ACELLULAR URETHRAL SCAFFOLD FROM SEX REASSIGMENT PATIENTS: DEVELOPMENT AND CRYOPRESERVATION EFFECTS

FRAUNHOFFER NICOLAS A.; BELINKY JAVIER; REY HORACIO; MEILERMAN ABUELAFIA ANALIA.; REY LOURDES; BLUMENGARTEN YAGO; SCHVARZMAN NIR; PACI MARINA; FERRARIS SERGIO; BARRIOS MARCELA

Mar del Plata

Congreso; LXI Reunión Anual de la Sociedad Argentina de Investigación Clínica; 2016

Sociedad Argentina de Investigación Clínica

Urethral reconstruction for both congenital and acquired causesremains challenge for urological surgeons. A wide variety of tissueshave been used in urethral repair. However, all of thesesubstitutes have limitations compared to the urethral tissue (UT).In this context, acellular scaffolds from human urethras would bethe best alternative, although the access to this tissue is limited.The objectives of this study were to develop a decellularizationmethod for UT and analyze UT cryopreservation effects on theacellular scaffold. 4 urethral samples from male patients withoutany urological pathology were used. Samples were obtained fromHospital Durand. Two decellularization protocols in 2 periods (3or 7 days) were analyzed: sodium deoxyxholate 1% (PR1) andTriton X-100 1% (PR2). Additionally, two freezing media withDMSO 0.7M (PRA) and 1.5M (PRB) were evaluated. Decellularizationand structural integrity were assessed by histologicalanalysis, actin WB, DNA levels and scanning electron microscopy(SEM). Extracellular matrix (EM) proteins (collagen I and IV,laminin, fibronectin and elastin) and VEGF were studied by IHCand dot blot. PR1 and PR2 applied for 3 days, maintained highcellular components, while PR2 applied for 7 days showed totaldecellularization, undetectable DNA and actin levels with highstructural integrity. EM proteins and VEGF were higher in PR2than PR1. Comparing the two freezing protocol, PRA presentedbetter integrity and protein levels than PRB, in combination withboth decellularization protocols. However, PRA/PR2 showed thehighest levels of EM proteins and VEGF, even better than PR2without freezing cycle. These results show that PR2 applied for7 days is the best decellularization protocol. Furthermore, ourresults suggest that a freezing cycle, previously to decellularizationpromotes the UT integrity. Therefore, these results suggestthat urethral scaffold from sex reassignment patients representsa feasible tissue for urethral repair.