It is unclear, however, whether a premature cervical ripening leads to PTB in cases of cervical insufficiency (CI), or a “weak cervix” 14, 15, 16, 17, 18. For instance, during a rapid process of cervical remodeling prior to delivery, called “ripening”, the human cervix undergoes a remarkable transformation from a rigid closed structure to a completely compliant organ 9, 10, 11, 12, 13. Multiple lines of evidence point to changes in the cervix as common factor that lead to delivery, both at term and preterm. Current methods to predict PTB are inadequate, which is due to the multifactorial etiology of this disease and significant gaps in our understanding of the physiological, biochemical and structural changes in the reproductive tract that lead to PTB. PTB affects 10.6% of pregnancies worldwide causing significant infant mortality and morbidities, and rates are continuing to increase 6, 7, 8. Premature opening of the cervix has been implicated in preterm birth (PTB), the delivery of a neonate prior to 37 weeks of gestation 4, 5. At term, the preparedness of the uterus (i.e., uterine smooth muscle, myometrium, and cervix) is crucial for successful delivery of a baby 1, 2, 3. Lasting for about 40 weeks, human pregnancy requires an intimate physiological interaction between mother and fetus. Diffusion MRI images detected changes in tissue organization as gestation progressed suggesting the potential application of this technique to non-invasively monitor cervical changes that precede the onset of labor in women at risk for preterm delivery. MRI detected three distinctive zones in pregnant mouse endocervix: (1) inner collagenous layer, (2) middle circular muscular layer, and (3) outer longitudinal muscular layer. With advanced gestation, endocervical changes, indicative of impending delivery, are manifested in fewer smooth muscle cells, expansion of the extracellular space, and lower presence of collagen fibers. Our results show that mouse endocervix has a higher proportion of smooth muscle cells and collagen fibers per area, with more compact tissue structure, than the ectocervix. We used a mouse model of pregnancy and term labor, to examine the cervical structure by histology (Masson Trichome and Picrosirius Red staining), immunohistochemistry (Hyaluronic Acid Binding Protein/HABP), and ex-vivo MRI (T 2-weighted and diffusion tensor imaging), focusing on two regions of the cervix (i.e., endocervix and ectocervix). Cervical insufficiency, or “weakness”, may lead to preterm birth, which causes infant morbidities and mortalities worldwide. In addition, the retrainable AI-module used in the present study allows for fully automated overnight batch processing of multiple annotated sections.The cervix is responsible for maintaining pregnancy, and its timely remodeling is essential for the proper delivery of a baby.
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We found that estimation of Oc.S/BS by the new AI-assisted method was considerably less time-consuming, while still providing similar results to the conventional manual method. Tibiae from Wistar rats were either enzymatically stained for TRAP or immunostained for cathepsin K to identify osteoclasts. We present a detailed step-by-step guide for the AI-based method. The present study directly compare estimates of osteoclast-covered surfaces (Oc.S/BS) obtained by the conventional manual method using a bright-field microscope to that obtained by a new AI-assisted method. However, most of the present alternatives depend on manual or semi-automatic color segmentation and do not take advantage of artificial intelligence (AI).
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Since the inception of bone histomorphometry and manual counting of osteoclasts using bright-field microscopy, several approaches have been proposed to accelerate the counting process using both free and commercially available software. Quantification of osteoclasts to assess bone resorption is a time-consuming and tedious process.
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