Bone consists of an organic and an inorganic matrix. During development, bone undergoes changes in its composition and structure. In this study we apply three different cluster analysis algorithms [K-means (KM), fuzzy C-means (FCM) and hierarchical clustering (HCA)], and discriminant analysis (DA) on infrared spectroscopic data from developing cortical bone with the aim of comparing their ability to correctly classify the samples into different age groups. Cortical bone samples from the mid-diaphysis of the humerus of New Zealand white rabbits from three different maturation stages (newborn (NB), immature (11 days–1 month old), mature (3–6 months old)) were used. Three clusters were obtained by KM, FCM and HCA methods on different spectral regions (amide I, phosphate and carbonate). The newborn samples were well separated (71–100% correct classifications) from the other age groups by all bone components. The mature samples (3–6 months old) were well separated (100%) from those of other age groups by the carbonate spectral region, while by the phosphate and amide I regions some samples were assigned to another group (43–71% correct classifications). The greatest variance in the results for all algorithms was observed in the amide I region. In general, FCM clustering performed better than the other methods, and the overall error was lower. The discriminate analysis results showed that by combining the clustering results from all three spectral regions, the ability to predict the correct age group for all samples increased (from 29–86% to 77–91%). This study is the first to compare several clustering methods on infrared spectra of bone. Fuzzy C-means clustering performed best, and its ability to study the degree of memberships of samples to each cluster might be beneficial in future studies of medical diagnostics.
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