Françoise Casanas-Roux

Histologic and ultrastructural evaluation of fresh and frozen-thawed human ovarian xenografts in nude mice

OBJECTIVEnTo compare histologic and ultrastructural characteristics of fresh and frozen-thawed human ovarian cortical tissue grafted into nude mice.nnnDESIGNnExperimental prospective study.nnnSETTINGnAn academic research environment.nnnPATIENT(S)nOvarian biopsy specimens were obtained from 13 women undergoing laparoscopy for tubal ligation or infertility.nnnANIMAL(S)nForty nude mice.nnnINTERVENTION(S)nA minilaparotomy was performed to place fresh and frozen-thawed ovarian grafts subcutaneously (sc) or intraperitoneally (ip). Removal of the ovarian grafts was performed at 24 days.nnnMAIN OUTCOME MEASURE(S)n[1] the follicular population, [2] fibrosis, [3] vascularization of the grafted tissue, and [4] ultrastructural evaluation.nnnRESULT(S)nA greater fibrosis relative surface area was noted in frozen-thawed transplanted tissue than in fresh transplants. Regardless of this fibrosis, a similar follicular density was observed in fresh and frozen-thawed ovarian tissue 24 days after transplantation. Active angiogenesis was proved by both immunohistochemical study of the vascular endothelial growth factor and morphometric study of the vascular network. Normal ultrastructural characteristics were noted in frozen-thawed ovarian biopsies.nnnCONCLUSION(S)nAngiogenesis allows implantation of the graft even if it has been cryopreserved and thawed similarly to implantation of fresh tissue. The greater fibrosis observed in grafts after cryopreservation and implantation does not seem to affect the primordial and primary ovocyte population and their ultrastructural characteristics, but further studies must be conducted to prove that after cryopreservation and transplantation, ovocytes may achieve full maturation and fertilization.

Oxidative stress and peritoneal endometriosis

OBJECTIVEnTo review the literature associating pelvic endometriosis with oxidative stress and to discuss the potential causes and consequences of a pro-oxidant environment in the peritoneal cavity.nnnDESIGNnLiterature survey.nnnRESULT(S)nSeveral studies suggest that oxidative stress is a component of the inflammatory reaction associated with endometriosis. Evidence includes the prevention of endometriosis induction in rabbits by the addition of antioxidants, an increase in reactive oxygen species release by macrophages, increased peritoneal levels of oxidized low-density lipoproteins and their by-products, altered expression of endometrial pro-oxidant and antioxidant enzymes, and consumption of peritoneal fluid vitamin E. Retrograde menstruation is likely to carry highly pro-oxidant factors, such as heme and iron, into the peritoneal cavity, as well as apoptotic endometrial cells, which are well-known inducers of oxidative stress. Reactive oxygen species may be involved in endometriosis-associated infertility and may play a role in the regulation of the expression of genes encoding immunoregulators, cytokines and cell adhesion molecules implicated in the pathogenesis of endometriosis.nnnCONCLUSION(S)nBetter understanding of the mechanisms of reactive oxygen species production and detoxification and further investigation of their effect on the peritoneal environment are essential to obtain new insight into this disease and eventually develop new diagnostic and therapeutic strategies.

Early-stage endometriosis: adhesion and growth of human menstrual endometrium in nude mice

OBJECTIVEnTo evaluate the implantation of menstrual endometrium and the early stages of evolution of endometriotic lesions.nnnDESIGNnExperimental prospective study.nnnSETTINGnAn academic research environment.nnnANIMALSnTen nude mice.nnnINTERVENTION(S)nA minilaparotomy was performed to place fresh human menstrual endometrial samples in the peritoneal cavity. Removal of the transplants was performed successively on days 1, 3, and 5 by laparotomy.nnnMAIN OUTCOME MEASURE(S)nAdhesion of endometrial fragments and early stages of endometrial lesions was morphologically and immunohistochemically studied.nnnRESULT(S)nAs early as day 1, stromal cells were found to be attached to the mesothelium. A progressive reorganization of epithelial and stromal cells into endometrial glands was observed. On day 5, cystic endometriotic lesions were characterized by more extensive proliferative activity in glandular cells and a higher VEGF score in stromal cells than that observed in previously removed transplants.nnnCONCLUSION(S)nMenstrual human endometrium is able to implant on intact mesothelium and to reorganize itself into structured glands and stroma under the influence of unknown factors. We suggest that stromal and glandular cells have two distinct roles: stromal cells are involved in the attachment process and glandular cells in the growth of the endometriotic lesion.

Peritoneal endometriosis and “endometriotic” nodules of the rectovaginal septum are two different entities *

OBJECTIVEnTo compare histologically and stereologically the endometriotic nodule of the rectovaginal septum to peritoneal endometriosis.nnnDESIGNnMorphometric investigation, cytokeratin and vimentin content, and steroid receptor evaluation were performed on endometriotic tissue from the peritoneum (n = 52) and rectovaginal nodules (n = 68).nnnSETTINGnAn academic teaching hospital.nnnPATIENTSnBiopsies were taken from 120 patients undergoing a laparoscopy for infertility and/ or pelvic pain (52 from typical black peritoneal endometriotic implants and 68 from endometriotic nodule of the rectovaginal septum). None of the patients were treated.nnnRESULTSnMitotic activity was found to be significantly different in peritoneal and rectovaginal endometriosis. The evaluation suggested that the stroma is not mandatory for the invasion of glandular epithelium in the rectovaginal nodule, which is, like a adenomyoma, a circumscribed nodular aggregate of smooth muscle and glandular elements. Cytokeratin and vimentin content as well as the estrogen receptor (ER) and P receptor (PR) content were significantly lower in both types of lesion when compared with eutopic endometrium. But vimentin immunoreactivity in epithelium, as well as the ER and PR content, were significantly lower in nodules when compared with black peritoneal lesions.nnnCONCLUSIONnIt is suggested that the rectovaginal endometriotic nodule is a different disease from peritoneal endometriosis and must be called rectovaginal adenomyosis or rectovaginal adenomyoma. Its histopathogenesis probably is not related to the implantation of regurgitated endometrial cells but to the metaplasia of Müllerian rests.

Iron overload in the peritoneal cavity of women with pelvic endometriosis.

OBJECTIVEnTo examine the possible involvement of iron in the physiopathology of endometriosis.nnnDESIGNnProspective study.nnnSETTINGnDepartment of gynecology in a university hospital.nnnPATIENT(S)nSeventy patients undergoing laparoscopy.nnnINTERVENTION(S)nCollection of peritoneal fluid (n = 57), blood samples, and biopsy samples from endometrium (n = 62) and from endometriotic (n = 33) and normal-appearing peritoneum (n = 53).nnnMAIN OUTCOME MEASURE(S)nMeasurement of iron and ferritin in serum and peritoneal fluid and staining of iron deposits with Prussian blue in tissues.nnnRESULT(S)nIron and ferritin concentrations were significantly higher in the peritoneal fluid of patients with endometriosis compared with controls during the secretory phase. Higher rates of ferritin and hemosiderin deposits were observed in the peritoneum adjacent to red (100%), black (57%), and white (62%) lesions compared with normal-appearing peritoneum (25%). Deposits were more frequent during the secretory phase than the proliferative phase in healthy peritoneum from controls, whereas they were found throughout the cycle in the vicinity of lesions in patients with endometriosis. Similar rates of iron deposition were observed in the stroma of black and white lesions and in eutopic endometrium from patients with endometriosis.nnnCONCLUSION(S)nIron overload was observed in the cellular and peritoneal fluid compartments of the peritoneal cavity of women with endometriosis. Iron deposits seem to be related to the presence of lesions, suggesting that iron may be involved in the pathogenesis of endometriosis.

Potential involvement of hemoglobin and heme in the pathogenesis of peritoneal endometriosis

OBJECTIVEnTo test whether hemoglobin may accumulate in the peritoneal cavity in case of endometriosis and to assess whether heme oxygenases (HO), detoxifying heme, are expressed in ectopic endometrium and peritoneal cells.nnnDESIGNnProspective study involving patients with and without endometriosis.nnnSETTINGnDepartment of gynecology in a university hospital.nnnPATIENT(S)nSeventy-six patients undergoing laparoscopy for tubal sterilization or infertility and/or pelvic pain.nnnINTERVENTION(S)nCollection of peritoneal fluid (PF), blood samples, and biopsies from endometrium and peritoneum.nnnMAIN OUTCOME MEASURE(S)nMeasurement of free hemoglobin and its byproduct, total and direct bilirubin, in serum and PF and analysis of HO-1 and HO-2 expression in biopsies by reverse transcription polymerase chain reaction and semiquantitative immunohistochemistry.nnnRESULT(S)nHigher levels of hemoglobin were found in the PF of patients with endometriosis. There was no concomitant increase in bilirubin concentrations in the PF, and HO-1 was poorly expressed in peritoneal mesothelium and macrophages. Heme oxygenase-1 and HO-2 were strongly expressed in ectopic endometrium, especially in red lesions.nnnCONCLUSION(S)nOur results suggest that heme may be involved in the pathogenesis and/or development of endometriosis and that the HO system, although expressed, might be insufficient to detoxify heme in women with endometriosis.

Morphometric, immunohistological and three-dimensional evaluation of the endometrium of menopausal women treated by oestrogen and Crinone, a new slow-release vaginal progesterone.

Recently advanced computerized technology was applied to the investigation of morphometric, immunohistological and three-dimensional changes of the endometrial mucosa in order to evaluate quantitatively the effects of three doses of a new slow-release vaginal progesterone on the endometrium in post-menopausal women. A total of 20 menopausal women, deprived of ovarian function, were given oestrogen for 12 days and a combined therapy of oestrogen (administered orally) and progesterone for another 12 day period. Progesterone was administered vaginally through a new gel (Crinone) utilizing a bioadhesive, biocompatible polymer as a base to achieve a sustained release effect. An endometrial biopsy was taken before treatment, after oestrogen-only treatment and after the oestro-progestogen therapy. Before treatment, all the patients exhibited an atrophic endometrium. After oestrogen-only treatment, typical proliferative changes occurred: an increase in the endometrium thickness, an increase in the mitotic index, numerous cylinder-like glands and no coiled glands, and high concentrations of oestrogen receptors (ER) and progesterone receptors (PR). After the oestro-progestogen therapy, whatever the dose of progesterone given, a secretory transformation of the endometrial mucosa occurred, mitotic activity decreased significantly, more ramified and coiled glands were observed, and a decrease in PR content was noted in epithelial and stromal nuclei, and a decrease in PR content was also observed in epithelial nuclei but not in stromal nuclei. Accurate new techniques of image analysis have shown that crinone therapy could eliminate the proliferative effects of oestrogen treatment in post-menopausal women, despite doses as low as 45 mg of progesterone administered vaginally every other day. The results suggest that the sustained release effects of Crinone are clinically relevant.

Typical and subtle atypical presentations of endometriosis.

PURPOSE OF REVIEWnTo review the concept of typical, subtle and invisible endometriosis and analyze the evolution and progression of the disease, as well as some factors possibly involved in the pathogenesis.nnnRECENT FINDINGSnAlthough rejected by Redwine, the concept of non-visible endometriosis has been proven to exist. Some new ideas on the implication of reactive oxygen species in the development of endometriosis and its early stages are described here.nnnSUMMARYnThe increased diagnosis of endometriosis can be explained not only by the increased experience and ability of the surgeon to detect typical and non-typical endometriotic lesions, but also by a better understanding of the pathogenesis.

Expression of inducible heme oxygenase in human endometrium

Heme oxygenase (HO) was initially described as the regulatory enzyme in the heme degradative pathways. It catalyzes the conversion of heme into carbon monoxide (CO), iron, and biliverdin, which acts as an antioxidant. Three isoforms of HO have been described to date: HO-1, which is inducible; HO-2, which is constitutively expressed; and the recently evidenced HO-3, probably involved in heme binding.

Histogenesis of Peritoneal Endometriosis

Endometriosis most commonly affects areas of the pelvic peritoneum close to the ovaries: the uterosacral ligaments, the ovarian fossa peritoneum, and the peritoneum of the cul-de-sac. The diagnosis of peritoneal endometriosis at the time of laparoscopy is often made by observation of typically puckered black or bluish lesions, but there are also numerous subtle appearances of peritoneal endometriosis.1–4 These subtle lesions, frequently nonpigmented, were first diagnosed as endometriosis following confirmation by biopsy in 1986.1 Experience has led to more frequent identification of subtle lesions, which increased in various reports from 15% in 1986 to 65% in 1988.1–6