C Berger1, N R Boggavarapu1, J Menezes2, P G L Lalitkumar3, K Gemzell-Danielsson3. 1. Department of Women's and Children's Health, Division of Obstetrics and Gynaecology, Karolinska Institutet/Karolinska University Hospital, S-171 76 Stockholm, Sweden. 2. Fertilitetscentrum, Stockholm, Sweden. 3. Department of Women's and Children's Health, Division of Obstetrics and Gynaecology, Karolinska Institutet/Karolinska University Hospital, S-171 76 Stockholm, Sweden kristina.gemzell@ki.se lalit.kumar@ki.se.
Abstract
STUDY QUESTION: Does ulipristal acetate (UPA) used for emergency contraception (EC) interfere with the human embryo implantation process? SUMMARY ANSWER: UPA, at the dosage used for EC, does not affect human embryo implantation process, in vitro. WHAT IS KNOWN ALREADY: A single pre-ovulatory dose of UPA (30 mg) acts by delaying or inhibiting ovulation and is recommended as first choice among emergency contraceptive pills due to its efficacy. The compound has also been demonstrated to have a dose-dependent effect on the endometrium, which theoretically could impair endometrial receptivity but its direct action on human embryo implantation has not yet been studied. STUDY DESIGN, SIZE, DURATION: Effect of UPA on embryo implantation process was studied in an in vitro endometrial construct. Human embryos were randomly added to the cultures and cultured for 5 more days with UPA (n = 10) or with vehicle alone (n = 10) to record the attachment of embryos. PARTICIPANTS/MATERIALS, SETTING, METHODS: Endometrial biopsies were obtained from healthy, fertile women on cycle day LH+4 and stromal and epithelial cells were isolated. A three-dimensional in vitro endometrial co-culture system was constructed by mixing stromal cells with collagen covered with a layer of epithelial cells and cultured in progesterone containing medium until confluence. The treatment group received 200 ng/ml of UPA. Healthy, viable human embryos were placed on both control and treatment cultures. Five days later the cultures were tested for the attachment of embryos and the 3D endometrial constructs were analysed for endometrial receptivity markers by real-time PCR. MAIN RESULTS AND THE ROLE OF CHANCE: There was no significant difference in the embryo attachment rate between the UPA treated group and the control group as 5 out of 10 human embryos exposed to UPA and 7 out of 10 embryos in the control group attached to the endometrial cell surface (P = 0.650). Out of 17 known receptivity genes studied here, only 2 genes, HBEGF (P = 0.009) and IL6 (P = 0.025) had a significant up-regulation and 4 genes, namely HAND2 (P = 0.003), OPN (P = 0.003), CALCR (P = 0.016) and FGF2 (P = 0.023) were down-regulated with the exposure of UPA, compared with control group. LIMITATIONS, REASONS FOR CAUTION: This proof of concept study was conducted with a few human embryos, as their availability was limited. Although the 3D model used for this study is well established and the artificial endometrial luminal epithelium shown to express progesterone regulated markers of endometrial receptivity it is still an in vitro model, lacking all cell types that constitute the receptive endometrium in vivo. WIDER IMPLICATIONS OF THE FINDINGS: This study provides new insights on the mechanism of action of UPA on human embryo implantation, demonstrating that UPA in a dosage used for EC does not affect embryo viability and the implantation process of embryo. Progesterone receptor modulators (PRMs) hold the potential to be attractive estrogen- and gestagen-free contraceptives and thus may be made available to a larger proportion of women globally due to these findings. STUDY FUNDING/COMPETING INTERESTS: Swedish Research Council (K2010-54X-14212-09-3) and support provided through the regional agreement on medical training and clinical research (ALF) between Stockholm County Council and Karolinska University Hospital.
STUDY QUESTION: Does ulipristal acetate (UPA) used for emergency contraception (EC) interfere with the human embryo implantation process? SUMMARY ANSWER: UPA, at the dosage used for EC, does not affect human embryo implantation process, in vitro. WHAT IS KNOWN ALREADY: A single pre-ovulatory dose of UPA (30 mg) acts by delaying or inhibiting ovulation and is recommended as first choice among emergency contraceptive pills due to its efficacy. The compound has also been demonstrated to have a dose-dependent effect on the endometrium, which theoretically could impair endometrial receptivity but its direct action on human embryo implantation has not yet been studied. STUDY DESIGN, SIZE, DURATION: Effect of UPA on embryo implantation process was studied in an in vitro endometrial construct. Human embryos were randomly added to the cultures and cultured for 5 more days with UPA (n = 10) or with vehicle alone (n = 10) to record the attachment of embryos. PARTICIPANTS/MATERIALS, SETTING, METHODS: Endometrial biopsies were obtained from healthy, fertile women on cycle day LH+4 and stromal and epithelial cells were isolated. A three-dimensional in vitro endometrial co-culture system was constructed by mixing stromal cells with collagen covered with a layer of epithelial cells and cultured in progesterone containing medium until confluence. The treatment group received 200 ng/ml of UPA. Healthy, viable human embryos were placed on both control and treatment cultures. Five days later the cultures were tested for the attachment of embryos and the 3D endometrial constructs were analysed for endometrial receptivity markers by real-time PCR. MAIN RESULTS AND THE ROLE OF CHANCE: There was no significant difference in the embryo attachment rate between the UPA treated group and the control group as 5 out of 10 human embryos exposed to UPA and 7 out of 10 embryos in the control group attached to the endometrial cell surface (P = 0.650). Out of 17 known receptivity genes studied here, only 2 genes, HBEGF (P = 0.009) and IL6 (P = 0.025) had a significant up-regulation and 4 genes, namely HAND2 (P = 0.003), OPN (P = 0.003), CALCR (P = 0.016) and FGF2 (P = 0.023) were down-regulated with the exposure of UPA, compared with control group. LIMITATIONS, REASONS FOR CAUTION: This proof of concept study was conducted with a few human embryos, as their availability was limited. Although the 3D model used for this study is well established and the artificial endometrial luminal epithelium shown to express progesterone regulated markers of endometrial receptivity it is still an in vitro model, lacking all cell types that constitute the receptive endometrium in vivo. WIDER IMPLICATIONS OF THE FINDINGS: This study provides new insights on the mechanism of action of UPA on human embryo implantation, demonstrating that UPA in a dosage used for EC does not affect embryo viability and the implantation process of embryo. Progesterone receptor modulators (PRMs) hold the potential to be attractive estrogen- and gestagen-free contraceptives and thus may be made available to a larger proportion of women globally due to these findings. STUDY FUNDING/COMPETING INTERESTS: Swedish Research Council (K2010-54X-14212-09-3) and support provided through the regional agreement on medical training and clinical research (ALF) between Stockholm County Council and Karolinska University Hospital.
Authors: Peter T Ruane; Chelsea J Buck; Phoebe A Babbington; Wedad Aboussahoud; Stéphane C Berneau; Melissa Westwood; Susan J Kimber; John D Aplin; Daniel R Brison Journal: Hum Reprod Open Date: 2020-02-28