Imaging cervical remodeling in pregnancy through polarization-based techniques

Heterogeneous Microstructural Changes of the Cervix Influence Cervical Funneling

Cervical insufficiency is one of the most frequent reasons for late miscarriage and preterm birth. Its rate is increased in women who conceived after the use of additional reproductive technologies. The objective: to assess the parameters of cervical elastography in women with cervical insufficiency and anovulatory infertility in a history. Materials and methods. 40 pregnant women with cervical insufficiency who conceived after treatment of infertility associated with anovulation were examined in the term of 16–20 weeks: in the I group (20 persons) pregnancy conceived after conservative treatment of infertility, in the II (20 patients) – after in vitro fertilization. 20 women with the physiological course of pregnancy without cervical insufficiency and with spontaneous gestation were controls. Cervical elastography index was used to assess the strain of cervical tissue. Results. According to the results of compression elastography all patients in the I and II groups had red or yellow color of the internal cervical os and cervical canal. In the persons without cervical insufficiency the cervical tissue was firm (purple and blue color). The number of women with purple color of the anterior labia of the cervix in the I and II groups was in 4.67 (р=0.001) and 2.33 (р=0.03) times less compared to the controls, of the posterior labia – in 3,2 times (р=0.001) in both groups. Conclusion. In women with cervical insufficiency and a history of anovulatory infertility there are considerable changes of strain in internal os and cervical canal of the cervix.

This article has been retracted. Background : Cancer is a disease characterized by a population of cells that grow and divide without respect to normal limits, invade and destroy adjacent tissues, and may spread to distant anatomic sites through a process called metastasis . These malignant properties of cancers differentiate them from benign tumors, which are self-limited in their growth and do not invade or metastasize for the development and progression of tumor cells. Cancer is a result of multistep process involving accumulation of genetic alternation, which results in loss of cell-cell interaction, increasing invasive migration, loss of control in cell division and abnormal matrix assembly. Carcinoma of the cervix is one of the most common malignancies. Cervical differentiation requires changes in the composition and structure of extracellular matrix. However, the biological roles of Hyaluronan (HA) in cervical remodeling are still under investigation and HA-receptors are not studied during cervical pathological remodeling. Hence, in the current study we investigated the expression and association of extracellular matrix components namely hyaluronan-hyaluronic acid binding protein (H 11 antigen) (HA-HABP) in normal and invasive lesion in cervical tissue section slides. Methods and Results : Cervical tissues from normal and suspected cancer patients from hospitals were examined. Immunohistochemistry was performed with specific HA probe, b. PG (biotinylated proteoglycan) and anti-HABP antibody H 11 B 2 C 2 for H 11 antigen. HA and H 11 antigen are overexpressed in well-differentiated tumors during progression. Conclusions : We conclude that the expression of HA and H 11 antigen and their interaction are very important in identifying cervical cancers and probably H 11 antigen can be used as a biomarker in identifying the tumor cells in cervical lesions. doi:10.4021/wjon287e

Longitudinal Assessment of Endocervical Canal Length Between 15 and 24 Weeks’ Gestation in Women at Risk for Pregnancy Loss or Preterm Birth

To explore the utility of cervical elastography in singleton pregnancies in the first and second trimesters to assess cervical remodelling and predict spontaneous preterm birth (sPTB). This prospective study enrolled pregnant women scheduled for first trimester ultrasound scans at our hospital. Cervical elastography was conducted by trained sonographers in both the first trimester (11-14 weeks) and the second trimester (20-24 weeks).Cervical elastographic parameters including cervical length (CL), elasticity contrast index (ECI), hardness ratio (HR), internal os stiffness(IOS), external os stiffness (EOS), and the ratio of IOS and EOS (RATIO) were measured using E-Cervix. A total of 587 pregnant women included in the study, 38 (6.47%) had sPTB at less than 37 weeks of gestation,43 cases (7.3%) had short cervix(CL ≤ 25 mm) in the second trimester. CL and HR were significantly lower in the second trimester compared to the first trimester, while ECI, IOS and RATIO were significantly higher(P < 0.001).The parameters in the first trimester showed no significant differences between the term and preterm birth groups (P > 0.05).In the second trimester, ECI was significantly higher and CL was significantly lower in the preterm birth group VS term birth group (P < 0.05). In the first trimester, ECI was significantly higher in the short cervix group, while HR and CL were significantly lower (P < 0.05) VS the normal group. The AUC for predicting preterm birth was 0.617 for CL, 0.603 for ECI alone, and 0.654 for the combination of ECI and CL. The AUC for predicting short cervix was 0.679 for CL alone and 0.703 for the combination of ECI and HR. Cervical Elastography can be used to visualise morphological and histological changes in the cervical remodelling. Combining CL with ECI in the second trimester can be used to predict the risk of sPTB. Although elastography parameters in the first pregnancy may not predict the risk of sPTB, combining CL with ECI and HR can be utilised to predict the risk of a short cervix.

Background . Cervical insufficiency, often associated with genital infections, remains one of the leading causes of extremely preterm and early preterm births. Since cervicovaginal infections change the antimicrobial properties of cervical mucus, studies of local immunity can offer new insights into the pathogenesis of this pregnancy complication and help predict preterm birth. Objective . To examine a local immune response in the cervical mucus of pregnant women with cervical insufficiency that leads to preterm birth. Methods . A retrospective cohort study of 106 pregnant women with cervical insufficiency was conducted. These women were diagnosed with cervical insufficiency according to the clinical guidelines (Cervical Insufficiency, 2021). A microscopic and biomolecular analysis of vaginal and cervical material was performed for all patients. Additionally, the following parameters were estimated in the cervical mucus of 58 randomly selected women: total white blood cell count and functional activity and viability of neutrophils, as well as the levels of macrophage inflammatory protein 1β, high-mobility group protein B1, and matrix metalloproteinase-8. A statistical analysis of the results was performed retrospectively, depending on obstetric outcomes: premature and term births. The obtained data were statistically processed using the SPSS software (version 25.0, IBM, USA). Differences in parameter values between the groups were considered statistically significant at p &lt; 0.05. Results . Cervicovaginal infections in women with cervical insufficiency were detected in 84.9% of cases in the preterm birth group and in 57.5% of cases in the term birth group (p = 0.002). In both groups, cervicitis was found to be prevalent. In the immunological study of cervical mucus, the group with cervical insufficiency leading to preterm birth differed most significantly in the number of neutrophil extracellular traps, as well as in the levels of macrophage inflammatory protein 1β and high-mobility group protein B1. Conclusion . Cervicovaginal infections constitute a risk factor for preterm birth. In patients with cervical insufficiency, changes in the local immunity markers of cervical mucus can serve as predictors of preterm birth.

Background:Estimation of age is an important aspect in determining a person's identity. Since teeth are resistant to decay and degradation, estimation of age using dental structures may be a valuable tool in human identification. Recent research suggests that tooth cemental annulation could be used as an effective method for age estimation than other morphologic or histological parameters.Materials and Methods:A total of 100 extracted teeth were chosen. Using a hard tissue microtome (SP 1600) longitudinal and transverse ground sections of 100 μm thickness of each tooth were prepared. It is further examined under light microscopy, polarized microscopy and phase-contrast microscopy. The images of longitudinal and transverse were magnified on a computer and the cemental lines were counted.Results:The present study showed is a strong positive correlation for cemental annulation between the calculated age and estimated age when phase-contrast microscopy was used in both longitudinal and transverse sections when compared with other microscopes. The correlation coefficient value was found more significant in the longitudinal section which implies that the phase-contrast microscope was highly reliable and had a significant correlation (P < 0.001) with the actual age compared to other microscopes.Conclusion:Cemental annulation by various microscopes improves accuracy in the prediction of age and also it allows for age estimation in cases where skeletal fragments are poorly preserved.

Preterm birth (PTB) is the primary cause of neonatal mortality worldwide. Infection and inflammation are considered to be the primary causes of PTB. Cervical remodeling is an important step in the process of preterm delivery, and the destruction of the cervical epithelial barrier and inflammation are important triggers of cervical remodeling. The aim of the present study was to determine the effect and underlying mechanism of microRNA (miR)-199a-3p/high-mobility group box 1 protein (HMGB1) signaling in cervical epithelial inflammation in PTB. The results of this study revealed that miR-199a-3p was significantly decreased in cervical epithelial tissue samples from patients in both the preterm labor and preterm premature rupture of membrane groups. This decrease was also observed in tissue samples from a lipopolysaccharide (LPS)-induced PTB mouse model and in LPS-induced ectocervical and endocervical cells. Whereas, the expression of HMGB1 and toll-like receptor 4 (TLR4) was significantly increased, which was associated with the upregulation of interleukin (IL)-1β and tumor necrosis factor (TNF)-α expression. Furthermore, overexpression of miR-199a-3p significantly suppressed the expression and activation of HMGB1 and TLR4/NF-κB signaling, and decreased the levels of IL-1β and TNF-α in vitro and in vivo. Additionally, overexpression of HMGB1 and/or TLR4 reversed the anti-inflammatory effects of miR-199a-3p mimics in vitro and in vivo. These results indicate that miR-199a-3p acts as a negative inflammatory regulator in PTB by targeting HMGB1 to regulate the TLR4/NF-κB pathway.

Preterm birth (PTB) remains a pressing global health concern associated with premature cervical ripening and weakened cervical mechanical strength. Second harmonic generation (SHG) microscopy has proved instrumental in tracking progressive changes in cervical collagen morphology during pregnancy. To translate this imaging modality into clinical practice, we have developed a flexible SHG endomicroscope for label-free visualization of cervical collagen architecture. This study aims to assess the feasibility of our SHG endomicroscope for non-invasive differentiation of normal and PTB mouse models, with the ultimate goal of enabling early diagnosis and risk assessment of PTB in vivo. in this pilot investigation, we conducted endomicroscopic SHG imaging on frozen cervical tissue sections and intact cervices resected from both normal pregnant mice and mifepristone-induced PTB mouse models, and then analyzed the acquired images to identify collagen morphology characteristics associated with abnormal cervical collagen remodeling. quantitative image analysis revealed significantly altered collage spatial distribution, larger collagen fiber diameter and pore size, along with reduced pore numbers in SHG endomicroscopy images from PTB mouse models compared to normal pregnant mice. Similar trends were consistent across SHG endomicroscopy images of subepithelial collagen fibers acquired directly from intact cervices. overall, the experiment results underscore the potential of SHG endomicroscopy, coupled with quantitative image analysis, for clinically evaluating cervical collagen remodeling and PTB risk.

Molecular mechanisms regulating preterm birth (PTB)-associated cervical remodeling remain unclear. Prior work demonstrated an altered miRNA profile, with significant increases in miR-143 and miR-145, in cervical cells of women destined to have a PTB. The study objective was to determine the effect of miR-143 and miR-145 on the cervical epithelial barrier and to elucidate the mechanisms by which these miRNAs modify cervical epithelial cell function. Ectocervical and endocervical cells transfected with miR-negative control, miR-143 or miR-145 were used in cell permeability and flow cytometry assays for apoptosis and proliferation. miR-143 and miR-145 target genes associated with cell adhesion, apoptosis and proliferation were measured. Epithelial cell permeability was increased in miR-143 and miR-145 transfected cervical epithelial cells. Cell adhesion genes, JAM-A and FSCN1, were downregulated with overexpression of miR-143 and miR-145. miR-143 and miR-145 transfection decreased cervical cell number by increasing apoptosis and decreasing cell proliferation through initiation of cell cycle arrest. Apoptosis genes, BCL2 and BIRC5, and proliferation genes, CDK1 and CCND2, were repressed by miR-143 and miR-145. These findings suggest that miR-143 and miR-145 play a significant role in cervical epithelial barrier breakdown through diverse mechanisms and could contribute to premature cervical remodeling associated with PTB.

Microenvironmental factors, including microbe-induced inflammation and immune-checkpoint proteins that modulate immune cells have been associated with both cervical insufficiency and preterm delivery. These factors are incompletely understood. This study aimed to explore and compare interactions among microbiome and inflammatory factors, such as cytokines and immune-checkpoint proteins, in patients with cervical insufficiency and preterm birth. In particular, factors related to predicting preterm birth were identified and the performance of the combination of these factors was evaluated. A total of 220 swab samples from 110 pregnant women, prospectively recruited at the High-Risk Maternal Neonatal Intensive Care Center, were collected between February 2020 and March 2021. This study included 63 patients with cervical insufficiency receiving cerclage and 47 control participants. Endo- and exocervical swabs and fluids were collected simultaneously. Shotgun metagenomic sequencing for the microbiome and the measurement of 34 immune-checkpoint proteins and inflammatory cytokines were performed. First, we demonstrated that immune-checkpoint proteins, the key immune-regulatory molecules, could be measured in endocervical and exocervical samples. Secondly, we identified significantly different microenvironments in cervical insufficiency and preterm birth, with precise cervical locations, to provide information about practically useful cervical locations in clinical settings. Finally, the presence of Moraxella osloensis (odds ratio = 14.785; P = 0.037) and chemokine CC motif ligand 2 levels higher than 73 pg/mL (odds ratio = 40.049; P = 0.005) in endocervical samples were associated with preterm birth. Combining M. osloensis and chemokine CC motif ligand 2 yielded excellent performance for predicting preterm birth (area under the receiver operating characteristic curve = 0.846, 95% confidence interval = 0.733-0.925). Multiple relationships between microbiomes, immune-checkpoint proteins, and inflammatory cytokines in the cervical microenvironment were identified. We focus on these factors to aid in the comprehensive understanding and therapeutic modulation of local microbial and immunologic compositions for the management of cervical insufficiency and preterm birth.

IntroductionTo prevent extreme preterm birth, women with cervical insufficiency are eligible for transabdominal cerclage in case of prior failure or technical impossibility for transvaginal cerclage. This study aimed to identify patient characteristics that affect the success rate of transabdominal cerclage to prevent extreme preterm birth in women with cervical insufficiency.Material and MethodsSingle‐center retrospective cohort study in 87 women who underwent transabdominal cerclage by open laparotomy during first and early second trimester of pregnancy over a 20‐year period. Participants were divided into subgroups according to indication for the intervention. Linear regression and meta‐regression‐analyses were performed to assess the effect of mean cervical length (before and after transabdominal cerclage placement) and gestational age of previous preterm birth, on gestational age at delivery. Kaplan–Meier analysis was performed to evaluate treatment effects on gestational age at delivery.ResultsOf 87 women, 62 women underwent a history‐indicated and 25 an ultrasound‐indicated transabdominal cerclage. Fetal survival was 92%: 91% in the history‐indicated and 96% in the ultrasound‐indicated group. Median gestational age at delivery was 37.3 weeks, with a median pregnancy prolongation of 163.0 days and with 92% of deliveries ≥34 weeks. Between groups, irrespective of singleton and twin pregnancies, outcomes were comparable. Gestational age at delivery was neither affected by cervical length before transabdominal cerclage, distance between transabdominal cerclage and external os, gestational age of previous preterm birth nor additional progesterone treatment.ConclusionsThe efficacy of transvaginal cerclage placement via open laparotomy during high‐risk pregnancy is favorable and relates to fetal survival of 92%. Regardless of indication, pregnancy outcomes after transabdominal cerclage are similar, and independent of gestational age at previous preterm birth, cervical length before transabdominal cerclage placement, distance between transabdominal cerclage and external os, and additional progesterone administration.

The mechanical response of the mouse cervix to tensile cyclic loading in term and preterm pregnancy

Few studies have investigated the roles of cytokines and chemokines in women with cervical insufficiency, and those that have done so evaluated only a limited number of cytokines in amniotic fluid. A retrospective cohort study enrolled 71 patients undergoing physical examination-indicated cerclage to determine whether expanded amniotic fluid cytokine levels predict spontaneous very pre-term birth (≤32weeks of gestation) in patients with cervical insufficiency. Analysis of multiple cytokines and chemokines was performed with the multiplex immunoassay. Sixty-seven amniotic fluid samples were available for analysis and assayed for 15 cytokines. Thirty-eight (56.7%) patients delivered pre-term. Of these, 26 (38.8%) were spontaneous very pre-term births. Most cytokine levels were significantly increased in the amniotic fluid from the study group when compared with those from controls. The levels of interleukin-1β (IL-1β), IL- 6, IL- 7, IL-15, IL-17α, tumour necrosis factor-α (TNF-α), MIP-1α, and MIP-1β were higher in patients with a very pre-term delivery than in those with a late pre-term delivery. IL-1β, IL-6, IL-7, IL-17α, TNF-α, and cervical dilation were independently associated with a very pre-term birth. Intra-amniotic inflammation may contribute to cervical insufficiency, and the severity of inflammation is associated with a very pre-term birth in women with cervical insufficiency.

BackgroundDespite the efforts to decrease the rate of preterm birth, preterm delivery is still the main cause of neonatal morbidity and mortality. Identifying patients threatened with preterm delivery remains one of the main obstetric challenges. The aim of this study was to estimate the potential value of elastographic evaluation of internal cervical os stiffness at 18-22 weeks of pregnancy in low risk, asymptomatic women in the prediction of spontaneous preterm delivery.MethodsThis prospective observational study included 333 low-risk, asymptomatic women presenting for the routine second trimester ultrasound scan according to the Polish Gynecological Society recommendation between 18-22 weeks of pregnancy. Ultrasound examinations of the cervix were performed transvaginally. The following data were recorded: elastographic color assessment of the internal os and ultrasound cervical length at 18-22 and 30 weeks of pregnancy; maternal age; obstetrical history; presence of cervical funneling at 30 weeks of pregnancy; gestational age at birth. Elastographic assessment of the internal os was performed using a color map: red (soft), yellow (medium soft), blue (medium hard) and purple (hard). If two colors were visible in the region of the internal os, the softer option was noted. Statistical analysis was performed using Statistica software (version 10, Statsoft Poland) using the following tests: chi square test to compare frequency of preterm deliveries in various categories of internal os assessment and Spearman correlation test to determine the correlation between elastographic assessment and cervical shortening. To determine the cut off category of internal os elastography assessment in selecting high preterm delivery risk patients we have calculated the sensivity, specifity, negative predictive value and positive predictive value.ResultsThe number of preterm deliveries (<37 weeks of pregnancy) was significantly higher in the red and yellow groups, than in the blue and purple groups. The sensivity, specifity, NPV and PPV for both red and yellow internal os assessment in predicting preterm delivery were 85.7%, 97.6%, 98.3% and 81.1% respectively.ConclusionsElastographic assessment of the internal cervical os at 18-22 weeks of pregnancy may identify patients with high risk of preterm delivery in low-risk, asymptomatic women.