title: Inosculation of blood vessels allows early perfusion and vitality of bladder grafts--implications for bioengineered bladder wall.
abstract: Bioengineered bladder tissue is needed for patients with neurogenic bladder disease as well as for cancer. Current technologies in bladder tissue engineering have been hampered by an inability to efficiently initiate blood supply to the graft, ultimately leading to complications that include graft contraction, ischemia, and perforation. To date, the biological mechanisms of vascularization on transplant have not been suitably investigated for urologic tissues. To better understand the mechanisms of neovascularization on bladder wall transplant, a chimeric mouse model was generated such that angiogenesis and vasculogenesis could be independently assessed in vivo. Green fluorescence protein (GFP) transgenic mice received bone marrow transplants from β-galactosidase (LacZ) transgenic animals and then subsequent bladder wall transplants from wild-type donor mice. Before euthanization, the aorta was infused with fluorescent microbeads (fluorospheres) to identify perfused vessels. The contributions of GFP (angiogenesis) and LacZ (vasculogenesis) to the formation of CD31-expressing blood vessels within the wild-type graft were evaluated by immunohistochemistry at different time points and locations within the graft (proximal, middle, and distal) to provide a spatiotemporal analysis of neovascularization. The GFP index, a measure of angiogenic host ingrowth, was significantly higher at proximal versus mid or distal regions in animals 2-16 weeks post-transplant. However, GFP index did not increase over time in any area. Within 7 days post-transplant, perfusion of primarily wild-type, donor blood vessels in the most distal areas of the graft was observed by intraluminal fluorospheres. In addition, chimeric host-donor (GFP-wild type) blood vessels were evident in proximal areas. The contribution of vasculogenesis to vascularization of the graft was limited, as LacZ cells were not specifically associated with the endothelial cells of blood vessels, but rather found primarily in areas of inflammation. The data suggest that angiogenesis of host blood vessels into the proximal region leads to inosculation between host and donor vessels and subsequent perfusion of the graft via pre-existing graft vessels within the first week after transplant. As such, the engineering of graft blood vessels and the promotion of inosculation might prevent graft contraction, thereby potentiating the use of bioengineered bladder tissue for transplantation.
authors: 1Department of Urology, University of California, Davis School of Medicine, Sacramento, California.|2Stem Cell Program, Institute for Regenerative Cures, University of California, Davis Medical Center, Sacramento, California.
authors: Stephanie L
authors: SL
authors: Osborn
authors: 1Department of Urology, University of California, Davis School of Medicine, Sacramento, California.|2Stem Cell Program, Institute for Regenerative Cures, University of California, Davis Medical Center, Sacramento, California.
authors: Michelle
authors: M
authors: So
authors: 1Department of Urology, University of California, Davis School of Medicine, Sacramento, California.|2Stem Cell Program, Institute for Regenerative Cures, University of California, Davis Medical Center, Sacramento, California.
authors: Shannon
authors: S
authors: Hambro
authors: 2Stem Cell Program, Institute for Regenerative Cures, University of California, Davis Medical Center, Sacramento, California.|3Department of Internal Medicine, University of California, Davis School of Medicine, Sacramento, California.
authors: Jan A
authors: JA
authors: Nolta
authors: 1Department of Urology, University of California, Davis School of Medicine, Sacramento, California.|2Stem Cell Program, Institute for Regenerative Cures, University of California, Davis Medical Center, Sacramento, California.
authors: Eric A
authors: EA
authors: Kurzrock
mesh_terms: D000818:Animals; D005260:Female; D005456:Fluorescent Dyes / Q000493:pharmacokinetics; D017930:Genes, Reporter; D006085:Graft Survival; D049452:Green Fluorescent Proteins / Q000032:analysis / Q000235:genetics; D007763:Lac Operon; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D008833:Microcirculation; D008863:Microspheres; D009130:Muscle, Smooth / Q000098:blood supply / Q000648:ultrastructure; D018919:Neovascularization, Physiologic / Q000502:physiology*; D019408:Platelet Endothelial Cell Adhesion Molecule-1 / Q000032:analysis; D011184:Postoperative Period; D011828:Radiation Chimera; D019737:Transplants / Q000098:blood supply* / Q000648:ultrastructure; D001743:Urinary Bladder / Q000098:blood supply / Q000637:transplantation*; D019459:Urothelium / Q000166:cytology
chemical_list: D005456:Fluorescent Dyes; D019408:Platelet Endothelial Cell Adhesion Molecule-1; D049452:Green Fluorescent Proteins