Evaluation of long-term efficacy of breastmilk exosome in experimental testicular torsion model

Aim: In our study, we aimed to investigate the effectiveness of exosomes obtained from breast milk on the long-term effects of ischemia reperfusion injury caused by testicular torsion. Materials and Methods: In our study, a total of 32 Sprague Dawley rats weighing between 300-350 g were divided into 4 equal groups. The experimental period was determined as 6 weeks. In Group 1 (control), a scrotal incision was made after anesthesia and closed without any other procedure. In Group 2 (trauma), after a scrotal incision, the left testis was rotated 720 degrees clockwise and fixed to the scrotum with 3/0 silk suture; 4 hours after this procedure, a re-incision was made and then detorsion was performed. In Group 3 (local), a scrotal incision was made and then the left testis was rotated 720 degrees clockwise and fixed to the scrotum with 3/0 silk suture; 4 hours after this procedure, incision and detorsion were performed again. In Group 3, 1 cc of exosome was administered locally to the left testis surface immediately after detorsion. In Group 4 (intraperitoneal), a scrotal incision was made and then the left testis was rotated 720 degrees clockwise and fixed to the scrotum with a 3/0 silk suture; 4 hours after this procedure, a new incision and detorsion were performed. Immediately after detorsion, 1 cc of exosome was administered intraperitoneally (ip) (systemically) to the rats in Group 4. The layers were closed in anatomical position in all groups. Orchiectomy was performed on the left testis 6 weeks after the detorsion procedure. Orchiectomy materials were divided into two for biochemical and histopathological examination and sent to the laboratory under appropriate conditions. All rats were euthanized by exsanguination. The effect of exosome in preventing reperfusion injury in testicular torsion was measured biochemically, tissue malondialdehyde (MDA), superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPX) levels were measured. It was also evaluated with Johnsen Testicular Biopsy Scoring (JTBS) to show its histopathological effect. Results: There was a significant difference in the MDA enzyme evaluation between the groups (p=0.042). The MDA value of the trauma group was found to be higher than the control group (p=0.025). When the SOD value was compared between the groups, no statistically significant difference was found (p=0.089). When the CAT value was compared between the groups, no statistically significant difference was found (p=0.828). There was a significant difference in the GPX enzyme evaluation between the groups (p=0.016). The GPX value of the trauma group was found to be statistically significant compared to the control group. (p1-2=0.019). The numerical decrease in the local group compared to the trauma group was also found to be statistically significant (p2-3=0.038). There was a significant difference between the groups in the Johnsen Testicular Biopsy Scoring evaluation (p<0.001). The numerical decrease between the control and trauma groups was found to be statistically significant (p1-2= <0.001). There was a significant difference between the control and local groups (p1-3=0.004). The numerical difference between the trauma and intraperitoneal groups was found to be statistically significant (p2-4= 0.003). Conclusion: In conclusion, this is the first study to investigate the long-term efficacy of breast milk exosomes in a testicular torsion model. All these results suggest that breast milk exosomes may be an effective therapeutic agent for patients with testicular torsion-detorsion. Key Words: Testicular torsion, exosome, breast milk, chronic