Cytokine levels before and after hysteroscopic polypectomy in infertile patients

Polip endometrijuma je najčešće benigna lokalizovana proliferacija žlezda i strome endometrijuma pokrivena epitelom koja se izdiže iznad ravni sluzokože. Najčešće se dijagnostikuju za vreme ispitivinja uzroka neurednog krvarenja iz materice ili neplodnosti. Citokini su biološki aktivne supstance sa glasničkom i regulatornom ulogom u imunim i inflamatornim reakcijama. Svi citokini se stvaraju i u endometrijumu.Citokini su uključeni u bitne reproduktivne funkcije kao što su ovulacija i implantacija. Cilj studije je bio da se utvrde promene na nivou endometrijuma pre i posle histeroskopske polipektomije kod infertilnih pacijentkinja sa endometrijalnim polipom i kod infertilnih pacijentkinja bez endometrijalnog polipa. Kod pacijentkinja sa polipom endometrijuma vrednosti ekspresije TNF-α su više u ispirku uterusa nego u serumu i pre i posle histeroskopije. Nakon uklanjanja polipa vrednosti TNF-α su više u serumu kod pacijentkinja bez polipa endometrijuma, nego kod pacijentkinja sa polipom endometrijuma. Kod pacijentkinja sa polipom endometrijuma vrednosti TNF-α su više u ispirku nego u serumu i pre i posle histeroskopije. Intezitet IHH ekspresija TNF-α u polipu endometrijuma i bioptatu endometrijuma pacijentkinja bez polipa pokazuje značajnu razliku, dok između bioptata endometrijuma pacijentkinja sa polipom i bioptata endometrijuma pacijentkinja bez polipa, ne pokazuje značajnu razliku intenziteta signala IHH ekspresija TNF-α . Vrednosti ekspresije INF-γ kod pacijentkinja sa polipom su više i u ispirku i u serumu i pre i posle histeroskopije, nego kod pacijentkinja bez polipa. Kod pacijentkinja sa polipom endometrijuma vrednosti INF-γ su niže u ispirku nego u serumu i pre i posle histeroskopije. Vrednosti INF-γ u serumu ovih pacijentkinja su više nakon uklanjanja polipa u serumu. Skor ekspresije INF-γ u bioptatu endometrijuma, kod pacijentkinja bez polipa, je značajno niži u odnosu na skor ekspresije INF-γ i u polipu i u bioptatu kod pacijentkinja sa polipom endometrijuma. Kada se u ispirku kavuma uterusa detektuju više vrednosti INF-γ imunohistohemijski možemo dokazati postojanje ekspresije INF-γ u endometrijumu. Vrednosti Glycodelina su bile više kod pacijentkinja sa polipom endometrijuma u ispirku uterusa, dok su vrednosti Glycodelina u serumu bile više kod pacijentkinja bez polipa. Nakon histeroskopije vrednosti Glycodelina u serumu kod pacijentkinja sa polipom endometrijuma su porasle, a kod pacijentkinja bez polipa endometrijuma vrednosti Glycodelina su pale nakon histeroskopije. Veći broj pacijentkinja sa polipom endometrijuma ne pokazuje ekspresiju Glycodelina, i to izraženije u bioptatu endometrijuma nego u polipu endometrijuma. Postoji značajna razlika skora IHH ekspresije Glycodelina između polipa i bioptata endometrijuma kod pacijentkinja sa polipom endometrijuma. Kod pacijentkinja sa polipom endometrijuma vrednost TNF-α u ispirku kavumu uterusa i serumu može biti dobar skrining test za otkrivanje endometrijalnog polipa. Kod pacijentkinja sa polipom endometrijuma vrednost INF-γ u serumu može biti skrining test za otkrivanje polipa endometrijuma. Kod vrednosti Glycodelina ne postoji prediktorska sposobnost testa u cilju otkrivanja polipa endometrijuma.An endometrial polyp is the most common benign localised proliferation of endometrial glands and stroma covered with the epithelium and extending above the level of the mucosa. Endometrial polyps are usually diagnosed while examining the cause of abnormal uterine bleeding or infertility. Cytokines are biologically active substances with a mediating and regulatory role in immune and inflammatory reactions. All cytokines are also generated in the endometrium. They are involved in important reproductive functions such as ovulation and implantation. The aim of this study was to determine endometrial changes prior to and following hysteroscopic polypectomy in infertile female patients with endometrial polyps and in infertile female patients without endometrial polyps. In the patients with endometrial polyps the values of TNF-α expression were higher in the uterine flushings than in the serum both prior to and following hysteroscopy. Once the polyps had been removed, serum TNF-α values were higher in the patients without endometrial polyps than in the patients with endometrial polyps. In the patients with endometrial polyps, TNF-α values were higher in the uterine flushings than in the serum both prior to and following hysteroscopy. The intensity of IHC expression of TNF-α in the endometrial polyps and endometrial biopsy specimens obtained from the patients without endometrial polyps revealed a significant difference, whereas there was no significant difference observed in the signal intensity of IHC expression of TNF-α in the biopsy specimens obtained from the patients with and without polyps. The values of INF-γ expression were higher in the uterine flushings and serum of the patients with polyps before and after hysteroscopy in comparison to the values obtained from the patients without polyps. The values INF-γ were lower in the uterine flushings than in the serum of the patients with endometrial polyps before and after hysteroscopy. In these patients, the serum values of INF-γ increased after the polyps had been removed. INF-γ expression score in endometrial biopsy specimens taken from the patients without endometrial polyps was significantly lower in comparison to INF-γ expression scores both in the polyps and biopsy specimens of the patients with endometrial polyps. When an analysis of uterine flushings reveals higher values of INF-γ, the immunohistochemical method is used to provide evidence for INF-γ expression in the endometrium. The values of glycodelin were higher in the uterine flushings of the patients with endometrial polyps, while higher values of glycodelin were detected in the serum of the patients without polyps. The values of glycodelin in the serum of the patients with endometrial polyps increased after hysteroscopy as opposed to the values of glycodelin in the serum of the patients without endometrial polyps, which decreased after hysteroscopy. Glycodelin expression was not detected in most patients, which was more obvious in the endometrial biopsy specimens than in the polyps. A significant difference in ICH scores of glycodelin expression was observed between the polyps and endometrial biopsy specimens in the patients with endometrial polyps. The values of TNF-α in the uterine flushings and serum of patients with endometrial polyps can be considered as a reliable screening test for the detection of endometrial polyps. In patients with endometrial polyps, the values of INF-γ in the serum can be used as a screening test for detecting endometrial polyps. The values of glycodelin do not have a predictive capacity of a test that could help detect endometrial polyps

A pre-conceptual endometrial test to predict pregnancy outcome in recurrent pregnancy loss

Recurrent pregnancy loss (RPL) and recurrent implantation failure (RIF) are major barriers to wanted pregnancies. Currently, there are few tests that reliably predict pregnancy outcome in these groups. Recent research demonstrated that an aetiological factor in early reproductive failure is impaired endometrial decidualisation due to accelerated senescence and loss of endometrial mesenchymal stem cells. Uterine Natural Killer (uNK) cells maintain homeostasis by clearing senescent decidual cells. I aimed to improve the ability of the uNK cell density test in predicting subsequent pregnancy outcomes, through a combined preconceptual test that includes cell senescence and proliferation markers. Methods This study was performed in a tertiary research ‘Implantation Clinic’, where the uNK cell density test is performed on mid-luteal endometrial biopsies in women with RPL and RIF. I began my investigation of a predictive test by assessing the predictive ability of the uNK test using the clinic’s large retrospective database (N=281). Next, immunohistochemistry was performed on endometrial serial sections of women with RPL using antibodies to CD56 (uNK), Ki67 (cell proliferation), HMGB2 and p16 (cell senescence) via an automated staining process. The images were analysed using Pannoramic Viewer and Image J software with colour de-convolution and thresholding technique. The findings of the exploratory test set (N=20) were used to develop a preconceptual combined test for outcome prediction in a larger study (N=89). Final validation of the combined test ‘uNK-glandular p16’ was conducted on samples from women with RPL recruited in the setting of a prospective, randomised-controlled trial (RCT). Results Combining the normalised uNK and glandular p16 results improved the prediction of pregnancy outcome (p=0.0163). The predictive ability of the combined test ‘uNK-glandular p16 centile’ was validated in the prospective RCT setting. Conclusion Glandular-stromal synchrony appears to be an important determinate of pregnancy outcome and may help focus future research or therapy