The ultrastructure of type I collagen at nanoscale: large or small D-spacing distribution?

D-Spacing is the most significant topographic feature of type I collagen fibril, and it is important for our understanding of the structure and function in collagens. Traditionally, the D-spacing of type I collagen fibril was shown to have a singular value of 67 nm, but recent works indicated that the D-spacing values have a large distribution of up to 10 nm when measured by atomic force microscopy. We found that this large distribution of D-spacing values mainly resulted from image drift during measurement. Note that the D-spacing was homogeneous in a single type I collagen fibril. Our statistical analysis indicated that the D-spacing values of type I collagen fibrils exhibited only a narrow distribution of 2.5 nm around the value of 67 nm. In addition, the D-spacing values of the collagen fibrils were nearly identical not only within a single fibril bundle, but also in different fibril bundles. The measurement of the D-spacing values of collagen may provide important structural information in many research areas such as collagen related diseases, construction of molecular model of collagen, and collagen fibrogenesis.