Estela Bevilacqua

To compare maternal serum CRIPTO‐1 concentrations in pregnancies complicated by placenta accreta spectrum (PAS) with those complicated by placenta previa (PP). We conducted a prospective, cross‐sectional, comparative study at a tertiary center in São Paulo, Brazil (August 2022–May 2025). Singleton pregnancies with ultrasound‐confirmed PP were included and classified as either PP without accreta or PAS (accreta/increta/percreta) according to standardized imaging criteria and, when applicable, surgical/histopathological confirmation. Maternal venous blood was collected predelivery, and serum CRIPTO‐1 concentrations were measured using ELISA (Human CRIPTO SimpleStep ELISA Kit, Abcam). Primary analyses used nonparametric tests for continuous variables and Fisher′s exact test for categorical variables. The study was approved by the Research Ethics Committee of the University of São Paulo (CAAE 58202122.8.0000.0065; CAAE 55121422.3.0000.0068). Forty‐three participants were analyzed (PP = 18; PAS = 25). Median serum CRIPTO‐1 did not differ significantly between groups (PP: 0.013 pg/mL, IQR: 0.001–0.067 vs. PAS: 0.079 pg/mL, IQR: 0.005–0.360; p = 0.156). PAS was associated with greater blood loss (median 2240 vs. 568 mL; p < 0.001), higher transfusion rate (88.0% vs. 16.7%; p < 0.001), more ICU admissions (68.0% vs. 16.7%; p = 0.002), longer ICU and hospital stays (both p < 0.01), and higher hysterectomy rates (84.0% vs. 5.6%; p < 0.001). Predelivery serum CRIPTO‐1 did not discriminate PAS from PP, possibly reflecting physiological variability at delivery.

Three-dimensional (3D) endothelium–stromal co-cultures were established using human endometrial cells from biopsy of healthy women (n = 13) and serum samples from both healthy and endometriotic women (n = 5). For 3D construction, stromal cells were mixed with extracellular matrix components, followed by endothelial cell seeding. Morphological analysis confirmed the organization of tissue-like structures. Immunofluorescence and flow cytometry verified the expression of specific stromal and endothelial markers (Cytokeratin, Vimentin, Insulin-like growth factor-binding protein 1, and von Willebrand factor). Cell viability and proliferation increased over time, with minimal cell death. To test functional responsiveness, these co-cultures were exposed to inflammatory serum from endometriotic patients. After 48 h, cytometric bead array showed elevated levels of IL-1β, IL-6, and IL-8 in cultures treated with inflammatory serum, indicating preserved functional activity and responsiveness. By allowing detailed investigation of functional endometrial states within a physiologically relevant cellular network, this approach provides a valuable organoid-like tool to explore conditions such as implantation failure and infertility and to study the cellular interactions underlying reproductive pathologies.

SARS-CoV-2, responsible for COVID-19, affects multiple bodily systems, including placenta. Understanding its effects on placental angioactive factors is vital due to its vascular implications. Herein, we hypothesized that maternal SARS-CoV-2 infection could impact placental morphology and the expression of angioactive and inflammatory factors. The study included 23 pregnant women tested for COVID-19 through reverse transcription polymerase chain reaction (RT-qPCR) and IgG serology around time of delivery and discriminated as: Viremia Group (VG, n = 6, maternal positive RT-qPCR, early infection), Serology Group (SG, n = 10, maternal positive serology, late infection), and Control Group (CG, n = 7, negative for SARS-CoV-2). PCR and immunolocalization of SARS-CoV-2 in the placenta tested negative. Placental morphology was examined using H&E-stained sections, along with immunostaining for vascular endothelial growth factor (VEGF), placental growth factor (PlGF), inducible and endothelial nitric oxide synthase (iNOS, eNOS), and cyclooxygenase 2 (COX2). SG showed a significant higher level of placental morphological changes compared to control samples, which included an increase in syncytial knots and fibrin deposits on the villous surface, and villous peripheral and central infarctions. In contrast, samples from VG exhibited more frequent vascular dilation, congestion, and chorioangiosis. Compared with the control group, immunohistochemical analysis showed significantly higher expression of VEGF ( P = 0.001), COX2 ( P = 0.03), and eNOS ( P = 0.02) in VG. Conversely, in the SG, PlGF levels decreased ( P = 0.01), while COX2 ( P = 0.01) and iNOS ( P = 0.01) levels were elevated. Our data suggests a temporal adaptation of the placenta to maternal SARS-CoV-2 infection, triggering structural, angioactive, and inflammatory responses, enhancing our understanding of potential placental pathways to cope with SARS-CoV-2 infection.

The functions of annexin A1 (ANXA1), which is expressed on membranes and in cytoplasmic granules, have been fully described. Nonetheless, the role of this protein in protecting against DNA damage in the nucleus is still emerging and requires further investigation. Here, we investigated the involvement of ANXA1 in the DNA damage response in placental cells. Placenta was collected from ANXA1 knockout mice (AnxA1^−/−) and pregnant women with gestational diabetes mellitus (GDM). The placental morphology and ANXA1 expression, which are related to the modulation of cellular response markers in the presence of DNA damage, were analyzed. The total area of AnxA1^−/− placenta was smaller due to a reduced labyrinth zone, enhanced DNA damage, and impaired base excision repair (BER) enzymes, which resulted in the induction of apoptosis in the labyrinthine and junctional layers. The placentas of pregnant women with GDM showed reduced expression of AnxA1 in the villous compartment, increased DNA damage, apoptosis, and a reduction of enzymes involved in the BER pathway. Our translational data provide valuable insights into the possible involvement of ANXA1 in the response of placental cells to oxidative DNA damage and represent an advancement in investigations into the mechanisms involved in placental biology.

The functions of annexin A1 (ANXA1) that is expressed on membranes and in cytoplasmic granules have been fully described. Nonetheless, the role of this protein in protecting against DNA damage in the nucleus is still emerging and requires further investigation. Here, we investigated the involvement of ANXA1 in DNA damage response in placental cells. Placenta was collected from ANXA1 knockout mice (AnxA1-/-) and pregnant women with gestational diabetes mellitus (GDM). The placental morphology and ANXA1 expression, which are related to the modulation of cellular response markers in the presence of DNA damage, were analyzed. Total area of AnxA1-/- placenta was smaller because of reduced labyrinth zone, enhanced DNA damage, and impaired base excision repair (BER) enzymes, which resulted in the induction of apoptosis in the labyrinthine and junctional layers. The placentas from pregnant women with GDM showed reduced expression of AnxA1 in the villous compartment, increased DNA damage, apoptosis, and reduction of enzymes involved in the BER pathway. Our translational data provide valuable insights into the possible involvement of ANXA1 in the response of placental cells to oxidative DNA damage and represent advancement in investigations on the mechanisms involved in placental biology.

Preeclampsia (PE) is a hypertensive disease of pregnancy-associated with placental cell death and endoplasmic reticulum (ER) stress. It is unknown whether systemic factors aggravate placental dysfunction. We investigated whether serum factors in pregnant women with PE activate ER stress and unfolded protein responses (UPRs) in placental explants and trophoblast cells lineage. We cultured placental explants from third-trimester term placentas from control non-preeclamptic (NPE) pregnant women with serum from women with PE or controls (NPE). In PE-treated explants, there was a significant increase in gene expression of GADD34, CHOP, and SDF2. At the protein level, GRP78, SDF2, p-eIF2α, and p-eIF2α/eIF2α ratio were also augmented in treated explants. Assays were also performed in HTR8/SV-neo trophoblast cell line to characterize the putative participation of trophoblast cells. In PE serum-treated protein levels of p-eIF2a and the ratio p-elF2 α/elF2α increased after 12 h of treatment, while the gene expression of GADD34, ATF4, and CHOP was greater than control. Increased expression of SDF2 was also detected after 24 h-cultured HTR8/SV-neo cells. PE serum increased sFLT1 gene expression and decreased PlGF gene expression in placental explants. Morphologically, PE serum increased the number of syncytial knots and reduced placental cell metabolism and viability. Analysis of the serum of pregnant women with PE through Raman spectroscopy showed changes in amino acids, carotenoids, lipids, and DNA/RNA, which may be associated with the induction of ER stress found in chorionic villi treated with this serum. In conclusion, this study provides evidence that the serum of pregnant women with PE may impact placental villi changing its morphology, viability, and secreted functional factors while triggers ER stress and an UPR. The differences between PE and control sera include molecules acting as inducing factors in these processes. In summary, the results obtained in our assays suggest that after the development of PE, the serum profile of pregnant women may be an additional factor that feeds a continuous imbalance of placental homeostasis. In addition, this study may expand the possibilities for understanding the pathogenesis of this disorder.

Introduction. Controlling the invasive activity of trophoblastic tissue has not been elucidated. In the accreta placenta, the invasion of placental tissue is directly related to the expression of CRIPTO‐1 at the maternal‐fetal interface. The aim of this study is to evaluate if the expression of the CRIPTO‐1 is related to different degrees of trophoblast invasion into the tube wall in ampullary pregnancy. Methods. Prospective study with 21 patients with ampullary tubal pregnancy undergoing salpingectomy. Anatomopathological evaluation determined the degree of invasion of trophoblast tissues into the tubal wall and grouped the samples into invasive degrees I, II, or III. The groups were compared for tissue expression of CRIPTO‐1 using the Kruskal‐Wallis nonparametric test. p values lower than 0.05 were considered significant. Results. Quantitative expression of CRIPTO‐1 differed in each of the three groups of trophoblast invasion in the tubal wall in ampullary pregnancies (p < 0.001). There is a difference between groups when grade I + grade II versus grade III (p < 0.001) and grade I versus grade II + grade III (p < 0.001). The tissue expression of CRIPTO‐1 in ectopic trophoblasts showed that deeper invasion of the tubal wall was associated with stronger expression than in shallow invasion (p < 0.001). Discussion. In ampullary pregnancies, the depth of penetration of trophoblast tissue in the tubal wall is related to CRIPTO‐1 tissue expression.