超声分子成像在妇产科应用

… Molecular imaging with contrast ultrasound relies on the detection of targeted microbubbles or … Targeted contrast ultrasound molecular imaging relies on a departure from this normal …

Phospholipid-coated microbubbles are ultrasound contrast agents that, when functionalized, adhere to specific biomarkers on cells. In this concise review, we highlight recent developments in strategies for targeting the microbubbles and their use for ultrasound molecular imaging (UMI) and therapy. Recently developed novel targeting strategies include magnetic functionalization, triple targeting, and the use of several new ligands. UMI is a powerful technique for studying disease progression, diagnostic imaging, and monitoring of therapeutic responses. Targeted microbubbles (tMBs) have been used for the treatment of cardiovascular diseases and cancer, with therapeutics either coadministered or loaded onto the tMBs. Regardless of which disease was treated, the use of tMBs always resulted in a better therapeutic outcome than non-tMBs when compared in vitro or in vivo.

… Echo from targeted bubbles allows visualization of the biomarker pattern. The purpose of this review is to describe trends in microbubble application for molecular imaging. Microbubble …

Ultrasound imaging has long demonstrated utility in the study and measurement of anatomic features and noninvasive observation of blood flow. Within the last decade, advances in molecular biology and contrast agents have allowed researchers to use ultrasound to detect changes in the expression of molecular markers on the vascular endothelium and other intravascular targets. This new technology, referred to as ultrasonic molecular imaging, is still in its infancy. However, in preclinical studies, ultrasonic molecular imaging has shown promise in assessing angiogenesis, inflammation, and thrombus. In this review, we discuss recent advances in microbubble-type contrast agent development, ultrasound technology, and signal processing strategies that have the potential to substantially improve the capabilities and utility of ultrasonic molecular imaging.

… on the production of microbubbles for molecular imaging. Tissue and cell specificity can be achieved by using passively or actively targeted microbubbles. Passive targeting refers to the …

… In this review, we highlight some of the recent examples on polymeric microbubbles and their applications in ultrasound molecular imaging and drug delivery. © Koninklijke Brill NV, …

… availability of ultrasound systems are advantages compared to other molecular imaging modalities. … and future directions of molecular ultrasound imaging, including different classes of …

… be used for molecular ultrasound imaging since some microbubbles change their acoustic … binding to the target [67]. As a consequence, immobilized and unbound microbubbles can be …

Targeted contrast agents are expanding the detectability and diagnosis of pathology from a strict anatomic to biochemical basis. Moreover, these new agents, in their various forms, offer …

… well as additional data such as molecular characteristics of the … and challenges of molecular ultrasound imaging and offers … of ultrasound and ease of use, molecular ultrasound imaging …

… for ultrasound molecular and cellular imaging using targeted microbubble contrast agents. … , a limitation is that with the current targeted microbubble agents, only intravascular events …

… microbubble is about a picogram; it can be clearly observed by ultrasound imaging of targeted microbubble in … Within a certain microbubble concentration range, ultrasound backscatter …

… Molecularly targeted ultrasound contrast agents are created by conjugating the microbubble shell with a peptide, antibody, or other ligand designed to target an endothelial biomarker …

Ultrasound imaging is a widely used, readily accessible and safe imaging modality. Molecularly-targeted microbubble- and nanobubble-based contrast agents used in conjunction with ultrasound imaging expand the utility of this modality by specifically targeting and detecting biomarkers associated with different pathologies including cancer. In this study, nanobubbles directed to a cancer biomarker derived from the Receptor Protein Tyrosine Phosphatase mu, PTPmu, were evaluated alongside non-targeted nanobubbles using contrast enhanced ultrasound both in vitro and in vivo in mice. In vitro resonant mass and clinical ultrasound measurements showed gas-core, lipid-shelled nanobubbles conjugated to either a PTPmu-directed peptide or a Scrambled control peptide were equivalent. Mice with heterotopic human tumors expressing the PTPmu-biomarker were injected with PTPmu-targeted or control nanobubbles and dynamic contrast-enhanced ultrasound was performed. Tumor enhancement was more rapid and greater with PTPmu-targeted nanobubbles compared to the non-targeted control nanobubbles. Peak tumor enhancement by the PTPmu-targeted nanobubbles occurred within five minutes of contrast injection and was more than 35% higher than the Scrambled nanobubble signal for the subsequent two minutes. At later time points, the signal in tumors remained higher with PTPmu-targeted nanobubbles demonstrating that PTPmu-targeted nanobubbles recognize tumors using molecular ultrasound imaging and may be useful for diagnostic and therapeutic purposes.

… biomarkers expressed on endothelial cells, to target, for … compressibility of the gas core of the microbubbles (MBs), they create … focuses on tUCAs for ultrasound molecular imaging (UMI) …

For preparation of ligand-decorated microbubbles for targeted ultrasound contrast imaging, it is important to maximize the amount of ligand associated with the bubble shell. We describe optimization of the use of a biocompatible cosurfactant in the microbubble formulation media to maximize the incorporation of targeting ligand-lipid conjugate into the microbubble shell, and thus reduce the fraction of ligand not associated with microbubbles, following amalgamation preparation. The influence of the concentration of a helper cosurfactant propylene glycol (PG) on the efficacy of microbubble preparation by amalgamation and on the degree of association of fluorescent PEG-lipid with the microbubble shell was tested. Three sets of targeted bubbles were then prepared: with VCAM-1-targeting peptide VHPKQHRGGSK(FITC)GC-PEG-DSPE, cyclic RGDfK-PEG-DSPE, selective for αVβ3, and control cRADfK-PEG-DSPE, without such affinity. Microbubbles were prepared by 45 s amalgamation, with DSPC and PEG stearate as the main components of the shell, with 15% PG in aqueous saline. In vitro microbubble targeting was assessed with a parallel plate flow chamber with a recombinant receptor coated surface. In vivo targeting was assessed in MC-38 tumor-bearing mice (subcutaneous tumor in hind leg), 10 min after intravenous bolus of microbubble contrast agent (20 million particles per injection). Ultrasound imaging of the tumor and control nontarget muscle tissue in a contralateral leg was performed with a clinical scanner. Amalgamation technique with PG cosurfactant produced microbubbles at concentrations exceeding 2 × 109 particles/mL, and ∼50-60% or more of the added fluorescein-PEG-DSPE or VCAM-1-targeted fluorescent peptide was associated with microbubbles, about 2 times higher than that in the absence of PG. After intravenous injection, peptide-targeted bubbles selectively accumulated in the tumor vasculature, with negligible accumulation in nontumor contralateral leg muscle, or with control nontargeted microbubbles (assessed by contrast ultrasound imaging). For comparison, administration of RGD-decorated microbubbles prepared by traditional sonication, and purified from free peptide-PEG-lipid by repeated centrifugation, resulted in the same accumulation pattern as for translatable amalgamated microbubbles. Following amalgamation in the presence of PG, efficient transfer of ligand-PEG-lipid to microbubble shell was achieved and quantified. Purification of microbubbles from free peptide-PEG-lipid was not necessary, as proven by in vitro and in vivo targeting studies, so PG cosurfactant amalgamation technique generated peptide-targeted microbubbles are amenable for bedside preparation and clinical translation. The pathway to clinical translation is simplified by the fact that most of the materials used in this study either are on the United States Food and Drug Administration GRAS list or can be procured as pharmaceutical grade substances.

… molecular imaging through the use of microbubbles targeted … , molecular acoustic angiography, to image fibrosarcoma … analyze the response to varying microbubble dose for both …

Ultrasound molecular imaging (UMI) has shown promise for assessing the expression levels of biomarkers for the early detection of various diseases. However, it remains difficult to simultaneously image multiple biomarkers in a single systemic administration, which is important for the accurate diagnosis of diseases and for understanding the dynamic intermolecular mechanisms that drive their malignant progression. The authors develop an ultrasound molecular imaging method by serial collapse of targeting microbubbles with distinct acoustic pressures for the simultaneous detection of two biomarkers. To test this, αv β3 -targeting lipid microbubbles (L-MBα ) and VEGFR2-targeting lipid-PLGA microbubbles (LP-MBv ) are fabricated and simultaneously injected into tumor-bearing mice at 7 and 14 days, followed by the low-intensity acoustic collapse of L-MBα and high-intensity acoustic collapse of LP-MBv . The UMI signals of L-MBα and LP-MBv are obtained by subtracting the first post-burst signals from the first pre-burst signals, and subtracting the second post-burst signals from the first post-burst signals, respectively. Interestingly, the signal intensities from UMI agree with the immunohistochemical staining results for αv β3 and VEGFR2. Importantly, they find a better fit for the invasive behavior of MDA-MB-231 breast tumors by analyzing the ratio of αv β3 integrin to VEGFR2, but not the single αv β3 or VEGFR2 levels.

… imaging sequences. In this article, we present an alternative approach for molecular imaging using ultrasound that relies on superharmonic signals produced by microbubble contrast …

… subject and probes are fixed, to awake humans. … of microbubble damping during molecular imaging was of particular interest to us because it is possible that the ligation of microbubbles …

In designing targeted contrast agent materials for imaging, the need to present a targeting ligand for recognition and binding by the target is counterbalanced by the need to minimize interactions with plasma components and to avoid recognition by the immune system. We have previously reported on a microbubble imaging probe for ultrasound molecular imaging that uses a buried-ligand surface architecture to minimize unwanted interactions and immunogenicity. Here we examine for the first time the utility of this approach for in vivo molecular imaging. In accordance with previous results, we showed a threefold increase in circulation persistence through the tumor of a fibrosarcoma model in comparison with controls. The buried-ligand microbubbles were then activated for targeted adhesion through the application of noninvasive ultrasound radiation forces applied specifically to the tumor region. Using a clinical ultrasound scanner, microbubbles were activated, imaged, and silenced. The results showed visually conspicuous images of tumor neovasculature and a twofold increase in ultrasound radiation force enhancement of acoustic contrast intensity for buried-ligand microbubbles, whereas no such increase was found for exposed-ligand microbubbles. We therefore conclude that the use of acoustically active buried-ligand microbubbles for ultrasound molecular imaging bridges the demand for low immunogenicity with the necessity of maintaining targeting efficacy and imaging conspicuity in vivo.

… a molecular target for delivery of imaging probes directly to … for molecular imaging by several imaging modalities. … The acoustic focal zone was placed at the center of the tumor …

… -3 tumors compared to CA-125 negative SKOV-3 tumors. Targeted nanobubbles also exhibited increased tumor … Data suggest that ultrasound molecular imaging using CA-125 antibody-…

… We performed a first-in-human clinical trial on ultrasound molecular imaging (USMI) in patients with breast and ovarian lesions using a clinical-grade contrast agent (kinase insert …

Background The current diagnostic methods of microinvasive cervical cancer lesions are imaging diagnosis and pathological evaluation. Pathological evaluation is invasive and imaging approaches are of extremely low diagnostic performance. There is a paucity of effective and noninvasive imaging approaches for these extremely early cervical cancer during clinical practice. In recent years, ultrasound molecular imaging (USMI) with vascular endothelial growth factor receptor type 2 (VEGFR2) targeted microbubble (MB_VEGFR2) has been reported to improve the early diagnosis rates of breast cancer (including ductal carcinoma in situ), pancreatic cancer and hepatic micrometastases. Herein, we aimed to assess the feasibility of MB_VEGFR2-based USMI in extremely early cervical cancer detection to provide an accurate imaging modality for microinvasive cervical cancer (International Federation of Gynecology and Obstetrics (FIGO) Stage IA1 and IA2). Results We found MB_VEGFR2-based USMI could successfully distinguish extremely early lesions in diameter < 3 mm from surrounding normal tissues (all P  < 0.05), and the sensitivity gradually decreased along with increasing tumor diameter. Moreover, normalized intensity difference (NID) values showed a good linear correlation with microvessel density (MVD) (R^2 = 0.75). In addition, all tumors could not be identified from surrounding muscles in subtracted ultrasound images when mice were administered MB_Con. Conclusions Overall, MB_VEGFR2-based USMI has huge potential for clinical application for the early detection of microinvasive cervical cancer (FIGO Stage IA1 and IA2), providing the foothold for future studies on the imaging screening of this patient population.

Purpose: To develop a mouse ovarian cancer model that allows modulating the expression levels of human vascular targets in mouse xenograft tumors and to test whether expression of CD276 during tumor angiogenesis can be visualized by molecularly targeted ultrasound in vivo. Experimental Design: CD276-expressing MILE SVEN 1 (MS1) mouse endothelial cells were engineered and used for coinjection with 2008 human ovarian cancer cells for subcutaneous xenograft tumor induction in 15 nude mice. Fourteen control mice were injected with 2008 cells only. After confirming their binding specificity in flow chamber cell attachment studies, anti-CD276 antibody-functionalized contrast microbubbles were used for in vivo CD276-targeted contrast-enhanced ultrasound imaging. Results: CD276-targeted ultrasound imaging signal was significantly higher (P = 0.006) in mixed MS1/2008 tumors than in control tumors. Compared with control microbubbles, the ultrasound signal using CD276-targeted microbubbles was significantly higher (P = 0.002), and blocking with purified anti-CD276 antibody significantly decreased (P = 0.0096) the signal in mixed MS1/2008 tumors. Immunofluorescence analysis of the tumor tissue confirmed higher quantitative immunofluorescence signal in mixed MS1/2008 tumors than in control 2008 only tumors, but showed not significantly different (P = 0.54) microvessel density. Conclusions: Our novel small animal model allows for modulating the expression of human tumor–associated vascular endothelial imaging targets in a mouse host and these expression differences can be visualized noninvasively by ultrasound molecular imaging. The animal model can be applied to other human vascular targets and may facilitate the preclinical development of new imaging probes such as microbubbles targeted at human vascular markers not expressed in mice. Clin Cancer Res; 20(5); 1313–22. ©2014 AACR.

Ovarian cancer is the fifth leading cause of cancer‐related deaths in women and the most lethal gynecologic cancer. It is curable when discovered at an early stage, but usually remains asymptomatic until advanced stages. It is crucial to diagnose the disease before it metastasizes to distant organs for optimal patient management. Conventional transvaginal ultrasound imaging offers limited sensitivity and specificity in the ovarian cancer detection. With molecularly targeted ligands addressing targets, such as kinase insert domain receptor (KDR), attached to contrast microbubbles, ultrasound molecular imaging (USMI) can be used to detect, characterize and monitor ovarian cancer at a molecular level. In this article, the authors propose a standardized protocol is proposed for the accurate correlation between in‐ vivo transvaginal KDR‐targeted USMI and ex vivo histology and immunohistochemistry in clinical translational studies. The detailed procedures of in vivo USMI and ex vivo immunohistochemistry are described for four molecular markers, CD31 and KDR with a focus on how to enable the accurate correlation between in vivo imaging findings and ex vivo expression of the molecular markers, even if not the entire tumor could can be imaged by USMI, which is not an uncommon scenario in clinical translational studies. This work aims to enhance the workflow and the accuracy of characterization of ovarian masses on transvaginal USMI using histology and immunohistochemistry as reference standards, which involves sonographers, radiologists, surgeons, and pathologists in a highly collaborative research effort of USMI in cancer.

… cancer types. This review describes the design of ultrasound molecular imaging contrast agents, imaging … , and potential future clinical applications of ultrasound molecular imaging. …

Purpose: Ultrasound (US) molecular imaging by examining the expression of vascular endothelial growth factor receptor 2 (VEGFR2) on uterus vascular endothelium was applied to evaluate the endometrial receptivity. Methods: VEGFR2-targeted ultrasound contrast agents (UCA) and the control UCA (without VEGFR2) were prepared and characterized. Adhesion experiment in vitro was performed with mouse microvascular endothelial cells (bEnd.3) and the ratio of the number of UCA to that of cells at the same field was compared. In vivo study, randomized boluses of targeted or control UCA were injected into the animals of non-pregnancy (D0), pregnancy on day 2 (D2) and day 4 (D4), respectively. Sonograms were acquired by an ultrasound equipment with a 40-MHz high-frequency transducer (Vevo 2100; VisualSonics, Toronto, Canada). The ultrasonic imaging signals were quantified as the video intensity amplitudes generated by the attachment of VEGFR2-targeted UCA. Immunoblotting and immunofluorescence assays were used for confirmation of VEGFR2 expression. Results: Our results showed that VEGFR2-targeted UCA could bind to bEnd.3 cells with significantly higher affinity than the control UCA (9.8 ± 1.0 bubbles/cell versus 0.7 ± 0.3 bubbles/cell, P < 0.01) in vitro. The mean video intensity from the US backscattering of the retained VEGFR2-targeted UCA was significantly higher than that of the control UCA in D2 and D4 mice (D2, 10.5 ± 2.5 dB versus 1.5 ± 1.1 dB, P < 0.01; D4, 15.7 ± 4.0 dB versus 1.5 ± 1.2 dB, P < 0.01), but not significantly different in D0 mice (1.0 ± 0.8 dB versus 0.9 ± 0.6 dB, P > 0.05). Moreover, D4 mice showed the highest video intensity amplitude, indicating the highest VEGFR2 expression when compared with D2 and D0 mice (P < 0.01). This was further confirmed by our immunoblotting and immunofluorescence experiments. Conclusion: Ultrasound molecular imaging with VEGFR2-targeted UCA may be used for noninvasive evaluation of endometrial receptivity in murine models.

To assess the severity of intrauterine adhesion (IUA) after endometrial injury via ultrasound molecular imaging (USMI) of vascular endothelial growth factor receptor 2 (VEGFR2) expression on the uterine endothelium.

Background Angiogenesis plays an important role in endometrial receptivity, determining the response of the endometrium to the blastocyst at the early stage of embryo implantation. During the application of assisted reproduction technologies, it is very important to evaluate the status of uterine angiogenesis before deciding on embryo implantation. Targeted microbubbles (MBs)-based ultrasound molecular imaging (UMI) can noninvasively detect the expression status of biomarkers at the molecular level, thereby being a potential diagnosis strategy for various diseases and their therapeutic evaluation. Methods The iRGD-lipopeptide (DSPE-PEG2000-iRGD) conjugate was prepared with iRGD peptides and DSPE-PEG2000-maleimide through the Michael-type addition reaction. Then, the magnetic iRGD-modified lipid-polymer hybrid MBs (Mag-iLPMs) were prepared with the double-emulsification-solvent-evaporation method. Magnetic targeting of Mag-iLPMs was confirmed under the microscope, followed by a rectangular magnet. Next, the in vitro targeted binding of MBs to murine brain-derived endothelial cells.3 (bEnd.3) and human umbilical vein endothelial cells (HUVEC) were evaluated. The ratio of MBs binding to bEnd.3 and HUVEC at the same field was also compared. For in vivo studies, bolus injections of targeted or control MBs were randomly administrated to non-pregnant or pregnant rats on day 5. Then, the uteri were imaged using a VisualSonics Vevo 2100 ultrasound system (Fujifilm VisualSonics Inc., Ontario, Canada) equipped with a high-frequency transducer. Ultrasonic imaging signals were acquired from Mag-iLPMs, and compared with Mag-LPMs, iLPMs, and LPMs. Results The Mag-iLPMs showed excellent performance in ultrasound contrast imaging and binding affinity to target cells. Using the magnetic field, 10.5- and 12.47-fold higher binding efficiency to bEnd.3 and HUVEC were achieved compared to non-magnetic iLPMs, respectively. Significantly enhanced UMI signals were also observed in the uteri of rats intravenously injected pregnant rats (6.58-fold higher than rats injected with iLPMs). Conclusion We provided a powerful ultrasonic molecular functional imaging tool for uterine angiogenesis evaluation before embryonic implantation.

OBJECTIVE To investigate the preparation of a new kind of targeted lipid ultrasound contrast agent with anti-KDR antibody based on biotin-avidin bridge (MB-BAB-KDR) which could combine specifically with KDR that increases during the time of embryo implantation. Then its binding capability in vitro was evaluated. MATERIALS AND METHODS The agitation of high-speed method was employed to prepare biotin-microbubbles (MB-B), and biotin-avidin mediated technique was used to produce MB-BAB-KDR. MB-BAB-KDR, MB-B, and biotin-microbubbles-KDR (MB-B-KDR) were incubated with fluorescein-conjugated affiniPure goat anti-rat IgG (H+L) to assess the linked condition. Second, MB-BAB-KDR and control groups (MB-B and MB-B-KDR) were incubated with human umbilical vein endothelial cell (HUVEC). Rosette formation rate was observed and calculated. Then, the parallel plate flow chamber technology was used to access attachment efficiency to KDR Fc. RESULTS The surface of bubbles could carry KDR antibody through "biotin-avidin" bridge. After incubated with second antibody, bright green fluorescence (II grade) could be observed in MB-BAB-KDR group, as compared with weak fluorescence in control groups of MB-B (0 grade) and MB-B-KDR (I grade). The surrounding rosette formation rate on HUVEC was 89.86% in MB-BAB-KDR group and that of control groups were 7.13% (MB-B-KDR) and 3.02% (MB-B) (p < 0.05). The number of MB-BAB-KDR bound to KDR Fc increased as the KDR Fc density increased (p < 0.05). Under the same concentration, the MB-BAB-KDR bound to KDR Fc increased as time extended. CONCLUSION The successful preparation of MB-BAB-KDR with anti-KDR antibody which shows specially targeting binding capability with HUVEC and stability in shear stress may be served as a noninvasive detection of endometrial vascular KDR expression and provide an experimental foundation for evaluating endometrial receptivity in vivo.

… A 40-fold increase in the spatial extent of microbubble adhesion occurred with targeted microbubbles, compared with a nontargeted control under static conditions. Additionally, …

INTRODUCTION Ovarian cancer (OC) is the deadliest gynecologic malignancy, with an overall 5-year survival rate of less than 30%. The existing paradigm for OC detection involves a serum marker, CA125, and ultrasound examination, neither of which is sufficiently specific for OC. This study addresses this deficiency through the use of a targeted ultrasound microbubble directed against tissue factor (TF). METHODS TF expression was examined in both OC cell lines and patient-derived tumor samples via western blotting and IHC. In vivo microbubble ultrasound imaging was analyzed using high grade serous ovarian carcinoma orthotopic mouse models. RESULTS While TF expression has previously been described on angiogenic, tumor-associated vascular endothelial cells (VECs) of several tumor types, this is first study to show TF expression on both murine and patient-derived ovarian tumor-associated VECs. Biotinylated anti-TF antibody was conjugated to streptavidin-coated microbubbles and in vitro binding assays were performed to assess the binding efficacy of these agents. TF-targeted microbubbles successfully bound to TF-expressing OC cells, as well as an in vitro model of angiogenic endothelium. In vivo, these microbubbles bound to the tumor-associated VECs of a clinically relevant orthotopic OC mouse model. CONCLUSION Development of a TF-targeted microbubble capable of successfully detecting ovarian tumor neovasculature could have significant implications towards increasing the number of early-stage OC diagnoses. This preclinical study shows potential for translation to clinical use, which could ultimately help increase the number of early OC detections and decrease the mortality associated with this disease.

… To evaluate the use of molecularly targeted microbubbles (MBs) and ultrasonography (US) in the … Figure 5a: Representative tumor slices from ovarian cancer xenograft confirm endoglin …

Abstract Background Ultrasound plays an important role in cancer diagnosis. B-mode imaging and contrast-enhanced ultrasound are routinely used to detect cancerous lesions in breast and liver. The use of ultrasound contrast agents (UCAs) such as microbubbles (MBs), which can be functionalized with targeting ligands, has further enabled ultrasound molecular imaging (USMI) of specific molecular markers in pre-clinical and the first clinical studies. As targeted MBs have a diameter of 1–4 μm, they are limited to the blood vasculature upon intravenous injection, and can bind to markers of the vascular endothelium. USMI with targeted MBs was applied for imaging of markers of inflammation, angiogenesis, and the tumor endothelium. Aim The present review provides an introduction to USMI and presents currently available UCAs, targeting strategies, pre-clinical targets, proposed applications, and the first clinical studies with USMI to guide novel users and assess the technique's potential for clinical use.

… The results of our study suggest that knottin peptide–based contrast microbubbles can be created and successfully used for targeted contrast-enhanced ultrasound imaging of integrins …

… Active targeting involves the use of biomarkers such as antibodies or small peptides to … might be detectable with ultrasound. For use as targeted contrast agents, microbubbles have …

OBJECTIVES The purpose of this review is to summarize literature pertaining to clinical roles of positron emission tomography (PET) or integrated PET and computed tomography (PET/CT) scans, magnetic resonance imaging (MRI) and emerging techniques of these two molecular imaging tools for gynecologic malignancies. METHODS PubMed and MEDLINE databases search for articles published before June 2014 was performed. Only English-language articles were considered. Search terms included "cervical cancer", "endometrial cancer", "uterine cancer", "uterine sarcoma", "ovarian cancer" and "vulvar cancer", in association with "FDG", "PET", "PET/CT", "MRI", "PET/MR", "diffusion", "spectroscopy" and "clinical trial". RESULTS Topics explored included PET, PET/CT and MRI for diagnosis of malignancy, prognostic implications, clinical staging of disease extent, monitoring treatment response, post-therapy surveillance, diagnosis of treatment failure and restaging, and follow-up after salvage therapy in gynecologic malignancies. CONCLUSIONS Molecular imaging (mainly PET and MRI) plays important roles in the management of gynecologic malignancies. Molecular imaging has various impacts in different clinical scenarios. Emerging technologies will continuously improve our practice. Prospective studies with defined endpoints are necessary to evaluate roles of these novel tools in management of gynecologic malignancies.

The aim of this review is to give an overview of the current status of molecular image–guided surgery in gynaecological malignancies, from both clinical and technological points of view. A narrative approach was taken to describe the relevant literature, focusing on clinical applications of molecular image–guided surgery in gynaecology, preoperative imaging as surgical roadmap, and intraoperative devices. The most common clinical application in gynaecology is sentinel node biopsy (SNB). Other promising approaches are receptor-target modalities and occult lesion localisation. Preoperative SPECT/CT and PET/CT permit a roadmap for adequate surgical planning. Intraoperative detection modalities span from 1D probes to 2D portable cameras and 3D freehand imaging. After successful application of radio-guided SNB and SPECT, innovation is leaning towards hybrid modalities, such as hybrid tracer and fusion of imaging approaches including SPECT/CT and PET/CT. Robotic surgery, as well as augmented reality and virtual reality techniques, is leading to application of these innovative technologies to the clinical setting, guiding surgeons towards a precise, personalised, and minimally invasive approach.

Gynecological pathologies account for approximately 4.5% of the overall global disease burden. Although cancers of the female reproductive system have understandably been the focus of a great deal of research, benign gynecological conditions—such as endometriosis, polycystic ovary syndrome, and uterine fibroids—have remained stubbornly understudied despite their astonishing ubiquity and grave morbidity. This historical inattention has frequently become manifested in flawed diagnostic and treatment paradigms. Molecular imaging could be instrumental in improving patient care on both fronts. In this Focus on Molecular Imaging review, we will examine recent advances in the use of PET, SPECT, MRI, and fluorescence imaging for the diagnosis and management of benign gynecological conditions, with particular emphasis on recent clinical reports, areas of need, and opportunities for growth.

… In summary, molecular imaging (mainly PET and MR imaging) has played important roles in gynecologic oncology, for tumor detection, primary staging, treatment planning, prediction of …

Simple Summary Gynecological malignancies are among the most common cancers with significant morbidity and mortality worldwide. Management and overall patient survival is reliant upon early detection, accurate staging and early detection of any recurrence. This article provides a comprehensive review of the recent advances in imaging of gynecologic malignancies with emphasis on cervical, endometrial, and ovarian neoplasms. Abstract Gynecologic malignancies are among the most common cancers in women worldwide and account for significant morbidity and mortality. Management and consequently overall patient survival is reliant upon early detection, accurate staging and early detection of any recurrence. Ultrasound, Computed Tomography (CT), Magnetic resonance imaging (MRI) and Positron Emission Tomography-Computed Tomography (PET-CT) play an essential role in the detection, characterization, staging and restaging of the most common gynecologic malignancies, namely the cervical, endometrial and ovarian malignancies. Recent advances in imaging including functional MRI, hybrid imaging with Positron Emission Tomography (PET/MRI) contribute even more to lesion specification and overall role of imaging in gynecologic malignancies. Radiomics is a neoteric approach which aspires to enhance decision support by extracting quantitative information from radiological imaging.

To describe the clinical and ultrasound characteristics of uterine sarcomas.

… Germ cell neoplasms comprise 15% to 20% of all ovarian tumors and 3% to 7% of malignant ovarian tumors. Germ cell neoplasms are further classified by the dominant cell type, with …

Gynecologic malignancies such as ovarian, endometrial, and cervical cancers are characterized by challenges in early diagnosis and high therapeutic complexity, creating an urgent need for more precise imaging techniques. Fluorescence molecular imaging, a modality with high sensitivity and high spatial resolution, has demonstrated considerable value in tumor diagnosis and therapy in recent years. Its application in gynecologic oncology is rapidly expanding. By using tumor-specific molecular probes to label neoplastic tissues, this technology enables real-time intraoperative navigation and visualization of tumor lesions and their local features, thereby significantly improving surgical accuracy and therapeutic outcomes. This review summarizes the basic principles of fluorescence imaging, recent advances in molecular probe design, and developments in imaging devices. It places particular emphasis on the value of fluorescence molecular imaging in the diagnosis and precision treatment of gynecologic tumors, aiming to provide systematic theoretical guidance and technical support for related research and clinical practice.

Different treatment options are available for patients with gynecological cancers. Imaging plays an important role in assessment of patients with common cancers involving uterine body, cervix and ovaries, from detection to evaluation of the extent of disease. The purpose of this review is to highlight the role of cross-sectional imaging techniques in treatment stratification and overall management of patients with endometrial, cervical and ovarian cancers. Several imaging techniques used are described, including ultrasound, computed tomography (CT), MRI and PET/CT. Specific imaging appearances of the most common uterine, cervical and ovarian cancers are discussed. Imaging findings corresponding to the 2009 revised International Federation of Gynecology and Obstetrics (FIGO) staging of gynecologic malignancies are also described. In the multidisciplinary evaluation of patients with gynecologic malignancies, the role of the radiologist has become central for accurate diagnosis and evaluation of extent of disease to achieve better treatment selection and planning.

To describe the clinical and sonographic characteristics of extragastrointestinal stromal tumors (eGISTs).

To describe the clinical and sonographic characteristics of benign, retroperitoneal, pelvic peripheral‐nerve‐sheath tumors (PNSTs).

Endometriosis is a highly prevalent, chronic gynecological disorder characterized by the ectopic presence of endometrial-like tissue, driving significant morbidity and chronic pelvic pain. Pathologically, it is increasingly recognized as a fibro-inflammatory condition involving extensive tissue remodeling and fibrosis. Current conventional imaging modalities, including ultrasound and MRI, are primarily morphological, while standard molecular imaging using Positron Emission Tomography (PET) tracers has shown limited diagnostic utility. [18F]Fluorodeoxyglucose (FDG) suffers from high physiological uptake in pelvic organs and inconsistent detection of lesions. Receptor-based tracers like [68Ga]Ga-DOTATATE have demonstrated uncertain efficacy. In contrast, radiopharmaceuticals targeting the Fibroblast Activation Protein (FAP) offer a promising molecular approach. FAP is specifically overexpressed by activated fibroblasts present in the stroma of endometriotic lesions, correlating significantly with tissue fibrosis (collagen content) and local immune infiltration (e.g., CD68 macrophages). This comprehensive review analyzes the landscape of radiopharmaceuticals for endometriosis imaging, contrasting the specific limitations of traditional metabolic and receptor agents with the molecular rationale and emerging evidence supporting the use of FAP Inhibitors (FAPI), positioning them as crucial, non-invasive tools for the future diagnosis and management of this challenging disease.

The paper aims to understand the expression of nerve growth factor receptor (TrkA) in the ectopic and entopic endometrium of patients with adenomyosis and dysmenorrhea, and at the same time explore and analyze the combination of hysteroscopy and ultrasound imaging based on multi-operator algorithms The clinical value of colour Doppler ultrasound in the diagnosis and treatment of endometrial polyps. The immunohistochemical avidin-peroxidase staining method (SP method) was used to detect 20 cases of ectopic endometrium and entopic endometrium in patients with dysmenorrhea of adenomyosis (study group) and 22 cases of uterine fibroids in the control group. The expression of nerve growth factor receptor in the endometrium of patients with dysmenorrhea and comparative analysis. At the same time, all patients were examined by transvaginal colour Doppler ultrasound and hysteroscopy. The scores of endometrial polyps, the diagnostic efficiency of the methods, and the ROC curve analysis of various diagnostic methods were observed and recorded in the two groups. There was no significant difference in the expression of TrkA in the ectopic endometrium and the entopic endometrium in the study group (P>0.05), but the expression intensity of TrkA in the two groups was significantly higher than that in the control group (P0.05). In the diagnosis of endometrial polyps, the area of ROC curve in the parallel combined diagnosis of ultrasound and hysteroscopy is significantly larger than that under the combined diagnosis of series. TrkA is widely expressed in the ectopic and entopic endometrium of patients with adenomyosis and dysmenorrhea, and may play an important role in the onset of adenomyosis and dysmenorrhea. Hysteroscopy combined with colour Doppler ultrasound can effectively improve the diagnostic accuracy of patients with endometrial polyps.

Objective. In this study, we utilized gonadotropin-releasing hormone analogue-modified indocyanine green (GnRHa-ICG) to improve the accuracy of intraoperative recognition and resection of endometriotic lesions. Methods. Gonadotropin-releasing hormone receptor (GnRHR) expression was detected in endometriosis tissues and cell lines via immunohistochemistry and western blotting. The in vitro binding capacities of GnRHa, GnRHa-ICG, and ICG were determined using fluorescence microscopy and flow cytometry. In vivo imaging was performed in mouse models of endometriosis using a near-infrared fluorescence (NIRF) imaging system and fluorescence navigation system. The ex vivo binding capacity was determined using confocal fluorescence microscopy. Results. GnRHa-ICG exhibited a significantly stronger binding capacity to endometriotic cells and tissues than ICG. In mice with endometriosis, GnRHa-ICG specifically imaged endometriotic tissues (EMTs) after intraperitoneal administration, whereas ICG exhibited signals in the intestine. GnRHa-ICG showed the highest fluorescence signals in the EMTs at 2 h and a good signal-to-noise ratio at 48 h postadministration. Compared with traditional surgery under white light, targeted NIRF imaging-guided surgery completely resected endometriotic lesions with a sensitivity of 97.3% and specificity of 77.8%. No obvious toxicity was observed in routine blood tests, serum biochemicals, or histopathology in mice. Conclusions. GnRHa-ICG specifically recognized and localized endometriotic lesions and guided complete resection of lesions with high accuracy.

Multifunctional small‐molecule theranostic agents hold significant clinical potential for non‐invasive endometriosis (EMS) management. Current EMS treatment faces challenges due to imprecise lesion localization and therapy‐associated side effects. Herein, an integrated theranostic probe, cRGDyK‐ICG‐Lys‐DTPA@Gd (cRGD‐ILD), designed for concurrent precise lesion localization and targeted phototherapy in EMS, is developed. This molecular probe integrates three key functional components: the DTPA@Gd complex provides contrast for magnetic resonance imaging, the ICG fragment enables fluorescence imaging and photoacoustic imaging while serving as the phototherapeutic agent, and the cRGD peptide targets integrin αvβ3 receptors to facilitate precise accumulation in ectopic endometrial tissue. Specifically, cRGD‐ILD leverages its small size and receptor‐mediated transcytosis to penetrate neovascularized tissue and target ectopic endometrial cells. Under 808 nm laser irradiation, the ICG moiety generates reactive oxygen species and heat, exerting combined photodynamic and photothermal therapeutic effects that effectively suppress lesion progression in both autografted EMS rats and xenografted nude mice. Notably, composed of clinically available and FDA‐approved constituents, the probe demonstrates favorable biocompatibility and biosafety for in vivo applications. Overall, this well‐designed molecular probe enables precise multimodal imaging guidance and localized phototherapy for EMS.

Endometrial cancer is the most common gynecologic malignancy in industrialized countries. Most patients are cured by surgery; however, about 15% of the patients develop recurrence with limited treatment options. Patient-derived tumor xenograft (PDX) mouse models represent useful tools for preclinical evaluation of new therapies and biomarker identification. Preclinical imaging by magnetic resonance imaging (MRI), positron emission tomography-computed tomography (PET-CT), single-photon emission computed tomography (SPECT) and optical imaging during disease progression enables visualization and quantification of functional tumor characteristics, which may serve as imaging biomarkers guiding targeted therapies. A critical question, however, is whether the in vivo model systems mimic the disease setting in patients to such an extent that the imaging biomarkers may be translatable to the clinic. The primary objective of this review is to give an overview of current and novel preclinical imaging methods relevant for endometrial cancer animal models. Furthermore, we highlight how these advanced imaging methods depict pathogenic mechanisms important for tumor progression that represent potential targets for treatment in endometrial cancer.

Imaging of clinically relevant preclinical animal models is critical to the development of personalized therapeutic strategies for endometrial carcinoma. Although orthotopic patient-derived xenografts (PDXs) reflecting heterogeneous molecular subtypes are considered the most relevant preclinical models, their use in therapeutic development is limited by the lack of appropriate imaging modalities. Here, we describe molecular imaging of a near-infrared fluorescently labeled monoclonal antibody targeting epithelial cell adhesion molecule (EpCAM) as an in vivo imaging modality for visualization of orthotopic endometrial carcinoma PDX. Application of this near-infrared probe (EpCAM-AF680) enabled both spatio-temporal visualization of development and longitudinal therapy monitoring of orthotopic PDX. Notably, EpCAM-AF680 facilitated imaging of multiple PDX models representing different subtypes of the disease. Thus, the combined implementation of EpCAM-AF680 and orthotopic PDX models creates a state-of-the-art preclinical platform for identification and validation of new targeted therapies and corresponding response predicting markers for endometrial carcinoma.

The success of gene therapy depends largely on the development of gene vectors and effective gene delivery systems. It has been demonstrated that cationic microbubbles can be loaded with negatively charged plasmid DNA and thus improve gene transfection efficiency. In this study, we developed dual-targeting cationic microbubbles conjugated with iRGD peptides(Cyclo(Cys-Arg-Gly-Asp-Lys-Gly-Pro-Asp-Cys)) and CCR2 (chemokine (C-C motif) receptor 2) antibodies (MBiRGD/CCR2) for ultrasound molecular imaging and targeted tumor gene therapy. The ultrasound molecular imaging experiments showed that there were significantly enhanced ultrasound molecular imaging signals in the tumor that received MBiRGD/CCR2, compared with those that received MBiRGD, MBCCR2, or MBcontrol. As a therapy plasmid, pGPU6/GFP/Neo-shAKT2, carrying an expression cassette for the human AKT2 RNA interference sequence, was used. Our results demonstrated that MBiRGD/CCR2 had a significantly higher gene transfection efficiency than MBiRGD, MBCCR2, or MBcontrol under ultrasound irradiation, resulting in much lower AKT2 protein expression and stronger tumor growth inhibition effects in vivo and in vitro. In conclusion, our study demonstrated a novel gene delivery system via MBiRGD/CCR2 for ultrasound molecular-imaging-guided gene therapy of breast cancer.

The commonly used ultrasound (US) molecular probes, such as targeted microbubbles and perfluorocarbon emulsions, present a number of inherent problems including the conflict between US visualization and particle penetration. This study describes the successful fabrication of phase changeable folate-targeted perfluoropentane nanodroplets (termed FA-NDs), a novel US molecular probe for tumor molecular imaging with US. Notably, these FA-NDs can be triggered by low-intensity focused US (LIFU) sonication, providing excellent US enhancement in B-mode and contrast-enhanced US mode in vitro. After intravenous administration into nude mice bearing SKOV3 ovarian carcinomas, 1,1′-dioctadecyl-3,3,3′,3′ -tetramethylindotricarbocya-nine iodide-labeled FA-NDs were found to accumulate in the tumor region. FA-NDs injection followed by LIFU sonication exhibited remarkable US contrast enhancement in the tumor region. In conclusion, combining our elaborately developed FA-NDs with LIFU sonication provides a potential protocol for US molecular imaging in folate receptor-overexpressing tumors.

… across various imaging modalities to assess the current status of clinical molecular imaging. The various physiologic and scientific bases of clinical molecular imaging are surveyed to …

Abstract Introduction Successful placentation is crucial for fetal development and maintaining a healthy pregnancy. Placental insufficiency can cause a variety of obstetric complications. Despite the many efforts to enhance diagnosing placental insufficiency, no imaging technique has proven satisfactory. A promising imaging technique is contrast-enhanced ultrasound (CEUS) using microbubbles which has proven capable of (micro)vascular imaging. Its use for placental vascularization assessment in human pregnancies remains constrained by limited evidence and safety concerns. This scoping review aims to demonstrate the safety of CEUS used in human pregnancy in the published literature to date. Material and Methods A systematic search using PubMed, Medline, Embase, and Cochrane databases was performed. All studies where contrast-enhanced ultrasound was used in pregnant humans were included. Studies, where there was a planned termination of pregnancy, were excluded. To assess the safety of CEUS during pregnancy, relevant outcomes were divided into the following 3 categories; fetal outcome, maternal outcome, and pregnancy and neonatal outcomes. Results A total of 13 articles were included, in which 256 women underwent CEUS during pregnancy. No clinically significant maternal or fetal adverse events or negative pregnancy or neonatal outcomes associated with CEUS were described. Conclusion Based on our findings, we consider expanding the knowledge of this promising diagnostic technique in future larger clinical studies to be safe and relevant.

… safety of ultrasound and contrast agents during pregnancy. … tion to the field of medical imaging. The review offers a … context of a scoping review aimed at synthesizing reliable evidence …

… microbubble contrast agents (MBCAs) were obtained in Asia, Europe, and Canada, with initial studies focused … for creation of a microbubble/contrast only image with superb detection of …

Objective To determine the influence of ultrasound/microbubble-mediated miR-424-5p delivery on trophoblast cells and the underlying mechanism. Methods Blood pressure and 24-h proteinuria of patients with preeclampsia (PE) were measured as well as the levels of miR-424-5p and amine oxidase copper containing 1 (AOC1) in placental tissues. HTR-8/Svneo and TEV-1 cells were subjected to cell transfection or ultrasonic microbubble transfection for determination of the expression of miR-424-5p, AOC1, β-catenin and c-Myc as well as cell proliferation, apoptosis, migration and invasiveness. The concentrations of placental growth factor (PLGF), human chorionic gonadotropin (β-hCG) and tumor necrosis factor-α (TNF-α) were measured in HTR-8/Svneo and TEV-1 cells. RNA immunoprecipitation (RIP) and dual luciferase reporter assay detected the binding of miR-424-5p to AOC1. A PE mouse model was induced by subcutaneous injection of L-NAME, where the influence of ultrasound/microbubble-mediated miR-424-5p delivery was evaluated. Results miR-424-5p was downregulated while AOC1 was upregulated in the placental tissues from PE patients. Overexpression of miR-424-5p activated Wnt/β-catenin signaling pathway and promoted the proliferation of HTR-8/Svneo and TEV-1 cells as well as enhanced the migratory and invasive behaviors. AOC1 overexpression partly eliminated the effects of miR-424-5p on HTR-8/Svneo and TEV-1 cells. Ultrasound and microbubble mediated gene delivery enhanced the transfection efficiency of miR-424-5p and further promoted the effects of miR-424-5p in trophoblast cells. Ultrasound/microbubble-mediated miR-424-5p delivery alleviated experimental PE in mice. Conclusion Ultrasound and microbubble-mediated miR-424-5p delivery targets AOC1 and activates Wnt/β-catenin signaling pathway, thus promoting the aggressive phenotype of trophoblast cells, which indicating that miR-424-5p/AOC1 axis might be involved with PE pathogenesis.

Ultrasound (US) and microbubble (MB) gene delivery has attracted growing interest as a clinically applicable gene therapy (GT). Though preclinical studies have investigated the system in various tissues, there is limited research in targeting the placenta. This is a potential therapeutic strategy for preeclampsia (PE), which has an underlying genetic basis and ineffective management strategies. Differentially expressed placental microRNAs (miRNAs) in PE may represent suitable targets for GT. Microbubbles (SonoVue) and plasmid (pGL3 or pGL4.13) were administered systemically to CD1 mice, followed by exposure of the heart to US (H14, 1.8 M.I., 1cm focal depth, 2 minutes), using Siemens Acuson Sequoia-512 system and 15L8 probe. Luciferase assays were performed to evaluate gene transfection. Significantly differentially expressed placental miRNAs in PE patients were identified as candidates based on detection by three or more screening studies. Expression of candidate miRNAs was measured by qRT-PCR in PE rat model placentas. In trial 1, low levels of luciferase activity were detected in the heart of treatment mouse 1, 2 and 3. In trial 2, luciferase activity was evident in the atria of treatment mouse 2. In trial 3, higher luciferase activity was detected in the ventricles of the treatment mouse and activity was also detected in the atria. The literature review identified eight candidate miRNAs. MiR-223 (1.46-fold increase) and miR-181a (0.81-fold decrease) were significantly differentially expressed in PE rat model placentas. MiR-223 and -181a may represent targets for US and MB gene delivery. Future studies will apply the US and MB gene delivery protocol for translation to targeting the placenta in our PE rodent model.

… To evaluate the clinical value of the contrast agent SonoVue in the treatment of uterine fibroids with ultrasound-guided high intensity focused ultrasound (HIFU) therapeutic ablation. …

… ultrasound imaging in practice is already over billions and all pregnant women have been screened with ultrasound … Spatially targeted drug delivery with ultrasound and microbubbles is …

Fetal growth restriction (FGR), preeclampsia, and other placental disorders are leading contributors to perinatal morbidity and mortality, primarily due to impaired uteroplacental perfusion. Existing imaging modalities, such as Doppler ultrasound and fetal MRI, provide indirect or limited functional insights into placental and fetal perfusion, constraining timely clinical intervention. To evaluate contrast-enhanced ultrasound (CEUS) as a promising, safe, and real-time tool for assessing placental perfusion and its potential application in maternal-fetal medicine through comprehensive analysis of methodological parameters, safety profiles, and emerging computational techniques. A comprehensive synthesis of preclinical and clinical studies was conducted, focusing on the safety, efficacy, and current use of CEUS in pregnancy. Key findings were drawn from animal models (rats, sheep, macaques) and human studies involving 256 pregnant individuals, with detailed analysis of imaging protocols, contrast agent characteristics, and quantification methods. CEUS utilizes intravascular microbubble contrast agents (1–8 μm diameter) that do not cross the placental barrier, enabling safe maternal imaging. However, size distribution analysis reveals sub-micron populations (8–20% by number) requiring careful evaluation. Preclinical models confirm CEUS ability to detect placental perfusion Changes with 54% reduction in perfusion index following uterine artery ligation (p < 0.001). Human studies demonstrate zero clinically significant adverse events among 256 cases, though critical gaps exist including absent biomarker monitoring and long-term follow-up. Emerging AI-enhanced analysis achieves 73–86% diagnostic accuracy using ensemble deep learning architectures. Current limitations include significant protocol heterogeneity (MI 0.05–0.19, frequency 2–9 MHz) and absence of standardization. CEUS presents a compelling solution for perfusion imaging in pregnancy, offering functional, bedside imaging without fetal exposure to contrast agents. However, methodological limitations, knowledge gaps regarding long-term outcomes, and the distinction between conventional microbubbles and emerging nanobubble formulations demand systematic research investment. Clinical translation requires standardized protocols, comprehensive safety monitoring including biomarker assessment, ethical oversight, and long-term outcome studies to support integration into routine obstetric care.

Microbubbles (MBs) have served as ultrasound (US) contrast agents for over 30 years in cardiac imaging and liver tumor characterization. Moreover, in recent years, molecularly targeted MBs are currently under clinical evaluation for oncological and inflammatory diseases. Beyond diagnostics, MBs are gaining attention as therapeutic tools, leveraging their strong acoustic properties for US-mediated drug delivery, sonopermeation of biological barriers (i.e., the blood-brain barrier), and targeted thrombolysis. Loading MB shells with magnetic or optical functionalities allows therapy monitoring using magnetic resonance or photoacoustic imaging, aligning with recent multimodal advances in (pre)clinical device developments. Therefore, this review summarizes and critically assesses advances in the use of multifunctional MBs for biomedical applications. A comprehensive overview of existing MB formulations is provided, analyzing the primary types of functional agents incorporated, including small molecules, nanomaterials, and targeting ligands, as well as the conjugation and functionalization strategies involved in constructing next-generation MBs. Current trends in multifunctional MBs for imaging and therapy are critically evaluated, along with the challenges in their clinical translation. Overall, this review highlights the potential of multifunctional MBs to address unmet biomedical needs that plain additives cannot fulfill, and it showcases promising future directions for the diagnostic and therapeutic use of next-generation MB formulations.

Intrauterine injection of naked DNA expressing luciferase, green fluorescent protein (GFP), or beta-galactosidase (beta-gal) and fluorescein isothiocyanate-labeled oligodeoxynucleotide (FITC-ODN), in combination with microbubble-enhanced ultrasound (US), referred to as the "shotgun method" (SGM), produced high-level protein expression in fetal mice. With the SGM, luciferase expression increased approximately 10(3)-fold in comparison with expression after injection of naked DNA alone. Electron microscopic analysis demonstrated transient formation of pores on the skin surface after intraamniotic (i.a.) injection with the SGM. Widespread expression of GFP and beta-gal and delivery of FITC-ODN were observed in multiple fetal tissues adjacent to the injection points. PCR analysis indicated that germline transfection was only transient following intraperitoneal (i.p) injection, and there was no evidence of transfer of the reporter gene to the offspring. Thus, SGM might provide a useful means to clarify the molecular mechanisms of genetic diseases in utero, as well as a tool to develop gene therapies in utero.

Background and objectives: Native ultrasound is the most common imaging modality in obstetrics. The use of contrast-enhanced ultrasound (CEUS) during pregnancy has not been officially approved by leading societies for obstetrics and ultrasound. The present study aims to monitor the safety and diagnostic performance of CEUS for assessing abdominal issues in five pregnant women. Materials and Methods: Five pregnant patients who underwent a total of 11 CEUS examinations between June 2020 and October 2020 were included (mean age: 34 years; mean time of pregnancy: 21 weeks). All CEUS scans were interpreted by one experienced consultant radiologist (EFSUMB Level 3). Results: Upon contrast application, no maternal nor fetal adverse effects were observed. Moreover, no fetal contrast enhancement was observed in any patient. CEUS helped to diagnose renal angiomyolipoma, pyelonephritis, necrotic uterine fibroid, gallbladder polyp, and superior mesenteric vein thrombosis. Conclusions: In our study, off-label use of CEUS showed an excellent safety profile allowing the avoidance of ionizing radiation exposure as well as contrast agents in case of CT or use of gadolinium-based contrast agents in case of MRI. CEUS is a promising diagnostic instrument for facilitating clinical decision-making and improving the management of pregnant women.

… Microbubbles provide image contrast through a combination of … Microbubbles efficiently reflect ultrasound energy, resulting … Targeting ligands can be incorporated onto the head groups …

Uterine disorders, including fibroids, polyps, and carcinomas, represent significant health concerns for women. Conventional diagnostic methods, including ultrasound and magnetic resonance imaging, often lack the sensitivity required to accurately delineate and evaluate these lesions. Consequently, there is a growing need for advanced imaging techniques to enhance both diagnostic precision and therapeutic decision-making. Standard imaging modalities typically provide low resolution, limiting their effectiveness in evaluating uterine pathology and determining optimal treatment strategies. In this context, microbubbles combined with contrast-enhanced ultrasonography offer a promising solution. This technique enhances the visualization of vascular patterns and tissue perfusion, facilitating a more accurate assessment of uterine lesions. Contrast-enhanced ultrasonography is less invasive, is more cost-effective, and offers real-time evaluation of vascular characteristics, making it a valuable tool for diagnosing uterine abnormalities. This review explores the clinical utility of contrast-enhanced ultrasonography with microbubbles in uterine diseases, focusing on its diagnostic accuracy, comparison with traditional imaging techniques, and its potential to improve the management of uterine pathology.

The application of serum biomarker to ovarian tumors for early stage detection and clinical diagnosis is a rapidly expanding research area. The problem with conventional markers is that they are often released too late or at too low a level to be detected in time to trigger effective treatment. Ultrasound has been used to influence bio-effects in living cells, but there is only one reported case of the use of ultrasound to enhance the release of a biomarker (Carcinoembryonic antigen CEA). In this study we report the use of ultrasound to enhance the release of a combination of ovarian cancer biomarkers (CA125 and CA19-9) to help in the diagnosis of ovarian cancer at an early stage. The results indicated that after 5 min sonication at a frequency of 1 MHz and intensity of 0.3 W cm(-2), the CA125 and CA19-9 levels were increased by 2.02 and 4.21-fold respectively. These findings suggest that ultrasonic treatment can be used to enhance the release of serum biomarkers from ovarian tumors.

According to the literature, the von Hippel-Lindau (VHL) gene has a certain correlation with ovarian cancer. In this study, we investigated the effect and mechanism of ultrasound microbubble-mediated VHL on the biological function of ovarian cancer cells. Non-targeting lipid microbubbles and targeted lipid microbubbles were prepared. OVCAR-3 cells were treated with VHL mediated by ultrasound and microbubbles alone or together. Expressions of VHL, Akt, epithelial-mesenchymal-transition-related proteins and apoptosis-related proteins were detected by Western blot and quantitative real-time polymerase chain reaction as needed. The effect of ultrasound microbubble-mediated VHL on the proliferation, apoptosis, cell cycle, migration and invasion of OVCAR-3 cells was examined by Cell Counting Kit-8, flow cytometry, wound-healing assay and Transwell. Compared with other treatment methods, ultrasound microbubble mediation enhanced VHL expression in OVCAR-3 cells. Overexpression of liposome-mediated VHL inhibited the proliferation and migration; caused cell-cycle arrest; promoted apoptosis: downregulated the expressions of MMP2, MMP9, E-cadherin, Akt and Bcl-2; and upregulated the expressions of VHL and BCL2-associated X protein (BAX) in OVCAR-3 cells. The effect of microbubble-treated VHL was similar to liposome-mediated regulation, while ultrasound treatment enhanced the effect of VHL on OVCAR-3 cells. More interestingly, ultrasound microbubble-mediated VHL had the most obvious regulatory effect on OVCAR-3 cells. Ultrasound microbubble technology increases the transfection efficiency of VHL into OVCAR-3 cells and enhances the effect of VHL gene on the biological function of OVCAR-3 cells.

… as an appropriate target for cancer treatment. We have … microbubbles (TOPLMBs) for ultrasound (US) mediated delivery for combination therapy in an intraperitoneal ovarian cancer …

… ) loaded lipid microbubbles (OPLMBs) for ultrasound mediated combination therapy in hypoxic ovarian cancer cells. Our experiments successfully demonstrated that ultrasound induced …

We have previously shown that low frequency ultrasound can release biomarkers from cells into the murine circulation enabling an amplification and localization of the released biomarker that could be used as a blood-based method to detect cancer earlier and monitor therapy. In this study, we further demonstrate that this technique could be used for characterization of tumors and/or identification of cellular masses of unknown origin due to the release of multiple protein and nucleic acid biomarkers in cells in culture, mice and patients. We sonicated colon (LS174T) and prostate (LNCaP) cancer cell lines in culture at a low frequency of 1 MHz and show that there were several-fold changes in multiple protein and microRNA (miRNA) abundance with treatment at various intensities and time. This release was dependent on the duration and intensity of the sonication for both cell lines. Significant increased release in biomarkers was also observed following tumor sonication in living mice bearing subcutaneous LS174T cell line xenografts (for proteins and nucleic acids) and in an experimental LS174T liver tumor model (for proteins only). Finally, we demonstrated this methodology of multiple biomarker release in patients undergoing ablation of uterine fibroids using MR guided high intensity focused ultrasound. Two protein biomarkers significantly increased in the plasma after the ultrasound treatment in 21 samples tested. This proof that ultrasound-amplification method works in soft tissue tumor models together with biomarker multiplexing, could allow for an effective non-invasive method for identification, characterization and localization of incidental lesions, cancer and other disease. Pre-treatment quantification of the biomarkers, allows for individualization of quantitative comparisons. This individualization of normal marker levels in this method allows for specificity of the biomarker-increase to each patient, tumor or organ being studied.

… Our experiment verifies the hypothesis that ultrasound mediation of ovarian cancer-targeted … -loaded microbubbles for ultrasound mediated delivery of paclitaxel to ovarian cancer cells. …

Ultrasound-targeted microbubble destruction (UTMD) has recently been developed as a promising noninvasive tool for organ- and tissue-specific gene or drug delivery. The aim of the present study was to explore the role of UTMD-mediated Sirtuin 3 (SIRT3) overexpression in the malignant behaviors of human ovarian cancer (HOC) cells. Reverse transcription-quantitative PCR was performed to detect SIRT3 mRNA expression levels in normal human ovarian epithelial cells and HOC cell lines; low SIRT3 expression was found in HOC cell lines, and the SKOV3 cell line was used in the following experiments. The SIRT3-microbubble (MB) was prepared, and the effects of ultrasound-treated SIRT3-MB on biological processes of SKOV3 cells were determined. The proliferation, migration, invasion and apoptosis of SKOV3 cells were measured after SIRT3 upregulation by UTMD. Xenograft tumors in nude mice were induced to observe tumor growth in vivo. Upregulation of SIRT3 inhibited the malignant behaviors of SKOV3 cells, whereas UTMD-mediated SIRT3 upregulation further inhibited proliferation, epithelial-mesenchymal transition, invasion and migration, and induced apoptosis of SKOV3 cells, and it also inhibited tumor formation and growth in vivo. Moreover, the present study identified hypoxia inducible factor-1α (HIF-1α) as a target of SIRT3. The present study provided evidence that UTMD-mediated overexpression of SIRT3 may suppress HOC progression through the inhibition of HIF-1α.

… and hinder efficacy as an ultrasound-liberated biomarker. We hypothesize that smaller … with ultrasound than larger molecules. Although investigation of large libraries of biomarkers …

Background Radiation therapy is regarded as an effective treatment for early ovarian cancer (OC). However, due to radiation resistance caused by DNA double-strand breaks (DSBs) and angiogenesis, the efficacy of radiotherapy for advanced OC is limited and controversial. Purpose To explore whether ultrasound-stimulated microbubbles (USMBs) can enhance the radiosensitivity of OC. Material and Methods OC cells (ES-2) were respectively irradiated with 5-Gy and 10-Gy radiation doses with or without exposure to USMB. Methyl thiazolyltetrazolium (MTT) and colony-formation assays were conducted to detect the viability and proliferation of ES-2 cells after USMBs and ionizing radiation (IR) treatment. Immunofluorescence assays were conducted to examine levels of gamma-H2A histone family member X (γ-H2AX), an indicator for DSBs. Flow cytometry analyses were carried out to assess the apoptosis of ES-2 cells. The angiogenic activity of human umbilical vein endothelial cells (HUVECs) was measured by tube formation assays. Results USMBs enhanced IR-induced suppressive effect on the viability and proliferation of OC cells. The protein levels of phosphorylated γ-H2AX and CHK1 were significantly upregulated after IR treatment and further enhanced by USMBs. In addition, USMBs enhanced the promotion of IR-mediated OC cell apoptosis. The inhibitory effect of IR on angiogenesis was further enhanced by USMBs, and protein levels of AT1R, VEGFA, and EGFR were downregulated by IR in a dose-dependent way and then enhanced by USMB treatment in HUVECs. Conclusions USMB exposure significantly enhances the radiosensitivity of OC by suppressing cell proliferation, promoting OC cell apoptosis, and inhibiting angiogenesis.

Early detection of ovarian cancer remains an important unmet medical need. Effective screening could reduce mortality by 10%–30%. Used individually, neither serum CA125 nor transvaginal sonography (TVS) is sufficiently sensitive or specific. Two-stage strategies have proven more effective, where a significant rise above a woman's baseline CA125 prompts TVS and an abnormal sonogram prompts surgery. Two major screening trials have documented that this strategy has adequate specificity, but sensitivity for early-stage (I–II) disease must improve to have a greater impact on mortality. To improve the first stage, different panels of protein biomarkers have detected cases missed by CA125. Autoantibodies against TP53 have detected 20% of early-stage ovarian cancers 8 months before elevation of CA125 and 22 months before clinical diagnosis. Panels of autoantibodies and antigen–autoantibody complexes are being evaluated with the goal of detecting >90% of early-stage ovarian cancers, alone or in combination with CA125, while maintaining 98% specificity in control subjects. Other biomarkers, including micro-RNAs, ctDNA, methylated DNA, and combinations of ctDNA alterations, are being tested to provide an optimal first-stage test. New technologies are also being developed with greater sensitivity than TVS to image small volumes of tumor. See all articles in this CEBP Focus section, “NCI Early Detection Research Network: Making Cancer Detection Possible.”

… biomarkers (eg, CA125), the primary benefit of ultrasound is to … located superiorly out of the range of TVU probe. When the … tubes by conventional ultrasound is necessary or microbubble …