Morphological and Functional Diagnosis of Precancerous Cervical Lesions in Premenopausal Women Using Advanced Diagnostic Methods

Vollständiger Abstract

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Background : Cervical cancer remains a major public health challenge in Uzbekistan, with 2,654 new cases diagnosed in 2022. The premenopausal period (40–55 years) represents a particularly difficult stage for cervical cancer detection, as epithelial atrophy and migration of the transformation zone into the endocervical canal reduce the sensitivity of conventional cytology. Objective: To characterize morphometric, ultrastructural, nanotopographic, and immunohistochemical features of precancerous cervical lesions in premenopausal women, and to identify, at an assessment of hypothesis-generating analysis, parameters that distinguish benign cervical lesions from cervical intraepithelial neoplasia (CIN). Materials and methods : A total of 402 premenopausal women aged 40–65 years were enrolled, including a benign cervical lesion Group I (n = 289), comprising cervical ectopy, squamous metaplasia, and Nabothian cysts, and a histologically confirmed CIN Group II (n = 113: CIN I, n = 48; CIN II, n = 35; CIN III, n = 30). Formalin-fixed, paraffin-embedded tissue specimens from the entire cohort underwent histomorphometric and immunohistochemical analyses (p16INK4a, Ki-67, p53). Separate, substantially smaller sub-samples of fresh or glutaraldehyde-fixed tissue were analyzed by scanning electron microscopy with energy-dispersive spectroscopy (SEM-EDS; 7 benign and 25 CIN specimens) and atomic force microscopy (AFM; 7 benign and 24 CIN specimens). Group comparisons were performed using the Mann–Whitney U test for continuous variables and the χ² test for categorical variables. p < 0.05 was considered statistically significant, 95% CI were reported for all group-level comparisons of continuous variables. Results: Epithelial thickness (64.2 ± 52.4 µm in Group I vs. 362.5 ± 35.0 µm in Group II), nuclear area (25.1 ± 5.3 vs. 206.0 ± 13.7 µm2) and the nuclear-to-cytoplasmic ratio (0.4 ± 0.1 vs. 0.8 ± 0.1) were significantly different between groups (p < 0.001) and increased progressively across CIN I to CIN III. In the SEM sub-sample, the distance of capillaries from the epithelial surface increased from 3.79 ± 2.00 µm in benign tissue to 352.14 ± 111.38 µm in CIN III. In the AFM sub-sample, surface relief height decreased from 1.13 ± 0.28 µm in benign tissue to 0.74 ± 0.27 µm in CIN III. EDS elemental microanalysis demonstrated calcium accumulation and phosphorus depletion in stromal tissue across CIN grades. Immunohistochemical markers (p16INK4a, Ki-67, and p53) differed significantly between Group I and pooled Group II (CIN I-III); however, grade-specific immunohistochemical analyses were not available, representing a limitation of the study. Conclusion : Morphometric, ultrastructural and nanotopographic parameters differ substantially between benign cervical lesions and CIN in this premenopausal cohort and warrant prospective validation. Further studies with histologically normal controls, grade-stratified immunohistochemical analyses, and larger nanoscale sub-samples, are required before diagnostic thresholds can be proposed for clinical application.