1. UKM Learning and Research Repository
2. PHD Thesis / Tesis PHD
3. Faculty of Medicine / Fakulti Perubatan

Title:	Urinary bladder reconstruction using human adipose stem cells and biocompatible scaffold via tissue engineering technique
Authors:	Salah Abood Salem (P49266)
Supervisor:	Ruszymah Idrus, Prof. Dr.
Issue Date:	4-Oct-2014
Description:	The urinary tract may be subjected to both congenital and acquired disorders, all of which may lead to organ damage or loss, requiring eventual reconstruction. The replacement of deficient tissues with functionally equivalent tissues would improve the outcome for these patients. The aim of the current study was to attain this goal with the use of regenerative techniques. Initially basic radiological, ultrasonic and urodynamic evaluations of normal experimental animal urinary bladder were performed respectively. Parameters such as bladder volume, dimensions, filling and voiding pressure were determined to be used as a reference. Human adipose-derived stem cells (HADSCs) were isolated from fat tissues donated by consented patients who underwent surgeries, then cultured and expanded in vitro. At passage 2, the stem cells were differentiated to smooth muscle lineage by incubating it in smooth muscle inductive media (SMIM) consisted of MCDB I31 medium, 1% FBS and 100 U/ml heparin. Differentiated HACSCs showed morphology, genes and protein expression of human smooth muscle cells (HSMCs). Differentiated HACSCs were then seeded on PLGA-collagen or PLGA-fibrin mesh to form a urinary bladder construct. Evaluation of cell biocompatibility to the PLGA-collagen or PLGA-fibrin composite were performed under light and scanning microscope. Mechanical properties, porosity and leaking properties were also evaluated. Eighteen male athymic rats with body weights (200-250) gm were prepared for urinary bladder reconstruction, three groups were prepared. The first control group had urinary bladder resection-anastamosis, the second group consists of urinary bladder reconstruction using PLGA with fibrin (without cells), the third group had urinary bladder reconstruction involve prefabricated PLGA scaffold. Radiography and ultrasonigraphy revealed that the rat bladder as hollow, oval organ, with wall thickness of 1mm at bladder dome and 2mm at bladder base. Urodynamic study showed low intravesical filling pressure of 5.5±0.71 cm H2O voiding pressure of 18.5±0.71 cm H2O and the bladder volume of 1.40 ±0.02 ml. Smooth muscle markers gene Alfa smooth muscle actin (ASMA), calponin, and Myocin heavy chain (MHC) were detected by PCR after 3 and 6 weeks differentiation. The protein expression of ASMA and Smoothelin were expressed after 3 and as well as 6 weeks differentiation. However MHC was expressed only after 6 weeks differentiation. PLGA-Collagen/-Fibrin composites represent a suitable composite for urinary bladder reconstruction in relation to biocompatibility, biodegradability, modulation, and leaking/porosity properties. PLGA-Fibrin composite was superior in relation to leaking /porosity property while PLGA-Collagen was better in terms of cell biocompatibility. Biomechanical test were performed for three layers PLGA with biological adhesive and 3 layers PLGA alone. The tensile stress at failure was 30.82±3.80 (MPa) and 34.36±2.57 (MPa), maximum tensile strain at failure was 19.42±2.24 (mm) and 23.06±2.47 (mm),and the maximum loud at break was 58.55±7.90 (N) and 65.29 ±4.89 (N) respectively. As a conclusion HADSCs could be differentiated effectively into smooth muscle cells, and PLGA collagen/fibrin composites represent a more suitable cell carrier for bladder reconstruction. Preliminary in vivo urinary bladder reconstruction in athymic rat models lead to reconstruction of urinary bladder wall with structural and functional properties comparable to the native bladder.,PhD
Publisher:	UKM, Kuala Lumpur
Appears in Collections:	Faculty of Medicine / Fakulti Perubatan

Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.