To determine whether cryopreserved trachea can be a substitute for tracheal replacement, we performed 3-step experimental study. First, we evaluated the viability of cryopreserved tracheal allografts in immunosuppressant free transplantation in dogs. Second, we investigated the origin of the epithelium in transplanted cryopreserved tracheal allografts in rats. Third, we examined histological changes and bacterial contamination of the human bronchus during the process of cryopreservation.
(1) Cryopreserved tracheal allotransplantation in dogs
Methods : In group A (n= 6), a cryopreserved tracheal allograft, which had been stored in liquid nitrogen (-196°C) after freezing to-80°C by a programmed freezer, in group B (n= 5), a fresh autograft, and in group C (n = 4), a fresh allograft were transplanted by replacing the thoracic segment of the trachea using an omental flap without immunosuppressive agents.
Results : In groups A and B, all dogs survived, but in group C, all died of airway obstruction between 1 to 2 months after the operation. Histologically, the cryopreserved allograft segment displayed normal epithelium and cartilage, but the fresh allograft segment showed chronic inflammatory changes with a lack of epithelium and cartilage.
(2) Epithelial regeneration in transplanted cryopreserved trachea in rats
Methods : Tracheal transplantation was performed (n=6) using PVG rats (allele at RT 1-locus : c) as a donor and ACI rats (allele at RT 1-locus. a) as a recipient. After resection of a five-ring segment of the cervical trachea, the trachea was reconstructed using the cryopreserved tracheal segment of a PVG rat (n = 6). No immunosuppressive agents including steroids were given to them.
Results : At 2 months after tracheal transplantation, 6 surviving ACI rats were killed. Histologically, the epithelium and tracheal cartilage of the transplanted cryopreserved segment displayed a normal structure. Immunohistochemical staining showed that the MHC Class I antigen of the ACI rat was expressed in the epithelium of the transplanted segment and that the MHC Class I antigen of the PVG rat was expressed in the cartilage of the transplanted segment.
(3) Cryopreservation of human bronchus
Methods : Six lobar bronchi were harvested from the resected lung of cancer patients and histological changes and bacterial contamination before and after cryopreservation were examined.
Results : No bacterial growth was detected after 2448 hours storage at 4°C or at thawing of the cryopreserved human bronchi. The epithelium of the human bronchi was almost deleted after cryopreservation.
[Conclusions] The epithelium, smooth muscle and cartilage were retained after implantation of cryopreserved tracheal allografts vascularized with the omentum and without immunosuppression. After transplantation of the cryopreserved trachea, the epithelium of the transplanted cryopreserved segment was originated from the recipient epithelium, while the donor's cartilage retained the structure of the trachea. The epithelium of the human bronchus was almost deleted after the process of cryopreservation. We believe that transplantation of a cryopreserved trachea leads to the growth of the recipient's epithelium over the donor's trachea, thereby reducing the antigenicity of the transplant.
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