The extracellular matrix of the human aortic wall: ultrastructural observations by FEG-SEM and by tapping-mode AFM

Fragments of human ascending aorta harvested during heart surgery were cryofractured and observed by scanning electron microscopy (SEM) and atomic force microscopy (AFM). Elastic fibers appear as irregular, undulated laminae of variable size and shape. Their surface shows an evident fibrous texture suggestive of a criss-crossed, delicate filamentous scaffold and is marked by a number of features such as ridges, holes and protruding ribs. At higher magnification, both SEM and AFM show the surface composed of a finely granular material, with a bead size of approximately 20 nm. However, the thickness of the metal coating in one case, and the tip convolution effect on the other, may equally result in an artifactual enlargement of the structures, so that the beads may be significantly smaller. The surfaces created by the fracture always appear smooth and compact and with this technique do not reveal significant detail. The collagen component is mostly represented by small, uniform fibrils gathered in flexuous bundles and following a wavy course not unlike that of the elastic laminae. An orthogonal lattice of small proteoglycans is readily evident even without a specific treatment. Occasionally, the fibrils appear encrusted or engulfed in a grainy matrix reminiscent of the elastic fiber surface. Fluid Tapping-Mode Atomic Force Microscopy simultaneously reveals the surface-bound proteoglycans and the inner architecture of the fibrils, composed of smaller subunits following a spiral course with a winding angle of approximately 17 degrees.