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Fabrication of Adipose-Derived Stem Cell-Based Self-Assembled Scaffold under Hypoxia and Mechanical Stimulation for Urethral Tissue Engineering

Long urethral strictures are often treated with autologous genital skin and buccal mucosa grafts; however, risk of hair ingrowth and donor site morbidity, restrict their application. To overcome this, we introduced a tissue-engineered human urethra comprising adipose-derived stem cell (ASC)-based se...

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Autores principales: Rashidbenam, Zahra, Jasman, Mohd Hafidzul, Tan, Guan Hee, Goh, Eng Hong, Fam, Xeng Inn, Ho, Christopher Chee Kong, Zainuddin, Zulkifli Md, Rajan, Reynu, Rani, Rizal Abdul, Nor, Fatimah Mohd, Shuhaili, Mohamad Aznan, Kosai, Nik Ritza, Imran, Farrah Hani, Ng, Min Hwei
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8036589/
https://www.ncbi.nlm.nih.gov/pubmed/33805910
http://dx.doi.org/10.3390/ijms22073350
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author Rashidbenam, Zahra
Jasman, Mohd Hafidzul
Tan, Guan Hee
Goh, Eng Hong
Fam, Xeng Inn
Ho, Christopher Chee Kong
Zainuddin, Zulkifli Md
Rajan, Reynu
Rani, Rizal Abdul
Nor, Fatimah Mohd
Shuhaili, Mohamad Aznan
Kosai, Nik Ritza
Imran, Farrah Hani
Ng, Min Hwei
author_facet Rashidbenam, Zahra
Jasman, Mohd Hafidzul
Tan, Guan Hee
Goh, Eng Hong
Fam, Xeng Inn
Ho, Christopher Chee Kong
Zainuddin, Zulkifli Md
Rajan, Reynu
Rani, Rizal Abdul
Nor, Fatimah Mohd
Shuhaili, Mohamad Aznan
Kosai, Nik Ritza
Imran, Farrah Hani
Ng, Min Hwei
author_sort Rashidbenam, Zahra
collection PubMed
description Long urethral strictures are often treated with autologous genital skin and buccal mucosa grafts; however, risk of hair ingrowth and donor site morbidity, restrict their application. To overcome this, we introduced a tissue-engineered human urethra comprising adipose-derived stem cell (ASC)-based self-assembled scaffold, human urothelial cells (UCs) and smooth muscle cells (SMCs). ASCs were cultured with ascorbic acid to stimulate extracellular matrix (ECM) production. The scaffold (ECM) was stained with collagen type-I antibody and the thickness was measured under a confocal microscope. Results showed that the thickest scaffold (28.06 ± 0.59 μm) was achieved with 3 × 10(4) cells/cm(2) seeding density, 100 μg/mL ascorbic acid concentration under hypoxic and dynamic culture condition. The biocompatibility assessment showed that UCs and SMCs seeded on the scaffold could proliferate and maintain the expression of their markers (CK7, CK20, UPIa, and UPII) and (α-SMA, MHC and Smootheline), respectively, after 14 days of in vitro culture. ECM gene expression analysis showed that the ASC and dermal fibroblast-based scaffolds (control) were comparable. The ASC-based scaffold can be handled and removed from the plate. This suggests that multiple layers of scaffold can be stacked to form the urothelium (seeded with UCs), submucosal layer (ASCs only), and smooth muscle layer (seeded with SMCs) and has the potential to be developed into a fully functional human urethra for urethral reconstructive surgeries.
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spelling pubmed-80365892021-04-12 Fabrication of Adipose-Derived Stem Cell-Based Self-Assembled Scaffold under Hypoxia and Mechanical Stimulation for Urethral Tissue Engineering Rashidbenam, Zahra Jasman, Mohd Hafidzul Tan, Guan Hee Goh, Eng Hong Fam, Xeng Inn Ho, Christopher Chee Kong Zainuddin, Zulkifli Md Rajan, Reynu Rani, Rizal Abdul Nor, Fatimah Mohd Shuhaili, Mohamad Aznan Kosai, Nik Ritza Imran, Farrah Hani Ng, Min Hwei Int J Mol Sci Article Long urethral strictures are often treated with autologous genital skin and buccal mucosa grafts; however, risk of hair ingrowth and donor site morbidity, restrict their application. To overcome this, we introduced a tissue-engineered human urethra comprising adipose-derived stem cell (ASC)-based self-assembled scaffold, human urothelial cells (UCs) and smooth muscle cells (SMCs). ASCs were cultured with ascorbic acid to stimulate extracellular matrix (ECM) production. The scaffold (ECM) was stained with collagen type-I antibody and the thickness was measured under a confocal microscope. Results showed that the thickest scaffold (28.06 ± 0.59 μm) was achieved with 3 × 10(4) cells/cm(2) seeding density, 100 μg/mL ascorbic acid concentration under hypoxic and dynamic culture condition. The biocompatibility assessment showed that UCs and SMCs seeded on the scaffold could proliferate and maintain the expression of their markers (CK7, CK20, UPIa, and UPII) and (α-SMA, MHC and Smootheline), respectively, after 14 days of in vitro culture. ECM gene expression analysis showed that the ASC and dermal fibroblast-based scaffolds (control) were comparable. The ASC-based scaffold can be handled and removed from the plate. This suggests that multiple layers of scaffold can be stacked to form the urothelium (seeded with UCs), submucosal layer (ASCs only), and smooth muscle layer (seeded with SMCs) and has the potential to be developed into a fully functional human urethra for urethral reconstructive surgeries. MDPI 2021-03-25 /pmc/articles/PMC8036589/ /pubmed/33805910 http://dx.doi.org/10.3390/ijms22073350 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) ).
spellingShingle Article
Rashidbenam, Zahra
Jasman, Mohd Hafidzul
Tan, Guan Hee
Goh, Eng Hong
Fam, Xeng Inn
Ho, Christopher Chee Kong
Zainuddin, Zulkifli Md
Rajan, Reynu
Rani, Rizal Abdul
Nor, Fatimah Mohd
Shuhaili, Mohamad Aznan
Kosai, Nik Ritza
Imran, Farrah Hani
Ng, Min Hwei
Fabrication of Adipose-Derived Stem Cell-Based Self-Assembled Scaffold under Hypoxia and Mechanical Stimulation for Urethral Tissue Engineering
title Fabrication of Adipose-Derived Stem Cell-Based Self-Assembled Scaffold under Hypoxia and Mechanical Stimulation for Urethral Tissue Engineering
title_full Fabrication of Adipose-Derived Stem Cell-Based Self-Assembled Scaffold under Hypoxia and Mechanical Stimulation for Urethral Tissue Engineering
title_fullStr Fabrication of Adipose-Derived Stem Cell-Based Self-Assembled Scaffold under Hypoxia and Mechanical Stimulation for Urethral Tissue Engineering
title_full_unstemmed Fabrication of Adipose-Derived Stem Cell-Based Self-Assembled Scaffold under Hypoxia and Mechanical Stimulation for Urethral Tissue Engineering
title_short Fabrication of Adipose-Derived Stem Cell-Based Self-Assembled Scaffold under Hypoxia and Mechanical Stimulation for Urethral Tissue Engineering
title_sort fabrication of adipose-derived stem cell-based self-assembled scaffold under hypoxia and mechanical stimulation for urethral tissue engineering
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8036589/
https://www.ncbi.nlm.nih.gov/pubmed/33805910
http://dx.doi.org/10.3390/ijms22073350
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