Urinary excretion of oxidative damage markers in a rat model of vascularized composite allotransplantation

BACKGROUND: Vascularized composite allotransplantation is an emerging field of transplantation that provides a potential treatment for complex tissue defects after traumatic loss or tumor resection and for the repair of congenital abnormalities. However, vascularized composite allotransplantation recipients have suffered from acute and chronic graft rejection that is associated with oxidative stress. This study investigated the oxidative damage in a rat vascularized composite allotransplantation model by measuring three urinary biomarkers, 8-oxo-7,8-dihydroguanine (8-oxoGua), 8-oxo-7,8-dihydro-2′- deoxyguanosine (8-oxodG), and malondialdehyde. METHODS: Rats received two different immunosuppressants, including cyclosporine A and mycophenolate mofetil after transplantation, with one group also receiving mesenchymal stem cells before transplantation. Urine was collected and analyzed for 8-oxo-7,8-dihydroguanine, 8-oxo-7,8-dihydro-2′-deoxyguanosine, and malondialdehyde by liquid chromatography coupled to tandem mass spectometry methods. RESULTS: Rats undergoing vascularized composite allotransplantation had higher urinary levels of 8-oxo-7,8-dihydroguanine, 8-oxo-7,8-dihydro-2′- deoxyguanosine, and malondialdehyde compared with rats undergoing syngeneic transplantation. Cyclosporine A/mycophenolate mofetil following treatment prolonged the allograft survival in a dose-dependent manner. Compared with rats undergoing vascularized composite allotransplantation with cyclosporine A/mycophenolate mofetil treatment alone, rats undergoing mesenchymal stem cell combined treatment showed the longest allograft survival, and had approximately 50 percent lower urinary levels of malondialdehyde together with approximately 2.7-times higher levels of 8-oxo-7,8-dihydroguanine. CONCLUSIONS: Mesenchymal stem cell combined treatment efficiently managed oxidative stress in rats undergoing vascularized composite allotransplantation, and urinary 8-oxo-7,8-dihydroguanine and malondialdehyde could be regarded as good responders to the mesenchymal stem cell therapy.