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Evaluation of Cold Ischemia for Preservation of Testicular Function during Partial Orchiectomy in the Rat Model.
Erin R. McNamara, MD, Ramiro J. Madden-Fuentes, MD, Elizabeth Masko, PhD, Jonathan C. Routh, MD, MPH, John F. Madden, MD, John S. Wiener, MD, Harry G. Rushton, Jr., MD, Sherry S. Ross, MD.
Duke University Medical Center, Durham, NC, USA.

Introduction
Multicenter studies have demonstrated that testicular tumors in prepubescent children are most often benign. This observation has led to an increasing practice of testis-sparing surgery in management of testicular tumors. Testicular viability and function are at the forefront of the urologists’ mind when performing testis sparing surgery. Cold ischemia in partial nephrectomy has been shown to correlate with improved renal preservation but has not been well described in partial orchiectomy. In this study, we aimed to determine if cold ischemia during partial orchiectomy resulted in improved serum testosterone levels, sperm count, and histological architecture when compared to warm ischemia in a prepubescent rat model.
Materials and Methods
Eighteen prepubescent male Sprague-Dawley rats were randomized to 3 different surgical groups. Rats in Group 1 underwent sham surgery (n=4 rats, 8 testicles), Group 2 underwent bilateral partial orchiectomy with 30 minutes of cord compression and cold ischemia via ice-slush (N=7 rats, 14 testicles) and Group 3 underwent bilateral partial orchiectomy with 30 minutes of cord compression and warm ischemia (no ice slush) (n=7 rats, 14 testicles). When animals reached puberty (approximately day of life 51), the animals were sacrificed. Sperm were collected via epididymal preparation and sperm count determined by an independent veterinarian. Histological evaluation of the testis was performed to evaluate interstitium:tubular ratio, interstitial fibrosis and germ cell maturation. Histological analysis was performed by a blinded uropathologist. Serum was collected at the time of sacrifice.
Results
All animals survived surgery and were included in the study. Serum testosterone levels were compared among the three groups. Mean serum testosterone levels were 1445+590 pg/ml for the sham group, 449+268 pg/ml for the cold ischemia group and 879+631 pg/ml for the warm ischemia group. Serum testosterone levels were higher in the warm ischemic group compared to the cold ischemic group; however, there was no statistically significant difference between groups (p=0.12). Mean sperm counts were 2.1x107 for the sham group, 4.4 x 106 for the cold ischemia group was, and 9.9 x 106 for the warm ischemia group. Sperm count were higher in the warm ischemic group compared to the cold ischemic group, but there was no statistically significant difference between groups (p=0.48). Histological evaluation and comparison revealed no difference in interstitium:tubular ratio, interstitial fibrosis and germ cell maturation between the cold and warm ischemic groups.
Conclusions
Serum testosterone, sperm count and histological findings were not statistically different between the cold ischemic and warm ischemic partial orchiectomy groups. Sham animals had higher serum testosterone and sperm count when compared to animals that had partial orchiectomy. In our preclinical rat model, there seemed to be no benefit for cold ischemia over warm ischemia at 30 minutes. The end points of testosterone production, sperm count post puberty and histology of the testes are relevant in the pediatric patient population undergoing partial orchiectomy.


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