Urethral Replacement Using Cell Seeded Tubularized Collagen Matrices (2002)

No AccessJournal of UrologyBasic Science1 Oct 2002Urethral Replacement Using Cell Seeded Tubularized Collagen Matrices Roger E. De Filippo, James J. Yoo, and Anthony Atala Roger E. De FilippoRoger E. De Filippo , James J. YooJames J. Yoo , and Anthony AtalaAnthony Atala View All Author Informationhttps://doi.org/10.1016/S0022-5347(05)64414-XAboutFull TextPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract Purpose: Acellular collagen matrices derived from bladder submucosa have been used successfully as an off-the-shelf biomaterial for urethral replacement, experimentally and clinically in an onlay fashion. We investigated whether collagen matrices, either alone or with autologous cells, could be used for tubularized urethral replacement. Materials and Methods: Acellular collagen matrices were processed and tubularized. Ten rabbits underwent an open bladder biopsy with subsequent cell expansion. Autologous bladder cells were grown and seeded onto the pre-configured tubular matrices. A 1 cm. long urethral segment was excised in 24 male rabbits. Urethroplasty was performed with the tubularized collagen matrices seeded with cells in 12 animals and without cells in 12. Serial urethrography was performed preoperatively and at 1, 2, 3 and 6 months postoperatively. Retrieved urethras were analyzed grossly, histologically, immunocytochemically and with Western blots. Contractility and the presence of neurotransmitter receptors were confirmed with organ bath studies. Results: Serial urethrography confirmed the maintenance of a wide urethral caliber without any signs of strictures in animals implanted with the cell seeded matrices. The urethral segments replaced with the collagen scaffolds without cells demonstrated strictures and graft collapse at all time points. The implanted cell seeded matrices had a normal urethral architecture by 1 month, consisting of a transitional cell layer surrounded by muscle cell fiber bundles with increasing cellular organization with time. Epithelial and smooth muscle phenotypes were confirmed immunocytochemically and with Western blot analyses using pancytokeratins AE1/AE3 and smooth muscle specific α-actin antibodies. Formation of a transitional cell layer was confirmed in the matrices implanted without cells but only scant unorganized muscle fiber bundles were present, mostly at the anastomotic sites. Organ bath studies demonstrated the capacity for contractility along with cholinergic and adrenergic specific receptors in the tissue engineered scaffolds compared to controls. Conclusions: These results show that collagen matrices seeded with cells form normal urethral tissue can be used for tubularized replacement, whereas tubularized collagen matrices alone without cells lead to poor tissue formation and strictures. The collagen matrices seeded with cells may offer a useful alternative in the future for patients requiring a tubularized urethral segment replacement. References 1 : A novel inert collagen matrix for hypospadias repair. J Urol1999; 162: 1148. Link, Google Scholar 2 : Experimental and clinical experience using tissue regeneration for urethral reconstruction. World J Urol2000; 18: 67. Google Scholar 3 Cilento B. Retik A. Atala A. Urethral reconstruction using polymer scaffolds seeded with urothelial and smooth muscle cells. J Urol, suppl., 155: 371A, abstract 571, 1996 Google Scholar 4 : Bladder augmentation using allogenic bladder submucosa seeded with cells. Urology1998; 51: 221. Google Scholar 5 : Dorsal onlay graft urethroplasty for repair of bulbar urethral stricture. J Urol1999; 161: 815. Link, Google Scholar 6 : Buccal mucosal grafts for urethral reconstruction. Urology1998; 51: 15. Google Scholar 7 : Free vascularized appendix transfer for reconstruction of penile urethras with severe fibrosis. Plast Reconstr Surg1999; 103: 964. Google Scholar 8 : Using buccal mucosa for urethral reconstruction in primary and re-operative surgery. Eur Urol1998; 34: 216. Google Scholar 9 : Split-thickness skin graft urethroplasty and tunica vaginalis flaps for failed hypospadias repairs. J Urol1996; 155: 131. Link, Google Scholar 10 : [Replacement of a urethral segment with a silicone elastomer tube in the male dog (preliminary note). ] J Urol Nephrol1971; 77: 562. Google Scholar 11 : [Urethral substitution with synthetic material. ] Actas Urol Esp2000; 24: 235. Google Scholar 12 : The use of fine double siliconised dacron in urethral replacement. Br J Urol1977; 49: 167. Google Scholar 13 : Urethral reconstruction with a new synthetic absorbable device. An experimental study. Scand J Urol Nephrol1992; 26: 323. Google Scholar 14 : Urethral tube graft in dogs: prosthesis of dacron lined silicone. Invest Urol1978; 15: 408. Google Scholar 15 : New treatment for urethral strictures. Urology1983; 21: 53. Google Scholar 16 : [Replacement of the urethra in man using a silicone elastomer prosthesis. 3 cases. Preliminary results.]. J Urol Nephrol1973; 79: 643. Google Scholar 17 : [Skin graft urethroplasty. ] Prog Urol1999; 9: 112. Google Scholar 18 : Dorsal free graft urethroplasty. J Urol1996; 155: 123. Link, Google Scholar 19 : Complications of paraexstrophy skin flaps in the reconstruction of classical bladder exstrophy. J Urol1993; 150: 627. Link, Google Scholar 20 : An evaluation of skin grafts for reconstruction of the penis and scrotum. Clin Plast Surg1988; 15: 411. Google Scholar 21 : Experience with free grafts in urethral reconstruction. J Urol1987; 137: 471. Link, Google Scholar 22 : Buccal mucosa urethroplasty for the treatment of bulbar urethral strictures. J Urol1999; 161: 1501. Link, Google Scholar 23 : Reconstruction or substitution of the pediatric urethra with buccal mucosa: indications, technical aspects, and results. Tech Urol1999; 5: 133. Google Scholar 24 : Coronal cuff: a problem site for buccal mucosal grafts. J Urol1999; 162: 1442. Link, Google Scholar 25 : [Regeneration of mouth mucosa in the buccal plane following graft procurement for reconstruction of bulbar urethral stenoses]. Mund Kiefer Gesichtshir1999; 3: 34. Google Scholar 26 : Long-term followup of bladder mucosa graft for male urethral reconstruction. J Urol1994; 151: 1056. Link, Google Scholar 27 : Free vascularized appendix transfer for reconstruction of penile urethras with severe fibrosis. Plast Reconstr Surg1999; 103: 964. Google Scholar 28 : Acellular collagen matrix as a possible "off the shelf"; biomaterial for urethral repair. Urology1999; 54: 407. Google Scholar 29 Kassaby E.A. Yoo J.J. Retik A.B. A novel inert collagen matrix for urethral stricture repair. J Urol, suppl., 163: 70, abstract 308, 2000 Google Scholar From the Laboratory for Tissue Engineering and Cellular Therapeutics, Department of Urology, Children's Hospital and Harvard Medical School, Boston, Massachusetts© 2002 by American Urological Association, Inc.FiguresReferencesRelatedDetailsCited byCanning D (2018) Re: Prepubertal Follow-up After Hypospadias Repair with Autologous In Vitro Cultured Urothelial CellsJournal of Urology, VOL. 189, NO. 5, (1904-1905), Online publication date: 1-May-2013.Mikami H, Kuwahara G, Nakamura N, Yamato M, Tanaka M and Kodama S (2018) Two-Layer Tissue Engineered Urethra Using Oral Epithelial and Muscle Derived CellsJournal of Urology, VOL. 187, NO. 5, (1882-1889), Online publication date: 1-May-2012.Li C, Xu Y, Song L, Fu Q, Cui L and Yin S (2018) Urethral Reconstruction Using Oral Keratinocyte Seeded Bladder Acellular Matrix GraftsJournal of Urology, VOL. 180, NO. 4, (1538-1542), Online publication date: 1-Oct-2008.SCHULTHEISS D, GABOUEV A, CEBOTARI S, TUDORACHE I, WALLES T, SCHLOTE N, WEFER J, KAUFMANN P, HAVERICH A, JONAS U, STIEF C and MERTSCHING H (2018) BIOLOGICAL VASCULARIZED MATRIX FOR BLADDER TISSUE ENGINEERING: MATRIX PREPARATION, RESEEDING TECHNIQUE AND SHORT-TERM IMPLANTATION IN A PORCINE MODELJournal of Urology, VOL. 173, NO. 1, (276-280), Online publication date: 1-Jan-2005. Volume 168Issue 4 Part 2October 2002Page: 1789-1793 Advertisement Copyright & Permissions© 2002 by American Urological Association, Inc.Keywordshypospadiasmusclesurotheliumurethral stricturetissue engineeringMetricsAuthor Information Roger E. De Filippo More articles by this author James J. Yoo More articles by this author Anthony Atala More articles by this author Expand All Advertisement PDF downloadLoading ...