title: Functional and histological changes after myoblast injections in the porcine rhabdosphincter.
abstract: OBJECTIVE
  Transurethral ultrasound-guided injection of autologous myoblasts has recently been shown to cure urinary stress incontinence. In the present study, the dose-dependent changes in maximal urethral closure pressures after application of myoblasts were investigated in a porcine animal model.
  
  
  METHODS
  Myoblast cultures were grown from a porcine muscle biopsy. The biopsy was enzymatically dissociated by using a modified cell dispersion technique. Single myoblasts in suspension were manually collected with a micropipette under microscopic control. Next a clonal myoblast culture was prepared. Before the cells were applied, fluorescence labelling (PKH) was used to assess integration of the injected myoblasts into the rhabdosphincter. With the help of a transurethral ultrasound probe (23 F, 11 MHz) and a special injection system, the myoblasts were injected into the rhabdosphincter of five pigs under direct sonographic control. Into two different areas of the rhabdosphincter, increasing different cell counts were injected (total volume 1.5 ml). At each area, 10 depots of 150 microl volume were injected all along the rhabdosphincter. The following cell counts were used: 1.5 x 10(6), 2.1 x 10(6), 4.2 x 10(6) (low range) 5.69 x 10(6), 8.1 x 10(6), 1.13 x 10(7), 1.6 x 10(7) (mid range) 2.26 x 10(7), 4.4 x 10(7), and 7.8 x 10(7) (high range). To avoid possible cell rejection, we immunosuppressed the pigs with daily cortisone (1g Solu Dacortin) because allogenic myoblasts were used. Urethral pressure profiles (UPPs) were measured before and 3 wk postoperatively before the pigs were put to sleep. The lower urinary tract was removed in all pigs for histological analysis.
  
  
  RESULTS
  Histological examination of the specimens revealed that the injected cells had survived at the injection site and had formed new myofibres. Overall the UPP curves revealed dose-dependent changes. Statistically significant increased pressure values of up to more than 300% could be observed in all cases in which higher concentrations of cells had been applied. Increases were also noted in mid range concentrations although not to such a high extent (approximately 150%). Pressure values had even diminished (approximately 50%) after injecting the three lowest concentrations (1.5 x 10(6), 2.1 x 10(6), 4.2 x 10(6)).
  
  
  CONCLUSIONS
  The present results show that the effects after application of myoblasts into the rhabdosphincter are dose-dependent.
authors.affiliation: Department of Urology, Medical University of Innsbruck, Innsbruck, Austria. michael.mitterberger@uibk.ac.at
authors.forename: Michael
authors.initials: M
authors.lastname: Mitterberger
authors.forename: Germar M
authors.initials: GM
authors.lastname: Pinggera
authors.forename: Rainer
authors.initials: R
authors.lastname: Marksteiner
authors.forename: Eva
authors.initials: E
authors.lastname: Margreiter
authors.forename: Raffael
authors.initials: R
authors.lastname: Plattner
authors.forename: Günter
authors.initials: G
authors.lastname: Klima
authors.forename: Georg
authors.initials: G
authors.lastname: Bartsch
authors.forename: Hannes
authors.initials: H
authors.lastname: Strasser
mesh_terms: D000818:Animals; D002478:Cells, Cultured; D009119:Muscle Contraction; D032446:Myoblasts / Q000637:transplantation*; D011312:Pressure; D013552:Swine; D014521:Urethra / Q000473:pathology / Q000503:physiopathology*; D014550:Urinary Incontinence, Stress / Q000628:therapy*