Quantitative Spatial Analysis of Matrix Remodelling in Wounds Using Digital Pathology and Fractal Analysis
Wound healing involves spatially heterogeneous changes in matrix composition and organisation that can be difficult to capture using qualitative histological assessment alone. We developed a reproducible digital pathology workflow to quantify matrix features in cutaneous wound sections, compare them with unwounded skin, and derive an exploratory Histological Wound Index (HWI). Four normal-skin…
Wound healing is a complex process characterized by spatially diverse alterations in matrix structure and organization, often challenging to assess through traditional histological methods. To address this issue, we established an accurate digital pathology workflow to measure matrix attributes in skin wound sections, contrast them against healthy skin, and create a preliminary Histological Wound Index (HWI).
Four sections of healthy skin and six segments of wounded skin underwent analysis using QuPath, ImageJ/FracLac, and Python programming.
Regions of interest (ROIs) with dimensions of approximately 500 x 500 micrometers were allocated to four categories: uninjured skin, wound center, proximal edge, and distant edge. Collagen, mucin, and necrotic tissue coverage were measured using image classifiers and further analyzed using linear mixed-effects models. Collagen architecture was evaluated from collagen-selective binary masks using fractal dimension and lacunarity.
Collagen coverage, fractal dimension, mucin coverage, necrosis, and lacunarity were subsequently standardized using z-score normalization and incorporated into the HWI calculation.
Healthy skin exhibited high collagen coverage and fractal dimension, along with low lacunarity, mucin, and necrosis. Conversely, wound center tissues displayed low collagen coverage and fractal dimension, accompanied by elevated mucin and necrosis levels. Regions situated at the wound's edges showed relatively normal-like characteristics, whereas edge regions nearest the wound's center displayed intermediate values.
The HWI reached its peak value in healthy skin and reached its minimum in the wound center. A descriptive ROI comparison of two wounded sections corroborated this regional pattern when all component measures were derived from identical ROI boundaries. This digital pathology workflow offers a reproducible approach for quantifying regional extracellular matrix features and collagen-mask architecture in wound histology.
The HWI serves as a descriptive, exploratory histological summary that necessitates further validation using larger longitudinal datasets.
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