Biomedical subjects

Anne Van Langendonckt

Publications and source records attributed to Anne Van Langendonckt.

At least 19 recordsLinked to original sources

PubMed2006

Iron overload enhances epithelial cell proliferation in endometriotic lesions induced in a murine model.

BACKGROUND: Iron deposits are characteristic of endometriotic lesions, and pelvic iron concentrations are higher in endometriosis patients than in women without endometriosis. In this study, the effect of iron overload and iron chelation on the development of endometriosis in a murine model was investigated. METHODS: Human menstrual endometrium was injected i.p. into nude mice, either alone (controls) or supplemented with erythrocytes or desferrioxamine (DFO), an iron chelator. After 5 days, the iron load of endometriosis-like lesions and peritoneal macrophages and fluid was evaluated. Lesions were quantified by immunohistochemical morphometry, and their proliferative activity was assessed. RESULTS: Injection of erythrocytes into the pelvic cavity caused iron overload in lesions (P < 0.025) and peritoneal macrophages (P < 0.01) and fluid (P < 0.05), whereas DFO effectively reduced iron status in lesions (P < 0.05) and macrophages (P < 0.01) compared with controls. No difference was observed in the number or surface area of lesions between the three groups. Erythrocytes increased (P < 0.05) and DFO significantly decreased (P < 0.01) the proliferative activity of lesions. CONCLUSIONS: Iron overload does not appear to affect lesion establishment but may contribute to the further growth of endometriosis by promoting cell proliferation of lesions. Iron chelator treatment could therefore be beneficial in endometriosis to prevent iron overload in the pelvic cavity and decrease cellular proliferation of lesions.

Animals↗

PubMed2006

Ovarian tissue cryopreservation and transplantation: a review.

The review covers current options for ovarian tissue cryopreservation and transplantation and provides a systematic review of the existing literature from the last 10 years, taking into account all previously published reviews on the subject. The different cryopreservation options available for fertility preservation in cancer patients are embryo cryopreservation, oocyte cryopreservation and ovarian tissue cryopreservation. The choice depends on various parameters: the type and timing of chemotherapy, the type of cancer, the patient's age and the partner status. The different options and their results are discussed, as well as their putative indications and efficacy. The review concludes that advances in reproductive technology have made fertility preservation techniques a real possibility for patients whose gonadal function is threatened by premature menopause, or by treatments such as radiotherapy, chemotherapy or surgical castration.

Antineoplastic Agents↗

PubMed2005

Quantification of endometriotic lesions in a murine model by fluorimetric and morphometric analyses.

BACKGROUND: In animal models of endometriosis, the identification and quantification of lesions originating from human endometrium is often hampered by the small size of the implants and their embedding in murine tissue. The purpose of the present study was to develop two new methods of quantifying endometriosis-like lesions in a nude mouse model: fluorimetry and morphometry. METHODS: Human menstrual endometrium was labelled using a fluorescent tracker, carboxyfluorescein diacetate, succinimidyl ester (CFDA-SE), and transplanted into the pelvic cavity of mice by injection through the peritoneum after performing a cutaneous incision. After 5 days, lesions were recovered by laparotomy. The fluorescence of the recovered endometriotic lesions was measured. Endometrial stroma and glands were immunostained in lesion sections with anti-CD10 and anti-CK22 antibodies, and their surface area was evaluated by morphometric analysis. RESULTS: Fluorescent labelling allows identification of lesions not visible macroscopically. A good correlation was observed between fluorimetry and morphometry (r=0.88) applied for lesion quantification. CONCLUSIONS: Fluorimetric evaluation combined with morphometric analysis of endometriosis-like lesions allows objective and reliable recording of endometriosis development in a nude mouse model. This quantification method could therefore be useful for future pharmacological and toxicological studies.

Animals↗

PubMed2005

Evaluation of Liberase, a purified enzyme blend, for the isolation of human primordial and primary ovarian follicles.

BACKGROUND: The purpose of this study is to evaluate the effectiveness of a standardized mixture of purified enzymes (Liberase), for the isolation of human ovarian follicles. METHODS: This is an experimental prospective study. Ovarian biopsies were obtained from eight young women undergoing laparoscopy for benign gynaecological disease. Follicles were isolated by Liberase or collagenase enzymatic digestion. Follicle quality was assessed by evaluating their general morphology and viability after fluorescent staining, and the ultrastructure by electron microscopy. RESULTS: The number of fully isolated follicles recovered from the Liberase-treated group was lower than from the collagenase group (156 versus 263) despite equal-sized biopsies being taken. A high proportion of follicles (98.6%, 70/71) were viable after Liberase isolation and most follicles were of good morphology with a complete granulosa cell layer (70.4%, 31/44). Ultrastructural studies indicated that Liberase-isolated follicles showed signs of atresia only occasionally and that the oolemma-follicular cell interface was well preserved. CONCLUSIONS: Liberase treatment allows the isolation of highly viable follicles from human ovarian tissue, with an unaltered morphology and ultrastructure. This purified endotoxin-free enzyme preparation is a promising alternative to impure collagenase preparations for the reproducible isolation of intact primordial and primary follicles for culture and grafting purposes.

Collagenases↗

PubMed2005

Time course of pelvic endometriotic lesion revascularization in a nude mouse model.

OBJECTIVE: To assess the timing of endometrial lesion revascularization in a murine model. DESIGN: Prospective experimental study. SETTING: An academic research environment. ANIMAL(S): Twenty-six nude mice. INTERVENTION(S): Endometriosis was induced in mice by intraperitoneal deposition of human menstrual endometrium. Endometrial implants were recovered on days 1, 3, 5, 8, 10, 15, and 21 after implantation. MAIN OUTCOME MEASURE(S): Sections from the endometrial implants were immunostained with species-specific antiplatelet endothelial cell adhesion molecule-1 (PECAM-1) antibodies and vessels of murine and human origin were counted. RESULT(S): Endothelial cells of human origin in the implant progressively disappeared between day 3 and day 10. Seventy-eight percent of the vessel sections were positive for human PECAM-1 on day 5, 40.1% on day 8, and only 14.1% on day 10. However, there was a marked increase in murine PECAM-1-expressing vessels in endometrial stroma between day 5 (1.4%) and day 8 (68.0%), 10 (69.5%), and 15 (87.2%). CONCLUSION(S): Our study demonstrates that PECAM-1 is a reliable endothelial cell marker to evaluate the role of angiogenesis in the nude mouse model. It also indicates that revascularization of human endometrial implants occurs between 5 and 8 days after implantation and involves the disappearance of native graft vessels, coinciding with the invasion of the interface and then the stroma by murine vessels.

Animals↗

PubMed2005

Orthotopic transplantation of fresh ovarian cortex: a report of two cases.

OBJECTIVE: To report two cases of orthotopic transplantation of fresh ovarian tissue. SETTING: Academic hospital. PATIENT(S): Two patients with severe endometriosis, who underwent left oophorectomy for recurrent endometriosis. INTERVENTION(S): Ovarian cortex was reimplanted in the heterolateral orthotopic site. RESULT(S): Biopsies of the grafted tissue were taken 3 months after reimplantation. Viable primordial follicles were found. The presence of a neovascular capillary network was demonstrated. CONCLUSION(S): Reimplantation of fresh ovarian cortex allows the survival of primordial follicles and may represent an alternative method for the preservation of ovarian cortex when oophorectomy is mandatory.

Adult↗

PubMed2005

The role of cryopreservation for women prior to treatment of malignancy.

PURPOSE OF REVIEW: The purpose of this review is to investigate recent advances in xenografting, as well as in orthotopic and heterotopic autotransplantation of human cryopreserved ovarian tissue. RECENT FINDINGS: The first livebirth after orthotopic transplantation of cryopreserved ovarian tissue was reported recently. We discuss this case and other cases of reimplantation of cryopreserved ovarian tissue, bearing in mind that many questions remain. SUMMARY: Finally, we report the latest developments in research on the transplantation of an intact ovary and the reimplantation of isolated follicles.

Animals↗

PubMed2005

Human endometrial epithelial cells (EEC) constitutively express more intercellular adhesion molecule (ICAM)-1 than endometrial stromal cells (ESC) in culture.

PROBLEM: Intercellular adhesion molecule-1 (ICAM-1) is thought to play an important role in pathophysiological processes in endometrial tissue. The aim of this study was to quantify and compare the expression of ICAM-1 mRNA and protein in cultured endometrial epithelial cells (EEC) versus endometrial stromal cells (ESC). METHOD OF STUDY: EEC and ESC were isolated from human endometrium and cultured. At confluency, ICAM-1 mRNA was measured by real-time reverse transcriptase-polymerase chain reaction, the membrane-bound form (mICAM-1) by immunocytodensitometry, and the soluble form (sICAM-1) by enzyme-linked immunosorbent assay. RESULTS: At the transcriptional level, we observed a 1.7-fold increase in ICAM-1 expression in EEC versus ESC. mICAM-1 immunostaining in EEC [cell-relative membrane-bound signal-specific optical density (CR-M-SOD): 0.056 +/- 0.05] was stronger (P < 0.05) than in ESC (CR-M-SOD: 0.009 +/- 0.07). EEC were found to secrete more (P < 0.01) sICAM-1 ([sICAM-1] = 15.59 +/- 2.96 ng/mL) than ESC ([sICAM-1] = 5.14 +/- 2.61 ng/mL). CONCLUSIONS: Cultured EEC constitutively express significantly more ICAM-1 mRNA and protein than ESC.

Adult↗

PubMed2004

Characterization of iron deposition in endometriotic lesions induced in the nude mouse model.

BACKGROUND: Pelvic endometriotic lesions are often laden with hemosiderin. In order to investigate the potential source of such iron deposits, we examined whether the seric and erythrocyte fractions of menstrual effluent might influence the occurrence of iron deposition within lesions in a murine model of endometriosis. METHODS: Endometriosis was induced in 57 nude mice by intraperitoneal injection of unfractionated human menstrual effluent, endometrial fragments plus serum, endometrial fragments plus erythrocytes or endometrial cell fraction alone. The number of implants, histologic aspect, proliferative activity and iron deposition in lesions was assessed. RESULTS: On day 5, lesions were evidenced in all 10 mice receiving menstrual effluent, in 9/13 of those injected with the cellular fraction, in 10/13 with the cellular fraction plus serum and in 10/12 with the cellular fraction plus erythrocytes. Iron conglomerates were observed at the interface between the lesion and peritoneum when menstrual effluent (47 deposits/mm(2)) and the cellular fraction with erythrocytes (20 deposits/mm(2)) were injected, but were scarce when the cellular fraction was injected without erythrocytes, either alone (4 deposits/mm(2)) or with serum (2 deposits/mm(2)) (P < 0.05). CONCLUSIONS: Iron conglomerates, typically found in the stroma of endometriotic lesions, were induced by erythrocytes present in menstrual effluent. This may be one of the factors triggering oxidative damage and chronic inflammation.

Adult↗

PubMed2004

Expression of platelet endothelial cell adhesion molecule-1 in red and black endometriotic lesions.

This immunohistochemical study shows that platelet endothelial cell adhesion molecule-1 (PECAM-1), known to play a role in neoangiogenesis and vascular development, is strongly and homogeneously expressed in endothelial cells lining blood vessels from red and black pelvic endometriotic lesions. The distribution of PECAM-1 within the stroma of the lesions was similar to that found in the corresponding eutopic endometrium, regardless of the phase of the cycle.

Endometriosis↗

PubMed2004

Ficoll density gradient method for recovery of isolated human ovarian primordial follicles.

OBJECTIVE: To develop a simple and efficient technique to allow rapid recovery of a maximum number of good quality isolated follicles. DESIGN: Prospective experimental study. SETTING: Academic research unit of the department of gynecology in a university hospital. PATIENT(S): Biopsies were obtained from five women (between 26 and 31 years of age). INTERVENTION(S): Biopsies were cut with a tissue sectioner. Enzymatic digestion was performed in a collagenase solution for 90 min at 37 degrees C. The follicles were recovered using a discontinuous Ficoll density gradient method. MAIN OUTCOME MEASURE(S): The number of follicles present in the interface layers of Ficoll gradient was quantified. Follicular viability of these recovered follicles was assessed with live-dead stains, using calcein-AM and ethidium homodimer-I. RESULT(S): Out of a total of 6,811 recovered follicles, we found 63% (n = 4,201) at the medium-1.06 Ficoll interface and 36.9% (n = 2,590) at the 1.06-1.09 Ficoll interface, which represents 99.9% of total recovered follicles. Analysis by vital fluorescent staining showed that 95.8% of the follicles treated with Ficoll were totally viable. CONCLUSION(S): The Ficoll density gradient method allows us to maximize the recovery of isolated human ovarian follicles and minimize the manipulation time while maintaining high follicular viability.

Adult↗

PubMed2004

Freeze-thawing intact human ovary with its vascular pedicle with a passive cooling device.

OBJECTIVE: To test the feasibility of freezing intact human ovary using a passive cooling device. DESIGN: Prospective experimental study. SETTING: Academic research unit of the department of gynecology in a university hospital. PATIENT(S): Ovaries were obtained from three women who were between 29 and 36 years of age. INTERVENTION(S): Ovarian perfusion with cryoprotective solution and slow freezing with a cryofreezing container (5100 Cryo 1 degrees C Freezing Container; Nalgene, VWR, Belgium). Rapid thawing of ovaries by perfusion and bathing with decreased sucrose gradient. MAIN OUTCOME MEASURE(S): Viability of follicles, stromal cells, and vascular components was assessed with live-dead stains in freshly removed ovary, after cryoprotectant exposure before freezing, and after thawing. Histological morphology was assessed at these three different times. RESULT(S): The percentage of live follicles was 99.4% in fresh tissue, 98.1% after cryoprotectant exposure, and 75.1% after thawing. Viability assessment showed live stromal cells and small vessels after thawing. On histological evaluation, the morphology of follicles and cortical and medullar tissue was similar in all three groups. CONCLUSION(S): We described the cryopreservation, using an accessible protocol, of intact human ovary with its vascular pedicle and proved high survival rates of follicles, small vessels, and stromal cells and a normal histological structure in all the ovarian components after thawing using our protocol.

Adult↗

PubMed2004

Typical and subtle atypical presentations of endometriosis.

PURPOSE OF REVIEW: To 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. RECENT FINDINGS: Although 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. SUMMARY: The 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.

Endometriosis↗

PubMed2003

Typical and subtle atypical presentations of endometriosis.

The diagnosis of peritoneal endometriosis at the time of laparoscopy is often made by the observation of typically puckered black or bluish lesions. There are also numerous subtle appearances of peritoneal endometriosis. The lesions are frequently non-pigmented. Red flame-like lesions, glandular excrescences, and subovarian adhesions must be considered as the most active lesions. Sometimes, however, subtle endometriotic lesions can be the only lesions seen at laparoscopy.

Diagnosis, Differential↗

PubMed2003

Effect of prooxidant agents added at the morula/blastocyst stage on bovine embryo development, cell death and glutathione content.

Two prooxidant agents, 2,2'-azobis(2-amidinopropane)dihydrochloride (AAPH), a generator of free radicals in the culture medium, and buthionine sulfoximine (BSO), an inhibitor of glutathione synthesis, were used to reinforce from the morula stage (day 5 post-insemination, p.i.) the oxidative stress encountered by bovine embryos in culture. Exposure to increasing concentrations of both prooxidants from the morula stage did not affect blastocyst formation but some blastocysts were found degenerated on day 8 in a dose-dependent manner (0, 0.001, 0.01, 0.1 mM AAPH gave respectively 0, 10%, 32%, 48% degeneration, while 0, 0.1, 0.2, 0.4 mM BSO led respectively to 0, 14%, 30%, 41% degeneration). Hatching rates and cell numbers of surviving blastocysts were not affected. Morulae and early blastocysts exposed from day 5 to day 6 p.i. appeared more resistant than expanded blastocysts (75-80% survival vs 20-65%; p < 0.05). Treatment with BSO significantly decreased the level of reduced glutathione in day 7 blastocysts (0.02 vs 0.42 pmol per embryo in the control) while AAPH had no effect (0.38 pmol per embryo). The proportion ofcells showing membrane lesions was increased in degenerated blastocysts from day 7.5 p.i. In AAPH-treated, but not in BSO-treated embryos, cell membrane permeabilisation seems to occur before blastocyst degeneration. DNA fragmentation evaluated by the TUNEL technique was increased in day 7 blastocysts by both prooxidants (2.8 +/- 0.4 in the control group vs 4.5 +/- 0.4 and 6.0 +/- 0.4 respectively in the AAPH- and BSO-treated groups). Addition of an inhibitor of caspase-3, DEVD-CHO, partially prevented DNA fragmentation, indicating that prooxidant treatment induced a caspase-dependent pathway of apoptosis.

Amidines↗

PubMed2002

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

OBJECTIVE: To 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. DESIGN: Prospective study involving patients with and without endometriosis. SETTING: Department of gynecology in a university hospital. PATIENT(S): Seventy-six patients undergoing laparoscopy for tubal sterilization or infertility and/or pelvic pain. INTERVENTION(S): Collection of peritoneal fluid (PF), blood samples, and biopsies from endometrium and peritoneum. MAIN OUTCOME MEASURE(S): Measurement 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. RESULT(S): Higher 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. CONCLUSION(S): Our 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.

Adult↗

PubMed2002

Oxidative stress and peritoneal endometriosis.

OBJECTIVE: To 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. DESIGN: Literature survey. RESULT(S): Several 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. CONCLUSION(S): Better 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.

Animals↗

PubMed2002

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

OBJECTIVE: To examine the possible involvement of iron in the physiopathology of endometriosis. DESIGN: Prospective study. SETTING: Department of gynecology in a university hospital. PATIENT(S): Seventy patients undergoing laparoscopy. INTERVENTION(S): Collection 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). MAIN OUTCOME MEASURE(S): Measurement of iron and ferritin in serum and peritoneal fluid and staining of iron deposits with Prussian blue in tissues. RESULT(S): Iron 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. CONCLUSION(S): Iron 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.

Adult↗