Forced Unfolding of Proteins Directs Biochemical Cascades.

Many proteins in cells and in the extracellular matrix assemble into force-bearing networks, and some proteins clearly transduce mechanical stimuli into biochemical signals. Although structural mechanisms remain poorly understood, the designs of such proteins enable mechanical forces to either inhibit or facilitate interactions of protein domains with other proteins, including small molecules and enzymes, including proteases and kinases. Here, we review some of the structural proteins and processes that exhibit distinct modes of force-dependent signal conversion.

Tissue mechanics coevolves with fibrillar matrisomes in healthy and fibrotic tissues.

Fibrillar proteins are principal components of extracellular matrix (ECM) that confer mechanical properties to tissues. Fibrosis can result from wound repair in nearly every tissue in adults, and it associates with increased ECM density and crosslinking as well as increased tissue stiffness. Such fibrotic tissues are a major biomedical challenge, and an emerging view posits that the altered mechanical environment supports both synthetic and contractile myofibroblasts in a state of persistent activation. Here, we review the matrisome in several fibrotic diseases, as well as normal tissues, with a focus on physicochemical properties. Stiffness generally increases with the abundance of fibrillar collagens, the major constituent of ECM, with similar mathematical trends for fibrosis as well as adult tissues from soft brain to stiff bone and heart development. Changes in expression of other core matrisome and matrisome-associated proteins or proteoglycans contribute to tissue stiffening in fibrosis by organizing collagen, crosslinking ECM, and facilitating adhesion of myofibroblasts. Understanding how ECM composition and mechanics coevolve during fibrosis can lead to better models and help with antifibrotic therapies.

Tension in fibrils suppresses their enzymatic degradation - A molecular mechanism for 'use it or lose it'.

Tissue homeostasis depends on a balance of synthesis and degradation of constituent proteins, with turnover of a given protein potentially regulated by its use. Extracellular matrix (ECM) is predominantly composed of fibrillar collagens that exhibit tension-sensitive degradation, which we review here at different levels of hierarchy. Past experiments and recent proteomics measurements together suggest that mechanical strain stabilizes collagen against enzymatic degradation at the scale of tissues and fibrils whereas isolated collagen molecules exhibit a biphasic behavior that depends on load magnitude. Within a Michaelis-Menten framework, collagenases at constant concentration effectively exhibit a low activity on substrate fibrils when the fibrils are strained by tension. Mechanisms of such mechanosensitive regulation are surveyed together with relevant interactions of collagen fibrils with cells.

Static and time-dependent mechanical response of organic matrix of bone.

Bone derives its mechanical strength from the complex arrangement of collagen fibrils (type-I primarily) reinforced with hydroxy-apatite (HAp) mineral crystals in extra- and intra-fibrillar compartments. This study demonstrates a novel approach to obtain organic matrix of bone through its demineralization as well as mechanically characterize it at small length scales using static and dynamic indentation techniques. Sample surface preparation protocol used in the present work maintained the surface integrity of demineralized bone samples which resulted sample surface of roughness (RMS) magnitude of approximately 14 nm (averaged over 1 × 1 µm2 area duly verified by atomic force microscope (AFM)). Elemental composition analysis via energy dispersive X-ray spectroscopy (EDX) (for probed depth upto 2 µm) confirmed the complete removal of HAp mineral from bone samples during their demineralization using EDTA leaving collagen molecule assemblies unaffected as represented by Second Harmonic Generation (SHG) imaging. The modulus magnitudes of organic matrix obtained using from quasistatic as well as dynamic indentations (at constant frequency of 30 Hz) as ∼2.6 GPa and 4.5 GPa respectively, demonstrated the influence of loading rate on the estimated mechanical properties. For indentation depth to surface roughness ratio greater than ∼5:1, interestingly, measured material properties of organic matrix were found to depend on increasing magnitude of indentation depth of up to ∼500 nm value which probed from few collagen fibrils to next level of hierarchy i.e. collagen fibers. These findings are very useful to accurately determine the elastic and visco-elastic response of organic matrices of mineralized tissues for various applications including tissue engineering, bio-mimetics, etc.

Mechanical response of collagen molecule under hydrostatic compression.

Proteins like collagen are the basic building blocks of various body tissues (soft and hard). Collagen molecules find their presence in the skeletal system of the body where they bear mechanical loads from different directions, either individually or along with hydroxy-apatite crystals. Therefore, it is very important to understand the mechanical behavior of the collagen molecule which is subjected to multi-axial state of loading. The estimation of strains of collagen molecule along different directions resulting from the changes in hydrostatic pressure magnitude, can provide us new insights into its mechanical behavior. In the present work, full atomistic simulations have been used to study global (volumetric) as well as local (along different directions) mechanical properties of the hydrated collagen molecule which is subjected to different hydrostatic pressure magnitudes. To estimate the local mechanical properties, the strains of collagen molecule along its longitudinal and transverse directions have been acquired at different hydrostatic pressure magnitudes. In spite of non-homogeneous distribution of atoms within the collagen molecule, the calculated values of local mechanical properties have been found to carry the same order of magnitude along the longitudinal and transverse directions. It has been demonstrated that the values of global mechanical properties like compressibility, bulk modulus, etc. as well as local mechanical properties like linear compressibility, linear elastic modulus, etc. are functions of magnitudes of applied hydrostatic pressures. The mechanical characteristics of collagen molecule based on the atomistic model have also been compared with that of the continuum model in the present work. The comparison showed up orthotropic material behavior for the collagen molecule. The information on collagen molecule provided in the present study can be very helpful in designing the future bio-materials.