Acellular Urethra Bioscaffold: Decellularization of Whole Urethras for Tissue Engineering Applications.
Patients with stress urinary incontinence mainly suffer from malfunction of the urethra closure mechanism. We established the decellularization of porcine urethras to produce acellular urethra bioscaffolds for future tissue engineering applications, using bioscaffolds or bioscaffold-derived soluble products. Cellular removal was evaluated by H&E, DAPI and DNA quantification. The presence of specific ECM proteins was assessed through immunofluorescence staining and colorimetric assay kits. Human skeletal muscle myoblasts, muscle progenitor cells and adipose-derived stromal vascular fractions were used to evaluate the recellularization of the acellular urethra bioscaffolds. The mechanochemical decellularization system removed ~93% of tissue's DNA, generally preserving ECM's components and microarchitecture. Recellularization was achieved, though methodological advances are required regarding cell seeding strategies and functional assessment. Through microdissection and partial digestion, different urethra ECM-derived coating substrates were formulated (i.e. containing smooth or skeletal muscle ECM) and used to culture MPCs in vitro. The skeletal muscle ECM substrates enhanced fiber formation leading to the expression of the main skeletal muscle-related proteins and genes, as confirmed by immunofluorescence and RT-qPCR. The described methodology produced a urethra bioscaffold that retained vital ECM proteins and was liable to cell repopulation, a crucial first step towards the generation of urethra bioscaffold-based Tissue Engineering products.
Department of Bioengineering and iBB - Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.|Wake Forest Institute for Regenerative Medicine, Winston-Salem, NC USA.|Laboratory for Tissue Engineering and Stem Cell Therapy, Department of Urology, University Hospital Zurich, Zurich, Switzerland.
Irina N
IN
Simões
Serviço Urologia, Hospital Garcia de Orta, Almada, Portugal.
Paulo
P
Vale
Wake Forest Institute for Regenerative Medicine, Winston-Salem, NC USA.
Shay
S
Soker
Wake Forest Institute for Regenerative Medicine, Winston-Salem, NC USA.
Anthony
A
Atala
Laboratory for Tissue Engineering and Stem Cell Therapy, Department of Urology, University Hospital Zurich, Zurich, Switzerland.
Daniel
D
Keller
Faculdade de Medicina Veterinária, The Interdisciplinary Centre of Research in Animal Health (CIISA), Universidade de Lisboa, Lisboa, Portugal.
Rute
R
Noiva
Faculdade de Medicina Veterinária, The Interdisciplinary Centre of Research in Animal Health (CIISA), Universidade de Lisboa, Lisboa, Portugal.
Sandra
S
Carvalho
Faculdade de Medicina Veterinária, The Interdisciplinary Centre of Research in Animal Health (CIISA), Universidade de Lisboa, Lisboa, Portugal.
Conceição
C
Peleteiro
Department of Bioengineering and iBB - Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.
Joaquim M S
JM
Cabral
Laboratory for Tissue Engineering and Stem Cell Therapy, Department of Urology, University Hospital Zurich, Zurich, Switzerland.
Daniel
D
Eberli
Department of Bioengineering and iBB - Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Lisboa, Portugal.
Cláudia L
CL
da Silva
Wake Forest Institute for Regenerative Medicine, Winston-Salem, NC USA.|Instituto de Investigacion Sanitaria de Aragón (IIS Aragon), Zaragoza, Spain.|CIBERehd, Zaragoza, Spain.
Pedro M
PM
Baptista
D000818:Animals; D005109:Extracellular Matrix / Q000502:physiology*; D016326:Extracellular Matrix Proteins / Q000378:metabolism; D005260:Female; D008297:Male; D013552:Swine; D023822:Tissue Engineering / Q000379:methods*; D054457:Tissue Scaffolds; D014521:Urethra / Q000166:cytology* / Q000502:physiology*
D016326:Extracellular Matrix Proteins