Mechanics of isolated individual collagen fibrils.

Collagen fibrils are the fundamental structural elements in vertebrate animals and compose a structural framework that provides mechanical support to load-bearing tissues. Understanding how these fibrils initially form and mechanically function has been the focus of a myriad of detailed investigations over the last few decades. From these studies a great amount of knowledge has been acquired as well as a number of new questions to consider. In this review, we examine the current state of our knowledge of the mechanical properties of extant fibrils. We emphasize on the mechanical response and related deformation of collagen fibrils upon tension, which is the predominant load imposed in most collagen-rich tissues. We also illuminate the gaps in knowledge originating from the intriguing results that the field is still trying to interpret. STATEMENT OF SIGNIFICANCE: Collagen is the result of millions of years of biological evolution and is a unique family of proteins, the majority of which provide mechanical support to biological tissues. Cells produce collagen molecules that self-assemble into larger structures, known as collagen fibrils. As simple as they appear under an optical microscope, collagen fibrils display a complex ultrastructural architecture tuned to the external forces that are imposed upon them. Even more complex is the way collagen fibrils deform under loading, and the nature of the mechanisms that drive their formation in the first place. Here, we present a cogent synthesis of the state-of-knowledge of collagen fibril mechanics. We focus on the information we have from in vitro experiments on individual, isolated from tissues, collagen fibrils and the knowledge available from in silico tests.

Instrument for tensile testing of individual collagen fibrils with facile sample coupling and uncoupling.

Collagen is the major structural protein in human bodies constituting about 30% of the entire protein mass. Through a self-assembly process, triple helical collagen molecules assemble into high aspect-ratio fibers of tens to hundreds of nanometer diameter, known as collagen fibrils (CFs). In the last decade, several methods for tensile testing these CFs emerged. However, these methods are either overly time-consuming or offer low data acquisition bandwidth, rendering dynamic investigation of tensile properties impossible. Here, we describe a novel instrument for tensile testing of individual CFs. CFs are furnished with magnetic beads using a custom magnetic tweezer. Subsequently, CFs are lifted by magnetic force, allowing them to be picked-up by a microgripper structure, which is mounted on a cantilever-based interferometric force probe. A piezo-lever actuator is used to apply tensile displacements and to perform tensile tests of tethered CFs, after alignment. Once the mechanical tests are finished, CFs are removed from the microgripper by application of a magnetic field. Our novel instrument enables tensile tests with at least 25-fold increased throughput compared to tensile testing with an atomic force microscope while achieving force resolution (p-p) of 10 nN at a strain resolution better than 0.1%.

Hydration and nanomechanical changes in collagen fibrils bearing advanced glycation end-products.

Accumulation of advanced glycation end-products (AGEs) in biological tissues occurs as a consequence of normal ageing and pathology. Most biological tissues are composed of considerable amounts of collagen, with collagen fibrils being the most abundant form. Collagen fibrils are the smallest discernible structural elements of load-bearing tissues and as such, they are of high biomechanical importance. The low turnover of collagen cause AGEs to accumulate within the collagen fibrils with normal ageing as well as in pathologies. We hypothesized that collagen fibrils bearing AGEs have altered hydration and mechanical properties. To this end, we employed atomic force and Brillouin light scattering microscopy to measure the extent of hydration as well as the transverse elastic properties of collagen fibrils treated with ribose. We find that hydration is different in collagen fibrils bearing AGEs and this is directly related to their mechanical properties. Collagen fibrils treated with ribose showed increased hydration levels and decreased transverse stiffness compared to controlled samples. Our results show that BLS and AFM yield complementary evidence on the effect of hydration on the nanomechanical properties of collagen fibrils.