MP22: Trauma/Reconstruction/Diversion: Urethral Reconstruction (including Stricture, Diverticulum) I
MP22-13: Off the Shelf Acellular Fetal Skin Scaffold as a Novel Alternative to Buccal Mucosa Graft: The Development and Characterization of Human Tissue Engineered Fetal Dermal Matrix in Rabbit Model of Hypospadiasis
Introduction: To introduce a novel alternative to buccal mucosal graft from the acellular human fetal skin for management of hypospadias in rabbit model. To optimize the acellularization protocol for development and characterization of human tissue engineered fetal dermal matrix as an “off the shelf” natural biomaterial
Methods: Human fetal skin were obtained (from the therapeutic abortion fetuses) with gestational age of 16-19 weeks with respect to a singed informed consent from parents following receiving the university ethical committee permission. The dissected full thickness fetal skin tissues were placed into SDS and Triton X-100 in different dosages to achieve the optimum decellularization protocol. Histopathology of the fetal acellular matrix were assess by hematoxcillin & eosin, DAPI staining for confirmation of removal of all cell materials, trichrome staining for collagen evaluation DNA quantification for confirmation of DNA content, scanning electron microscopy for evaluation of scaffold microstructure. Specific dermal markers (Vimentin, type I collagen, CK14) were depict by IHC. The prepared dermal scaffolds were then grafted into the induced model of hypospadias in 8 male rabbits. The rabbits were followed up for 1, 2, 3 and 6 months post operatively.
Results: In order to show the success of decellularization process, we characterized it with H&E, trichrome-Masson staining, DAPI and SEM which confirmed the significant removal of cells; meanwhile the ECM was completely preserved. At the time of biopsy after 2, 4 and 6 moths, no evidence of inflammation, fibrosis, necrosis or rejection was observed and the grafted dermal scaffolds appeared histologically and anatomically normal histologic appearance by recellularization of scaffold by circulating CD 34 + bone marrow stem cells “represent the body as a natural bioreactor”
Conclusions: The use of this novel “off the shelf” natural fetal skin scaffold without cell seeding is an excellent alterative to buccal mucosal graft and pave the road for decreasing surgical time (at least one hour) in complex hypospadias reconstruction. This method is safe, feasible with excellent angioneogenesis and cell seeding in due course without cell culture and cell seeding or shrinkage of the graft, no creation of oral mucosal wound and in addition, there will be no limitations of graft size for staged hypospadias cripple surgery.
Source of Funding: There was no source of funding for this study