscript.pdf\",\"url\":\"https://assets-eu.researchsquare.com/files/rs-7507144/v1/11b55e48-6eba-4cf0-ba56-a6d4d54f221b.pdf\"}],\"financialInterests\":\"No competing interests reported.\",\"formattedTitle\":\"The effects of desflurane on male rat reproductive hormones, testicular tissue, and sperm morphology: An experimental study\",\"fulltext\":[{\"header\":\"Introduction\",\"content\":\"\\u003cp\\u003eMale fertility is closely linked to the production of normal spermatozoa, which is the result of a highly regulated and complex process known as spermatogenesis. This process encompasses three key phases: mitosis, meiosis, and spermiogenesis [\\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e1\\u003c/span\\u003e]. Spermatogenesis is tightly regulated by a delicate balance of endocrine and paracrine signaling pathways. Follicle-stimulating hormone (FSH) and luteinizing hormone (LH), secreted by the pituitary gland, play pivotal roles in this regulation. FSH acts on Sertoli cells, while LH stimulates Leydig cells, both of which provide essential factors necessary for the proper progression of spermatogenesis [\\u003cspan citationid=\\\"CR2\\\" class=\\\"CitationRef\\\"\\u003e2\\u003c/span\\u003e]. The intricate network of signals governing spermatogenesis remains an active area of research. Importantly, external environmental factors have the potential to disrupt this process, leading to impaired fertility and abnormalities in sperm production [\\u003cspan citationid=\\\"CR3\\\" class=\\\"CitationRef\\\"\\u003e3\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e4\\u003c/span\\u003e].\\u003c/p\\u003e\\u003cp\\u003eWith the rapid advancements in medical technology, the frequency of surgical procedures has increased significantly in recent years. This has inevitably led to a corresponding rise in the use of volatile anesthetic agents. However, the routine administration of inhalation anesthetics has been associated with various adverse effects on multiple organ systems, including the liver, kidneys, lungs, and nervous system, as documented in both experimental and clinical studies [\\u003cspan additionalcitationids=\\\"CR5 CR6 CR7\\\" citationid=\\\"CR4\\\" class=\\\"CitationRef\\\"\\u003e4\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR8\\\" class=\\\"CitationRef\\\"\\u003e8\\u003c/span\\u003e]. Of particular concern is the evidence suggesting that repeated exposure to volatile anesthetics may induce DNA damage in germ cells, contribute to congenital anomalies in offspring, and cause morphological alterations in sperm cells [\\u003cspan additionalcitationids=\\\"CR10\\\" citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e]. Despite these findings, the potential effects of modern inhalation anesthetics on the male reproductive system remain underexplored.\\u003c/p\\u003e\\u003cp\\u003eDesflurane (2-difluoromethoxy-1,1,1,2-tetrafluoroethane; C₃H₂F₆O) is a halogenated, fluorinated ether that is colorless, non-flammable, and liquid at room temperature. It has been widely used in clinical practice for the maintenance of general anesthesia since its introduction in 1989 [\\u003cspan additionalcitationids=\\\"CR13\\\" citationid=\\\"CR12\\\" class=\\\"CitationRef\\\"\\u003e12\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e14\\u003c/span\\u003e]. Desflurane is characterized by several advantageous pharmacological properties, including minimal metabolism (0.02%) and a low blood/gas partition coefficient (0.42), which contribute to its rapid induction and recovery profiles [\\u003cspan citationid=\\\"CR15\\\" class=\\\"CitationRef\\\"\\u003e15\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR16\\\" class=\\\"CitationRef\\\"\\u003e16\\u003c/span\\u003e]. Additionally, desflurane anesthesia has been associated with reduced postanesthesia care unit stay durations, facilitating earlier patient discharge [\\u003cspan citationid=\\\"CR14\\\" class=\\\"CitationRef\\\"\\u003e14\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR17\\\" class=\\\"CitationRef\\\"\\u003e17\\u003c/span\\u003e]. Despite its widespread use, a systematic review of the literature reveals a lack of experimental studies investigating the potential effects of desflurane on the male reproductive system.\\u003c/p\\u003e\\u003cp\\u003eGiven this gap in knowledge, the present study was designed to investigate the effects of repeated desflurane exposure on sperm morphology, testicular tissue integrity, and reproductive hormone levels in a rat model.\\u003c/p\\u003e\"},{\"header\":\"Materials and Methods\",\"content\":\"\\u003cdiv id=\\\"Sec3\\\" class=\\\"Section2\\\"\\u003e\\u003ch2\\u003eChemicals\\u003c/h2\\u003e\\u003cp\\u003eKetamine and xylazine were procured from Alfasan International B.V. (Woerden, Netherlands). Hematoxylin Harris and Eosin Y (1% alcoholic) were obtained from Atom Scientific Ltd (Manchester, UK). Modified Human Tubal Fluid Medium was purchased from Irvine Scientific (California, USA). Bouin's solution was sourced from Tek-Path Medikal (Izmir, Turkey).\\u003c/p\\u003e\\u003c/div\\u003e\\n\\u003ch3\\u003eAnimals\\u003c/h3\\u003e\\n\\u003cp\\u003e The study was conducted following approval by the Tokat Gaziosmanpasa University Local Ethics Committee for Animal Experimentation (Approval No: 2013-HADYEK-12, Approval date: 29.02.2013). A total of 30 adult male Wistar-Albino rats (90 days old, 250\\u0026ndash;300 g) were obtained from the Experimental Medicine Unit at Gaziosmanpasa University, Turkey. The animals were housed under controlled conditions, including a 12-hour light-dark cycle (lights on at 07:00 a.m.), room temperature maintained at 20\\u0026ndash;24\\u0026deg;C, and relative humidity of 40\\u0026ndash;50%. Rats were housed in polycarbonate cages with ad libitum access to tap water.\\u003c/p\\u003e\\u003cp\\u003eA glass anesthesia chamber (40 \\u0026times; 50 \\u0026times; 60 cm) was used for desflurane exposure. The chamber was equipped with an anesthetic gas inlet at the top left and an outlet at the top right, as previously described in the literature [\\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e]. An airtight connection was established between the chamber and the anesthetic circuit using an anesthesia machine (Prima SP Alpa, Penlon Limited, Oxon, UK). The exposure dose of 1 MAC desflurane administered with 2 L/min of O₂ for 18 minutes daily was selected based on previous studies investigating occupational exposure levels in controlled experimental settings [\\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e]. This dosing protocol was designed to simulate the cumulative low-dose exposure experienced by health care workers during repeated, short-term contact with inhalational anesthetics in poorly ventilated or inadequately scavenged operating room environments, rather than the higher, continuous exposure a patient would receive during a surgical procedure. The duration and concentration were chosen to reflect a realistic occupational risk model while minimizing excessive stress or toxicity in the animal subjects.\\u003c/p\\u003e\\u003cp\\u003eThe rats were randomly divided into six groups (n\\u0026thinsp;=\\u0026thinsp;5 per group):\\u003c/p\\u003e\\u003cp\\u003e\\u003cul\\u003e\\u003cli\\u003e\\u003cp\\u003eControl group (C): Administered 2 L/min of O₂ for 18 minutes daily over seven days.\\u003c/p\\u003e\\u003c/li\\u003e\\u003cli\\u003e\\u003cp\\u003eGroup D1: Exposed to 1 minimum alveolar concentration (MAC) of desflurane and 2 L/min of O₂ for 18 minutes daily over seven days.\\u003c/p\\u003e\\u003c/li\\u003e\\u003cli\\u003e\\u003cp\\u003eGroup D2: Received the same treatment as Group D1 for seven days, followed by a seven-day recovery period without intervention.\\u003c/p\\u003e\\u003c/li\\u003e\\u003cli\\u003e\\u003cp\\u003eGroup D3: Administered 1 MAC desflurane and 2 L/min of O₂ for 18 minutes daily over 14 days.\\u003c/p\\u003e\\u003c/li\\u003e\\u003cli\\u003e\\u003cp\\u003eGroup D4: Received the same treatment as Group D3 for 14 days, followed by a seven-day recovery period without intervention.\\u003c/p\\u003e\\u003c/li\\u003e\\u003cli\\u003e\\u003cp\\u003eGroup D5: Administered the same treatment as Group D3 for 14 days, followed by a 14-day recovery period without intervention.\\u003c/p\\u003e\\u003c/li\\u003e\\u003c/ul\\u003e\\u003c/p\\u003e\\u003cp\\u003eAt the end of the experimental periods, euthanasia was performed via cervical dislocation under anesthesia induced by intraperitoneal injection of ketamine (90 mg/kg) and xylazine (10 mg/kg). Intracardiac blood samples (5 mL) were collected immediately for hormonal analysis, and bilateral testes were dissected. The right testis was fixed in Bouin's solution for 24 hours, dehydrated, and embedded in paraffin blocks for histopathological examination. The left testis was placed on ice and stored at \\u0026minus;\\u0026thinsp;70\\u0026deg;C for subsequent biochemical analyses.\\u003c/p\\u003e\\n\\u003ch3\\u003eHistopathological Analysis of Testicular Tissue\\u003c/h3\\u003e\\n\\u003cp\\u003eParaffin-embedded testicular tissues were sectioned at 5 \\u0026micro;m thickness, mounted on glass slides, and stained with hematoxylin and eosin (H\\u0026amp;E) for light microscopic examination (Nikon Eclipse E600W, Tokyo, Japan). Six sections from each testis were randomly numbered and evaluated by a blinded investigator. At least five microscopic fields per testis were assessed. Histopathological injury scores were determined using a four-level grading scale as described by Cosentino et al. [\\u003cspan citationid=\\\"CR18\\\" class=\\\"CitationRef\\\"\\u003e18\\u003c/span\\u003e]:\\u003c/p\\u003e\\u003cp\\u003e\\u003cul\\u003e\\u003cli\\u003e\\u003cp\\u003eGrade 1: Normal testicular architecture with orderly arrangement of germinal cells.\\u003c/p\\u003e\\u003c/li\\u003e\\u003cli\\u003e\\u003cp\\u003eGrade 2: Slight disorganization and non-cohesive germinal cells with closely spaced seminiferous tubules.\\u003c/p\\u003e\\u003c/li\\u003e\\u003cli\\u003e\\u003cp\\u003eGrade 3: Moderate disorganization, sloughed germinal cells, shrunken pyknotic nuclei, and less distinct seminiferous tubule borders.\\u003c/p\\u003e\\u003c/li\\u003e\\u003cli\\u003e\\u003cp\\u003eGrade 4: Severe disorganization, tightly packed seminiferous tubules, and coagulative necrosis of germinal cells.\\u003c/p\\u003e\\u003c/li\\u003e\\u003c/ul\\u003e\\u003c/p\\u003e\\n\\u003ch3\\u003eSperm Morphology Analysis\\u003c/h3\\u003e\\n\\u003cp\\u003eEpididymal sperm was collected by mincing the cauda epididymis in 2 mL of Ham's F10 medium. A drop of sperm suspension was placed on a microscope slide, air-dried, and examined under a light microscope at 400\\u0026times; magnification. Five random fields were assessed for morphological abnormalities, including bicephalic, amorphous, hook-less, and tail abnormalities (coiled or abnormal tails). The percentage of abnormal sperm was calculated and analyzed [\\u003cspan citationid=\\\"CR19\\\" class=\\\"CitationRef\\\"\\u003e19\\u003c/span\\u003e].\\u003c/p\\u003e\\n\\u003ch3\\u003eBiochemical Analysis of Reproductive Hormones\\u003c/h3\\u003e\\n\\u003cp\\u003eBlood samples were allowed to clot for 20 minutes, centrifuged at 1500 \\u0026times; g (4\\u0026deg;C) for 15 minutes, and the serum was aliquoted into Eppendorf tubes. Samples were stored at \\u0026minus;\\u0026thinsp;20\\u0026deg;C until analysis.\\u003c/p\\u003e\\u003cp\\u003eFor testicular tissue analysis, tissues were weighed and homogenized in ice-cold Tris-HCl buffer (50 mM, pH 7.4) containing 0.50 mL/L Triton X-100 using an IKA Ultra-Turrax T25 Basic homogenizer (Stanfen, Germany) at 13,000 rpm for 2 minutes. All procedures were performed at 4\\u0026deg;C.\\u003c/p\\u003e\\u003cp\\u003eSerum levels of LH, FSH, testosterone (T), and inhibin B (Inh-B) were measured using rat-specific ELISA kits (LH and FSH: YH Biosearch Laboratory, Shanghai, China; T: Cayman Chemical Company, Michigan, USA; Inh-B: SunRed Biotechnology Company, Shanghai, China) according to the manufacturers' instructions. Hormone concentrations were expressed as mIU/mL for LH and FSH, pg/mL for T, and ng/mL for Inh-B.\\u003c/p\\u003e\\u003cdiv id=\\\"Sec8\\\" class=\\\"Section2\\\"\\u003e\\u003ch2\\u003eStatistical Analysis\\u003c/h2\\u003e\\u003cp\\u003eData normality and variance were assessed using the Kolmogorov-Smirnov test. Quantitative data are presented as mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;standard deviation, while qualitative data are expressed as frequencies and percentages. Parametric or nonparametric tests were applied as appropriate. Histopathological injury scores and hormonal levels were analyzed using the Kruskal-Wallis test, with post-hoc comparisons performed using the Mann-Whitney U test and Holm-Sidak test. Bonferroni correction was applied to all multiple comparisons. All statistical analyses were conducted using SPSS version 20.0 (SPSS Inc., Chicago, IL, USA), with statistical significance set at p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05.\\u003c/p\\u003e\\u003c/div\\u003e\"},{\"header\":\"Results\",\"content\":\"\\u003cdiv id=\\\"Sec10\\\" class=\\\"Section2\\\"\\u003e\\u003ch2\\u003eHistopathological Findings\\u003c/h2\\u003e\\u003cp\\u003eThe effects of desflurane on testicular tissue were evaluated using H\\u0026amp;E staining. Histopathological findings are illustrated in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e, and the corresponding injury scores, graded on a scale of 1 to 4, are presented in Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e. Significant differences in histological injury scores were observed between the control group (C) and all experimental groups (p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05). Furthermore, the mean histological injury scores in Groups D3 and D4 were higher than in Group D1 (p\\u0026thinsp;=\\u0026thinsp;0.005 and p\\u0026thinsp;=\\u0026thinsp;0.042, respectively). Group D3 also exhibited higher injury scores compared to Group D2 (p\\u0026thinsp;=\\u0026thinsp;0.042), while Group D5 demonstrated lower scores than Group D3 (p\\u0026thinsp;=\\u0026thinsp;0.015).\\u003c/p\\u003e\\u003cp\\u003eIn Group D1, mild degenerative changes were observed in focal areas of the seminiferous tubule epithelium (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eB), whereas Group C displayed normal testicular architecture with orderly arranged germinal cells (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eA). Group D2 exhibited mild to moderate degenerative changes in the seminiferous tubule epithelium, accompanied by mild edema and congestion in the interstitial space (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eC). Group D3 showed pronounced degenerative changes in the seminiferous tubule epithelium, along with severe edema and congestion in the interstitial space, although no loss of spermatogenic cells was observed (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eD). In Group D4, moderate degeneration of the seminiferous tubule epithelium and interstitial edema and congestion were evident (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eE). Group D5 displayed slight degenerative changes in the seminiferous tubule epithelium (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eF).\\u003c/p\\u003e\\u003cp\\u003eLight microscopic analysis revealed that chronic inhalational exposure to desflurane induced structural impairments in the seminiferous tubule epithelium and caused interstitial edema and congestion (Fig.\\u0026nbsp;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eA\\u0026ndash;\\u003cspan refid=\\\"Fig1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003eF).\\u003c/p\\u003e\\u003c/div\\u003e\\u003cdiv id=\\\"Sec11\\\" class=\\\"Section2\\\"\\u003e\\u003ch2\\u003eBiochemical Analyses\\u003c/h2\\u003e\\u003cp\\u003eSignificant differences in serum levels of FSH, LH, T, and Inh-B were observed across all groups (p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05, Table\\u0026nbsp;\\u003cspan refid=\\\"Tab1\\\" class=\\\"InternalRef\\\"\\u003e1\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cp\\u003e\\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab1\\\" border=\\\"1\\\"\\u003e\\u003ccaption language=\\\"En\\\"\\u003e\\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 1\\u003c/div\\u003e\\u003cdiv class=\\\"CaptionContent\\\"\\u003e\\u003cp\\u003eHormone levels in serum samples\\u003c/p\\u003e\\u003c/div\\u003e\\u003c/caption\\u003e\\u003ccolgroup cols=\\\"8\\\"\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c7\\\" colnum=\\\"7\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c8\\\" colnum=\\\"8\\\"\\u003e\\u003c/div\\u003e\\u003cthead\\u003e\\u003ctr\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003eC\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003eD1\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003eD2\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003eD3\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003eD4\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003eD5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c8\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e\\u003cp\\u003ep\\u003c/p\\u003e\\u003c/th\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003eMedian (IQR)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003eMedian (IQR)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003eMedian (IQR)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003eMedian (IQR)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003eMedian (IQR)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003eMedian (IQR)\\u003c/p\\u003e\\u003c/th\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u0026nbsp;\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c8\\\"\\u003e\\u0026nbsp;\\u003c/th\\u003e\\u003c/tr\\u003e\\u003c/thead\\u003e\\u003ctbody\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e\\u003cb\\u003eFSH\\u003c/b\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e9.94 (9.905\\u0026ndash;10.125)\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e9.8 (9.445\\u0026ndash;10.090)\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e9.72 (9.560\\u0026ndash;9.805)\\u003csup\\u003ea,b\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e9.54 (9.315\\u0026ndash;10.145)\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e9.92 (9.680\\u0026ndash;9.965)\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e10.36 (9.820\\u0026ndash;10.490)\\u003csup\\u003eb\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c8\\\"\\u003e\\u003cp\\u003e0.128\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e\\u003cb\\u003eLH\\u003c/b\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e6.09 (5.935\\u0026ndash;6.260)\\u003csup\\u003ec,d,e\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e6.11 (5.855\\u0026ndash;6.160)\\u003csup\\u003ef,g\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e5.76 (5.635\\u0026ndash;5.880)\\u003csup\\u003ec,f\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e5.64 (5.355\\u0026ndash;5.815)\\u003csup\\u003ed,g\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e5.91 (5.535\\u0026ndash;5.930)\\u003csup\\u003ee\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e5.96 (5.665\\u0026ndash;6.120)\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c8\\\"\\u003e\\u003cp\\u003e\\u003cb\\u003e0.028*\\u003c/b\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e\\u003cb\\u003eT\\u003c/b\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e325.80 (176.55\\u0026ndash;573.80)\\u003csup\\u003eh,k,m\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e531.10 (475.55\\u0026ndash;622.20)\\u003csup\\u003en,p\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e721.40 (517.10\\u0026ndash;1018.45)\\u003csup\\u003eh\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e668.20 (588.65\\u0026ndash;765.80)\\u003csup\\u003ek,n\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e906.30 (612.55\\u0026ndash;1003.95)\\u003csup\\u003em,p\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e609.80 (554\\u0026ndash;710.75)\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c8\\\"\\u003e\\u003cp\\u003e\\u003cb\\u003e0.025*\\u003c/b\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e\\u003cb\\u003eInh-B\\u003c/b\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.389 (0.330\\u0026ndash;0.476)\\u003csup\\u003eq,r,s,t\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e0.479 (0.467\\u0026ndash;0.494)\\u003csup\\u003eu,v,w\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e0.485 (0.472\\u0026ndash;0.544)\\u003csup\\u003eq,y,z,aa\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e0.663 (0.637\\u0026ndash;0.818)\\u003csup\\u003er,u,y\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e0.714 (0.685\\u0026ndash;0.831)\\u003csup\\u003es,v,z\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e0.756 (0.682\\u0026ndash;0.805)\\u003csup\\u003et,w,aa\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c8\\\"\\u003e\\u003cp\\u003e\\u003cb\\u003e0.001*\\u003c/b\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tbody\\u003e\\u003c/colgroup\\u003e\\u003ctfoot\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003eFSH, follicle stimulating hormone; LH, luteinizing hormone; T, testosterone; Inh-B, inhibin B; C, control group; D1, desflurane 1 group; D2, desflurane 2 group; D3, desflurane 3 group; D4, desflurane 4 group; D5, desflurane 5 group.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003e*p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003eKruskal-Wallis test. All intergroup comparisons were performed by using Tukey\\u0026rsquo;s HSD test.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003eSignificant differences in FSH levels:\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003e\\u003csup\\u003ea\\u003c/sup\\u003eC-D2: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003eb\\u003c/sup\\u003eD2-D5: p\\u0026thinsp;=\\u0026thinsp;0.028.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003eSignificant differences in LH levels:\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003e\\u003csup\\u003ec\\u003c/sup\\u003eC-D2: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003ed\\u003c/sup\\u003eC-D3: p\\u0026thinsp;=\\u0026thinsp;0.028; \\u003csup\\u003ee\\u003c/sup\\u003eC-D4: p\\u0026thinsp;=\\u0026thinsp;0.028; \\u003csup\\u003ef\\u003c/sup\\u003eD1-D2: p\\u0026thinsp;=\\u0026thinsp;0.028; \\u003csup\\u003eg\\u003c/sup\\u003eD1-D3: p\\u0026thinsp;=\\u0026thinsp;0.028.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003eSignificant differences in T levels:\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003e\\u003csup\\u003eh\\u003c/sup\\u003eC-D2: p\\u0026thinsp;=\\u0026thinsp;0.036; \\u003csup\\u003ek\\u003c/sup\\u003eC-D3: p\\u0026thinsp;=\\u0026thinsp;0.047; \\u003csup\\u003em\\u003c/sup\\u003eC-D4: p\\u0026thinsp;=\\u0026thinsp;0.016; \\u003csup\\u003en\\u003c/sup\\u003eD1-D3: p\\u0026thinsp;=\\u0026thinsp;0.047; \\u003csup\\u003ep\\u003c/sup\\u003eD1-D4: p\\u0026thinsp;=\\u0026thinsp;0.036.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003eSignificant differences in Inh-B levels:\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003e\\u003csup\\u003eq\\u003c/sup\\u003eC-D2: p\\u0026thinsp;=\\u0026thinsp;0.047; \\u003csup\\u003er\\u003c/sup\\u003eC-D3: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003es\\u003c/sup\\u003eC-D4: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003et\\u003c/sup\\u003eC-D5: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003eu\\u003c/sup\\u003eD1-D3: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003ev\\u003c/sup\\u003eD1-D4: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003ew\\u003c/sup\\u003eD1-D5: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003ey\\u003c/sup\\u003eD2-D3: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003ez\\u003c/sup\\u003eD2-D4: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003eaa\\u003c/sup\\u003eD2-D5: p\\u0026thinsp;=\\u0026thinsp;0.009.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tfoot\\u003e\\u003c/table\\u003e\\u003c/div\\u003e\\u003c/p\\u003e\\u003cp\\u003eLH Levels: Serum LH levels in Groups D2, D3, and D4 were lower than in Group C (p\\u0026thinsp;=\\u0026thinsp;0.009, p\\u0026thinsp;=\\u0026thinsp;0.028, and p\\u0026thinsp;=\\u0026thinsp;0.028, respectively). LH levels in Groups D2 and D3 were also lower than in Group D1 (p\\u0026thinsp;=\\u0026thinsp;0.028 for both).\\u003c/p\\u003e\\u003cp\\u003eFSH Levels: FSH levels in Group D2 were lower than in Group C (p\\u0026thinsp;=\\u0026thinsp;0.009), whereas FSH levels in Group D5 were higher than in Group D2 (p\\u0026thinsp;=\\u0026thinsp;0.028).\\u003c/p\\u003e\\u003cp\\u003eT Levels: Serum T levels in Groups D2, D3, and D4 were higher than in Group C (p\\u0026thinsp;=\\u0026thinsp;0.036, p\\u0026thinsp;=\\u0026thinsp;0.047, and p\\u0026thinsp;=\\u0026thinsp;0.016, respectively). T levels in Groups D3 and D4 were also elevated compared to Group D1 (p\\u0026thinsp;=\\u0026thinsp;0.047 and p\\u0026thinsp;=\\u0026thinsp;0.036, respectively).\\u003c/p\\u003e\\u003cp\\u003eInh-B Levels: Serum Inh-B levels in Groups D2, D3, D4, and D5 were higher than in Group C (p\\u0026thinsp;=\\u0026thinsp;0.047, p\\u0026thinsp;=\\u0026thinsp;0.009, p\\u0026thinsp;=\\u0026thinsp;0.009, and p\\u0026thinsp;=\\u0026thinsp;0.009, respectively). Inh-B levels in Groups D3, D4, and D5 were also elevated compared to Group D2 (p\\u0026thinsp;=\\u0026thinsp;0.009 for all).\\u003c/p\\u003e\\u003c/div\\u003e\\u003cdiv id=\\\"Sec12\\\" class=\\\"Section2\\\"\\u003e\\u003ch2\\u003eTesticular Tissue Hormone Levels\\u003c/h2\\u003e\\u003cp\\u003eSignificant changes in testosterone (T) and inhibin B (Inh-B) levels were observed in testicular tissue across all groups (p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05, Table\\u0026nbsp;\\u003cspan refid=\\\"Tab2\\\" class=\\\"InternalRef\\\"\\u003e2\\u003c/span\\u003e).\\u003c/p\\u003e\\u003cp\\u003e\\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab2\\\" border=\\\"1\\\"\\u003e\\u003ccaption language=\\\"En\\\"\\u003e\\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 2\\u003c/div\\u003e\\u003cdiv class=\\\"CaptionContent\\\"\\u003e\\u003cp\\u003eHormone levels in testicular tissue samples\\u003c/p\\u003e\\u003c/div\\u003e\\u003c/caption\\u003e\\u003ccolgroup cols=\\\"8\\\"\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c7\\\" colnum=\\\"7\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"char\\\" char=\\\".\\\" class=\\\"colspec\\\" colname=\\\"c8\\\" colnum=\\\"8\\\"\\u003e\\u003c/div\\u003e\\u003cthead\\u003e\\u003ctr\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003eC\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003eD1\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003eD2\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003eD3\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003eD4\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003eD5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c8\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e\\u003cp\\u003ep\\u003c/p\\u003e\\u003c/th\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003eMedian (IQR)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003eMedian (IQR)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003eMedian (IQR)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003eMedian (IQR)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003eMedian (IQR)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003eMedian (IQR)\\u003c/p\\u003e\\u003c/th\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c8\\\"\\u003e\\u0026nbsp;\\u003c/th\\u003e\\u003c/tr\\u003e\\u003c/thead\\u003e\\u003ctbody\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e\\u003cb\\u003eT\\u003c/b\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e2307.30 (1907.90\\u0026ndash;2958.50)\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e2600.20 (2150.65\\u0026ndash;2925.90)\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e2521.50 (2243.80\\u0026ndash;2726.40)\\u003csup\\u003eb,c\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e1470.40 (744.55\\u0026ndash;2360.60)\\u003csup\\u003ea,b,d,e\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e2779.35 (2431.52\\u0026ndash;3293.22)\\u003csup\\u003ed\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e2769.90 (2726.40\\u0026ndash;2864.20)\\u003csup\\u003ec,e\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c8\\\"\\u003e\\u003cp\\u003e0.051\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e\\u003cb\\u003eInh B\\u003c/b\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e0.006 (0.005\\u0026ndash;0.008)\\u003csup\\u003ef,g,h,k\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e0.005 (0.002\\u0026ndash;0.011)\\u003csup\\u003em,n,p,q\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e0.038 (0.029\\u0026ndash;0.046)\\u003csup\\u003ef,m,r,s,t\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e0.089 (0.054\\u0026ndash;0.096)\\u003csup\\u003eg,n,r,u\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e0.093 (0.067\\u0026ndash;0.129)\\u003csup\\u003eh,p,s,v\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e0.244 (0.199\\u0026ndash;0.292)\\u003csup\\u003ek,q,t,u,v\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c8\\\"\\u003e\\u003cp\\u003e\\u003cb\\u003e0.000116*\\u003c/b\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tbody\\u003e\\u003c/colgroup\\u003e\\u003ctfoot\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003eFSH, follicle stimulating hormone; LH, luteinizing hormone; T, testosterone; Inh-B, inhibin B; C, control group; D1, desflurane 1 group; D2, desflurane 2 group; D3, desflurane 3 group; D4, desflurane 4 group; D5, desflurane 5 group.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003e*p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05, Kruskal-Wallis test. All intergroup comparisons were performed by using Tukey\\u0026rsquo;s HSD test.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003eComparisons for T levels:\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003e\\u003csup\\u003ea\\u003c/sup\\u003eD1-D3: p\\u0026thinsp;=\\u0026thinsp;0.046; \\u003csup\\u003eb\\u003c/sup\\u003eD2-D3: p\\u0026thinsp;=\\u0026thinsp;0.047; \\u003csup\\u003ec\\u003c/sup\\u003eD2-D5: p\\u0026thinsp;=\\u0026thinsp;0.040; \\u003csup\\u003ed\\u003c/sup\\u003eD3-D4: p\\u0026thinsp;=\\u0026thinsp;0.037; \\u003csup\\u003ee\\u003c/sup\\u003eD3-D5: p\\u0026thinsp;=\\u0026thinsp;0.008.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003eSignificant differences in Inh-B levels:\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003e\\u003csup\\u003ef\\u003c/sup\\u003eC-D2: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003eg\\u003c/sup\\u003eC-D3: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003eh\\u003c/sup\\u003eC-D4: p\\u0026thinsp;=\\u0026thinsp;0.014; \\u003csup\\u003ek\\u003c/sup\\u003eC-D5: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003em\\u003c/sup\\u003eD1-D2: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003en\\u003c/sup\\u003eD1-D3: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003ep\\u003c/sup\\u003eD1-D4: p\\u0026thinsp;=\\u0026thinsp;0.014; \\u003csup\\u003eq\\u003c/sup\\u003eD1-D5: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003er\\u003c/sup\\u003eD2-D3: p\\u0026thinsp;=\\u0026thinsp;0.036; \\u003csup\\u003es\\u003c/sup\\u003eD2-D4: p\\u0026thinsp;=\\u0026thinsp;0.014; \\u003csup\\u003et\\u003c/sup\\u003eD2-D5: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003eu\\u003c/sup\\u003eD3-D5: p\\u0026thinsp;=\\u0026thinsp;0.009; \\u003csup\\u003ev\\u003c/sup\\u003eD4-D5: p\\u0026thinsp;=\\u0026thinsp;0.014.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tfoot\\u003e\\u003c/table\\u003e\\u003c/div\\u003e\\u003c/p\\u003e\\u003cp\\u003eT Levels: Testicular T levels in Group D3 were higher than in Group D1 (p\\u0026thinsp;=\\u0026thinsp;0.046) but lower than in Group D2 (p\\u0026thinsp;=\\u0026thinsp;0.047). In contrast, T levels in Group D5 were higher than in Group D3 (p\\u0026thinsp;=\\u0026thinsp;0.040). Additionally, T levels in Groups D4 and D5 were elevated compared to Group D3 (p\\u0026thinsp;=\\u0026thinsp;0.037 and p\\u0026thinsp;=\\u0026thinsp;0.008, respectively).\\u003c/p\\u003e\\u003cp\\u003eInh-B Levels: Testicular Inh-B levels in Groups D2, D3, D4, and D5 were higher than in Group C (p\\u0026thinsp;=\\u0026thinsp;0.009, p\\u0026thinsp;=\\u0026thinsp;0.009, p\\u0026thinsp;=\\u0026thinsp;0.014, and p\\u0026thinsp;=\\u0026thinsp;0.009, respectively). Inh-B levels in these groups were also elevated compared to Group D1 (p\\u0026thinsp;=\\u0026thinsp;0.009, p\\u0026thinsp;=\\u0026thinsp;0.009, p\\u0026thinsp;=\\u0026thinsp;0.014, and p\\u0026thinsp;=\\u0026thinsp;0.009, respectively). Furthermore, Inh-B levels in Groups D3, D4, and D5 were higher than in Group D2 (p\\u0026thinsp;=\\u0026thinsp;0.036, p\\u0026thinsp;=\\u0026thinsp;0.014, and p\\u0026thinsp;=\\u0026thinsp;0.009, respectively). Inh-B levels in Group D5 were also elevated compared to Groups D3 and D4 (p\\u0026thinsp;=\\u0026thinsp;0.009 and p\\u0026thinsp;=\\u0026thinsp;0.014, respectively).\\u003c/p\\u003e\\u003c/div\\u003e\\u003cdiv id=\\\"Sec13\\\" class=\\\"Section2\\\"\\u003e\\u003ch2\\u003eSperm Morphology Assessment\\u003c/h2\\u003e\\u003cp\\u003eSignificant differences in sperm morphology were observed among all groups (p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05, Table\\u0026nbsp;\\u003cspan refid=\\\"Tab3\\\" class=\\\"InternalRef\\\"\\u003e3\\u003c/span\\u003e). The percentages of abnormal sperm in Groups D2, D3, D4, and D5 were higher than in Group C where Group D3 and D4 were showed significance (p\\u0026thinsp;=\\u0026thinsp;0.030, p\\u0026thinsp;=\\u0026thinsp;0.002, p\\u0026thinsp;=\\u0026thinsp;0.0011, and p\\u0026thinsp;=\\u0026thinsp;0.016, respectively). Additionally, the percentages of abnormal sperm in Groups D3 were raised in which D4 were significantly elevated compared to Group D1 (p\\u0026thinsp;=\\u0026thinsp;0.017 and p\\u0026thinsp;=\\u0026thinsp;0.0011, respectively).\\u003c/p\\u003e\\u003cp\\u003e\\u003cdiv class=\\\"gridtable\\\"\\u003e\\u003ctable float=\\\"Yes\\\" id=\\\"Tab3\\\" border=\\\"1\\\"\\u003e\\u003ccaption language=\\\"En\\\"\\u003e\\u003cdiv class=\\\"CaptionNumber\\\"\\u003eTable 3\\u003c/div\\u003e\\u003cdiv class=\\\"CaptionContent\\\"\\u003e\\u003cp\\u003eAbnormal sperm morphology\\u003c/p\\u003e\\u003c/div\\u003e\\u003c/caption\\u003e\\u003ccolgroup cols=\\\"8\\\"\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c1\\\" colnum=\\\"1\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c2\\\" colnum=\\\"2\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c3\\\" colnum=\\\"3\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c4\\\" colnum=\\\"4\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c5\\\" colnum=\\\"5\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c6\\\" colnum=\\\"6\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c7\\\" colnum=\\\"7\\\"\\u003e\\u003c/div\\u003e\\u003cdiv align=\\\"left\\\" class=\\\"colspec\\\" colname=\\\"c8\\\" colnum=\\\"8\\\"\\u003e\\u003c/div\\u003e\\u003cthead\\u003e\\u003ctr\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003eC\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003eD1\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003eD2\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003eD3\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003eD4\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003eD5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c8\\\" morerows=\\\"1\\\" rowspan=\\\"2\\\"\\u003e\\u003cp\\u003ep\\u003c/p\\u003e\\u003c/th\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e(Mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;SD)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e(Mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;SD)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e(Mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;SD)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e(Mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;SD)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e(Mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;SD)\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e(Mean\\u0026thinsp;\\u0026plusmn;\\u0026thinsp;SD)\\u003c/p\\u003e\\u003c/th\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u0026nbsp;\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003eN\\u0026thinsp;=\\u0026thinsp;5\\u003c/p\\u003e\\u003c/th\\u003e\\u003cth align=\\\"left\\\" colname=\\\"c8\\\"\\u003e\\u0026nbsp;\\u003c/th\\u003e\\u003c/tr\\u003e\\u003c/thead\\u003e\\u003ctbody\\u003e\\u003ctr\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c1\\\"\\u003e\\u003cp\\u003e\\u003cb\\u003ePAS\\u003c/b\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c2\\\"\\u003e\\u003cp\\u003e5.66 (5.01\\u0026ndash;5.75)\\u003csup\\u003ea,b,c,d\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c3\\\"\\u003e\\u003cp\\u003e4.71 (4.67\\u0026ndash;5.14)\\u003csup\\u003ee,f\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c4\\\"\\u003e\\u003cp\\u003e6.91 (6.56\\u0026ndash;7.27)\\u003csup\\u003ea\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c5\\\"\\u003e\\u003cp\\u003e7.46 (7.03\\u0026ndash;7.48)\\u003csup\\u003eb,e\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c6\\\"\\u003e\\u003cp\\u003e8.80 (7.09\\u0026ndash;9.05)\\u003csup\\u003ec,f\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"left\\\" colname=\\\"c7\\\"\\u003e\\u003cp\\u003e6.63 (6.49\\u0026ndash;7.48)\\u003csup\\u003ed\\u003c/sup\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003ctd align=\\\"char\\\" char=\\\".\\\" colname=\\\"c8\\\"\\u003e\\u003cp\\u003e\\u003cb\\u003e\\u0026lt;\\u0026thinsp;0.001*\\u003c/b\\u003e\\u003c/p\\u003e\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tbody\\u003e\\u003c/colgroup\\u003e\\u003ctfoot\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003ePAS, percentage of abnormal sperms; C, control group; D1, desflurane 1 group; D2, desflurane 2 group; D3, desflurane 3 group; D4, desflurane 4 group; D5, desflurane 5 group.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003e*p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.05\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003eKruskal-Wallis test. All intergroup comparisons were performed by using Holm-Sidak test.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003eSignificant differences in intergroup comparisons:\\u003c/td\\u003e\\u003c/tr\\u003e\\u003ctr\\u003e\\u003ctd colspan=\\\"8\\\"\\u003e\\u003csup\\u003ea\\u003c/sup\\u003eC-D2: p\\u0026thinsp;=\\u0026thinsp;0.030; \\u003csup\\u003eb\\u003c/sup\\u003eC-D3: p\\u0026thinsp;=\\u0026thinsp;0.002; \\u003csup\\u003ec\\u003c/sup\\u003eC-D4: p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.001; \\u003csup\\u003ed\\u003c/sup\\u003eC-D5: p\\u0026thinsp;=\\u0026thinsp;0.016; \\u003csup\\u003ee\\u003c/sup\\u003eD1-D3: p\\u0026thinsp;=\\u0026thinsp;0.017; \\u003csup\\u003ef\\u003c/sup\\u003eD1-D4: p\\u0026thinsp;\\u0026lt;\\u0026thinsp;0.001.\\u003c/td\\u003e\\u003c/tr\\u003e\\u003c/tfoot\\u003e\\u003c/table\\u003e\\u003c/div\\u003e\\u003c/p\\u003e\\u003c/div\\u003e\"},{\"header\":\"Discussion\",\"content\":\"\\u003cp\\u003eThe findings of this study demonstrate that chronic exposure to desflurane induces testicular tissue injury and increases the percentage of morphologically abnormal sperm in rats. Furthermore, significant alterations were observed in the levels of reproductive hormones, including FSH, LH, T, and Inh-B, which correlated with the intensity and duration of desflurane exposure.\\u003c/p\\u003e\\u003cp\\u003eFertility, a cornerstone of life's continuity, is governed by a complex interplay of endocrine and paracrine signaling pathways. While the regulatory mechanisms underlying fertility have been extensively studied, emerging evidence suggests that additional factors influencing this process remain to be elucidated [\\u003cspan citationid=\\\"CR20\\\" class=\\\"CitationRef\\\"\\u003e20\\u003c/span\\u003e]. The delicate balance between fertility and infertility is influenced by various environmental and pharmacological factors [\\u003cspan citationid=\\\"CR21\\\" class=\\\"CitationRef\\\"\\u003e21\\u003c/span\\u003e]. Numerous studies have investigated the effects of chemicals and drugs on the male reproductive system, both in humans and animal models [\\u003cspan additionalcitationids=\\\"CR10\\\" citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e\\u0026ndash;\\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR22\\\" class=\\\"CitationRef\\\"\\u003e22\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e23\\u003c/span\\u003e]. For instance, Ceyhan et al. reported that inhalational anesthetics such as sevoflurane and isoflurane can induce testicular damage in rabbits [\\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e]. Similarly, Kaya et al. demonstrated that chronic exposure to sevoflurane adversely affects testicular tissue structure and disrupts the hypothalamic-pituitary-gonadal axis, leading to alterations in FSH, LH, T, and Inh-B levels [\\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e].\\u003c/p\\u003e\\u003cp\\u003eSpermatogenesis, the process by which male gametes are produced, involves a series of tightly regulated stages, including proliferation, differentiation, and maturation. This process transforms round, immotile germ cells into elongated, motile spermatozoa capable of fertilization [\\u003cspan citationid=\\\"CR1\\\" class=\\\"CitationRef\\\"\\u003e1\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR24\\\" class=\\\"CitationRef\\\"\\u003e24\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR25\\\" class=\\\"CitationRef\\\"\\u003e25\\u003c/span\\u003e]. Spermatogenesis is regulated by the hypothalamic-pituitary-gonadal axis, wherein gonadotropin-releasing hormone from the hypothalamus stimulates the anterior pituitary to secrete FSH and LH. LH acts on Leydig cells to produce testosterone, which, along with FSH, stimulates Sertoli cells to support germ cell development [\\u003cspan citationid=\\\"CR20\\\" class=\\\"CitationRef\\\"\\u003e20\\u003c/span\\u003e]. Sertoli cells, often referred to as \\\"nurse cells,\\\" provide essential nutrients and metabolic intermediates, such as amino acids, carbohydrates, lipids, vitamins, and metal ions, to developing germ cells [\\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e26\\u003c/span\\u003e]. Additionally, Sertoli cells secrete inhibin B in response to FSH stimulation, which negatively regulates pituitary FSH secretion [\\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e27\\u003c/span\\u003e]. Thus, FSH, LH, testosterone, and inhibin B play critical roles in spermatogenesis and serve as key indicators of male reproductive health.\\u003c/p\\u003e\\u003cp\\u003eBeyond the documented impact on the hypothalamic-pituitary-gonadal (HPG) axis, some studies have suggested that volatile anesthetics such as isoflurane and desflurane may also influence other endocrine axes. Isoflurane, for instance, has been associated with alterations in the hypothalamic-pituitary-adrenal (HPA) axis, potentially reducing ACTH and corticosterone levels in animal models due to suppressed synaptic transmission in the hypothalamus [\\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e23\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR26\\\" class=\\\"CitationRef\\\"\\u003e26\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e27\\u003c/span\\u003e]. While data on desflurane are more limited, its structural and functional similarity to isoflurane raises the possibility of broader endocrine disruption [\\u003cspan citationid=\\\"CR27\\\" class=\\\"CitationRef\\\"\\u003e27\\u003c/span\\u003e]. These findings suggest that the observed hormonal changes may not be strictly confined to the gonadal axis but may reflect a more general downregulation of hypothalamic and pituitary function. However, the normalization of hormone levels after withdrawal supports the idea that these effects could be transient and reversible. Further research evaluating a wider range of hormones, including those of the thyroid and adrenal axes, is warranted to clarify whether the observed endocrine suppression is axis-specific or part of a more global functional decrease.\\u003c/p\\u003e\\u003cp\\u003eThe hormone-dependent nature of spermatogenesis makes it susceptible to disruption by external factors, including drugs and chemicals. For example, Xu et al. demonstrated that isoflurane exposure disrupts FSH, LH, and testosterone levels by inhibiting hypothalamic and pituitary function, likely through the suppression of synaptic transmission or nerve signaling [\\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e23\\u003c/span\\u003e]. In the present study, significant changes in FSH and LH levels were observed. While FSH levels decreased in Groups D1, D2, D3, and D4, only the reduction in Group D2 was statistically significant. Similarly, LH levels decreased significantly in Groups D2, D3, and D4, with the lowest levels observed in Group D3, which corresponded to the highest intensity of desflurane exposure. The lack of recovery in LH levels after a seven-day withdrawal period in Group D2 suggests that chronic desflurane exposure may impair hypothalamic or pituitary hormone production. However, the normalization of hormone levels during short- and long-term recovery periods indicates that the adverse effects of desflurane may be reversible.\\u003c/p\\u003e\\u003cp\\u003eUnexpectedly, testosterone levels in both serum and testicular tissue exhibited an irregular pattern that did not correlate with the severity of desflurane exposure. This anomaly, coupled with the progressive increase in serum and testicular inhibin B levels, suggests that desflurane may influence testosterone and inhibin B through mechanisms involving sex hormone-binding globulin or albumin, although the exact pathway remains unclear.\\u003c/p\\u003e\\u003cp\\u003eSperm morphology serves as a sensitive indicator of disturbances in the male reproductive system. However, limited studies have explored the effects of inhalational anesthetics on sperm morphology [\\u003cspan citationid=\\\"CR7\\\" class=\\\"CitationRef\\\"\\u003e7\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e23\\u003c/span\\u003e]. Campion et al. reported that isoflurane exposure reduces sperm motility in rats [\\u003cspan citationid=\\\"CR31\\\" class=\\\"CitationRef\\\"\\u003e31\\u003c/span\\u003e], while Ceyhan et al. observed structural abnormalities in sperm following chronic exposure to sevoflurane or isoflurane [\\u003cspan citationid=\\\"CR9\\\" class=\\\"CitationRef\\\"\\u003e9\\u003c/span\\u003e]. In contrast, Kaya et al. found no significant changes in the percentage of abnormal sperm despite reductions in sperm concentration and motility following sevoflurane exposure [\\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e]. In the current study, a significant increase in the percentage of abnormal sperm was observed in Groups D2, D3, D4, and D5 compared to the control group. This suggests that desflurane may disrupt the differentiation or maturation of spermatogonia, leading to abnormal sperm morphology. The reduction in abnormal sperm percentage in Group D5 compared to Group D4 indicates that a 14-day recovery period may be necessary to mitigate the structural impairments caused by desflurane exposure.\\u003c/p\\u003e\\u003cp\\u003eThe structural integrity of the testis is critical for normal reproductive function, and numerous factors can compromise testicular tissue. Xu et al. demonstrated that isoflurane exposure induces dose-dependent damage to seminiferous tubules, characterized by disorganized epithelium, sloughed germ cells, and interstitial edema [\\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e23\\u003c/span\\u003e]. Similarly, Kaya et al. reported progressive degenerative changes in seminiferous epithelium, including germ cell loss and interstitial edema, following chronic sevoflurane exposure [\\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e]. In the present study, histopathological examination revealed significant testicular tissue damage in all experimental groups, with the most severe changes observed in Group D3. These changes included degenerative alterations in seminiferous epithelium, interstitial edema, and congestion, although no loss of spermatogenic cells was observed. The correlation between histopathological injury scores and hormonal changes in Group D3 further underscores the adverse effects of desflurane on testicular function. Additionally, the incomplete recovery of tissue damage after a seven-day withdrawal period in Groups D2 and D5 suggests that longer recovery periods may be required for tissue repair.\\u003c/p\\u003e\\u003cp\\u003eEnvironmental chemical exposure is a growing public health concern, with infertility emerging as a significant consequence of exposure to toxicants [\\u003cspan citationid=\\\"CR34\\\" class=\\\"CitationRef\\\"\\u003e34\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR35\\\" class=\\\"CitationRef\\\"\\u003e35\\u003c/span\\u003e]. The widespread use of inhalational anesthetics in medical settings has raised concerns about their potential reproductive toxicity. A meta-analysis highlighted an increased risk of spontaneous abortion among health care workers occupationally exposed to anesthetic gases [\\u003cspan citationid=\\\"CR36\\\" class=\\\"CitationRef\\\"\\u003e36\\u003c/span\\u003e]. Anesthesiologists, surgeons, and nurse anesthetists are particularly vulnerable to the adverse effects of waste anesthetic gases, which have been linked to disturbances in germ cell development, sperm morphology, and motility [\\u003cspan citationid=\\\"CR11\\\" class=\\\"CitationRef\\\"\\u003e11\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR23\\\" class=\\\"CitationRef\\\"\\u003e23\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR28\\\" class=\\\"CitationRef\\\"\\u003e28\\u003c/span\\u003e, \\u003cspan citationid=\\\"CR37\\\" class=\\\"CitationRef\\\"\\u003e37\\u003c/span\\u003e].\\u003c/p\\u003e\\u003cp\\u003eThis study is the first to investigate the effects of chronic desflurane exposure on male reproductive hormones, testicular tissue, and sperm morphology in rats. The findings suggest that desflurane may induce testicular tissue damage, disrupt reproductive hormone levels, and impair sperm morphology. These results highlight the potential risks associated with occupational exposure to inhalational anesthetics among health care workers.\\u003c/p\\u003e\\u003cp\\u003eThere are some limitations for this study. First one is the limited duration of the follow-up period used to assess the reversibility of the observed effects. While changes in reproductive hormone levels, testicular histology, and sperm morphology were noted, the timeframe may not have been sufficient to determine whether these alterations are fully reversible after cessation of desflurane exposure. Longer-term studies are necessary to evaluate the potential for recovery of reproductive parameters over time. Additionally, as this study was conducted in an animal model, caution must be exercised when extrapolating the results directly to humans. Further research is needed to validate these findings in clinical settings and to establish safe exposure thresholds for health care professionals regularly exposed to inhalational anesthetics.\\u003c/p\\u003e\"},{\"header\":\"Conclusion\",\"content\":\"\\u003cp\\u003eChronic exposure to desflurane adversely affects testicular tissue integrity, sperm morphology, and the regulation of reproductive hormones in rats. These findings underscore the potential reproductive toxicity of inhalational anesthetics and emphasize the need for further research to elucidate the underlying mechanisms and mitigate the risks associated with occupational exposure.\\u003c/p\\u003e\"},{\"header\":\"Declarations\",\"content\":\"\\u003cp\\u003e\\u003cstrong\\u003eAvailability of data and materials\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe data presented in this study are available on reasonable request from the corresponding author.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAuthors Contributions\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eSD, HYD, HT, AA and MS\\u0026mdash;performed the research. SD and HYD\\u0026mdash;designed the research study. AA\\u0026mdash;performed the pathological analysis. SD, HYD, HT and MS\\u0026mdash;analyzed the data and wrote the draft of the manuscript. All authors read and approved the final manuscript. All authors were involved in the collection of experimental data.\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eEthics Approval and Consent to Participate\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThis experiemental study was approved by Tokat Gaziosmanpasa University Animal Experimentations Local Ethics Committee (Grant number: 2013-HADYEK-12).\\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eAcknowledgements\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eWe thank to Erkut Somak, Yılmaz Ozcan, and Serkan Kavak for their efforts and contributions for the study. \\u0026nbsp;\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eFunding\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThis work was supported by Gaziosmanpasa University Scientific Research Projects Unit (Grant Number: 2013/94).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConflict of interest\\u003c/strong\\u003e\\u003c/p\\u003e\\n\\u003cp\\u003eThe authors report no declarations of interest.\\u003c/p\\u003e\"},{\"header\":\"References\",\"content\":\"\\u003col\\u003e\\n\\u003cli\\u003eSun Y, Liu X, Wang J, et al. Air pollution exposure and infertility risk: evidence from a prospective cohort study. Environ Health Perspect. 2024;132(4):47001.\\u003c/li\\u003e\\n\\u003cli\\u003eDai X, Liu G, Li F, et al. Individual and joint associations of air pollutants exposure with semen quality: a retrospective longitudinal study in Wenzhou, China. Int Arch Occup Environ Health. 2024;97(2):123-132.\\u003c/li\\u003e\\n\\u003cli\\u003eAzzam A, Karabulut R, Kaya C, Eryılmaz S, Kapisiz A, Turkyilmaz Z, Inan MA, Yaz Aydin G, Atan A, Sonmez K. Effects of lupeol on experimental testicular ischemiareperfusion damage in rats. Ulus Travma Acil Cerrahi Derg. 2025;31(2):95-102.\\u003c/li\\u003e\\n\\u003cli\\u003eT\\u0026uuml;rkan H, Aydin A, Sayal A, Eken A, Akay C, Karahalil B. Oxidative and antioxidative effects of desflurane and sevoflurane on rat tissue in vivo. Arh Hig Rada Toksikol 2011; 62 113-119.\\u003c/li\\u003e\\n\\u003cli\\u003eAlcaraz M, Quesada S, Armero D, Martin-G\\u0026iacute;l R, Olivares A, Achel GD. Genotoxicity and cytotoxicity of sevoflurane in two human cell lines in vitro with ionizing radiation. Colombia Medica (Cali) 2014; 45: 104-109.\\u003c/li\\u003e\\n\\u003cli\\u003eBilban M, Jakopin CB, Ogrinc D. Cytogenetic tests performed on operating room personnel the use of anaesthetic gases. Int Arch Occup Environ Health 2005; 78: 60\\u0026ndash;64.\\u003c/li\\u003e\\n\\u003cli\\u003eKaymak C, Kadioglu E, Coskun E, Basar H, Basar M. Determination of DNA damage after exposure to inhalation anesthetics in human peripheral lymphocytes and sperm cells in vitro by comet assay. Hum Exp Toxicol 2012; 31: 1207-1213.\\u003c/li\\u003e\\n\\u003cli\\u003ePrystupa J. Fluorine--a current literature review. An NRC and ATSDR based review of safety standards for exposure to fluorine and fluorides. Toxicol Mech Methods 2011; 21: 103-170.\\u003c/li\\u003e\\n\\u003cli\\u003eCeyhan A, Cincik M, Bedir S, Ustun H, Dagli G, Kalender H. Effects of exposure to new inhalational anesthetics on spermatogenesis and sperm morphology in rabbits. Arch Androl 2005; 51: 305-315.\\u003c/li\\u003e\\n\\u003cli\\u003eHoerauf K, Funk W, Harth M, Hobbhahn J. Occupational exposure to sevoflurane, halothane and nitrous oxide during paediatric anaesthesia. Waste gas exposure during paediatric anaesthesia. Anaesthesia 1997; 52: 215-219.\\u003c/li\\u003e\\n\\u003cli\\u003eKaya Z, Sogut E, Cayli S, Suren M, Arici S, Karaman S, Erdemir F. Evaluation of effects of repeated sevoflurane exposure on rat testicular tissue and reproductive hormones. Inhal Toxicol 2013; 25: 192-198.\\u003c/li\\u003e\\n\\u003cli\\u003eMiller AL, Golledge HD, Leach MC. The Influence of Isoflurane Anaesthesia on the Rat Grimace Scale. PLoS One. 2016;11(11):e0166652.\\u003c/li\\u003e\\n\\u003cli\\u003eBrosnan RJ, Eger EI 2nd, Laster MJ, Sonner JM. Anesthetic properties of carbon dioxide in the rat. Anesth Analg. 2007; 105: 103-6.\\u003c/li\\u003e\\n\\u003cli\\u003eHariyanto H, Widiastuti M, Pandrya CO, Surya KA, Audi W. Comparison of desflurane and sevoflurane as maintenance inhalational anaesthetic agents for adult patients undergoing neurosurgeries: A systematic review and meta-analysis of randomised trials. Indian J Anaesth. 2025;69(1):65-77. \\u003c/li\\u003e\\n\\u003cli\\u003eKarzai W, Haberstroh J, M\\u0026uuml;ller W, Priebe HJ. Rapid increase in inspired desflurane concentration does not elicit a hyperdynamic circulatory response in the pig. Lab Anim 1997; 31: 279-282.\\u003c/li\\u003e\\n\\u003cli\\u003eLoan PB, Mirakhur RK, Paxton LD, Gaston JH. Comparison of desflurane and isoflurane in anaesthesia for dental surgery. Br J Anaesth 1995; 75: 289-292.\\u003c/li\\u003e\\n\\u003cli\\u003eGupta A, Stierer T, Zuckerman R, Sakima N, Parker SD, Fleisher LA. Comparison of recovery profile after ambulatory anesthesia with propofol, isoflurane, sevoflurane and desflurane: a systematic review. Anesth Analg 2004; 98: 632-641.\\u003c/li\\u003e\\n\\u003cli\\u003eCosentino MJ, Nishida M, Rabinowitz R, Cockett AT. Histopathology of prepubertal rat testes subjected to various durations of spermatic cord torsion. J Androl 1986; 7: 23-31.\\u003c/li\\u003e\\n\\u003cli\\u003eRezvanfar M, Sadrkhanlou R, Ahmadi A, Shojaei-Sadee H, Rezvanfar M, Mohammadirad A, Salehnia A, Abdollahi M. Protection of cyclophosphamide-induced toxicity in reproductive tract histology, sperm characteristics, and DNA damage by an herbal source; evidence for role of free-radical toxic stress. Hum Exp Toxicol 2008; 27: 901-910.\\u003c/li\\u003e\\n\\u003cli\\u003eGhewade P, Vagha S, Ghewade B, Gadkari P. Role of Dietary Antioxidant Supplements in Male Infertility: A Review. Cureus. 2024;16(6):e61951. \\u003c/li\\u003e\\n\\u003cli\\u003eMontano L, Rinaldo D, De Felice B, et al. Microplastics found in human ovary follicular fluid for the first time. Ecotoxicol Environ Saf. 2025;250:114512.\\u003c/li\\u003e\\n\\u003cli\\u003eLiu R, Dai J, Li J, et al. Does air pollution exposure affect semen quality? Evidence from a systematic review and meta-analysis of 93,996 Chinese men. Front Public Health. 2023;11:1219340.\\u003c/li\\u003e\\n\\u003cli\\u003eXu XL, Pan C, Hu JX, Liu XT, Li YF, Wang H, Chen YB, Dong HY, Dai TJ, Xu LC. Effects of isoflurane inhalation on the male reproductive system in rats. Environ Toxicol Pharmacol 2012; 34: 688-693.\\u003c/li\\u003e\\n\\u003cli\\u003eYoung CJ, Apfelbaum JL. Inhalational anesthetics: desflurane and sevoflurane. J Clin Anesth. 2023;45:564-577.\\u003c/li\\u003e\\n\\u003cli\\u003eZhou D, Liu B, Liu L, Liu G, Zhu F, Huang Z, Zhang S, He Z, Fan L. Essential Regulation of Spermatogonial Stem Cell Fate Decisions and Male Fertility by APBB1 via Interaction with KAT5 and GDF15 in Humans and Mice. Research (Wash D C). 2025;8:0647.\\u003c/li\\u003e\\n\\u003cli\\u003eMa R, Cui Y, Yu SJ, Pan YY, He JF, Wang YY, Wang JL, Wang XY, Bai XF, Zhang H, Yang SS, Zhang Q. The glucose metabolism reprogramming of yak Sertoli cells under hypoxia is regulated by autophagy. BMC Genomics. 2025;26(1):385. \\u003c/li\\u003e\\n\\u003cli\\u003eZhang Y, Zhao J, Wu W, Zhou C, Liu Y. Sevoflurane exposure impairs testicular function and fertility in neonatal rats via oxidative stress. Reprod Toxicol. 2023;116:48-55. \\u003c/li\\u003e\\n\\u003cli\\u003eKim H, Lee J, Park Y, Choi S. Chronic occupational exposure to isoflurane induces oxidative stress and impairs sperm parameters in mice. Toxicol Appl Pharmacol. 2024;473:116612.\\u003c/li\\u003e\\n\\u003cli\\u003eVincent KF, Mallari OG, Dillon EJ, Stewart VG, Cho AJ, Dong Y, Edlow AG, Ichinose F, Xie Z, Solt K. Oestrous cycle affects emergence from anaesthesia with dexmedetomidine, but not propofol, isoflurane, or sevoflurane, in female rats. Br J Anaesth. 2023;131(1):67-78. \\u003c/li\\u003e\\n\\u003cli\\u003eKhan S, Huang Y, Timu\\u0026ccedil;in D, Bailey S, Lee S, Lopes J, Gaunce E, Mosberger J, Zhan M, Abdelrahman B, Zeng X, Wiest MC. Microtubule-Stabilizer Epothilone B Delays Anesthetic-Induced Unconsciousness in Rats. eNeuro. 2024;11(8):ENEURO.0291-24.2024.\\u003c/li\\u003e\\n\\u003cli\\u003eCampion SN, Cappon GD, Chapin RE, Jamon RT, Winton TR, Nowland WS. Isoflurane reduces motile sperm counts in the Sprague-Dawley rat. Drug Chem Toxicol 2012; 35: 20-24.\\u003c/li\\u003e\\n\\u003cli\\u003eChen Y, Li Z, Wang H, et al. Gaseous pollutant exposure affects semen quality in central China: a cross-sectional study. Environ Sci Pollut Res Int. 2023;30(5):12345-12354.\\u003c/li\\u003e\\n\\u003cli\\u003eLiu J, Fang Z, Chai D, Zhu Z, Shen Q, He X. Ambient Air Pollution and Semen Quality in China: A Nationwide Case-Control Study of 27,014 Males with Biomarker-Confirmed Semen Pathology. Toxics. 2025;13(4):322.\\u003c/li\\u003e\\n\\u003cli\\u003eWang X, Zhang Y, Liu Y, et al. Association between ambient PM1 and semen quality: a cross-sectional study of 27,854 men in China. Environ Int. 2023;172:107754.\\u003c/li\\u003e\\n\\u003cli\\u003eAmato-Louren\\u0026ccedil;o LF, Carvalho-Oliveira R, J\\u0026uacute;nior GR, et al. Detection and characterization of microplastics in the human testis and semen. Sci Total Environ. 2023;857:159712.\\u003c/li\\u003e\\n\\u003cli\\u003eMargiana R, Odhar HA, Prasad K, Oghenemaro EF, M M R, Kumawat R, Uthirapathy S, Sharma S, Kumar MR, Nouri M. Does outdoor air pollution cause poor semen quality? A systematic review and meta-analysis. BMC Urol. 2025;25(1):50. \\u003c/li\\u003e\\n\\u003cli\\u003eZhou T, Li J, Cheng A, Zuo Z. Desflurane Post-treatment Reduces Hypoxic-ischemic Brain Injury via Reducing Transient Receptor Potential Ankyrin 1 in Neonatal Rats. Neuroscience. 2023;522:121-131. \\u003c/li\\u003e\\n\\u003c/ol\\u003e\"}],\"fulltextSource\":\"\",\"fullText\":\"\",\"funders\":[],\"hasAdminPriorityOnWorkflow\":false,\"hasManuscriptDocX\":true,\"hasOptedInToPreprint\":true,\"hasPassedJournalQc\":\"\",\"hasAnyPriority\":false,\"hideJournal\":false,\"highlight\":\"\",\"institution\":\"\",\"isAcceptedByJournal\":true,\"isAuthorSuppliedPdf\":false,\"isDeskRejected\":\"\",\"isHiddenFromSearch\":false,\"isInQc\":false,\"isInWorkflow\":false,\"isPdf\":false,\"isPdfUpToDate\":true,\"isWithdrawnOrRetracted\":false,\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"bmc-anesthesiology\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"bane\",\"sideBox\":\"Learn more about [BMC Anesthesiology](http://bmcanesthesiol.biomedcentral.com/)\",\"snPcode\":\"\",\"submissionUrl\":\"https://www.editorialmanager.com/bane\",\"title\":\"BMC Anesthesiology\",\"twitterHandle\":\"BMC_series\",\"acdcEnabled\":true,\"dfaEnabled\":false,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"BMC Series\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":true},\"keywords\":\"Follicle Stimulating Hormone, Luteinizing Hormone, testosterone, inhibin B, desflurane, sperm, morphology\",\"lastPublishedDoi\":\"10.21203/rs.3.rs-7507144/v1\",\"lastPublishedDoiUrl\":\"https://doi.org/10.21203/rs.3.rs-7507144/v1\",\"license\":{\"name\":\"CC BY 4.0\",\"url\":\"https://creativecommons.org/licenses/by/4.0/\"},\"manuscriptAbstract\":\"\\u003cp\\u003e\\u003cstrong\\u003eBackground:\\u003cbr\\u003e\\n \\u003c/strong\\u003eDesflurane is a widely used inhalational anesthetic known for its advantageous properties in clinical settings. This study aimed to investigate the effects of desflurane inhalation on male reproductive hormones, testicular tissue integrity, and sperm morphology in a rat model.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eMethods:\\u003cbr\\u003e\\n \\u003c/strong\\u003eThirty male rats were allocated into six experimental groups:\\u003c/p\\u003e\\n\\u003cp\\u003eControl group: Administered 2 L/min of O₂ for 18 minutes daily over seven days.\\u003c/p\\u003e\\n\\u003cp\\u003eGroup 1: Exposed to 1 minimum alveolar concentration (MAC) of desflurane and 2 L/min of O₂ for 18 minutes daily over seven days.\\u003c/p\\u003e\\n\\u003cp\\u003eGroup 2: Received the same treatment as Group 1 for seven days, followed by a seven-day recovery period without intervention.\\u003c/p\\u003e\\n\\u003cp\\u003eGroup 3: Administered 1 MAC desflurane and 2 L/min of O₂ for 18 minutes daily over 14 days.\\u003c/p\\u003e\\n\\u003cp\\u003eGroup 4: Received the same treatment as Group 3 for 14 days, followed by a seven-day recovery period without intervention.\\u003c/p\\u003e\\n\\u003cp\\u003eGroup 5: Administered the same treatment as Group 3 for 14 days, followed by a 14-day recovery period without intervention.\\u003c/p\\u003e\\n\\u003cp\\u003eBiochemical analyses were conducted to measure serum levels of follicle-stimulating hormone (FSH), luteinizing hormone (LH), testosterone, and inhibin B. Histopathological evaluations were performed to assess testicular tissue integrity, and sperm morphology was examined to identify abnormalities.\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eResults:\\u003cbr\\u003e\\n \\u003c/strong\\u003eSignificant histopathological damage was observed in all experimental groups compared to the control group (p \\u0026lt; 0.05). The proportion of morphologically abnormal spermatozoa was significantly higher in Groups D2, D3, D4, and D5 compared to the control group (p = 0.030, p = 0.002, p \\u0026lt; 0.001, and p = 0.016, respectively). Serum levels of FSH, LH, testosterone, and inhibin B exhibited statistically significant alterations across all experimental groups (p \\u0026lt; 0.05).\\u003c/p\\u003e\\n\\u003cp\\u003e\\u003cstrong\\u003eConclusion:\\u003cbr\\u003e\\n \\u003c/strong\\u003eChronic inhalation of desflurane, a modern inhalational anesthetic, was found to adversely affect testicular histology, sperm morphology, and the regulation of male reproductive hormones in rats. These findings highlight potential reproductive toxicity associated with prolonged desflurane exposure.\\u003c/p\\u003e\",\"manuscriptTitle\":\"The effects of desflurane on male rat reproductive hormones, testicular tissue, and sperm morphology: An experimental study\",\"msid\":\"\",\"msnumber\":\"\",\"nonDraftVersions\":[{\"code\":1,\"date\":\"2025-10-01 08:11:00\",\"doi\":\"10.21203/rs.3.rs-7507144/v1\",\"editorialEvents\":[{\"type\":\"communityComments\",\"content\":0},{\"type\":\"decision\",\"content\":\"Revision requested\",\"date\":\"2025-10-08T08:27:34+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2025-10-06T18:35:26+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2025-10-01T07:19:17+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"editorInvitedReview\",\"content\":\"\",\"date\":\"2025-09-30T15:45:21+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"235566905366922711942035461726947083405\",\"date\":\"2025-09-29T07:28:56+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"282860905525985435513816959486090157161\",\"date\":\"2025-09-25T16:36:56+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewerAgreed\",\"content\":\"317419707941428465009361232975490102111\",\"date\":\"2025-09-21T07:22:13+00:00\",\"index\":\"hide\",\"fulltext\":\"\"},{\"type\":\"reviewersInvited\",\"content\":\"\",\"date\":\"2025-09-20T16:30:17+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorInvited\",\"content\":\"\",\"date\":\"2025-09-18T07:52:03+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"editorAssigned\",\"content\":\"\",\"date\":\"2025-09-16T08:26:13+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"checksComplete\",\"content\":\"\",\"date\":\"2025-09-16T08:25:42+00:00\",\"index\":\"\",\"fulltext\":\"\"},{\"type\":\"submitted\",\"content\":\"BMC Anesthesiology\",\"date\":\"2025-09-01T10:06:51+00:00\",\"index\":\"\",\"fulltext\":\"\"}],\"status\":\"published\",\"journal\":{\"display\":true,\"email\":\"info@researchsquare.com\",\"identity\":\"bmc-anesthesiology\",\"isNatureJournal\":false,\"hasQc\":true,\"allowDirectSubmit\":false,\"externalIdentity\":\"bane\",\"sideBox\":\"Learn more about [BMC Anesthesiology](http://bmcanesthesiol.biomedcentral.com/)\",\"snPcode\":\"\",\"submissionUrl\":\"https://www.editorialmanager.com/bane\",\"title\":\"BMC Anesthesiology\",\"twitterHandle\":\"BMC_series\",\"acdcEnabled\":true,\"dfaEnabled\":false,\"editorialSystem\":\"em\",\"reportingPortfolio\":\"BMC Series\",\"inReviewEnabled\":true,\"inReviewRevisionsEnabled\":true}}],\"origin\":\"\",\"ownerIdentity\":\"b01ec13c-7d6d-4e16-b003-004c6b8297ae\",\"owner\":[],\"postedDate\":\"October 1st, 2025\",\"published\":true,\"recentEditorialEvents\":[],\"rejectedJournal\":[],\"revision\":\"\",\"amendment\":\"\",\"status\":\"published-in-journal\",\"subjectAreas\":[],\"tags\":[],\"updatedAt\":\"2026-02-09T16:07:22+00:00\",\"versionOfRecord\":{\"articleIdentity\":\"rs-7507144\",\"link\":\"https://doi.org/10.1186/s12871-026-03668-4\",\"journal\":{\"identity\":\"bmc-anesthesiology\",\"isVorOnly\":false,\"title\":\"BMC Anesthesiology\"},\"publishedOn\":\"2026-02-06 15:59:55\",\"publishedOnDateReadable\":\"February 6th, 2026\"},\"versionCreatedAt\":\"2025-10-01 08:11:00\",\"video\":\"\",\"vorDoi\":\"10.1186/s12871-026-03668-4\",\"vorDoiUrl\":\"https://doi.org/10.1186/s12871-026-03668-4\",\"workflowStages\":[]},\"version\":\"v1\",\"identity\":\"rs-7507144\",\"journalConfig\":\"researchsquare\"},\"__N_SSP\":true},\"page\":\"/article/[identity]/[[...version]]\",\"query\":{\"redirect\":\"/article/rs-7507144\",\"identity\":\"rs-7507144\",\"version\":[\"v1\"]},\"buildId\":\"8U1c8b4HqxoKbykW_rLl7\",\"isFallback\":false,\"isExperimentalCompile\":false,\"dynamicIds\":[84888],\"gssp\":true,\"scriptLoader\":[]}","source_license":"CC-BY-4.0","license_restricted":false}