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Bladder smooth muscle cell differentiation and induction of mesenchymal stem cell differentiation in a three dimensional hydrogel matrix
ESPU Meeting 2007
S55
urethral plate is divided into 2 flaps in an oblique fashion (16 patients) or with an only lateral flap (7 patients). We resect the fibrous ventral tissue after mobilization of the plate and we achieve penile straightening. The ventral side of the neo-urethra is harvested from the prepuce with an onlay island flap (18 patients) or penile skin with the Mathieu ‘‘flip-flap’’ procedure (5 patients).
developed during division of the plate and detethering. 8 post operative complications developed (34.7%): Urethrocutaneos fistulae developed in 6 patients, and 2 meatal retractions that required a GAP procedure. No residual curvature or urethral stricture was noted in our series. All 21 patients had excellent cosmetic results.
RESULTS
CONCLUSIONS
Mean follow up was 4.2 years (Range 0.6 to 6.8). No intraoperative complications
We believe that the lengthening of the urethral plate could be a useful procedure
in select cases of primary hypospadias with severe chordee. This technique achieves satisfactory penile straightening and excellent cosmetic results, with a similar complications rate to others procedures for this anomaly.
REFERENCE Vela et al . JUrol 2002; 167: 306-8.
S10: Augmentation / Diversion # S10-1 (PP)
BLADDER SMOOTH MUSCLE CELL DIFFERENTIATION AND INDUCTION OF MESENCHYMAL STEM CELL DIFFERENTIATION IN A THREE DIMENSIONAL HYDROGEL MATRIX ¨ W* and Jeffrey.a. HUBBELL* Peter FREY, Tatiana SEGURA*, Catharina ADELO CHUV, Pediatric Urology, Lausanne, SWITZERLAND - * EPFL, IBI, Lausanne, SWITZERLAND
PURPOSE To allow cell attachment and proliferation an adequate temporary scaffold must be provided, while synthesis of extra-cellular matrix proteins and tissue re-organisation take place. Bladder tissue engineering suffers from the formation of scar tissue partly due to the phenotypic switch of smooth muscle cells from a quiescent contractile to a synthetic proliferative phenotype.
MATERIAL AND METHODS Human bladder smooth muscle and mesenchymal stem cells were successfully cultured three-dimensionally in enzymatically degradable polyethyleneglycol hydrogels, modified with integrin
binding peptides (e.g. RGD peptide sequence). Cell viability was analyzed using the the live/dead staining method (Rodamin reaction).
Furthermore an up-regulation of alpha1 beta1 integrin subunits was noted in favour of differentiation into quiescent contractile smooth muscle cells.
RESULTS CONCLUSIONS On zymography (FACS) the smooth muscle cells showed an up-regulation of matrix metalloproteinase 2 (MMP-2) speaking in favour of differentiation into the quiescent, contractile type of smooth muscle cells. 3-dimensional growth of mesenchymal stem cells in the hydrogel induced up-regulation of smooth muscle alpha-actin and myosin and a downregulation of CD 90, a marker for undifferentiated mesenchymal stem cells. All these findings are in favour of stem cell differentiation into smooth muscle cells.
We have demonstrated the feasibility of a model for three-dimensional growth of human bladder smooth muscle cells, differentiating into the quiescent contractile type and of mesenchymal stem cells differentiating into functional smooth muscle cells. This model can be used to study reconstructive processes and can also contribute to the understanding of underlying basic mechanisms of human smooth muscle cell differentiation.
# S10-2 (PP)
INVOLVEMENT OF CD34D PROGENITOR CELLS IN REGENERATION OF THE BLADDER ACELLULAR MATRIX GRAFT (BAMG): TIME-DEPENDENT NEOVASCULOGENESIS AND REGENERATION OF DIFFERENT BLADDER WALL COMPONENTS Azadeh ELMI, Abdol-mohammad KAJBAFZADEH*, Seyedmehdi PAYABVASHy, Amirali H SALMASIy, Zhina SADEGHI, Seyed Mohammad TAVANGARz and Fatemeh MAHJOUB{ Pediatric Urology Research Centre, Department of Urology, Tehran, IRAN (ISLAMIC REPUBLIC OF) - * Tehran, IRAN (ISLAMIC REPUBLIC OF) - y Children’s Hospital Medical Center, Department of Urology, Tehran, IRAN (ISLAMIC REPUBLIC OF) - z Shariati Hospital, Department of Pathology, Tehran, IRAN (ISLAMIC REPUBLIC OF) - { Children’s Hospital Medical Center, Department of Pathology, Tehran, IRAN (ISLAMIC REPUBLIC OF)
S55
urethral plate is divided into 2 flaps in an oblique fashion (16 patients) or with an only lateral flap (7 patients). We resect the fibrous ventral tissue after mobilization of the plate and we achieve penile straightening. The ventral side of the neo-urethra is harvested from the prepuce with an onlay island flap (18 patients) or penile skin with the Mathieu ‘‘flip-flap’’ procedure (5 patients).
developed during division of the plate and detethering. 8 post operative complications developed (34.7%): Urethrocutaneos fistulae developed in 6 patients, and 2 meatal retractions that required a GAP procedure. No residual curvature or urethral stricture was noted in our series. All 21 patients had excellent cosmetic results.
RESULTS
CONCLUSIONS
Mean follow up was 4.2 years (Range 0.6 to 6.8). No intraoperative complications
We believe that the lengthening of the urethral plate could be a useful procedure
in select cases of primary hypospadias with severe chordee. This technique achieves satisfactory penile straightening and excellent cosmetic results, with a similar complications rate to others procedures for this anomaly.
REFERENCE Vela et al . JUrol 2002; 167: 306-8.
S10: Augmentation / Diversion # S10-1 (PP)
BLADDER SMOOTH MUSCLE CELL DIFFERENTIATION AND INDUCTION OF MESENCHYMAL STEM CELL DIFFERENTIATION IN A THREE DIMENSIONAL HYDROGEL MATRIX ¨ W* and Jeffrey.a. HUBBELL* Peter FREY, Tatiana SEGURA*, Catharina ADELO CHUV, Pediatric Urology, Lausanne, SWITZERLAND - * EPFL, IBI, Lausanne, SWITZERLAND
PURPOSE To allow cell attachment and proliferation an adequate temporary scaffold must be provided, while synthesis of extra-cellular matrix proteins and tissue re-organisation take place. Bladder tissue engineering suffers from the formation of scar tissue partly due to the phenotypic switch of smooth muscle cells from a quiescent contractile to a synthetic proliferative phenotype.
MATERIAL AND METHODS Human bladder smooth muscle and mesenchymal stem cells were successfully cultured three-dimensionally in enzymatically degradable polyethyleneglycol hydrogels, modified with integrin
binding peptides (e.g. RGD peptide sequence). Cell viability was analyzed using the the live/dead staining method (Rodamin reaction).
Furthermore an up-regulation of alpha1 beta1 integrin subunits was noted in favour of differentiation into quiescent contractile smooth muscle cells.
RESULTS CONCLUSIONS On zymography (FACS) the smooth muscle cells showed an up-regulation of matrix metalloproteinase 2 (MMP-2) speaking in favour of differentiation into the quiescent, contractile type of smooth muscle cells. 3-dimensional growth of mesenchymal stem cells in the hydrogel induced up-regulation of smooth muscle alpha-actin and myosin and a downregulation of CD 90, a marker for undifferentiated mesenchymal stem cells. All these findings are in favour of stem cell differentiation into smooth muscle cells.
We have demonstrated the feasibility of a model for three-dimensional growth of human bladder smooth muscle cells, differentiating into the quiescent contractile type and of mesenchymal stem cells differentiating into functional smooth muscle cells. This model can be used to study reconstructive processes and can also contribute to the understanding of underlying basic mechanisms of human smooth muscle cell differentiation.
# S10-2 (PP)
INVOLVEMENT OF CD34D PROGENITOR CELLS IN REGENERATION OF THE BLADDER ACELLULAR MATRIX GRAFT (BAMG): TIME-DEPENDENT NEOVASCULOGENESIS AND REGENERATION OF DIFFERENT BLADDER WALL COMPONENTS Azadeh ELMI, Abdol-mohammad KAJBAFZADEH*, Seyedmehdi PAYABVASHy, Amirali H SALMASIy, Zhina SADEGHI, Seyed Mohammad TAVANGARz and Fatemeh MAHJOUB{ Pediatric Urology Research Centre, Department of Urology, Tehran, IRAN (ISLAMIC REPUBLIC OF) - * Tehran, IRAN (ISLAMIC REPUBLIC OF) - y Children’s Hospital Medical Center, Department of Urology, Tehran, IRAN (ISLAMIC REPUBLIC OF) - z Shariati Hospital, Department of Pathology, Tehran, IRAN (ISLAMIC REPUBLIC OF) - { Children’s Hospital Medical Center, Department of Pathology, Tehran, IRAN (ISLAMIC REPUBLIC OF)