Image-based clustering analysis of calcification patterns in bioprosthetic heart valves

Objective: to identify key patterns of calcification in explanted bioprosthetic heart valves (BHVs) using cluster analysis of computed tomography-derived graphical data. Materials and methods. The study included 11 UniLine BHVs that were routinely explanted during reoperations for structural valve dysfunction. Computed tomography was used to obtain DICOM images of each sample, followed by generation of maximum intensity projections and segmentation of the valves into individual leaflets (n = 33). The images were pre-processed using binary thresholding to differentiate calcified regions from non-calcified biological tissue. Cluster analysis was performed using various algorithms: Gaussian mixture models, Ordering Points To Identify the Clustering Structure (OPTICS), k-means clustering, agglomerative (hierarchical) clustering, and spectral clustering. A basic quantitative method assessing the proportion of pixels corresponding to calcified areas was used for comparison. The performance of clustering algorithms was evaluated using the silhouette score. The presence of calcium deposits in the valves and the accuracy of binary thresholding were further verified histologically by alizarin red S staining of valve cryosections. Results. Data preprocessing based on image binarization yielded a maximum silhouette score of 0.55. Among the clustering algorithms, the highest silhouette scores were achieved with the agglomerative (0.55) and k-means (0.54) methods; however, both demonstrated substantial data imbalance, with up to 85% of samples grouped within a single cluster, limiting their practical applicability. The most balanced clustering was achieved using spectral clustering (silhouette score 0.45) and the basic quantitative approach (0.44). Both methods identified three distinct patterns of bioprosthetic valve leaflet calcification: (1) non-calcified leaflets, (2) partial calcification, and (3) total calcification. Conclusion. Three key calcification patterns were identified in explanted BHVs – absence of calcium, partial calcification, and total calcification. Spectral clustering and the basic quantitative method demonstrated the most balanced results, while other algorithms showed pronounced cluster imbalance. Heat map analysis revealed that in partial calcification, mineral deposition typically begins in the commissural and dome regions of the leaflets, near the free edge, and in total calcification, extends across the entire dome and leaflet base.