{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,23]],"date-time":"2026-10-23T21:42:57Z","timestamp":1792791777151,"version":"4.5.0"},"reference-count":41,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2024,11,18]],"date-time":"2024-11-18T00:00:00Z","timestamp":1731888000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Integrated Infrastructure Operational Program","award":["313011AFG5"],"award-info":[{"award-number":["313011AFG5"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>This study evaluates the effects of two decellularization protocols, enzyme-detergent (ED) and detergent-detergent (DD), on the structural and biomechanical properties of human urethral tissue. Urethral samples from 18 individuals were divided into ED (n = 7) and DD (n = 11) groups, with native samples (n = 3) serving as controls. Histological and ultrastructural analyses confirmed that both protocols effectively removed cellular content while preserving essential extracellular matrix (ECM) elements, such as collagen and elastic fibers. Immunohistochemical staining for collagen IV and fibronectin revealed no significant differences between decellularized and native tissues, indicating intact ECM structure. Biomechanical testing demonstrated that DD-treated tissues had significantly lower Cauchy stress (1494.8 \u00b1 518.4 kPa) when compared to native tissues (2439.7 \u00b1 578.7 kPa, p = 0.013), while ED-treated tissues were similar to both groups. Both decellularized groups exhibited reduced stretch at failure and elastic modulus compared to native tissues. Cytotoxicity assays using adipose-derived stem cells demonstrated no signs of toxicity in either protocol. Overall, both ED and DD protocols effectively preserved the urethral ECM structure and mechanical properties, making them suitable for potential use in tissue-engineered grafts and for biobanking purposes. Further research is needed to refine and optimize decellularization methods to improve scaffold recellularization and ensure clinical safety and efficacy.<\/jats:p>","DOI":"10.3390\/ijms252212361","type":"journal-article","created":{"date-parts":[[2024,11,18]],"date-time":"2024-11-18T04:07:09Z","timestamp":1731902829000},"page":"12361","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":9,"title":["Effects of Two Decellularization Protocols on the Mechanical Behavior and Structural Properties of the Human Urethra"],"prefix":"10.3390","volume":"25","author":[{"given":"Marcela","family":"Kuniakova","sequence":"first","affiliation":[{"name":"Institute of Medical Biology, Genetics and Clinical Genetics, Faculty of Medicine, Comenius University in Bratislava, Sasinkova 4, 811 08 Bratislava, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zuzana Varchulova","family":"Novakova","sequence":"additional","affiliation":[{"name":"Institute of Medical Biology, Genetics and Clinical Genetics, Faculty of Medicine, Comenius University in Bratislava, Sasinkova 4, 811 08 Bratislava, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Daniel","family":"Haspinger","sequence":"additional","affiliation":[{"name":"Division of Macroscopic and Clinical Anatomy, Gottfried Schatz Research Center, Medical University of Graz, Auenbruggerplatz 25, A-8036 Graz, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Justyna Anna","family":"Niestrawska","sequence":"additional","affiliation":[{"name":"Division of Macroscopic and Clinical Anatomy, Gottfried Schatz Research Center, Medical University of Graz, Auenbruggerplatz 25, A-8036 Graz, Austria"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7145-7206","authenticated-orcid":false,"given":"Martin","family":"Klein","sequence":"additional","affiliation":[{"name":"Institute of Histology and Embryology, Faculty of Medicine, Comenius University in Bratislava, Sasinkova 4, 811 08 Bratislava, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Paulina","family":"Galfiova","sequence":"additional","affiliation":[{"name":"Institute of Histology and Embryology, Faculty of Medicine, Comenius University in Bratislava, Sasinkova 4, 811 08 Bratislava, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jan","family":"Kovac","sequence":"additional","affiliation":[{"name":"Institute of Medical Biology, Genetics and Clinical Genetics, Faculty of Medicine, Comenius University in Bratislava, Sasinkova 4, 811 08 Bratislava, Slovakia"},{"name":"National Institute of Rheumatic Diseases, Nabr. I. Krasku 4, 921 12 Piestany, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Michal","family":"Palkovic","sequence":"additional","affiliation":[{"name":"Institute of Pathological Anatomy, Faculty of Medicine, Comenius University in Bratislava, Sasinkova 4, 811 08 Bratislava, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5074-9621","authenticated-orcid":false,"given":"Lubos","family":"Danisovic","sequence":"additional","affiliation":[{"name":"Institute of Medical Biology, Genetics and Clinical Genetics, Faculty of Medicine, Comenius University in Bratislava, Sasinkova 4, 811 08 Bratislava, Slovakia"},{"name":"National Institute of Rheumatic Diseases, Nabr. I. Krasku 4, 921 12 Piestany, Slovakia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8230-9383","authenticated-orcid":false,"given":"Niels","family":"Hammer","sequence":"additional","affiliation":[{"name":"Division of Macroscopic and Clinical Anatomy, Gottfried Schatz Research Center, Medical University of Graz, Auenbruggerplatz 25, A-8036 Graz, Austria"},{"name":"Department of Orthopedic and Trauma Surgery, University of Leipzig, Liebigstra\u00dfe 20, 04103 Leipzig, Germany"},{"name":"Division of Biomechatronics, Fraunhofer Institute for Forming Tools, 01187 Dresden, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stanislav","family":"Ziaran","sequence":"additional","affiliation":[{"name":"National Institute of Rheumatic Diseases, Nabr. I. 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Urol."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1111\/bju.14875","article-title":"Trends in Urethral Stricture Management over Two Decades","volume":"124","author":"McGeorge","year":"2019","journal-title":"BJU Int."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"481","DOI":"10.1016\/j.urology.2010.05.055","article-title":"Trends in Stricture Management Among Male Medicare Beneficiaries: Underuse of Urethroplasty?","volume":"77","author":"Anger","year":"2011","journal-title":"Urology"},{"key":"ref_4","first-page":"656459","article-title":"Visual Internal Urethrotomy for Adult Male Urethral Stricture Has Poor Long-Term Results","volume":"2015","author":"Seyam","year":"2015","journal-title":"Adv. Urol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1016\/S0022-5347(01)65942-1","article-title":"Internal Urethrotomy in the Management of Anterior Urethral Strictures: Long-Term Followup","volume":"156","author":"Pansadoro","year":"1996","journal-title":"J. Urol."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1859","DOI":"10.1016\/j.juro.2010.01.020","article-title":"Urethrotomy Has a Much Lower Success Rate Than Previously Reported","volume":"183","author":"Santucci","year":"2010","journal-title":"J. Urol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1097\/01.ju.0000132156.76403.8f","article-title":"Repeat Urethrotomy and Dialation for the Treatment of Urethral Stricture are Neither Clinically Effective Nor Cost-Effective","volume":"172","author":"Greenwell","year":"2004","journal-title":"J. Urol."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"98","DOI":"10.1016\/S0022-5347(01)65296-0","article-title":"Internal Urethrotomy versus Dilation as Treatment for Male Urethral Strictures: A Prospective, Randomized Comparison","volume":"157","author":"Steenkamp","year":"1997","journal-title":"J. Urol."},{"key":"ref_9","unstructured":"EAU (2021, January 9\u201313). EAU Guidelines. Proceedings of the Edn. Presented at the EAU Annual Congress Milan 2021, Milan, Italy."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"797","DOI":"10.1016\/j.eururo.2011.02.010","article-title":"A Systematic Review of Graft Augmentation Urethroplasty Techniques for the Treatment of Anterior Urethral Strictures","volume":"59","author":"Mangera","year":"2011","journal-title":"Eur. Urol."},{"key":"ref_11","doi-asserted-by":"crossref","unstructured":"Sinha, R.J., Singh, V., Sankhwar, S., and Dalela, D. (2009). Donor Site Morbidity in Oral Mucosa Graft Urethroplasty: Implications of Tobacco Consumption. BMC Urol., 9.","DOI":"10.1186\/1471-2490-9-15"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"716","DOI":"10.1016\/j.urology.2005.04.045","article-title":"Comparison of Donor Site Intraoral Morbidity after Mucosal Graft Harvesting for Urethral Reconstruction","volume":"66","author":"Jang","year":"2005","journal-title":"Urology"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"154564","DOI":"10.1155\/2013\/154564","article-title":"Tissue Engineering of Urinary Bladder and Urethra: Advances from Bench to Patients","volume":"2013","author":"Orabi","year":"2013","journal-title":"Sci. World J."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"89","DOI":"10.1038\/aja.2012.91","article-title":"Tissue Engineering in Urethral Reconstruction\u2014An Update","volume":"15","author":"Mangera","year":"2013","journal-title":"Asian J. Androl."},{"key":"ref_15","first-page":"3","article-title":"Collagen Biosynthesis in Lichen Sclerosus et Atrophicus Studied by Biochemical and in Situ Hybridization Techniques","volume":"162","author":"Oikarinen","year":"1991","journal-title":"Acta Derm.-Venereol. Suppl."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1123","DOI":"10.1007\/s00383-020-04722-z","article-title":"Tissue Engineering: From the Bedside to the Bench and Back to the Bedside","volume":"36","author":"Sahakyants","year":"2020","journal-title":"Pediatr. Surg. Int."},{"key":"ref_17","doi-asserted-by":"crossref","unstructured":"Farzamfar, S., Richer, M., Rahmani, M., Naji, M., Aleahmad, M., Chabaud, S., and Bolduc, S. (2023). Biological Macromolecule-Based Scaffolds for Urethra Reconstruction. Biomolecules, 13.","DOI":"10.3390\/biom13081167"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1186\/s13287-022-03164-9","article-title":"Clinical Application of a Double-Modified Sulfated Bacterial Cellulose Scaffold Material Loaded with FGFR2-Modified Adipose-Derived Stem Cells in Urethral Reconstruction","volume":"13","author":"Zhu","year":"2022","journal-title":"Stem Cell Res. Ther."},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Fang, W., Yang, M., Jin, Y., Zhang, K., Wang, Y., Liu, M., Wang, Y., Yang, R., and Fu, Q. (2023). Injectable Decellularized Extracellular Matrix-Based Bio-Ink with Excellent Biocompatibility for Scarless Urethra Repair. Gels, 9.","DOI":"10.3390\/gels9110913"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1016\/j.msec.2006.05.007","article-title":"Development of Poly (Lactic-Co-Glycolic Acid)-Collagen Scaffolds for Tissue Engineering","volume":"27","author":"Wen","year":"2007","journal-title":"Mater. Sci. Eng. C"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"114413","DOI":"10.1016\/j.addr.2022.114413","article-title":"Insights into the Use of Genetically Modified Decellularized Biomaterials for Tissue Engineering and Regenerative Medicine","volume":"188","author":"Zhu","year":"2022","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"233","DOI":"10.1016\/j.jbiotec.2020.11.001","article-title":"Adhesion of Fibroblast Cells on Thin Films Representing Surfaces of Polymeric Scaffolds of Human Urethra Rationalized by Molecular Models of Integrin Binding","volume":"324","author":"Braccini","year":"2020","journal-title":"J. Biotechnol."},{"key":"ref_23","doi-asserted-by":"crossref","unstructured":"Abbas, T.O., Yalcin, H.C., and Pennisi, C.P. (2019). From Acellular Matrices to Smart Polymers: Degradable Scaffolds That Are Transforming the Shape of Urethral Tissue Engineering. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20071763"},{"key":"ref_24","doi-asserted-by":"crossref","unstructured":"Aodi, J., Ying, L., Chengyang, S., and Hongfeng, Z. (2024). Acellular Dermal Matrix in Urethral Reconstruction. Front. Pediatr., 12.","DOI":"10.3389\/fped.2024.1342906"},{"key":"ref_25","first-page":"98","article-title":"Decellularized Extracellular Matrix Biomaterials for Regenerative Therapies: Advances, Challenges and Clinical Prospects","volume":"32","author":"Golebiowska","year":"2024","journal-title":"Bioact. Mater."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"109524","DOI":"10.1016\/j.compositesb.2021.109524","article-title":"Extracellular Matrix Scaffold-Immune Microenvironment Modulates Tissue Regeneration","volume":"230","author":"Yu","year":"2022","journal-title":"Compos. Part B Eng."},{"key":"ref_27","doi-asserted-by":"crossref","unstructured":"Gilpin, A., and Yang, Y. (2017). Decellularization Strategies for Regenerative Medicine: From Processing Techniques to Applications. BioMed Res. Int., 2017.","DOI":"10.1155\/2017\/9831534"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Sim\u00f5es, I.N., Vale, P., Soker, S., Atala, A., Keller, D., Noiva, R., Carvalho, S., Peleteiro, C., Cabral, J.M.S., and Eberli, D. (2017). Acellular Urethra Bioscaffold: Decellularization of Whole Urethras for Tissue Engineering Applications. Sci. Rep., 7.","DOI":"10.1038\/srep41934"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"1034","DOI":"10.1177\/15353702231162092","article-title":"Decellularization of the Human Urethra for Tissue Engineering Applications","volume":"248","author":"Kuniakova","year":"2023","journal-title":"Exp. Biol. Med."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1002\/term.1647","article-title":"Penile Urethra Replacement with Autologous Cell-Seeded Tubularized Collagen Matrices: Collagen Matrices for Urethral Replacement","volume":"9","author":"Kornitzer","year":"2015","journal-title":"J. Tissue Eng. Regen. Med."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"301","DOI":"10.3346\/jkms.2015.30.3.301","article-title":"Urethroplasty Using Autologous Urethral Tissue-Embedded Acellular Porcine Bladder Submucosa Matrix Grafts for the Management of Long-Segment Urethral Stricture in a Rabbit Model","volume":"30","author":"Chun","year":"2015","journal-title":"J. Korean Med. Sci."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2133","DOI":"10.1007\/s00345-019-02836-1","article-title":"Placental Membrane Grafts for Urethral Replacement in a Rabbit Model: A Pilot Study","volume":"38","author":"Pusateri","year":"2020","journal-title":"World J. Urol."},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"E101","DOI":"10.1002\/term.1767","article-title":"A Dynamic Distention Protocol for Whole-Organ Bladder Decellularization: Histological and Biomechanical Characterization of the Acellular Matrix: Whole Bladder Dynamic Decellularization","volume":"10","author":"Consolo","year":"2016","journal-title":"J. Tissue Eng. Regen. Med."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1201","DOI":"10.1089\/ten.tec.2009.0311","article-title":"Development of a Porcine Bladder Acellular Matrix with Well-Preserved Extracellular Bioactive Factors for Tissue Engineering","volume":"16","author":"Yang","year":"2010","journal-title":"Tissue Eng. Part C Methods"},{"key":"ref_35","first-page":"38","article-title":"Acellular Matrix in Urethral Reconstruction","volume":"82\u201383","author":"Sievert","year":"2015","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"843","DOI":"10.1016\/j.jpurol.2024.06.002","article-title":"Assessing the Effects of Bladder Decellularization Protocols on Extracellular Matrix (ECM) Structure, Mechanics, and Biology","volume":"20","author":"Yiu","year":"2024","journal-title":"J. Pediatr. Urol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2100160","DOI":"10.1002\/adhm.202100160","article-title":"Renal Regeneration: The Role of Extracellular Matrix and Current ECM-Based Tissue Engineered Strategies","volume":"10","author":"Quinteira","year":"2021","journal-title":"Adv. Healthc. Mater."},{"key":"ref_38","doi-asserted-by":"crossref","unstructured":"Schleifenbaum, S., Prietzel, T., Aust, G., Boldt, A., Fritsch, S., Keil, I., Koch, H., M\u00f6bius, R., Scheidt, H.A., and Wagner, M.F.X. (2016). Acellularization-Induced Changes in Tensile Properties Are Organ Specific\u2014An In-Vitro Mechanical and Structural Analysis of Porcine Soft Tissues. PLoS ONE, 11.","DOI":"10.1371\/journal.pone.0151223"},{"key":"ref_39","doi-asserted-by":"crossref","unstructured":"Scholze, M., Singh, A., Lozano, P.F., Ondruschka, B., Ramezani, M., Werner, M., and Hammer, N. (2018). Utilization of 3D Printing Technology to Facilitate and Standardize Soft Tissue Testing. Sci. Rep., 8.","DOI":"10.1038\/s41598-018-29583-4"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"e00159","DOI":"10.1016\/j.ohx.2020.e00159","article-title":"Standardized Tensile Testing of Soft Tissue Using a 3D Printed Clamping System","volume":"8","author":"Scholze","year":"2020","journal-title":"HardwareX"},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Lozano, P.F., Scholze, M., Babian, C., Scheidt, H., Vielmuth, F., Waschke, J., Ondruschka, B., and Hammer, N. (2019). Water-Content Related Alterations in Macro and Micro Scale Tendon Biomechanics. Sci. Rep., 9.","DOI":"10.1038\/s41598-019-44306-z"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/22\/12361\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T16:34:22Z","timestamp":1760114062000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/25\/22\/12361"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2024,11,18]]},"references-count":41,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2024,11]]}},"alternative-id":["ijms252212361"],"URL":"https:\/\/doi.org\/10.3390\/ijms252212361","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2024,11,18]]}}}