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Revista Brasileira de Ginecologia e Obstetrícia. 2005;27(11):642-649
DOI 10.1590/S0100-72032005001100002
PURPOSE: to evaluate follicular preservation and histologic characteristics of the cryopreserved ovarian tissue and to compare with the fresh one, in rabbits. METHODS: ten adult female white rabbits were submitted to right oophorectomy. The dried ovary was dissected and the cortex was maintained with approximately 1.5 millimeter thickness. The tissue was fractionated into small sections, some reserved for control histologic study and others destined for cryopreservation. Six weeks later the ovarian tissue was thawed and evaluated histologically. After histologic processing, the control and the experimental samples were stained with hematoxylin and eosin and treated immunohistochemically by the PCNA technique for evaluation of DNA preservation. Histologic alterations present in the fresh and in the cryopreserved tissues were identified, and cryopreserved tissue viability was evaluated. RESULTS: in the cryopreserved tissue only primordial follicles persisted. Reversible alterations were identified: cytoplasmatic vacuolation (p=0,039), stromal lysis (p=0.648) and oocytes with irregular contours (p=0.007). Irreversible alterations: (hyalin degeneration and pyknosis) were found, but not at significant levels (p=0.210). The immunohistochemical analysis showed PCNA staining of follicles at different stages of development in the fresh tissue and primordial follicles in the cryopreserved tissue, indicating the presence of active DNA in both tissues. CONCLUSION: in the cryopreserved ovarian tissue the following were observed: survival of only primordial follicles; significant reversible histologic alterations (cytoplasmic vacuolation, stromal lysis and oocytes with irregular contours); irreversible alterations (hyalin degeneration and pyknosis), and PCNA staining of all follicles.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2005;27(11):642-649
DOI 10.1590/S0100-72032005001100002
PURPOSE: to evaluate follicular preservation and histologic characteristics of the cryopreserved ovarian tissue and to compare with the fresh one, in rabbits. METHODS: ten adult female white rabbits were submitted to right oophorectomy. The dried ovary was dissected and the cortex was maintained with approximately 1.5 millimeter thickness. The tissue was fractionated into small sections, some reserved for control histologic study and others destined for cryopreservation. Six weeks later the ovarian tissue was thawed and evaluated histologically. After histologic processing, the control and the experimental samples were stained with hematoxylin and eosin and treated immunohistochemically by the PCNA technique for evaluation of DNA preservation. Histologic alterations present in the fresh and in the cryopreserved tissues were identified, and cryopreserved tissue viability was evaluated. RESULTS: in the cryopreserved tissue only primordial follicles persisted. Reversible alterations were identified: cytoplasmatic vacuolation (p=0,039), stromal lysis (p=0.648) and oocytes with irregular contours (p=0.007). Irreversible alterations: (hyalin degeneration and pyknosis) were found, but not at significant levels (p=0.210). The immunohistochemical analysis showed PCNA staining of follicles at different stages of development in the fresh tissue and primordial follicles in the cryopreserved tissue, indicating the presence of active DNA in both tissues. CONCLUSION: in the cryopreserved ovarian tissue the following were observed: survival of only primordial follicles; significant reversible histologic alterations (cytoplasmic vacuolation, stromal lysis and oocytes with irregular contours); irreversible alterations (hyalin degeneration and pyknosis), and PCNA staining of all follicles.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2001;23(8):507-513
DOI 10.1590/S0100-72032001000800005
Purpose: to evaluate the effect of hormone replacement therapy on breast cell proliferation and on collagen and elastic fiber formation and to analyze the changes occurring in the breast parenchyma as a whole. Method: a total of 61 adult Wistar rats were divided into 5 groups. The standard group (12 rats) represented the normal hormonal ovarian status. The remaining 49 rats were oophorectomized and, starting on the 96th P.O. day, received the specific drug for 30 days. The CEE group received 50 mg/day conjugated equine estrogens (13 rats); the MPA group, 2.0 mg/day medroxyprogesterone acetate (12 rats); the CEE + MPA group, both drugs (12 rats), and the DW group, distilled water (12 rats). On the 31st day of medication, the animals were sacrificed and the inguinal mammary glands were removed for histological analysis. Cell proliferation was assessed at the ductal and acinar levels using anti-PCNA antibody. Mature collagen (type I) and immature collagen (type III) were quantified by Sirius-Red staining, and elastic fiber formation was quantified by Weigert staining. Anatomopathological analysis was performed by hematoxylin-eosin staining, with the determination of number of acini per terminal duct, number of ducts per field, presence of intraductal secretion, and intensity of intracytoplasmic vacuolization. Results: the CEE + MPA group presented a smaller percentage of proliferating ductal cells (46.1%) (p<0.0001) and a greater proliferation of acinar cells (66.3%), similar to those detected in the MPA group (p=0.075) but differing from those detected in the remaining groups (p<0.004). The CEE group showed the largest amount of immature collagen (33.6%) (p<0.01) and the MPA group showed the highest concentration of elastic fibers (11.7%) (p<0.0001). The CEE + MPA and MPA groups showed secretory acinar hyperplasia that was intense (91.7%) in the CEE + MPA group and mild (41.7%) or moderate (58.3%) in the MPA group, but differering in both cases from the remaining groups (p<0,097). Conclusions: conjugated equine estrogens in combination with medroxyprogesterone inhibit ductal cell proliferation and stimulate acinar cell proliferation causing secretory acinar hyperplasia; conjugated horse estrogens intensify the formation of immature (type III) collagen, and medroxyprogesterone acetate increases the formation of elastic fibers.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2001;23(8):507-513
DOI 10.1590/S0100-72032001000800005
Purpose: to evaluate the effect of hormone replacement therapy on breast cell proliferation and on collagen and elastic fiber formation and to analyze the changes occurring in the breast parenchyma as a whole. Method: a total of 61 adult Wistar rats were divided into 5 groups. The standard group (12 rats) represented the normal hormonal ovarian status. The remaining 49 rats were oophorectomized and, starting on the 96th P.O. day, received the specific drug for 30 days. The CEE group received 50 mg/day conjugated equine estrogens (13 rats); the MPA group, 2.0 mg/day medroxyprogesterone acetate (12 rats); the CEE + MPA group, both drugs (12 rats), and the DW group, distilled water (12 rats). On the 31st day of medication, the animals were sacrificed and the inguinal mammary glands were removed for histological analysis. Cell proliferation was assessed at the ductal and acinar levels using anti-PCNA antibody. Mature collagen (type I) and immature collagen (type III) were quantified by Sirius-Red staining, and elastic fiber formation was quantified by Weigert staining. Anatomopathological analysis was performed by hematoxylin-eosin staining, with the determination of number of acini per terminal duct, number of ducts per field, presence of intraductal secretion, and intensity of intracytoplasmic vacuolization. Results: the CEE + MPA group presented a smaller percentage of proliferating ductal cells (46.1%) (p<0.0001) and a greater proliferation of acinar cells (66.3%), similar to those detected in the MPA group (p=0.075) but differing from those detected in the remaining groups (p<0.004). The CEE group showed the largest amount of immature collagen (33.6%) (p<0.01) and the MPA group showed the highest concentration of elastic fibers (11.7%) (p<0.0001). The CEE + MPA and MPA groups showed secretory acinar hyperplasia that was intense (91.7%) in the CEE + MPA group and mild (41.7%) or moderate (58.3%) in the MPA group, but differering in both cases from the remaining groups (p<0,097). Conclusions: conjugated equine estrogens in combination with medroxyprogesterone inhibit ductal cell proliferation and stimulate acinar cell proliferation causing secretory acinar hyperplasia; conjugated horse estrogens intensify the formation of immature (type III) collagen, and medroxyprogesterone acetate increases the formation of elastic fibers.
