Romain Boissier1, Jeremy Magalon2, Florence Sabatier2, Julie Veran3, Laurent Giraudo3, Sophie Giusiano4, Stephane Garcia5, Françoise Dignat-George6, Laurent Arnaud6, Guy Magalon7, Eric Lechevallier8, Stephane Berdah9, Gilles Karsenty8. 1. Aix-Marseille University, 13284, Marseille, France; Department of Urology and Kidney Transplantation, 13285, Assistance Publique Hôpitaux de Marseille, Marseille, France. Electronic address: romain.boissier@ap-hm.fr. 2. Institut national de la santé et de la recherche médicale Unités mixtes de recherche 1076, Aix-Marseille University, 13284, Marseille, France; Department of Cell Therapy, Institut national de la santé et de la recherche médicale Unités mixtes de recherche 1076, 13285, Assistance Publique Hôpitaux de Marseille, Marseille, France; Center for Research and Cliniques en biothérapies 1409, 13285, Assistance Publique Hôpitaux de Marseille, Marseille, France. 3. Center for Research and Cliniques en biothérapies 1409, 13285, Assistance Publique Hôpitaux de Marseille, Marseille, France. 4. Aix-Marseille University, 13284, Marseille, France; Center for Research and Cliniques en biothérapies 1409, 13285, Assistance Publique Hôpitaux de Marseille, Marseille, France; Department of Pathology, 13015, Assistance Publique Hôpitaux de Marseille, Nord University Hospital, Marseille, France. 5. Aix-Marseille University, 13284, Marseille, France. 6. Department of Biology and Hematology, 13285, Assistance Publique Hôpitaux de Marseille, Hospital Conception, Marseille, France. 7. Aix-Marseille University, 13284, Marseille, France; Department of Plastic and Reconstructive Surgery, 13285, Assistance Publique Hôpitaux de Marseille, Marseille, France. 8. Aix-Marseille University, 13284, Marseille, France; Department of Urology and Kidney Transplantation, 13285, Assistance Publique Hôpitaux de Marseille, Marseille, France. 9. Center for Research and Teaching in Surgery, Aix-Marseille University, 13284, Marseille, France; Aix-Marseille University, 13284, Marseille, France.
Abstract
PURPOSE: To evaluate the healing abilities of autologous stem cell therapy (stromal vascular fraction) prepared from adipose tissue we used an automated system without an ex vivo culture phase in a pig model of intrinsic sphincteric deficiency. MATERIALS AND METHODS: A total of 15 pigs underwent endoscopic section of the urethral sphincter. Animals were then randomly assigned to 3 groups, including 1) controls without stromal vascular fraction injection, 2) early injection with stromal vascular fraction 2 to 3 days after section and 3) late stromal vascular fraction injection delivery 30 days after injury. Extraction and stromal vascular fraction injection were performed as a single procedure. The stromal vascular fraction was characterized by flow cytometry. Mesenchymal stem cell-like cells were enumerated by clonogenicity (cfu fibroblast) assay. Study end points included histological assessment of the urethral injury surface and urodynamics to determine maximum urethral pressure. RESULTS: Flow cytometry analysis revealed a mesenchymal stem cell-like phenotype in a mean ± SD of 47.3% ± 11.8% of stromal vascular fraction cells. The cfu fibroblast frequency was 1.3 to 6.6/100 stromal vascular fraction cells (1.3% to 6.6%). Stromal vascular fraction injection was associated with a reduction of the urethral injury surface in the early and late injection groups compared with the respective controls (7% vs 17% and 1% vs 13%, p = 0.050 and 0.029, respectively). On day 30 after injection maximum urethral pressure was significantly higher in the injected groups than in the control group, that is 64% vs 50% of maximum urethral pressure on day 0 (p = 0.04). CONCLUSIONS: These data demonstrate the ability of an autologous stromal vascular fraction to improve the urethral healing process in a large animal model of intrinsic sphincteric deficiency.
PURPOSE: To evaluate the healing abilities of autologous stem cell therapy (stromal vascular fraction) prepared from adipose tissue we used an automated system without an ex vivo culture phase in a pig model of intrinsic sphincteric deficiency. MATERIALS AND METHODS: A total of 15 pigs underwent endoscopic section of the urethral sphincter. Animals were then randomly assigned to 3 groups, including 1) controls without stromal vascular fraction injection, 2) early injection with stromal vascular fraction 2 to 3 days after section and 3) late stromal vascular fraction injection delivery 30 days after injury. Extraction and stromal vascular fraction injection were performed as a single procedure. The stromal vascular fraction was characterized by flow cytometry. Mesenchymal stem cell-like cells were enumerated by clonogenicity (cfu fibroblast) assay. Study end points included histological assessment of the urethral injury surface and urodynamics to determine maximum urethral pressure. RESULTS: Flow cytometry analysis revealed a mesenchymal stem cell-like phenotype in a mean ± SD of 47.3% ± 11.8% of stromal vascular fraction cells. The cfu fibroblast frequency was 1.3 to 6.6/100 stromal vascular fraction cells (1.3% to 6.6%). Stromal vascular fraction injection was associated with a reduction of the urethral injury surface in the early and late injection groups compared with the respective controls (7% vs 17% and 1% vs 13%, p = 0.050 and 0.029, respectively). On day 30 after injection maximum urethral pressure was significantly higher in the injected groups than in the control group, that is 64% vs 50% of maximum urethral pressure on day 0 (p = 0.04). CONCLUSIONS: These data demonstrate the ability of an autologous stromal vascular fraction to improve the urethral healing process in a large animal model of intrinsic sphincteric deficiency.