Uterine transplant: a successful porcine model.

Wednesday, October 24, 2001 3:00 P.M. O-279 Uterine transplant: a successful porcine model. F. Zakaria, D. Boyle, G. Del Priore, D. Corless, L. Ungars, J. Smith. Chelsea and Westminster Hospital, London, UK. Objectives: To develop a sustainable uterine transplantation model. Design: Case reports and accompanying video. Material and Methods: Human cadaveric dissections were performed in anticipation of developing an animal model. Using this preliminary data, ethics board approval was obtained for investigations in animal models. Both rat (Lewis) and pig transplantation investigations have been undertaken. Initially individual animals were used as both donor and recipient. The porcine model has proven to be more suitable as an anatomic correlate to the human condition. The rat model is currently being developed as a better method for assessment of assisted reproductive technologies in the uterine transplant patient. Variations in operative technique have included different methods of organ harvest (e.g. intact versus sacrificed ovarian vessels and/or uterine pelvic vessels). Other techniques explored included in-situ re-anastomosis (i.e., the donor vaginal cuff attached to the recipient vaginal cuff) or ligated endometrial cavity (i.e., intraperitoneal placement, closed vaginal cuff). Vascular permutations were also investigated including bilateral ovarian and pelvic vascular anastomosis versus bilateral pelvic (i.e., uterine) vessels only. Both end-to-end and end-to-side techniques were investigated. Standard human transplantation protocols were followed including donor heparinization and storage at 4°C. Additional details of the transplant will be demonstrated in the video. Results: After the above described permutations and techniques, we have achieved a stable, long term (⬎6 months) porcine uterine transplantation. It is a suitable model for investigating ART before human transplantation. Conclusions: Uterine transplantation is technically feasible in this animal model. Additional investigations must continue before human transplantation should be considered.

REPRODUCTIVE SURGERY Wednesday, October 24, 2001 3:45 P.M. O-280 Does surgical management of endometriosis (ENDO) within 6 months of in vitro fertilization-embryo transfer (IVF-ET) improve cycle outcome? E. S. Surrey, W. B. Schoolcraft. Colorado Ctr for Reproductive Medicine, Englewood, CO. Objective: Previous investigators have reported that surgical therapy for ENDO may improve short-term spontaneous pregnancy rates (PR) (N Engl J Med 1997;337:217–22). The objectives of this study were to assess the effect of surgical management of ENDO performed in the 6 months prior to an IVF-ET cycle. Design: Retrospective chart review of all patients with a primary diagnosis of ENDO undergoing IVF-ET during a 12 month period in a tertiary care assisted reproductive technology program. Materials/Methods: All patients with a primary diagnosis of ENDO who were candidates for autologous IVF-ET with early follicular phase FSH levels ⬍12 mIU/ml and hysteroscopically normal uterine cavities who had undergone surgical resection/ablation of ENDO (SURG) within 60 months of oocyte retrieval (OPU) and had no evidence of an endometrioma ⬎3 cm were evaluated. Patients who had received prolonged GnRH agonist courses within 9 months of OPU were excluded. Patients were divided into two groups based on the interval between the most recent SURG and OPUGroup I: no more than 6 months’ interval (19 cycles); Group II: 6 months’ interval (76 cycles). Response to ovarian stimulation (COH) and IVF-ET cycle outcome were evaluated. PR and implantation rates (IR) were defined as cardiac activity/ET procedure.Data analysis: Student’s group t-tests, Chi square and regression analysis as appropriate with results expressed as mean ⫾ SEM unless otherwise indicated. Results: The mean SURG-OPU intervals were 3.7 ⫾ 0.3 mos. (Group I) and 21.8 ⫾ 1.6 mos. (Group II) (p ⬍ 0.001). There were no significant differences between the groups with regards to age (I:34.1 ⫾ 0.8 vs. II:

S106

Abstracts

34.1 ⫾ 0.4 years), ENDO score (I: 18.7 ⫾ 5.1 vs. II: 15.8 ⫾ 1.6), COH dose (I: 41.1 ⫾ 3.4 vs. II: 43.5 ⫾ 1.8 ampules), oocytes retrieved (I: 17.8 ⫾ 2.2 vs. II: 17.1 ⫾ 1.1), fertilization rate (I: 62.8 ⫾ 5.3% vs. II: 59.8 ⫾ 2.3%), PR (I: 63.2% vs. II: 60.5%), or IR/ET (I: 34.6 ⫾ 7.9% vs II: 36.2 ⫾ 4.3%). Regression analysis revealed no correlation between IR/ET and either ENDO score (r ⫽ ⫺0.09) or SURG-OPU interval (r ⫽ ⫺0.13). Conclusions: 1) The time interval between surgical management on ENDO and OPU had no significant effect on IVF-ET cycle outcome regardless of the extent of the disease 2) Any potential benefit derived by ENDO surgery in achieving spontaneous conception may be overcome by the inherently higher PR and IR associated with IVF-ET 3) The specific effect of precycle resection of large endometriomata (3 cm) was not addressed in this trial in that all such masses were resected prior to cycle initiation.

Wednesday, October 24, 2001 4:00 P.M. O-281 Significance of the effect of hypoxia on the rate of apoptosis of human peritoneal and adhesion fibroblasts for postoperative adhesion development. G. M. Saed, B. Boytchev, K. Collins, M. P. Diamond. Wayne State Univ, Detroit, MI; Dept of Obstetrics and Gynecology, Wayne State Univ, Detroit, MI. Objective: Clinically, adhesion reformation has been observed to occur more frequently than de novo adhesion formation. Tissue modeling during the wound healing process is governed by a dynamic equilibrium between growth and programmed cell death (apoptosis). Growth control and apoptosis are intimately associated, and a disturbance of the balance between these two processes often leads to pathological situations, such as cell accumulations in cancer and tissue fibrosis. Design: To test the hypothesis that tissue hypoxia, which results from tissue injury during surgery, lowers the rate of apoptosis of peritoneal fibroblasts during peritoneal healing. These fibroblasts remain in the proliferative stage and do not die through apoptosis and therefore continue to produce extracellular molecules (ECM), resulting in adhesion development. Materials/Methods: We have obtained fibroblast primary cultures from normal peritoneal (NF) and adhesion tissues (ADF) of the same patient. Fibroblasts were cultured under normal and hypoxic condition for 24 h before evaluating apoptosis by the Tunel assay. Tunel assay measures the fragmented DNA of apoptotic cells by incorporating fluorescein-12-dUTP at 3⬘-OH DNA ends using the enzyme Terminal Deoxynucleotidyl Transferase (TdT). The fluorescein-labeled DNA was visualized directly by fluorescence microscopy and quantitated by flow cytometry (FACS). Results: Hypoxia resulted in an increase in the rate of apoptosis in peritoneal fibroblasts but decreased apoptosis of adhesion fibroblasts. FACS analysis showed that apoptosis was higher in fibroblasts of normal peritoneum (MFC ⫽ 18.32) than adhesions (MFC ⫽ 14.11) of the same patient in response to hypoxia treatment. Conclusions: Our data suggests that adhesion fibroblasts respond to hypoxia by decreasing their rate of apoptosis, which contribute to the reformation of peritoneal adhesions.

Wednesday, October 24, 2001 4:15 P.M. O-282 Increased transforming growth factor beta 1 (TGF-␤1)/TGF-␤2 ratio following surgically induced peritoneal injury. M. L. Freeman, G. M. Saed, M. P. Diamond. Wayne State Univ, Detroit, MI. Objective: Transforming growth factor beta (TGF-␤) plays a key role in orchestrating wound repair in both the epithelium and mesothelium. Its three isoforms, TGF-␤1, TGF-␤2, and TGF-␤3, have direct roles in regulating the deposition and turnover of extracellular matrix components, vascular in-growth, and fibroblast activation. Previous epidermal studies in murine and human models have shown that an increased TGF-␤1/␤2 ratio in healing tissue is associated with increased fibrosis and scarring. Our

Vol. 76, No. 3, Suppl. 1, September 2001