Recovery of Cryo-injured Rabbit Urethras by Biofabricated C-shaped Adipose-derived Mesenchymal Cell Structures

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Abstract

Background : Urethral tissue damage can cause stress urinary incontinence or urinary retention, and there are few long-term, effective treatments. For structural and functional recovery of urethral tissues, we designed and constructed C-shaped structures composed of adipose-derived mesenchymal cells (AMCs) by using a three-dimensional (3D)-bioprinter system, which could be transplanted in damaged urethras without obstructing the lumen.<br>Methods : AMCs were harvested from New Zealand White rabbits, labeled with PKH26 for later identification, and cultured to form spheroids. The spheroids were assembled on a custom-designed C-shaped support by a three-dimensional bioprinter, and perfusion cultured for 7 days. Urethras of female rabbits were cryo-injured by spraying with liquid nitrogen for 20 seconds. The damaged region was incised, and the biofabricated structure was autologously transplanted into the cryo-injured region (n＝4). Control rabbits were treated similarly but without transplantation of a C-shaped structure (n＝4). Two and four weeks after surgery, the urethras were harvested for analyses.<br>Results : Prior to transplantation, the AMCs within the biofabricated structures expressed mesenchymal cell markers and extracellular matrices. Two and four weeks after surgery, the control urethras were partially constricted ; however, the structure-transplanted urethras were patent. The AMCs within the structures had differentiated into skeletal muscle, smooth muscle, or nerve cells. Some cells contained growth factors and cytokines. In addition, they expressed decreased fibroblast markers, hypoxia markers, and apoptotic markers.<br>Conclusions : The transplantation of biofabricated C-shaped AMC structure has the potential to be an effective treatment for urethral recovery due to enhanced reconstruction of muscle, and nerve cells.

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