The interplay between surface micro-topography and -mechanics of type I collagen fibrils in air and aqueous media: An atomic force microscopy study

Abstract

Calf skin type I collagen fibrils were regenerated from acidic solution and imaged with contact mode atomic force microscopy in air, water, and buffer solution. When imaged in air at a contact force of 20-150 nN, collagen fibrils exhibited a distinct transverse banding pattern with a period of 65 nm, consisting of high ridges and shallow grooves. The force dependence of the images suggests that such banding pattern is attributed to the transverse contraction of the fibril upon dehydration during sample preparation, which reflects the tangential mass density across the fibril. Imaging in water and phosphate buffer solution at a contact force of 15-80 nN revealed hydrated collagen fibrils with smooth surfaces. The rigidity of the collagen fibrils decreased considerably upon hydration. Scanning the cantilever tip in an aqueous medium at a contact force of 90-280 nN enabled us to probe subunit arrangement in the bulk region of the collagen fibril. The results indicate that the molecular assembly in the hydrated fibril is akin to that in the intact form. The image resolution was improved by stabilizing the collagen molecules through crosslinking with glutaraldehyde, which served to resolve microfibril-like structure on the fibril surface.