When activated muscle in the muscle bath is subjected to progressively increasing load fluctuations approaching the magnitude and frequency expected during normal breathing, the muscle lengthens appreciably in response . But when load fluctuations are progressively reduced, the muscle reshortens somewhat but fails to return to its original length. Incomplete reshortening after exposure to tidal loading is not accounted for by muscle injury; the original operating length can be recovered simply by removing the contractile agonist and allowing the muscle a short interval before recontracting. Neither can incomplete reshortening be accounted for by myosin dynamics; myosin dynamics by themselves predict complete reshortening when the load fluctuations are removed . Thus, the failure of activated muscle to reshorten completely is evidence of a plasticity of the contractile response. During a sustained contraction, the operational length of the muscle for a given loading, or the force at a given length, can be reset by loading and the history of that loading [1,24,25,27,49,70,171-174]. In healthy individuals this plasticity seems to work in a favorable direction, allowing activated muscle to be reset to a longer length. The asthmatic, it has been argued, never manages to melt the contractile domain in the airway smooth muscle and, as such, the benefits of this plastic response are not attained.
It is now firmly established that airway smooth muscle can somehow adapt its contractile machinery, as well as the cytoskeletal scaffolding on which that machinery operates, in such a way that the muscle can maintain the same high force over an extraordinary range of muscle length [1,24,25,27,70,173-178]; ASM is characterized by its ability to disassemble its contractile apparatus when an appropriate stimulus is given, and its ability to reassemble that apparatus when accommodated at a fixed length. When exposed to contractile agonists, airway smooth muscle cells in culture reorganize cytoskeletal polymers, especially actin , and become stiffer . Although cell stiffening is attributable largely to activation of the contractile machinery, an intact actin lattice has been shown to be necessary but not sufficient to account for the stiffening response .
Malleability of the cell and its mechanical consequences have been called by various authors mechanical plasticity, remodeling, accommodation or adaptation. Even though the force generating capacity varies little with length in the fully adapted muscle, the unloaded shortening velocity and the muscle compliance vary with muscle length in such a way as to suggest that the muscle cell adapts by adding or subtracting contractile units that are mechanically in series. The mechanisms by which these changes come about and the factors that control the rate of plastic adaptation are unknown, however.
Several hypotheses have been advanced to explain smooth muscle plasticity. Ford and colleagues have suggested that the architecture of the myosin fibers themselves may change , while Gunst and colleagues have argued that it is the connection of the actin filament to the focal adhesion plaque at the cell boundary that is influenced by loading history [70,171,172]. Alternatively, another notion is that secondary but important molecules stabilize the cytoskeleton, and as the contractile domain melts under the influence of imposed load fluctuations, those loads must be borne increasingly by the scaffolding itself, and thus reflects malleability of the cytoskeletal domain [23,54,70,181]. In that connection a role for the Rho-A pathway has been suggested [54,182] and some evidence now suggests that the p38 MAP kinase pathway may be involved . Airway smooth muscle incubated with an inhibitor of the p38 MAP kinase pathway demonstrates a greater degree of fluctuation-driven muscle lengthening than does control muscle, and upon removal of the force fluctuations it remains at a greater length. Moreover, force fluctuations themselves activate the p38 MAP kinase pathway. It is noteworthy in that connection that heat shock protein 27 has been implicated as an essential element in the motility of airway smooth muscle cells and is a downstream target of rho and p38 [118,183-187]. These findings are consistent with the hypothesis that stress response pathways may somehow stabilize the airway smooth muscle cytoskeleton and limit the bronchodilating effects of deep inspirations.
Regardless of the specific molecules and mechanisms invoked to explain the plasticity of the contractile response, the melting of the contractile domain would appear to be a necessary (or permissive) event, but one that by itself is not sufficient to explain the effects of the history of tidal loading. This brings us, finally, to the notion of the cytoskeleton acting as a glassy material.
An emerging question: are we built of glass?
The abilities of the cytoskeleton (CSK) to deform, to flow, and to remodel (i.e., mechanical plasticity) come into play in a wide variety of situations, from cell division, crawling and extravasation to invasion, contraction and wound healing.
Deformation, flow and remodeling of the airway smooth muscle cell can be described at the molecular level by specific modes of molecular motion and interactions between specific molecular species, at least in principle. The straight-forward approach would be to use direct numerical simulation to describe these molecular interactions within an integrated cytoskeletal lattice and then go on to compute the macroscale integrative properties that result. Direct numerical simulation faces three daunting problems, however. The first is shear complexity; the number of cytoskeletal molecular species is counted in the scores. Moreover, integrated multi-molecular assemblies in airway smooth muscle comprise a messy microstructural geometry, one characterized by a degree of long range order that is far less than that observed in ordinary solids but far greater than that found in fluids. The second is that the list of species remains incomplete and the nature of most protein-protein interactions has yet to be characterized biophysically, with the acto-myosin interaction being the exception that proves the rule. And third, there is good evidence to suggest that in several regards these systems exist far away from local thermodynamic equilibrium (LTE); elemental components do not enjoy a fixed spatial address and, instead, are closely packed and continuously jostling one another in a never-ending search for a minimum energy configuration but never managing to find one. Accordingly, these configurations are adaptable, being in a continuous state of remodeling and, in the process, consuming energy on an ongoing basis via the hydrolysis of ATP. If the first two problems are not enough to saturate the most powerful present-day computers, the non-LTE nature of the problem is particularly thorny because it adds a crucial dimension to the biophysics at the same time that it invalidates a major class of computational approaches, namely, those based on principles of energy minimization.
All three problems are addressed by the approach of Fabry et al. [[187,189]. Using airway smooth muscle cells in a culture system, Fabry discovered that integrative statistical properties of the cytoskeleton, such as measures of its ability to deform, to flow and to remodel, conform at the macroscale to a universal empirical framework, namely, that of glassy systems. Findings to date suggest that these processes depend mainly on a putative energy level in the cytoskeletal lattice, where that energy is representative of the amount of molecular agitation, or jostling, present in the lattice relative to the depth of energy wells that constrain molecular motions. This energy level can be expressed as an effective lattice temperature – as distinct from the familiar thermodynamic temperature. Even while the thermodynamic temperature is held fixed this effective temperature can change, can be manipulated, and can be measured. The higher the effective temperature, the more frequently do elemental structures trapped in one energy well manage to hop out of that well only to fall into another. The hop, therefore, can be thought of as the fundamental molecular remodeling event.
Among the many consequences of these findings, one can easily show that the rate of cytoskeletal remodeling (plastic adaptation) must scale as this effective temperature; when the matrix is "hot", as it is early an a contractile event for example, the rate of remodeling can be fast. But later in the contractile event, as the effective temperature falls and the matrix "cools", the rate of remodeling slows [188-190]. If the effective temperature falls enough that remodeling virtually comes to a standstill, the matrix behaves as if it were frozen. This latter state is consistent with and subsumes the latch state, as described above, while load fluctuations driven by the action of breathing impinge on the cell and represent another source of agitation whose action is consistent with an elevated effective temperature [[187,189]: Gunst, 2003 #1051]. Taken together, these features describe a soft glass, and the effective temperature at which all remodeling ceases is called the glass transition temperature. Data available to date conform in many ways to predictions based upon the idea of traps and hops, and conform as well to closely similar behavior that is shared by other soft materials found in nature including foams, pastes, slurries, colloids and some clays. Taken together, these are referred to in the literature as the class of soft glassy materials.