Quantitative description of collagen fibre network on trabecular bone surfaces based on AFM imaging

The collagen fibre network is an important part of extracellular matrix (ECM) on trabecular bone surface. The geometry features of the network can provide us insights into its physical and physiological properties. However, previous researches have not focused on the geometry and the quantitative description of the collagen fibre network on trabecular bone surface. In this study,we developed a procedure to quantitatively describe the network and verified the validity of the procedure. The experiment proceeds as follow. Atomic force microscopy (AFM) was used to acquire submicron resolution images of the trabecular surface. Then, an image analysing procedure was built to extract important parameters, including, fibre orientation, fibre density, fibre width, fibre crossing numbers, the number of holes formed by fibre s, and the area of holes from AFM images. In order to verify the validity of the parameters extracted by image analysing methods, we adopted two other methods, which are statistical geometry model and computer simulation, to calculate those same parameters and check the consistency of the three methods' results. Statistical tests indicate that there is no significant difference between three groups. We conclude that, (a) the ECM on trabecular surface mainly consists of random collagen fibre network with oriented fibres; (b) our method based on image analysing can be used to characterize quantitative geometry features of the collagen fibre network effectively. This method may provide a basis for quantitative investigating the architecture and function of collagen fibre network.