Volume 59, Issue 1, Pages (January 2011)

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Volume 59, Issue 1, Pages 155-163 (January 2011) Periurethral Injection of Autologous Adipose-Derived Stem Cells with Controlled- Release Nerve Growth Factor for the Treatment of Stress Urinary Incontinence in a Rat Model Weiming Zhao, Cheng Zhang, Chengjun Jin, Zijian Zhang, Deling Kong, Wanhai Xu, Youcheng Xiu European Urology Volume 59, Issue 1, Pages 155-163 (January 2011) DOI: 10.1016/j.eururo.2010.10.038 Copyright © 2010 European Association of Urology Terms and Conditions

Fig. 1 (A) Expression of nerve growth factor (NGF) receptor Ark-A in rat adipose-derived stem cells (ADSC) was determined by real-time polymerase chain reaction and (B) confirmed by western blotting. (C) NGF resulted in increased proliferation of ADSCs in a dose-dependent manner in vitro. ADSC=adipose-derived stem cells; NGF=nerve growth factor. European Urology 2011 59, 155-163DOI: (10.1016/j.eururo.2010.10.038) Copyright © 2010 European Association of Urology Terms and Conditions

Fig. 2 Transverse sections through urethras collected 2 wk after autologous transplantation of adipose-derived stem cells (ADSC) transfected with the β-galactosidase gene and nerve growth factor (NGF): (A) ADSCs and 10 ng of NGF, (B) ADSCs alone, and (C) negative control. Bars represent 50μm. (D) Quantified exogenous β-galactosidase (derived from transplanted cells) in collected urethras. ADSC=adipose-derived stem cells; NGF=nerve growth factor. * p<0.01 vs ADSCs alone. European Urology 2011 59, 155-163DOI: (10.1016/j.eururo.2010.10.038) Copyright © 2010 European Association of Urology Terms and Conditions

Fig. 3 Urodynamic abdominal leak point pressure (ALPP) measurements (A) on the day before pudendal nerve transection (PNT). (B) On day 7 after PNT, there was a large reduction in ALPP compared with the preoperative value. After these measurements, the adipose-derived stem cells (ADSC) and poly(lactic-co-glycolic acid)/nerve growth factor were transplanted. (C) Time-dependent changes in ALPP. ALPP=abdominal leak point pressure; PBS=phosphate-buffered saline; ADSC=adipose-derived stem cells; PLGA=poly(lactic-co-glycolic acid); NGF=nerve growth factor. * p<0.01 vs PBS and ADSC+PLGA groups. Δ p<0.05 vs the ADSC and ADSC+NGF groups. European Urology 2011 59, 155-163DOI: (10.1016/j.eururo.2010.10.038) Copyright © 2010 European Association of Urology Terms and Conditions

Fig. 4 Urodynamic retrograde urethral perfusion pressure (RUPP) measurements (A) on the day before pudendal nerve transection (PNT). (B) On day 7 after PNT, there was a large reduction in RUPP compared with the preoperative value. After these measurements, the adipose-derived stem cells and poly(lactic-co-glycolic acid)/nerve growth factor were transplanted. (C) Time-dependent changes in RUPP. RUPP=retrograde urethral perfusion pressure; PBS=phosphate-buffered saline; ADSC=adipose-derived stem cells; PLGA=poly(lactic-co-glycolic acid); NGF=nerve growth factor. * p<0.01 vs the PBS group. Δ p<0.05 vs the ADSC, ADSC+NGF, and ADSC+PLGA groups. European Urology 2011 59, 155-163DOI: (10.1016/j.eururo.2010.10.038) Copyright © 2010 European Association of Urology Terms and Conditions

Fig. 5 Masson trichrome stain for representative sections and quantified muscle components of the urethras in the five groups. Green represents collagen; red represents muscle. Images were acquired at×10 magnification. PBS=phosphate-buffered saline; ADSC=adipose-derived stem cells; PLGA=poly(lactic-co-glycolic acid); NGF=nerve growth factor. * p<0.01 vs the PBS and ADSC+NGF groups. Δ p<0.05 vs the ADSC and ADSC+PLGA groups. European Urology 2011 59, 155-163DOI: (10.1016/j.eururo.2010.10.038) Copyright © 2010 European Association of Urology Terms and Conditions

Fig. 6 PGP9.5 immunoflourescence acquired at×20 magnification for representative sections of the five groups. Area of positive PGP9.5 staining for the entire urethral cross-section. PBS=phosphate-buffered saline; ADSC=adipose-derived stem cells; PLGA=poly(lactic-co-glycolic acid); NGF=nerve growth factor. * p<0.01 vs the ADSC+NGF group. Δ p<0.05 vs the PBS, ADSC, and ADSC+PLGA groups. European Urology 2011 59, 155-163DOI: (10.1016/j.eururo.2010.10.038) Copyright © 2010 European Association of Urology Terms and Conditions