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

International Journal of Molecular Sciences

RizalAbdul Rani¹  Zahra Rashidbenam²  MinHwei Ng²  MohdHafidzul Jasman³  Reynu Rajan⁴  MohamadAznan Shuhaili⁴  NikRitza Kosai⁴  FatimahMohd Nor⁵  FarrahHani Imran⁵  ChristopherChee Kong Ho⁶  EngHong Goh⁷  GuanHee Tan⁷  XengInn Fam⁷  ZulkifliMd Zainuddin⁷ 

[1] Arthoplasty Unit, Department of Orthopaedics and Traumatology Surgery, Universiti Kebangsaan Malaysia Medical Centre, 9th Floor, Clinical Block, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, Kuala Lumpur 56000, Malaysia;Centre for Tissue Engineering and Regenerative Medicine, Universiti Kebangsaan Malaysia Medical Centre, 12th Floor, Clinical Block, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, Kuala Lumpur 56000, Malaysia;Clinical Skills Learning and Simulation Unit, Department of Medical Education, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, Kuala Lumpur 56000, Malaysia;Minimally Invasive Upper Gastrointestinal and Bariatric Surgery Unit, Department of Surgery, Universiti Kebangsaan Malaysia Medical Centre, 8th Floor, Clinical Block, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, Kuala Lumpur 56000, Malaysia;Plastic and Reconstructive Surgery Unit, Department of Surgery, Universiti Kebangsaan Malaysia Medical Centre, Clinical Block, 8th Floor, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, Kuala Lumpur 56000, Malaysia;School of Medicine, Taylor’s University, No. 1 Jalan Taylor’s, Subang Jaya 47500, Selangor Darul Ehsan, Malaysia;Urology Unit, Department of Surgery, Universiti Kebangsaan Malaysia Medical Centre, 8th Floor, Clinical Block, Jalan Yaacob Latif, Bandar Tun Razak, Cheras, Kuala Lumpur 56000, Malaysia;

关键词: adipose-derived stem cell;    self-assembled scaffold;    extracellular matrix;    hypoxic condition;    tissue engineering;    autologous urethral graft;   

DOI  :  10.3390/ijms22073350

来源: DOAJ

【 摘 要 】

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 × 10⁴ cells/cm² 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.

【 授权许可】

Unknown