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Objective: To explore the relationship between serum 25-hydroxycholecalciferol (25[OH]D3) and pressurepain thresholds, as measured by algometer, in advance of a main study to determine whether PPT is a potentially cost-effective proxy measure of 25[OH]D3 status in the general population. Methods: The cross-sectional pilot study involved a convenience sample of twenty-two subjects (10 males, 12 females), aged 18 to 67 years. All subjects consented to three trials of pressure-pain threshold readings on both tibiae and the manubrium. Serum 25[OH]D3 levels were determined from blood samples drawn post-algometry. Results: The average pressure pain thresholds were 14.92 (±6.03), 15.07(±6.07), 11.10 (±6.68) for the left and right tibia and sternum, respectively. The stability between the measurements was very high with the interclass correlation coefficient (95% CI) calculated as 0.94 (0.62-1.00), 0.9 (0.81-1.00), 0.96(0.93-1.00). The Pearson correlation coefficients were 0.03 for the left tibia, 0.17 for the right tibia and 0.20 for the sternum, showing a negligible correlation for the left and right tibia, but a low positive correlation for the sternum. Conclusion: We did not find preliminary evidence of a strong or otherwise clinically meaningful correlation between bone tenderness and manual algometry in this pilot study. Only a weak linear relationship between PPT in the sternum and serum 25[OH]D3 concentrations was found. Replication of this study is warranted in larger and more representative study populations of interest. Discussion on a number of feasibility issues is provided to inform those future studies. [ABSTRACT FROM AUTHOR]