Evaulation the effect of the testis fixation is performed through the epididiymo-testicular junction on enflamation, oxidative stress and spermatogenesis parameters in rats

Background and Purpose: Among the pediatric urology surgeries, undescended testis and testicular torsion operations are frequently being performed. In both cases, testis may be fixed to the scrotum depending on the preference of the surgeons. However, the vast majority of the experimental studies suggest that if fixation is performed through transparancymal site, the sutures may lead to histopathological alterations and deterioration in spermatogenesis. In this study we hypothesized that if the fixation suture is performed through the epididymo-testicular junction (where no seminifery tubules and a vascular structure exist) we may cause less trauma to the testis. We tested this hypothesis by assesing the spermatogenetic functions, histopathology and biochemical parameters. Materials and Methods: 28 Sprague-Dawley rats were randomly assigned into 4 groups. In sham group, after the scrotal exploration, nothing was performed and the incision line was sutured with 5-0 vicryl. In the second group (transparenchymal sutur fixation = TPF) following the scrotal exploration, testis fixation was performed by passing the suture through the testis parenchyma. In the third group (transepididymo-testicular fixation = TETF) following the scrotal exploration, fixation was performed by passing the suture through the epididymo-testicular junction. In the last group (dartos pouch fixation = DPO) following the scrotal exploration, testis was placed into the dartos pouch without fixation. Rats were sacrificed after 1 month following the surgery. Testicular tissues were examined histologically and biochemically. Suture environment inflammation, spermatogenesis (Johnsen criteria), microlithiasis were criteria of histopathological examination. The biochemical markers of inflammation (IL-1B, IL-6, TNF-α) were evaulated by ELISA kits. Oxidative stress parameters (TAS, TOS, OSI) were evaulated by Erel method. For statistical evaluation SPSS 23.0 (Statistical Package for the Social Sciences) program was used. Data are expressed as median (minimum-maximum). Kruskal-Wallis test was used for comparison of all values. p < 0.05 was considered significant. Results: In comparison of testis weights, no statistically difference was detected among all groups (p = 0.324). The evaluation of spermatogenesis by Johnsen scoring system revealed that sham group was significantly better than DPO and TETF groups (p = 0.001). When assesing inflammation parameters histologically, the results were statistically comparable among all groups. In the sham group, no inflammation was detected. In the TPF group 6 testicles (%85.7) had no inflammation while 1 testis (%14.2) had moderate inflammation. In the TETF group 5 testicles (%71.4) had no inflammation while 1 testis (%14.2) had moderate inflammation and 1 testis (%14.2) had severe inflammation. In the DPO group 6 testicles (%85.7) had no inflammation while 1 testis (%14.2) had moderate inflammation. When assessing microlithiasis, groups demonstrated a heterogeneous distribution. In the sham group no microlithiasis was detected. In the TPF group, at 6 testicles (%85.7) microlithiasis was detected whereas in the TETF group, 2 testicles (%28.5) demonstrated microlithiasis. In the DPO group only at 1 testis (%14.2) microlithiazis was detected. The rate of microlithiasis was significantly higher in TPF group when compared to the other groups (p= 0.046). The assessment of the inflammation markers in the testis tissue revealed that IL-1B values for TPF and TETF groups were significantly higher than sham group (p = 0.002). For IL-6, TPF group values were significantly higher than sham and DPO groups (p = 0.001). For TNF-α, TPF and DPO groups values were significantly higher than sham group (p = 0.008). In the testis tissues, TAS, TOS and OSI values were calculated. According to these results, TOS and TAS values found similar among all groups (p = 0.487, 0.522; respectively). However, for OSI, sham group value was significantly lower than TPF and TETF groups (p = 0,001). Conclusions: The findings of our experimental study revealed that, fixation of the testis, regardless of the technique used, cause histopathological and biochemical damages to the testicles when compared to sham group. 1. In TETF and DPO groups, it was shown that spermatogenesis was significantly decreased. 2. Histopathologically, except sham group, moderate or severe inflamation was depicted in all groups. 3. Microlithiasis was observed at the highest rate within the TPF group. 4. Biochemical markers of inflammation were identified with the most significant elevation in the TPF group. 5. While TAS and TOS values were found similar between all groups, OSI value was found to be significantly increased in the TPF group.