P-393 Three-dimensional contrast-enhanced micro-computed tomography of ovaries: a novel method to visualize the different stages of folliculogenesis and ovarian vasculature
J Jansen, S Lie Fong, P Neven, J Vriens, J Van Rheenen, H Hristova, A Vanhie, K De ClercqAbstract
Study question
Can contrast-enhanced micro-focus computed tomography (CE-CT) be used to evaluate the 3D ovarian tissue morphology and is this approach better compared to conventional histology?
Summary answer
CE-CT is a nondestructive technique that provides high-resolution 3D visualization of ovarian tissue, enabling detailed differentiation of follicles and stromal structures across species.
What is known already
Conventional histology provides valuable insight into ovarian tissue architecture but is limited by its two-dimensional and time-consuming nature, labor intensity and possible tissue shrinkage. Advanced imaging modalities, including magnetic resonance imaging and ultrasound, offer spatial information but lack sufficient resolution for detailed follicular analysis. CE-CT has emerged as a high-throughput and high-resolution imaging tool for soft tissues, yet its application to ovarian tissue from multiple species remains underexplored. Studies suggest CE-CT can enhance visualization of follicular and stromal structures, facilitating better understanding of ovarian physiology and pathology, particularly in the context of fertility preservation and oncology.
Study design, size, duration
This cross-species methods study analyzed ovarian tissue of 4 mice, 1 baboon and 5 humans. Tissue samples underwent CE-CT imaging with a resolution of 3,2-5,4μm voxel size using the contrast agent zirconium-substituted Keggin polyoxome-talate (Zr-POM). We were able to visualize the 3D morphology of the ovarian tissue (i.e. different layers, follicles and its vasculature) and quantify ovarian tissue parameters (i.e. follicle and corpus luteum volumes) that are of importance for comparison studies.
Participants/materials, setting, methods
Mouse ovaries were obtained from 4 healthy mice. The baboon ovary was harvested from an olive baboon (Papio anubis). The 5 human ovarian tissue samples were relinquished for scientific research after (partial) oophorectomy was performed for fertility preservation purposes since the subjects needed to undergo chemo- or radiotherapy for various types of malignancies and conditions.
Main results and the role of chance
CE-CT using Zr-POM is a high-throughput technique that allows evaluation of 3D ovarian tissue morphology in mice, baboons and humans with high resolution. It was successfully used to visualize the different stages of folliculogenesis in all species studied and provided the possibility to quantify ovarian tissue parameters. Investigation of the ovarian vasculature was also possible. This technique allowed nondestructive sampling and preserving tissues for further histological and molecular analyses. Tissue processing protocols (fixation and staining) were standardized during the study to reduce artifacts. We used the Bruker X-ray micro-CT and nano-CT software CTAn (CT-analyser) for analysis of the images by surface rendering.
Limitations, reasons for caution
CE-CT is a novel, nondestructive method awaiting histological validation. In addition, CE-CT can currently only be used in a preclinical phase.
Wider implications of the findings
In the future, CE-CT may provide valuable insights in the pathogenesis of accelerated depletion of follicles, e.g. after gonadotoxic treatment which may disrupt ovarian architecture. Also, in individuals at risk of early menopause or premature ovarian insufficiency, such as BRCA-carriers or mosaic Turner syndrome, altered folliculogenesis may be identified.
Trial registration number
No