Clinical Groups

Clinical Groups

Inclusion criteria: Patients and controls were enrolled in the Hospital General Universitarioand in the University Hospital La Fe (Valencia, Spain) from 2003 to 2016.

Endometriosis patients:

All women underwent laparoscopic surgical examination of the abdominal cavity and complete excision of endometriotic tissue. The presence of the disease was suspected either clinically or by ultrasonography and confirmed by surgical findings and postoperative pathological examination. Laparoscopic examination of the abdominal cavity excluded the presence of any other pelvic pathology that could potentially confound data.

Control Non-endometriosis group:

The absence of endometriosis was confirmed by meticulous examination of the pelvic and extrapelvic peritoneum, ovaries, intestine, and diaphragm to detect typical or atypical endometriotic lesions.

Exclusion criteria

Women affected by menorrhagia or hypermenorrhea or women who had been pregnant or breastfeeding during the previous 6 months were excluded from the study. None of the women had received any form of hormone therapy for at least 3 months prior to the study.

ELISAs kits descriptions

For VEGF-A determinations, Human VEGF ELISA kit (IBL International, Hamburg, Germany) was employed. No cross-reactivity or interference with platelet-derived growth factor (PDGF) was observed. This assay recognizes both human VEGF-A165 and VEGF-A121 isoforms. The intra-assay and inter-assay variability were 4–6% and 7–10%, respectively. For TSP-1 determinations, Human TSP-1ELISA kit from Development System (DuoSet, RD systems, Minneapolis, USA) was employed. No cross-reactivity or interference with TSP-2 or TSP-4 was observed. The intra-assay and inter-assay variability were 5–6% and 8–11%, respectively.uPA determinations were performed employing a commercial ELISA from Zymutest (Hyphen Biomed, Neuville-sur-Oise, France), which measures both single-chain urokinase (scuPA) and the high weight molecular form of uPA (HMW-uPA) with similar efficiency. The intra-assay and inter-assay variation coefficients were 3-5% and 8-11%, respectively.PAI-1 determinations were performed employing a commercially available ELISA kit from Immubind (America Diagnostica, Pfungstadt, Germany). The assay detects free and complexed PAI-1 forms, remaining insensitive to PAI-2. The intra-assay and inter-assay variation coefficients were 3-4% and 6-8%, respectively.MMP-3 levels were determined by means of a commercial ELISA kit from Calbiochem (San Diego, USA). The assay detects MMP-3, but does not recognize MMP-3-TIMP complexes. The intra- and inter-assay variability were 5 and 9%, respectively. ELISA kit for TIMP-1 determinations was also purchased from Calbiochem (San Diego, USA).The assay recognizes both free and complexed forms of TIMP-1. The intra- and inter-assay variability were 4 and 7%, respectively.