Common pitfalls in some of the experimental studies in erectile function and dysfunction: A consensus article

Journal of Sexual Medicine

Volumn 9, Issue 11, 2012, Pages 2770-2784

  Cellek, Selim ^a   Bivalacqua, Trinity J ^b   Burnett, Arthur L ^b   Chitaley, Kanchan ^c   Lin, Ching Shwun ^d  

^a CRANFIELD UNIVERSITY   (United Kingdom)

^b DEPARTMENT OF PATHOLOGY   (United States)

^c UNIVERSITY OF WASHINGTON MEDICAL CENTER   (United States)

^d UNIVERSITY OF CALIFORNIA   (United States)

Author keywords

Erectile; Experimental; Nitric Oxide

Indexed keywords

EID: 84868215243     PISSN: 17436095     EISSN: 17436109     Source Type: Journal    
DOI: 10.1111/j.1743-6109.2012.02916.x     Document Type: Article

References (97)

1. Brioni JD, Moreland RB, Cowart M, Hsieh GC, Stewart AO, Hedlund P, Donnelly-Roberts DL, Nakane M, Lynch JJ III, Kolasa T, Polakowski JS, Osinski MA, Marsh K, Andersson KE, Sullivan JP. Activation of dopamine D4 receptors by ABT-724 induces penile erection in rats. Proc Natl Acad Sci U S A 2004;101:6758-63.
2. Mizusawa H, Hedlund P, Hakansson A, Alm P, Andersson KE. Morphological and functional in vitro and in vivo characterization of the mouse corpus cavernosum. Br J Pharmacol 2001;132:1333-41.
3. Bivalacqua TJ, Armstrong JS, Biggerstaff J, Abdel-Mageed AB, Kadowitz PJ, Hellstrom WJ, Champion HC. Gene transfer of extracellular SOD to the penis reduces O2- and improves erectile function in aged rats. Am J Physiol Heart Circ Physiol 2003;284:H1408-21.
4. Bivalacqua TJ, Usta MF, Champion HC, Leungwattanakij S, Dabisch PA, McNamara DB, Kadowitz PJ, Hellstrom WJ. Effect of combination endothelial nitric oxide synthase gene therapy and sildenafil on erectile function in diabetic rats. Int J Impot Res 2004;16:21-9.
5. Montorsi F, Adaikan G, Becher E, Giuliano F, Khoury S, Lue TF, Sharlip I, Althof SE, Andersson KE, Brock G, Broderick G, Burnett A, Buvat J, Dean J, Donatucci C, Eardley I, Fugl-Meyer KS, Goldstein I, Hackett G, Hatzichristou D, Hellstrom W, Incrocci L, Jackson G, Kadioglu A, Levine L, Lewis RW, Maggi M, McCabe M, McMahon CG, Montague D, Montorsi P, Mulhall J, Pfaus J, Porst H, Ralph D, Rosen R, Rowland D, Sadeghi-Nejad H, Shabsigh R, Stief C, Vardi Y, Wallen K, Wasserman M. Summary of the recommendations on sexual dysfunctions in men. J Sex Med 2010;7:3572-88.
6. Quinlan DM, Nelson RJ, Partin AW, Mostwin JL, Walsh PC. The rat as a model for the study of penile erection. J Urol 1989;141:656-61.
7. Zhang X, Hu L, Yin J, Mo Z, Chen J. Rat model of erectile dysfunction caused by cavernous nerve ablation. Chin Med J (Engl) 2002;115:1179-82.
8. Burnett AL, Becker RE. Immunophilin ligands promote penile neurogenesis and erection recovery after cavernous nerve injury. J Urol 2004;171:495-500.
9. Mullerad M, Donohue JF, Li PS, Scardino PT, Mulhall JP. Functional sequelae of cavernous nerve injury in the rat: Is there model dependency. J Sex Med 2006;3:77-83.
10. Kaufmann O, Claro J, Cury J, Andrade E, Longo B, Aguiar W, Mello L, Srougi M. The development of a rat model of erectile dysfunction after radical prostatectomy: Preliminary findings. BJU Int 2008;102:1026-8.
11. Burnett AL. Strategies to promote recovery of cavernous nerve function after radical prostatectomy. World J Urol 2003;20:337-42.
12. Burnett AL, Lue TF. Neuromodulatory therapy to improve erectile function recovery outcomes after pelvic surgery. J Urol 2006;176:882-7.
13. Bella AJ, Lin G, Cagiannos I, Lue TF. Emerging neuromodulatory molecules for the treatment of neurogenic erectile dysfunction caused by cavernous nerve injury. Asian J Androl 2008;10:54-9.
14. Mulhall JP. Exploring the potential role of neuromodulatory drugs in radical prostatectomy patients. J Androl 2009;30:377-83.
15. Nielsen ME, Schaeffer EM, Marschke P, Walsh PC. High anterior release of the levator fascia improves sexual function following open radical retropubic prostatectomy. J Urol 2008;180:2557-64.
16. Burnett AL. Rationale for cavernous nerve restorative therapy to preserve erectile function after radical prostatectomy. Urology 2003;61:491-7.
17. Hatzimouratidis K, Burnett AL, Hatzichristou D, McCullough AR, Montorsi F, Mulhall JP. Phosphodiesterase type 5 inhibitors in postprostatectomy erectile dysfunction: A critical analysis of the basic science rationale and clinical application. Eur Urol 2009;55:334-47.
18. Klein LT, Miller MI, Buttyan R, Raffo AJ, Burchard M, Devris G, Cao YC, Olsson C, Shabsigh R. Apoptosis in the rat penis after penile denervation. J Urol 1997;158:626-30.
19. User HM, Hairston JH, Zelner DJ, McKenna KE, McVary KT. Penile weight and cell subtype specific changes in a post-radical prostatectomy model of erectile dysfunction. J Urol 2003;169:1175-9.
20. Ferrini MG, Davila HH, Kovanecz I, Sanchez SP, Gonzalez-Cadavid NF, Rajfer J. Vardenafil prevents fibrosis and loss of corporal smooth muscle that occurs after bilateral cavernosal nerve resection in the rat. Urology 2006;68:429-35.
21. Lagoda G, Jin L, Lehrfeld TJ, Liu T, Burnett AL. FK506 and sildenafil promote erectile function recovery after cavernous nerve injury through antioxidative mechanisms. J Sex Med 2007;4:908-16.
22. Mulhall JP, Muller A, Donohue JF, Mullerad M, Kobylarz K, Paduch DA, Tal R, Li PS, Cohen-Gould L, Scardino PT. The functional and structural consequences of cavernous nerve injury are ameliorated by sildenafil citrate. J Sex Med 2008;5:1126-36.
23. Canguven O, Burnett A. Cavernous nerve injury using rodent animal models. J Sex Med 2008;5:1776-85.
24. Pastelin CF, Pacheco P, Camacho M, Cruz Y. Another component of the pelvic plexus that innervates the penis in the rat. Urology 2011;78:232.e7-13.
25. Sezen SF, Hoke A, Burnett AL, Snyder SH. Immunophilin ligand FK506 is neuroprotective for penile innervation. Nat Med 2001;7:1073-4.
26. El-Sakka AI, Hassan MU, Selph C, Perinchery G, Dahiya R, Lue TF. Effect of cavernous nerve freezing on protein and gene expression of nitric oxide synthase in the rat penis and pelvic ganglia. J Urol 1998;160:2245-52.
27. Shin D, Pregenzer G Jr, Gardin JM. Erectile dysfunction: A disease marker for cardiovascular disease. Cardiol Rev 2011;19:5-11.
28. Toblli JE, Cao G, Lombrana A, Rivero M. Functional and morphological improvement in erectile tissue of hypertensive rats by long-term combined therapy with phosphodiesterase type 5 inhibitor and losartan. J Sex Med 2007;4:1291-303.
29. Behr-Roussel D, Chamiot-Clerc P, Bernabe J, Mevel K, Alexandre L, Safar ME, Giuliano F. Erectile dysfunction in spontaneously hypertensive rats: Pathophysiological mechanisms. Am J Physiol Regul Integr Comp Physiol 2003;284:R682-8.
30. Behr-Roussel D, Gorny D, Mevel K, Compagnie S, Kern P, Sivan V, Bernabe J, Bedigian MP, Alexandre L, Giuliano F. Erectile dysfunction: An early marker for hypertension? A longitudinal study in spontaneously hypertensive rats. Am J Physiol Regul Integr Comp Physiol 2005;288:R276-83.
31. Fibbi B, Morelli A, Marini M, Zhang XH, Mancina R, Vignozzi L, Filippi S, Chavalmane A, Silvestrini E, Colli E, Adorini L, Vannelli GB, Maggi M. Atorvastatin but not elocalcitol increases sildenafil responsiveness in spontaneously hypertensive rats by regulating the RhoA/ROCK pathway. J Androl 2008;29:70-84.
32. Toblli JE, Cao G, Casas G, Mazza ON. In vivo and in vitro effects of nebivolol on penile structures in hypertensive rats. Am J Hypertens 2006;19:1226-32.
33. Ushiyama M, Morita T, Kuramochi T, Yagi S, Katayama S. Erectile dysfunction in hypertensive rats results from impairment of the relaxation evoked by neurogenic carbon monoxide and nitric oxide. Hypertens Res 2004;27:253-61.
34. Yono M, Yamamoto Y, Yoshida M, Ueda S, Latifpour J. Effects of doxazosin on blood flow and mRNA expression of nitric oxide synthase in the spontaneously hypertensive rat genitourinary tract. Life Sci 2007;81:218-22.
35. Wilkes N, White S, Stein P, Bernie J, Rajasekaran M. Phosphodiesterase-5 inhibition synergizes rho-kinase antagonism and enhances erectile response in male hypertensive rats. Int J Impot Res 2004;16:187-94.
36. Jiang R, Chen JH, Jin J, Shen W, Li QM. Ultrastructural comparison of penile cavernous tissue between hypertensive and normotensive rats. Int J Impot Res 2005;17:417-23.
37. Sohn DW, Kim HY, Kim SD, Lee EJ, Kim HS, Kim JK, Hwang SY, Cho YH, Kim SW. Elevation of intracavernous pressure and NO-cGMP activity by a new herbal formula in penile tissues of spontaneous hypertensive male rats. J Ethnopharmacol 2008;120:176-80.
38. Ushiyama M, Kuramochi T, Katayama S. Treatment with hypotensive agents affects the impaired relaxation of the penile corpus cavernosum in hypertensive rats. Hypertens Res 2006;29:523-32.
39. Ushiyama M, Kuramochi T, Yagi S, Katayama S. Antioxidant treatment with alpha-tocopherol improves erectile function in hypertensive rats. Hypertens Res 2008;31:1007-13.
40. Carneiro FS, Nunes KP, Giachini FR, Lima VV, Carneiro ZN, Nogueira EF, Leite R, Ergul A, Rainey WE, Clinton WR, Tostes RC. Activation of the ET-1/ETA pathway contributes to erectile dysfunction associated with mineralocorticoid hypertension. J Sex Med 2008;5:2793-807.
41. Chitaley K, Webb RC, Dorrance AM, Mills TM. Decreased penile erection in DOCA-salt and stroke prone-spontaneously hypertensive rats. Int J Impot Res 2001;13(suppl 5):S16-20.
42. Carneiro FS, Giachini FR, Lima VV, Carneiro ZN, Nunes KP, Ergul A, Leite R, Tostes RC, Webb RC. DOCA-salt treatment enhances responses to endothelin-1 in murine corpus cavernosum. Can J Physiol Pharmacol 2008;86:320-8.
43. Martinez AC, Stankevicius E, Jakobsen P, Simonsen U. Blunted non-nitric oxide vasodilatory neurotransmission in penile arteries from renal hypertensive rats. Vascul Pharmacol 2006;44:354-62.
44. Ahn TY, Gómez-Coronado D, Martínez V, Cuevas P, Goldstein I, Sáenz de Tejada I. Enhanced contractility of rabbit corpus cavernosum smooth muscle by oxidized low density lipoproteins. Int J Impot Res 1999;11:9-14.
45. Linder AE, Dorrance AM, Mills TM, Webb RC, Leite R. Erectile function in two-kidney, one-clip hypertensive rats is maintained by a potential increase in nitric oxide production. J Sex Med 2009;6(suppl 3):279-85.
46. Claudino MA, Priviero FB, Camargo EA, Teixeira CE, De Nucci G, Antunes E, Zanesco A. Protective effect of prior physical conditioning on relaxing response of corpus cavernosum from rats made hypertensive by nitric oxide inhibition. Int J Impot Res 2007;19:189-95.
47. Claudino MA, Franco-Penteado CF, Priviero FB, Camargo EA, Teixeira SA, Muscara MN, De Nucci G, Zanesco A, Antunes E. Upregulation of gp91phox subunit of NAD(P)H oxidase contributes to erectile dysfunction caused by long-term nitric oxide inhibition in rats: Reversion by regular physical training. Urology 2010;75:961-7.
48. Claudino MA, da Silva FH, Monica FZ, Rojas-Moscoso JA, De Nucci G, Antunes E. Long-term oral treatment with BAY 41-2272 ameliorates impaired corpus cavernosum relaxations in a nitric oxide-deficient rat model. BJU Int 2011;108:116-22.
49. Tostes RC, Carneiro FS, Lee AJ, Giachini FR, Leite R, Osawa Y, Webb RC. Cigarette smoking and erectile dysfunction: Focus on NO bioavailability and ROS generation. J Sex Med 2008;5:1284-95.
50. McVary KT, Carrier S, Wessells H. Smoking and erectile dysfunction: Evidence based analysis. J Urol 2001;166:1624-32.
51. Elhanbly S, Abdel-Gaber S, Fathy H, El-Bayoumi Y, Wald M, Niederberger CS. Erectile dysfunction in smokers: A penile dynamic and vascular study. J Androl 2004;25:991-5.
52. Rosen MP, Greenfield AJ, Walker TG, Grant P, Dubrow J, Bettmann MA, Fried LE, Goldstein I. Cigarette smoking: An independent risk factor for atherosclerosis in the hypogastric-cavernous arterial bed of men with arteriogenic impotence. J Urol 1991;145:759-63.
53. Kupelian V, Link CL, McKinlay JB. Association between smoking, passive smoking, and erectile dysfunction: Results from the Boston Area Community Health (BACH) Survey. Eur Urol 2007;52:416-22.
54. Corona G, Mannucci E, Petrone L, Ricca V, Mansani R, Cilotti A, Balercia G, Chiarini V, Giommi R, Forti G, Maggi M. Psychobiological correlates of smoking in patients with erectile dysfunction. Int J Impot Res 2005;17:527-34.
55. Sighinolfi MC, Mofferdin A, De Stefani S, Micali S, Cicero AF, Bianchi G. Immediate improvement in penile hemodynamics after cessation of smoking: Previous results. Urology 2007;69:163-5.
56. Juenemann KP, Lue TF, Luo JA, Benowitz NL, Abozeid M, Tanagho EA. The effect of cigarette smoking on penile erection. J Urol 1987;138:438-41.
57. Xie Y, Garban H, Ng C, Rajfer J, Gonzalez-Cadavid NF. Effect of long-term passive smoking on erectile function and penile nitric oxide synthase in the rat. J Urol 1997;157:1121-6.
58. Bivalacqua TJ, Sussan TE, Gebska MA, Strong TD, Berkowitz DE, Biswal S, Burnett AL, Champion HC. Sildenafil inhibits superoxide formation and prevents endothelial dysfunction in a mouse model of secondhand smoke induced erectile dysfunction. J Urol 2009;181:899-906.
59. Gocmez SS, Utkan T, Duman C, Yildiz F, Ulak G, Gacar MN, Erden F. Secondhand tobacco smoke impairs neurogenic and endothelium-dependent relaxation of rabbit corpus cavernosum smooth muscle: Improvement with chronic oral administration of L-arginine. Int J Impot Res 2005;17:437-44.
60. Imamura M, Waseda Y, Marinova GV, Ishibashi T, Obayashi S, Sasaki A, Nagai A, Azuma H. Alterations of NOS, arginase, and DDAH protein expression in rabbit cavernous tissue after administration of cigarette smoke extract. Am J Physiol Regul Integr Comp Physiol 2007;293:R2081-9.
61. Toda N, Ayajiki K, Okamura T. Nitric oxide and penile erectile function. Pharmacol Ther 2005;106:233-66.
62. Cellek S. Let's make NO mistake! Int J Impot Res 2005;17:388-9.
63. Moncada S, Higgs A, Furchgott R. International Union of Pharmacology Nomenclature in Nitric Oxide Research. Pharmacol Rev 1997;49:137-42.
64. Burnett AL. Novel nitric oxide signaling mechanisms regulate the erectile response. Int J Impot Res 2004;16(suppl 1):S15-19.
65. Moncada S, Palmer RM, Higgs EA. Nitric oxide: Physiology, pathophysiology, and pharmacology. Pharmacol Rev 1991;43:109-42.
66. Musicki B, Ross AE, Champion HC, Burnett AL, Bivalacqua TJ. Posttranslational modification of constitutive nitric oxide synthase in the penis. J Androl 2009;30:352-62.
67. Knowles RG. Nitric oxide biochemistry. Biochem Soc Trans 1997;25:895-901.
68. Kalyanaraman B, Darley-Usmar V, Davies KJ, Dennery PA, Forman HJ, Grisham MB, Mann GE, Moore K, Roberts LJ, Ischiropoulos H. Measuring reactive oxygen and nitrogen species with fluorescent probes: Challenges and limitations. Free Radic Biol Med 2012;52:1-6.
69. Kalyanaraman B. Oxidative chemistry of fluorescent dyes: Implications in the detection of reactive oxygen and nitrogen species. Biochem Soc Trans 2011;39:1221-5.
70. Manning P, McNeil CJ. Electrochemical and optical sensing of reactive oxygen species: Pathway to an integrated intracellular and extracellular measurement platform. Biochem Soc Trans 2011;39:1288-92.
71. Vernet D, Nolazco G, Cantini L, Magee TR, Qian A, Rajfer J, Gonzalez-Cadavid NF. Evidence that osteogenic progenitor cells in the human tunica albuginea may originate from stem cells: Implications for Peyronie disease. Biol Reprod 2005;73:1199-210.
72. Ichim TE, Zhong Z, Mikirova NA, Jackson JA, Hunninghake R, Mansilla E, Marin G, Nunez L, Patel AN, Angle N, Murphy MP, Dasanu CA, Alexandrescu DT, Bogin V, Riordan NH. Circulating endothelial progenitor cells and erectile dysfunction: Possibility of nutritional intervention? Panminerva Med 2010;52:75-80.
73. La Vignera S, Condorelli R, Vicari E, D'Agata R, Calogero AE. Circulating endothelial progenitor cells and endothelial microparticles in patients with arterial erectile dysfunction and metabolic syndrome. J Androl 2012;33:202-9.
74. Prokopi M, Pula G, Mayr U, Devue C, Gallagher J, Xiao Q, Boulanger CM, Westwood N, Urbich C, Willeit J, Steiner M, Breuss J, Xu Q, Kiechl S, Mayr M. Proteomic analysis reveals presence of platelet microparticles in endothelial progenitor cell cultures. Blood 2009;114:723-32.
75. Yoder MC. Is endothelium the origin of endothelial progenitor cells? Arterioscler Thromb Vasc Biol 2010;30:1094-103.
76. Lin CS, Xin ZC, Wang Z, Deng C, Huang YC, Lin G, Lue TF. Stem cell therapy for erectile dysfunction: A critical review. Stem Cells Dev 2012;21:343-51.
77. Qiu X, Fandel TM, Ferretti L, Albersen M, Zhang H, Lin G, Lin CS, Schroeder T, Lue TF. Both immediate and delayed intracavernous injection of autologous adipose-derived stromal vascular fraction enhances recovery of erectile function in a rat model of cavernous nerve injury. Eur Urol 2012 Feb 18 [Epub ahead of print] doi: 10.1016/j.eururo.2012.02.003.
78. Nishimatsu H, Suzuki E, Kumano S, Nomiya A, Liu M, Kume H, Homma Y. Adrenomedullin mediates adipose tissue-derived stem cell-induced restoration of erectile function in diabetic rats. J Sex Med 2012;9:482-93.
79. Qiu X, Villalta J, Ferretti L, Fandel TM, Albersen M, Lin G, Dai Y, Lue TF, Lin CS. Effects of intravenous injection of adipose-derived stem cells in a rat model of radiation therapy-induced erectile dysfunction. J Sex Med 2012;9:1834-41.
80. Fandel TM, Albersen M, Lin G, Qiu X, Ning H, Banie L, Lue TF, Lin CS. Recruitment of intracavernously injected adipose-derived stem cells to the major pelvic ganglion improves erectile function in a rat model of cavernous nerve injury. Eur Urol 2012;61:201-10.
81. Lin G, Qiu X, Fandel T, Banie L, Wang G, Lue TF, Lin CS. Tracking intracavernously injected adipose-derived stem cells to bone marrow. Int J Impot Res 2011;23:268-75.
82. Song YS, Lee HJ, Park IH, Kim WK, Ku JH, Kim SU. Potential differentiation of human mesenchymal stem cell transplanted in rat corpus cavernosum toward endothelial or smooth muscle cells. Int J Impot Res 2007;19:378-85.
83. Song YS, Lee HJ, Park IH, Lim IS, Ku JH, Kim SU. Human neural crest stem cells transplanted in rat penile corpus cavernosum to repair erectile dysfunction. BJU Int 2008;102:220-4.
84. Weiss DJ, Liggitt D, Clark JG. Histochemical discrimination of endogenous mammalian beta-galactosidase activity from that resulting from lac-Z gene expression. Histochem J 1999;31:231-6.
85. Billinton N, Knight AW. Seeing the wood through the trees: A review of techniques for distinguishing green fluorescent protein from endogenous autofluorescence. Anal Biochem 2001;291:175-97.
86. Castanheira P, Torquetti LT, Magalhas DR, Nehemy MB, Goes AM. DAPI diffusion after intravitreal injection of mesenchymal stem cells in the injured retina of rats. Cell Transplant 2009;18:423-31.
87. Brulport M, Schormann W, Bauer A, Hermes M, Elsner C, Hammersen FJ, Beerheide W, Spitkovsky D, Hartig W, Nussler A, Horn LC, Edelmann J, Pelz-Ackermann O, Petersen J, Kamprad M, von Mach M, Lupp A, Zulewski H, Hengstler JG. Fate of extrahepatic human stem and precursor cells after transplantation into mouse livers. Hepatology 2007;46:861-70.
88. Kruyt MC, De Bruijn J, Veenhof M, Oner FC, Van Blitterswijk CA, Verbout AJ, Dhert WJ. Application and limitations of chloromethyl-benzamidodialkylcarbocyanine for tracing cells used in bone tissue engineering. Tissue Eng 2003;9:105-15.
89. Li N, Yang H, Lu L, Duan C, Zhao C, Zhao H. Comparison of the labeling efficiency of BrdU, DiI and FISH labeling techniques in bone marrow stromal cells. Brain Res 2008;1215:11-9.
90. Onifer SM, White LA, Whittemore SR, Holets VR. In vitro labeling strategies for identifying primary neural tissue and a neuronal cell line after transplantation in the CNS. Cell Transplant 1993;2:131-49.
91. Schormann W, Hammersen FJ, Brulport M, Hermes M, Bauer A, Rudolph C, Schug M, Lehmann T, Nussler A, Ungefroren H, Hutchinson J, Fandrich F, Petersen J, Wursthorn K, Burda MR, Brustle O, Krishnamurthi K, von Mach M, Hengstler JG. Tracking of human cells in mice. Histochem Cell Biol 2008;130:329-38.
92. Salic A, Mitchison TJ. A chemical method for fast and sensitive detection of DNA synthesis in vivo. Proc Natl Acad Sci U S A 2008;105:2415-20.
93. Rakic P. Neurogenesis in adult primate neocortex: An evaluation of the evidence. Nat Rev Neurosci 2002;3:65-71.
94. Lin G, Huang YC, Shindel AW, Banie L, Wang G, Lue TF, Lin CS. Labeling and tracking of mesenchymal stromal cells with EdU. Cytotherapy 2009;11:864-73.
95. Albersen M, Fandel TM, Lin G, Wang G, Banie L, Lin CS, Lue TF. Injections of adipose tissue-derived stem cells and stem cell lysate improve recovery of erectile function in a rat model of cavernous nerve injury. J Sex Med 2010;7:3331-40.
96. Huang YC, Ning H, Shindel AW, Fandel TM, Lin G, Harraz AM, Lue TF, Lin CS. The effect of intracavernous injection of adipose tissue-derived stem cells on hyperlipidemia-associated erectile dysfunction in a rat model. J Sex Med 2010;7:1391-400.
97. Lin G, Albersen M, Harraz AM, Fandel TM, Garcia M, McGrath MH, Konety BR, Lue TF, Lin CS. Cavernous nerve repair with allogenic adipose matrix and autologous adipose-derived stem cells. Urology 2011;77:1509-8.