Vascular Remodeling Across the Spectrum of Cervical Intraepithelial Neoplasia and Squamous Cell Carcinoma: A Digital Pathology-Based Morphometric and Immunohistochemical Analysis

Angiogenesis is a critical component of tumour progression, yet its role in precancerous lesions of the cervix remains underexplored. Cervical intraepithelial neoplasia (CIN) grades 1 to 3 represent a histological continuum toward invasive squamous cell carcinoma (SCC), primarily driven by persistent high-risk human papillomavirus (HPV) infection. This study investigates vascular remodelling across the CIN spectrum and SCC using morphometric and immunohistochemical (IHC) approaches. Methods: Archival cervical tissue samples (n=5; normal cervix, CIN1–CIN3, invasive SCC) were retrieved from the Tbilisi State Medical University laboratory. Clinical data included patient age, HPV status, and menstrual status. IHC for CD34 and CD31 was performed. Digital pathology analysis using Motic EasyScan, QuPath, and ImageJ assessed microvessel density (MVD), vessel diameter, vascular orientation, perivascular localisation, inflammation, and fibrosis. Results: CD34-based MVD increased progressively from the normal cervix (90 vessels/mm²) to invasive SCC (250 vessels/mm²). Vessel diameter expanded from 4 µm in normal tissue to 20 µm in CIN3. Vascular orientation shifted from parallel to chaotic. Perivascular clustering and VEGF expression also intensified with lesion grade. All vascular features showed a strong positive correlation with CIN severity (Spearman ρ > 0.89; p < 0.01). Kruskal-Wallis testing confirmed significant differences across groups (p < 0.01 for MVD, diameter). This study highlights progressive vascular remodelling as a key feature of CIN advancement and transition to invasive carcinoma. Combined morphometric and IHC analysis reveals early and measurable changes in the cervical stromal microvasculature, with implications for early diagnosis, biomarker development, and therapeutic targeting of angiogenesis in cervical neoplasia.