Produção Nacional
Revisado por pares
Natural pregnancy in rabbits that underwent oophorectomy and orthotopic allogeneic or autologous ovarian transplantation
2002; Elsevier BV; Volume: 77; Issue: 6 Linguagem: Inglês
10.1016/s0015-0282(02)03107-2
ISSN1556-5653
AutoresAndy Petroianu, Leonardo de Souza Vasconcellos, Luiz Ronaldo Alberti, Lúcia Porto Fonseca de Castro, Juliana Moysés Leite,
Tópico(s)Renal and related cancers
ResumoTo date, most studies on the preservation of ovarian tissue have been limited to experiments with autografts (1Corleta H.E. Corleta O. Capp E. Edelweiss M.I. Subcutaneous autologous ovarian transplantation in Wistar rats maintains hormone secretion.Fertil Steril. 1998; 70: 16-19Abstract Full Text PDF PubMed Scopus (37) Google Scholar, 2Theobald P. Roffé J.L. Berrocal J. Porrier M. Lévy G. Muller G. Autotransplantation ovarienne hétérotopique chez la femme.Presse Med. 1987; 26: 1239-1241Google Scholar, 3Aubard Y. Piver P. Cognié Y. Fermeaux V. Poulin N. Driancourt M.A. Orthotopic and heterotopic autografts of frozen-thawed ovarian cortex sheep.Hum Reprod. 1999; : 2149-2154Crossref PubMed Scopus (197) Google Scholar, 4Baird D.T. Webb R. Campbell B.K. Harkness L.M. Gosden R.G. Long-term ovarian function in sheep after ovariectomy and transplantation of autografts stored at −196°C.Endocrinology. 1999; 140: 462-471Crossref PubMed Scopus (399) Google Scholar, 5Salle B. Cornoge J. Franck M. Isoard L. Rudigoz R.C. Guerin J.F. Freezing, thawing, and autograft of ovarian fragments in sheep.Fertil Steril. 1998; 70: 124-128Abstract Full Text Full Text PDF PubMed Scopus (50) Google Scholar, 6Sztein J. Sweet H. Farley J. Mobraaten L. Cryopreservation and orthotopic transplantation in mouse ovaries.Biol Reprod. 1998; 58: 1071-1074Crossref PubMed Scopus (141) Google Scholar). In contrast, allogeneic or xenogeneic grafts have been little investigated.Successful pregnancies have been established in animals that are immunoincompetent or genetically similar to the donor (7Cox S.L. Shaw J. Jenkin G. Transplantation of cryopreserved fetal ovarian tissue to adult recipients in mice.J Reprod Fertil. 1996; 107: 315-322Crossref PubMed Scopus (138) Google Scholar, 8Green C.J. Simpkin S. Grimaldi G. Johnson A. Pregnancy after autografting and allografting vascularized ovaries and en bloc vascularized ovaries with adnexa in rabbits.Br J Obstet Gynaecol. 1982; 89: 645-651Crossref PubMed Scopus (26) Google Scholar, 9Gunasena K.T. Villines P.M. Critser E.S. Critser J.K. Live births after autologous transplant of cryopreserved mouse ovaries.Hum Reprod. 1997; 12: 101-106Crossref PubMed Scopus (201) Google Scholar). We sought to evaluate the fertility of rabbits after autologous and allogeneic ovary transplantation without a vascular pedicle in the setting of immunosuppression with cyclosporine.We studied 18 sexually mature California or New Zealand White female rabbits and five New Zealand male rabbits that were known to be fertile. The female rabbits were randomized to one of two procedures: removal and orthotopic replacement of both ovaries (8 rabbits) or removal and orthotopic transplantation of both ovaries (10 rabbits). On one side the implanted ovary was intact; on the other side, three sections were made in the ovary before implantation. Ovaries were transplanted from California rabbits (n = 5) to New Zealand White rabbits (n = 5) and vice versa.The postgraduate surgery course review board approved our study.One hour before the operation, all animals undergoing transplantation were weighed and given cyclosporine, 10.0 mg/kg of body weight (Sigmasporin; Natures Plus-Sigma Pharma, São Paulo, Brazil) and cefadroxil, 50.0 mg/kg (Cefamox; Bristol-Myers-Squibb, São Paulo, Brazil). Both substances were dissolved in 10 mL milk and administered through a 12-French orogastric tube. Anesthesia was induced by intramuscular injection of ketamine hydrochloride, 1.0 g/kg (Ketalar; Parke-Davis, São Paulo, Brazil).Each animal undergoing transplantation was weighed weekly to adjust the cyclosporine dose. These rabbits received the cyclosporine solution daily for 9 months. For the first 3 days after surgery, all rabbits undergoing transplantation also received the cefadroxil solution through the orogastric tube.Animals in which the ovaries were replaced received no medication before or after the operation.Three months after the operation, the female rabbits were paired with a sexually mature male almost daily during the other 6 months of the study.At the end of follow-up, blood samples were obtained and E2, P, FSH, and LH were measured. All animals were killed with an ether overdose. Uterus, ovaries, tubes, lungs, liver, spleen and kidney were withdrawn for gross and microscopic assessment. Part of the samples was placed in Bouin’s fixative and part was fixed in formalin and prepared as paraffin-embedded wax blocks. Serial sections were cut at a thickness of 5 μm and stained with hematoxylin and eosin.We used the χ2 test with Yates correction and the Student’s t-test to compare the number of litters and hormones levels of the two groups. P<.05 was considered statistically significant.One rabbit died at the end of surgery. The other rabbits had an uneventful postoperative course and survived for the entire 9-month study.Rabbits receiving autologous implantation and those receiving allogeneic ovary transplantation did not differ in fertility. Live births were recorded in four New Zealand White rabbits that underwent autologous implantation. The deliveries occurred 2 months after the females were paired with a male. The uterus of one New Zealand White rabbit contained two embryos at the end of the experiment. The groups did not differ in the latency period between pairing and birth of litters (Table 1). TABLE 1Hormone levels and litters in rabbits that underwent ovarian autologous implantation or allogeneic transplantation.RabbitNo. of littersE2 level (pg/mL)P level (pg/mL)FSH level (IU/L)LH level (IU/L)Autologous implantationA1011,110103,0000.200.1A2010,400101,0000.300.1A3010,800102,0006.600.1A4314,40091,1000.080.2A5011,000105,0000.100.2A6110,900106,0000.200.1A7aThe blood of these rabbits was hemolized.1————A8aThe blood of these rabbits was hemolized.2————Allogeneic transplantationB1210,900105,0000.10.3B2aThe blood of these rabbits was hemolized.2———B3114,60090,3000.20.1B4012,20099,0000.90.3B5073391,0002.70.1B6011,100105,0002.20.1B7011,100108,0001.00.2B8383729,5000.20.1B914,09037,7001.20.1Groups did not differ significantly for any variable.Petroianu. Rabbit fertility after autologous or allogeneic ovarian transplantation. Fertil Steril 2002.a The blood of these rabbits was hemolized. Open table in a new tab When the rabbits underwent a second laparotomy, moderate adhesions were seen in all animals. The ovaries and tubes of the rabbits without litters were surrounded by more adhesions than were those in animals with litters. On gross examination, all organs were normal, except for the uterus of pregnant rabbits. In both groups, histologic characteristics of all ovaries were normal, with graafian follicles in different phases of maturation. Intact and sliced ovaries did not differ. The tubes of one rabbit that underwent transplantation and did not have a litter were atrophic.Table 1 shows the hormone values at the end of the experiment.In our study, most rabbits became pregnant, but undetected pregnancies may have occurred in both groups. We could not verify abortions or other gestational abnormalities. The absence of pregnancy in some rabbits may have resulted from tube atrophy or thick adhesions surrounding ovaries and tubes.In conclusion, fertility after autogenous or allogeneic ovary transplantation in a rabbit model was successful. A vascular pedicle was not necessary for maintenance of ovarian viability and function (1Corleta H.E. Corleta O. Capp E. Edelweiss M.I. Subcutaneous autologous ovarian transplantation in Wistar rats maintains hormone secretion.Fertil Steril. 1998; 70: 16-19Abstract Full Text PDF PubMed Scopus (37) Google Scholar, 2Theobald P. Roffé J.L. Berrocal J. Porrier M. Lévy G. Muller G. Autotransplantation ovarienne hétérotopique chez la femme.Presse Med. 1987; 26: 1239-1241Google Scholar, 3Aubard Y. Piver P. Cognié Y. Fermeaux V. Poulin N. Driancourt M.A. Orthotopic and heterotopic autografts of frozen-thawed ovarian cortex sheep.Hum Reprod. 1999; : 2149-2154Crossref PubMed Scopus (197) Google Scholar, 10Callejo J. Jáuregui M.T. Valls C. Fernandez M.E. Cabré S. Lailla J.M. Heterotopic ovarian transplantation without vascular pedicle in synergic Lewis rats.Fertil Steril. 1999; 72: 513-517Abstract Full Text Full Text PDF PubMed Scopus (33) Google Scholar). These findings may be useful in future studies of ovarian endocrine function and fertility. To date, most studies on the preservation of ovarian tissue have been limited to experiments with autografts (1Corleta H.E. Corleta O. Capp E. Edelweiss M.I. Subcutaneous autologous ovarian transplantation in Wistar rats maintains hormone secretion.Fertil Steril. 1998; 70: 16-19Abstract Full Text PDF PubMed Scopus (37) Google Scholar, 2Theobald P. Roffé J.L. Berrocal J. Porrier M. Lévy G. Muller G. Autotransplantation ovarienne hétérotopique chez la femme.Presse Med. 1987; 26: 1239-1241Google Scholar, 3Aubard Y. Piver P. Cognié Y. Fermeaux V. Poulin N. Driancourt M.A. Orthotopic and heterotopic autografts of frozen-thawed ovarian cortex sheep.Hum Reprod. 1999; : 2149-2154Crossref PubMed Scopus (197) Google Scholar, 4Baird D.T. Webb R. Campbell B.K. Harkness L.M. Gosden R.G. Long-term ovarian function in sheep after ovariectomy and transplantation of autografts stored at −196°C.Endocrinology. 1999; 140: 462-471Crossref PubMed Scopus (399) Google Scholar, 5Salle B. Cornoge J. Franck M. Isoard L. Rudigoz R.C. Guerin J.F. Freezing, thawing, and autograft of ovarian fragments in sheep.Fertil Steril. 1998; 70: 124-128Abstract Full Text Full Text PDF PubMed Scopus (50) Google Scholar, 6Sztein J. Sweet H. Farley J. Mobraaten L. Cryopreservation and orthotopic transplantation in mouse ovaries.Biol Reprod. 1998; 58: 1071-1074Crossref PubMed Scopus (141) Google Scholar). In contrast, allogeneic or xenogeneic grafts have been little investigated. Successful pregnancies have been established in animals that are immunoincompetent or genetically similar to the donor (7Cox S.L. Shaw J. Jenkin G. Transplantation of cryopreserved fetal ovarian tissue to adult recipients in mice.J Reprod Fertil. 1996; 107: 315-322Crossref PubMed Scopus (138) Google Scholar, 8Green C.J. Simpkin S. Grimaldi G. Johnson A. Pregnancy after autografting and allografting vascularized ovaries and en bloc vascularized ovaries with adnexa in rabbits.Br J Obstet Gynaecol. 1982; 89: 645-651Crossref PubMed Scopus (26) Google Scholar, 9Gunasena K.T. Villines P.M. Critser E.S. Critser J.K. Live births after autologous transplant of cryopreserved mouse ovaries.Hum Reprod. 1997; 12: 101-106Crossref PubMed Scopus (201) Google Scholar). We sought to evaluate the fertility of rabbits after autologous and allogeneic ovary transplantation without a vascular pedicle in the setting of immunosuppression with cyclosporine. We studied 18 sexually mature California or New Zealand White female rabbits and five New Zealand male rabbits that were known to be fertile. The female rabbits were randomized to one of two procedures: removal and orthotopic replacement of both ovaries (8 rabbits) or removal and orthotopic transplantation of both ovaries (10 rabbits). On one side the implanted ovary was intact; on the other side, three sections were made in the ovary before implantation. Ovaries were transplanted from California rabbits (n = 5) to New Zealand White rabbits (n = 5) and vice versa. The postgraduate surgery course review board approved our study. One hour before the operation, all animals undergoing transplantation were weighed and given cyclosporine, 10.0 mg/kg of body weight (Sigmasporin; Natures Plus-Sigma Pharma, São Paulo, Brazil) and cefadroxil, 50.0 mg/kg (Cefamox; Bristol-Myers-Squibb, São Paulo, Brazil). Both substances were dissolved in 10 mL milk and administered through a 12-French orogastric tube. Anesthesia was induced by intramuscular injection of ketamine hydrochloride, 1.0 g/kg (Ketalar; Parke-Davis, São Paulo, Brazil). Each animal undergoing transplantation was weighed weekly to adjust the cyclosporine dose. These rabbits received the cyclosporine solution daily for 9 months. For the first 3 days after surgery, all rabbits undergoing transplantation also received the cefadroxil solution through the orogastric tube. Animals in which the ovaries were replaced received no medication before or after the operation. Three months after the operation, the female rabbits were paired with a sexually mature male almost daily during the other 6 months of the study. At the end of follow-up, blood samples were obtained and E2, P, FSH, and LH were measured. All animals were killed with an ether overdose. Uterus, ovaries, tubes, lungs, liver, spleen and kidney were withdrawn for gross and microscopic assessment. Part of the samples was placed in Bouin’s fixative and part was fixed in formalin and prepared as paraffin-embedded wax blocks. Serial sections were cut at a thickness of 5 μm and stained with hematoxylin and eosin. We used the χ2 test with Yates correction and the Student’s t-test to compare the number of litters and hormones levels of the two groups. P<.05 was considered statistically significant. One rabbit died at the end of surgery. The other rabbits had an uneventful postoperative course and survived for the entire 9-month study. Rabbits receiving autologous implantation and those receiving allogeneic ovary transplantation did not differ in fertility. Live births were recorded in four New Zealand White rabbits that underwent autologous implantation. The deliveries occurred 2 months after the females were paired with a male. The uterus of one New Zealand White rabbit contained two embryos at the end of the experiment. The groups did not differ in the latency period between pairing and birth of litters (Table 1). Groups did not differ significantly for any variable. Petroianu. Rabbit fertility after autologous or allogeneic ovarian transplantation. Fertil Steril 2002. When the rabbits underwent a second laparotomy, moderate adhesions were seen in all animals. The ovaries and tubes of the rabbits without litters were surrounded by more adhesions than were those in animals with litters. On gross examination, all organs were normal, except for the uterus of pregnant rabbits. In both groups, histologic characteristics of all ovaries were normal, with graafian follicles in different phases of maturation. Intact and sliced ovaries did not differ. The tubes of one rabbit that underwent transplantation and did not have a litter were atrophic. Table 1 shows the hormone values at the end of the experiment. In our study, most rabbits became pregnant, but undetected pregnancies may have occurred in both groups. We could not verify abortions or other gestational abnormalities. The absence of pregnancy in some rabbits may have resulted from tube atrophy or thick adhesions surrounding ovaries and tubes. In conclusion, fertility after autogenous or allogeneic ovary transplantation in a rabbit model was successful. A vascular pedicle was not necessary for maintenance of ovarian viability and function (1Corleta H.E. Corleta O. Capp E. Edelweiss M.I. Subcutaneous autologous ovarian transplantation in Wistar rats maintains hormone secretion.Fertil Steril. 1998; 70: 16-19Abstract Full Text PDF PubMed Scopus (37) Google Scholar, 2Theobald P. Roffé J.L. Berrocal J. Porrier M. Lévy G. Muller G. Autotransplantation ovarienne hétérotopique chez la femme.Presse Med. 1987; 26: 1239-1241Google Scholar, 3Aubard Y. Piver P. Cognié Y. Fermeaux V. Poulin N. Driancourt M.A. Orthotopic and heterotopic autografts of frozen-thawed ovarian cortex sheep.Hum Reprod. 1999; : 2149-2154Crossref PubMed Scopus (197) Google Scholar, 10Callejo J. Jáuregui M.T. Valls C. Fernandez M.E. Cabré S. Lailla J.M. Heterotopic ovarian transplantation without vascular pedicle in synergic Lewis rats.Fertil Steril. 1999; 72: 513-517Abstract Full Text Full Text PDF PubMed Scopus (33) Google Scholar). These findings may be useful in future studies of ovarian endocrine function and fertility.
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