title: Local injection of mesenchymal stem cells protects testicular torsion-induced germ cell injury.
abstract: BACKGROUND
  Testicular torsion is a urological emergency and infertility is a common complication due to ischemic injury. Surgical reduction and orchiopexy is indicated, but to date there is no effective method for restoration of spermatogenesis. The effects of mesenchymal stem cells (MSCs) on acute tissue injury have been demonstrated, and the abilities of paracrine support, differentiation and immune-modulation may benefit to testicular torsion-induced infertility. We investigate the therapeutic efficacy and the mechanisms of MSCs in testicular torsion-induced germ cell injury when injected locally.
  
  
  METHODS
  Six to eight-week-old Sprague-Dawley rats received surgical 720 degree torsion for 3 hours, followed by detorsion on the left testis. 20 μl of phosphate-buffered saline (PBS) without or with 3 x 10(4) MSCs from human orbital fat tissues (OFSCs) were given for 10 rats, respectively, via local injection into the left testis 30 minutes before detorsion. 20 μl of PBS injection for 6 rats with surgical exposure without torsion served as sham control. Histopathology with Johnsen's score analysis, Western blot analysis for superoxide dismutase 2, Bax, Caspase-3, human insulin growth factor-1 and human stem cell factor, malondialdehyde (MDA) assay in testis and plasma, hormones level including testosterone, follicle-stimulating hormone (FSH) and luteinizing hormone (LH) by ELISA Kits, terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay and fluorescence staining for P450, Sox-9 and VASA were performed.
  
  
  RESULTS
  Animals were sacrificed and bilateral orchiectomy was performed 7 days after torsion-detorsion. Local injections of OFSCs prevented torsion-induced infertility judging from Johnsen's score. TUNEL assay and Western blot analysis on caspase 3 and Bax demonstrated that OFSCs prevented ischemic/reperfusion induced intrinsic apoptosis. MDA assay revealed that OFSCs significantly reduced the oxidative stress in the damaged testicular tissues. After the OFSC injection, serum testosterone secretion was increased, while the elevation of FSH triggered by testicular injury was balanced. OFSCs also produced stem cell factor in the damaged testis. Immunofluorescence staining revealed that most transplanted cells surrounded the Leydig cells. Some of transplanted cells differentiated into p450 expressing cells within 7 days.
  
  
  CONCLUSIONS
  Local injection of allogenic MSCs before surgical detorsion is a simple, clinical friendly procedure to rescue torsion-induced infertility.
authors.affiliation: Department of Urology, Wan Fang Hospital, Taipei Medical University, #111, Section 3, Hsing-Long Road, Taipei, 116, Taiwan. 93062@w.tmu.edu.tw.|Graduate Institute of Clinical Medicine, Taipei Medical University, #250 Wu-Hsing Street, Taipei, 110, Taiwan. 93062@w.tmu.edu.tw.
authors.forename: Chi-Hao
authors.initials: CH
authors.lastname: Hsiao
authors.affiliation: Center for Stem Cell Research, Wan Fang Hospital, Taipei Medical University, #111, Section 3, Hsing-Long Road, Taipei, 116, Taiwan. yoshenhsnu@hotmail.com.
authors.forename: Andrea Tung-Qian
authors.initials: AT
authors.lastname: Ji
authors.affiliation: Graduate Institute of Clinical Medicine, Taipei Medical University, #250 Wu-Hsing Street, Taipei, 110, Taiwan. changpredictor@gmail.com.|Department of Pulmonary Medicine, Shuang Ho Hospital, Taipei Medical University, #291, Zhongzheng Road, Zhonghe District, New Taipei, 235, Taiwan. changpredictor@gmail.com.
authors.forename: Chih-Cheng
authors.initials: CC
authors.lastname: Chang
authors.affiliation: Department of Pathology, College of Medicine, Taipei Medical University, #250, Wu-Hsing Street, Taipei, Taiwan. cjcheng@tmu.edu.tw.|Department of Pathology, Taipei Medical University Hospital, Taipei Medical University, #250, Wu-Hsing Street, Taipei, 110, Taiwan. cjcheng@tmu.edu.tw.
authors.forename: Chien-Jui
authors.initials: CJ
authors.lastname: Cheng
authors.affiliation: Department of Urology, Wan Fang Hospital, Taipei Medical University, #111, Section 3, Hsing-Long Road, Taipei, 116, Taiwan. 86092@w.tmu.edu.tw.
authors.forename: Liang-Ming
authors.initials: LM
authors.lastname: Lee
authors.affiliation: Graduate Institute of Clinical Medicine, Taipei Medical University, #250 Wu-Hsing Street, Taipei, 110, Taiwan. wh9801@yahoo.com.tw.|Center for Stem Cell Research, Wan Fang Hospital, Taipei Medical University, #111, Section 3, Hsing-Long Road, Taipei, 116, Taiwan. wh9801@yahoo.com.tw.
authors.forename: Jennifer Hui-Chun
authors.initials: JH
authors.lastname: Ho
mesh_terms: D000273:Adipose Tissue / Q000166:cytology; D000818:Animals; D017209:Apoptosis; D053148:Caspase 3 / Q000378:metabolism; D005640:Follicle Stimulating Hormone / Q000097:blood; D005854:Germ Cells / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007334:Insulin-Like Growth Factor I / Q000378:metabolism; D007986:Luteinizing Hormone / Q000097:blood; D008297:Male; D008315:Malondialdehyde / Q000378:metabolism; D045164:Mesenchymal Stem Cell Transplantation; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000378:metabolism; D018384:Oxidative Stress; D051381:Rats; D017207:Rats, Sprague-Dawley; D013086:Spermatic Cord Torsion / Q000628:therapy*; D013482:Superoxide Dismutase / Q000378:metabolism; D013737:Testis / Q000473:pathology*; D013739:Testosterone / Q000097:blood; D014183:Transplantation, Heterologous; D000099156:Superoxide Dismutase 2
chemical_list: D053148:Caspase 3; D005640:Follicle Stimulating Hormone; D007334:Insulin-Like Growth Factor I; D007986:Luteinizing Hormone; D008315:Malondialdehyde; D013482:Superoxide Dismutase; D013739:Testosterone; D000099156:Superoxide Dismutase 2; C583557:IGF1 protein, human