OBJECTIVE: Our previous study demonstrated that expression levels of E-cadherin were distinctive among different forms of endometriosis. We further investigated several markers of epithelial-mesenchymal transition (EMT) (N-cadherin, vimentin, S100A4, dephosphorylated form of beta-catenin) in endometriosis.DESIGN: Prospective study.MATERIALS AND METHODS: Samples of deep infiltrating endometriosis (n= 61), ovarian endometriosis (n=48) and superficial peritoneal endometriosis (red lesions: n=30, black lesions: n = 46) were collected during laparoscopic surgery. In addition, samples of menstrual endometrium, abdominal wall endometriosis, other benign ovarian cysts, carcinoma arising in ovarian endometriosis, colon endometriosis and scar endometriosis, were included in the present study for comparison. Immunohistochemical staining was performed on paraffin sections and quantitation of immunostained cells (percentage of immunostained surface × mean staining intensity) was obtained using a computerized image analysis system.RESULTS: Vimentin expression was detected in the menstrual endometrium, whereas no expression of N-cadherin, S100A4 and dephosphorylated beta-catenin was detected. S100A4 expression was significantly lower in deep infiltrating endometriosis compared to that of ovarian endometriosis and red peritoneal lesions. Vimentin expression was significantly lower in ovarian endometriosis compared to that of deep infiltrating endometriosis, red and black peritoneal lesions. In addition, vimentin expression in deep infiltrating endometriosis, red and black peritoneal lesions was significantly higher than that of menstrual endometrium. Dephosphorylated beta-catenin expression was significantly higher in deep infiltrating endometriosis compared to that of ovarian endometriosis, red and black peritoneal lesions.CONCLUSION: The present findings suggested that markers of EMT might be gained depending on the microenvironment during the evolution of endometriotic implants. OBJECTIVE: Our previous study demonstrated that expression levels of E-cadherin were distinctive among different forms of endometriosis. We further investigated several markers of epithelial-mesenchymal transition (EMT) (N-cadherin, vimentin, S100A4, dephosphorylated form of beta-catenin) in endometriosis. DESIGN: Prospective study. MATERIALS AND METHODS: Samples of deep infiltrating endometriosis (n= 61), ovarian endometriosis (n=48) and superficial peritoneal endometriosis (red lesions: n=30, black lesions: n = 46) were collected during laparoscopic surgery. In addition, samples of menstrual endometrium, abdominal wall endometriosis, other benign ovarian cysts, carcinoma arising in ovarian endometriosis, colon endometriosis and scar endometriosis, were included in the present study for comparison. Immunohistochemical staining was performed on paraffin sections and quantitation of immunostained cells (percentage of immunostained surface × mean staining intensity) was obtained using a computerized image analysis system. RESULTS: Vimentin expression was detected in the menstrual endometrium, whereas no expression of N-cadherin, S100A4 and dephosphorylated beta-catenin was detected. S100A4 expression was significantly lower in deep infiltrating endometriosis compared to that of ovarian endometriosis and red peritoneal lesions. Vimentin expression was significantly lower in ovarian endometriosis compared to that of deep infiltrating endometriosis, red and black peritoneal lesions. In addition, vimentin expression in deep infiltrating endometriosis, red and black peritoneal lesions was significantly higher than that of menstrual endometrium. Dephosphorylated beta-catenin expression was significantly higher in deep infiltrating endometriosis compared to that of ovarian endometriosis, red and black peritoneal lesions. CONCLUSION: The present findings suggested that markers of EMT might be gained depending on the microenvironment during the evolution of endometriotic implants.

OBJECTIVE: Hyaluronan is used in anti adhesive products to prevent postoperative adhesions. The objective of the present study was to investigate whether peritoneal hyaluronan synthase (HAS) mRNA transcription is affected by the intraperitoneal pressure (IPP) and duration of a CO2 pneumoperitoneum. DESIGN: Prospective, randomized study. MATERIALS AND METHODS: Patients undergoing laparoscopic hysterectomy for benign gynecological diseases were recruited for this study. Patients with previous history for pelvic surgeries, endometriosis and/or infection were excluded from the present study. Patients were divided into two groups: CO2 pneumoperitoneum at a low (8mmHg) (n=28) or high (12mmHg) (n= 30) IPP. Macroscopically normal peritoneum was collected from right anterior parietal wall at the beginning (immediately after the insertion of the trocars) and each 60 min afterward. Samples were immediately collected in RNAlater and HAS1, HAS2 and HAS3 mRNA expression levels were measured using real-time PCR. The Wilcoxon sign rank test was performed to compare differences in gene expression levels in paired samples (0, 1 hour and/or 2 hour). Statistical significance was defined as a P-value of <0.05. RESULTS: HAS1 mRNA expression levels were significantly increased after 1 hour and 2 hour in the group of 8 mmHg. However, there was no significant difference in the group of 12mmHg after either 1 hour or 2 hour. There was no significant difference in HAS2 mRNA expression levels in both groups. HAS3 mRNA expression levels were significantly decreased after 1 hour in the group of 12mmHg. However, there was no significant difference in the 8mmHg group after either 1 hour or 2 hour. CONCLUSION: The present findings suggest that the synthesis of hyaluronan might be altered during a CO2 pneumoperitoneum at a high IPP.

OBJECTIVE: To investigate peritoneal fibrinolytic system over time under various surgical peritoneal environments in a mouse model. DESIGN: Prospective, controlled study MATERIALS AND METHODS: Protocol 1: On day 0, C57BJ6 mice were divided into four groups: anesthesia alone (control), CO2 pneumoperitoneum at a low (2mmHg) or high (8mmHg) IPP, or laparotomy. Groups were further sub-divided into eight groups of 5 animals each and a laparotomy was performed to collect peritoneal tissue at 0, 4, 8, 24, 48, 72 hours, 5 and 7 days after surgery. Protocol 2: On day 0, C57BJ6 mice were divided into three groups of 10 animals each: CO2 pneumoperitoneum at a low or high IPP, or laparotomy. The bilateral caudal epigastric arteries and uterine horns were then coagulated using a 1.4 mm bipolar. On day 7, pimonidazole hydrochloride was administered intraperitoneally before samples collection to detect tissue hypoxia. Peritoneal tissue samples were collected for real-time RT-PCR and immunohistochemistry. RESULTS: Protocol 1: Laparotomy: PAI-1 mRNA expression was elevated at 0h and maintained at a higher level up to POD 7, whereas tPA mRNA expression was elevated at 4h and decreased to near normal level at 24h. High IPP: PAI-1 mRNA expression was elevated at 4h and then decreased; but maintained at a higher level up to POD 5, whereas tPA mRNA expression was increased at 0h and reached the highest level at 4h and then decreased, but maintained at a higher level up to POD 7. Low IPP: No significant change was detected in expression of either tPA or PAI-1 mRNA expression over time. Protocol 2: Hypoxic mesothelial cells and fibroblasts were detected in distant non-injured normal peritoneum in the high IPP group. CONCLUSIONS: The present findings suggest that a CO2 pneumoperitoneum at a low IPP has few effects on peritoneal fibrinolytic system in non-injured peritoneum. A high IPP during a CO2 pneumoperitoneum might cause hypoxia in distant non injured normal peritoneum.

OBJECTIVE: To investigate expression of matrix metalloproteinase-7 (MMP-7), E-cadherin and beta catenin in different forms of endometriosis. DESIGN: Prospective study MATERIALS AND METHODS: Samples of deep infiltrating endometriosis (n= 46), ovarian endometriosis (n=38) and superficial peritoneal endometriosis (red lesions: n=24, black lesions: n = 30) were collected during laparoscopic surgery. In addition, benign ovarian cyst wall was collected from patients with serous (n=10) and mucinous cyst (n=10). None of the patients received hormonal treatments for ≥6 months before surgery. Immunohistochemical staining was performed on paraffin sections with mouse monoclonal antibodies to MMP-7, E-cadherin and beta catenin. The percentage of immunostained surface (PI) was obtained using a computerized image analysis system. RESULTS: The PI for MMP-7 in epithelial cells was significantly higher in red peritoneal lesions (72.1 ± 29.5, mean ± SEM, P<.0001) compared to that of deep infiltrating endometriosis (1.6 ± 0.7), ovarian endometriosis (2.4 ± 1.2) and black peritoneal lesions (8.2 ± 2.3). MMP-7 expression was detected in stromal and endothelial cells as well as fibrotic tissues in superficial peritoneal lesions. The PI for E cadherin was significantly lower in red peritoneal lesions (9.5 ± 2.4, p<.001) and ovarian endometriosis (17.7 ± 4.4, p<.01) compared to that of black peritoneal lesions (46.3 ± 4.5) and deep infiltrating endometriosis (40.1 ± 4.7). There was a significant negative correlation between MMP-7 and E-cadherin expression in superficial peritoneal lesions (p<.0001). Intense pre-nuclear staining of beta catenin in fibroblasts was detected in fibrotic tissue of ovarian endometriosis. CONCLUSIONS: Red peritoneal lesions might have the invasive phenotype. Beta catenin signaling pathway might be involved in fibrogenesis of ovarian endometriosis. The present findings suggested that strategies for treatment of endometriosis might differ among different forms of endometriosis.