Mechano-biological and bio-mechanical pathways in cutaneous wound healing.

Injuries to the skin heal through coordinated action of fibroblast-mediated extracellular matrix (ECM) deposition, ECM remodeling, and wound contraction. Defects involving the dermis result in fibrotic scars featuring increased stiffness and altered collagen content and organization. Although computational models are crucial to unravel the underlying biochemical and biophysical mechanisms, simulations of the evolving wound biomechanics are seldom benchmarked against measurements. Here, we leverage recent quantifications of local tissue stiffness in murine wounds to refine a previously-proposed systems-mechanobiological finite-element model. Fibroblasts are considered as the main cell type involved in ECM remodeling and wound contraction. Tissue rebuilding is coordinated by the release and diffusion of a cytokine wave, e.g. TGF-ß, itself developed in response to an earlier inflammatory signal triggered by platelet aggregation. We calibrate a model of the evolving wound biomechanics through a custom-developed hierarchical Bayesian inverse analysis procedure. Further calibration is based on published biochemical and morphological murine wound healing data over a 21-day healing period. The calibrated model recapitulates the temporal evolution of: inflammatory signal, fibroblast infiltration, collagen buildup, and wound contraction. Moreover, it enables in silico hypothesis testing, which we explore by: (i) quantifying the alteration of wound contraction profiles corresponding to the measured variability in local wound stiffness; (ii) proposing alternative constitutive links connecting the dynamics of the biochemical fields to the evolving mechanical properties; (iii) discussing the plausibility of a stretch- vs. stiffness-mediated mechanobiological coupling. Ultimately, our model challenges the current understanding of wound biomechanics and mechanobiology, beside offering a versatile tool to explore and eventually control scar fibrosis after injury.

Nonaffine Mechanics of Entangled Networks Inspired by Intermediate Filaments.

Inspired by massive intermediate filament (IF) reorganization in superstretched epithelia, we examine computationally the principles controlling the mechanics of a set of entangled filaments whose ends slide on the cell boundary. We identify an entanglement metric and threshold beyond which random loose networks respond nonaffinely and nonlinearly to stretch by self-organizing into structurally optimal star-shaped configurations. A simple model connecting cellular and filament strains links emergent mechanics to cell geometry, network topology, and filament mechanics. We identify a safety net mechanism in IF networks and provide a framework to harness entanglement in soft fibrous materials.

Evaluation of global load sharing and shear-lag models to describe mechanical behavior in partially lacerated tendons.

The mechanical effect of a partial thickness tear or laceration of a tendon is analytically modeled under various assumptions and results are compared with previous experimental data from porcine flexor tendons. Among several fibril-level models considered, a shear-lag model that incorporates fibril-matrix interaction and a fibril-fibril interaction defined by the contact area of the interposed matrix best matched published data for tendons with shallow cuts (less than 50% of the cross-sectional area). Application of this model to the case of many disrupted fibrils is based on linear superposition and is most successful when more fibrils are incorporated into the model. An equally distributed load sharing model for the fraction of remaining intact fibrils was inadequate in that it overestimates the strength for a cut less than half of the tendon's cross-sectional area. In a broader sense, results imply that shear-lag contributes significantly to the general mechanical behavior of tendons when axial loads are nonuniformly distributed over a cross section, although the predominant hierarchical level and microstructural mediators for this behavior require further inquiry.

Activin-mediated alterations of the fibroblast transcriptome and matrisome control the biomechanical properties of skin wounds.

Matrix deposition is essential for wound repair, but when excessive, leads to hypertrophic scars and fibrosis. The factors that control matrix deposition in skin wounds have only partially been identified and the consequences of matrix alterations for the mechanical properties of wounds are largely unknown. Here, we report how a single diffusible factor, activin A, affects the healing process across scales. Bioinformatics analysis of wound fibroblast transcriptome data combined with biochemical and histopathological analyses of wounds and functional in vitro studies identify that activin promotes pro-fibrotic gene expression signatures and processes, including glycoprotein and proteoglycan biosynthesis, collagen deposition, and altered collagen cross-linking. As a consequence, activin strongly reduces the wound and scar deformability, as identified by a non-invasive in vivo method for biomechanical analysis. These results provide mechanistic insight into the roles of activin in wound repair and fibrosis and identify the functional consequences of alterations in the wound matrisome at the biomechanical level.

Tear resistance of soft collagenous tissues.

Fracture toughness characterizes the ability of a material to maintain a certain level of strength despite the presence of a macroscopic crack. Understanding this tolerance for defects in soft collagenous tissues (SCT) has high relevance for assessing the risks of fracture after cutting, perforation or suturing. Here we investigate the peculiar toughening mechanisms of SCT through dedicated experiments and multi-scale simulations, showing that classical concepts of fracture mechanics are inadequate to quantify and explain the high defect tolerance of these materials. Our results demonstrate that SCT strength is only modestly reduced by defects as large as several millimeters. This defect tolerance is achieved despite a very narrow process zone at the crack tip and even for a network of brittle fibrils. The fracture mechanics concept of tearing energy fails in predicting failure at such defects, and its magnitude is shown to depend on the chemical potential of the liquid environment.

On the compressibility and poroelasticity of human and murine skin.

A total of 37 human and 33 murine skin samples were subjected to uniaxial monotonic, cyclic, and relaxation experiments. Detailed analysis of the three-dimensional kinematic response showed that skin volume is significantly reduced as a consequence of a tensile elongation. This behavior is most pronounced in monotonic but persists in cyclic tests. The dehydration associated with volume loss depends on the osmolarity of the environment, so that tension relaxation changes as a consequence of modifying the ionic strength of the environmental bath. Similar to ex vivo observations, complementary in vivo stretching experiments on human volar forearms showed strong in-plane lateral contraction. A biphasic homogenized model is proposed which allows representing all relevant features of the observed mechanical response.

Publisher Correction: Tear resistance of soft collagenous tissues.

The original version of this Article contained errors in the 'Computational material models' section of the Methods in which some values were displayed with incorrect units. As a result of this, a number of changes have been made to both the PDF and the HTML versions of the Article. A full description of these changes is available online and can be accessed via a link at the top of the Article.

The mechanical fingerprint of murine excisional wounds.

A multiscale mechanics approach to the characterization of murine excisional wounds subjected to uniaxial tensile loading is presented. Local strain analysis at a physiological level of tension uncovers the presence of two distinct regions within the wound: i) a very compliant peripheral cushion and ii) a core area undergoing modest deformation. Microstructural visualizations of stretched wound specimens show negligible engagement of the collagen located in the center of a 7-day old wound; fibers remain coiled despite the applied tension, confirming the existence of a mechanically isolated wound core. The compliant cushion located at the wound periphery appears to protect the newly-formed tissue from excessive deformation during the phase of new tissue formation. The early remodeling phase (day 14) is characterized by a restored mechanical connection between far field and wound center. The latter remains less deformable, a characteristic possibly required for cell activities during tissue remodeling. The distribution of fibrillary collagens at these two time points corresponds well to the identified heterogeneity of mechanical properties of the wound region. This novel approach provides new insight into the mechanical properties of wounded skin and will be applicable to the analysis of compound-treated wounds or wounds in genetically modified tissue. STATEMENT OF SIGNIFICANCE: Biophysical characterization of healing wounds is crucial to assess the recovery of the skin barrier function and the associated mechanobiological processes. For the first time, we performed highly resolved local deformation analysis to identify mechanical characteristics of the wound and its periphery. Our results reveal the presence of a compliant cushion surrounding a stiffer wound core; we refer to this heterogeneous mechanical behavior as "mechanical fingerprint" of the wound. The mechanical response is shown to progress towards that of the intact skin as healing takes place. Histology and multiphoton microscopy suggest that wounded skin recovers its mechanical function via progressive reconnection of the newly-deposited collagen fibers with the surrounding intact matrix.

Location-specific mechanical response and morphology of facial soft tissues.

The facial tissue of 9 healthy volunteers (m/f; age: 23-60y) is characterized at three different locations using a procedure combining suction measurements and 18MHz ultrasound imaging. The time-dependent and multilayered nature of skin is accounted for by adopting multiple loading protocols which differ with respect to suction probe opening size and rate of tissue deformation. Over 700 suction measurements were conducted and analyzed according to location-specific mechanical and morphological characteristics. All corresponding data are reported and made available for facial tissue analysis and biomechanical modeling. Higher skin stiffness is measured at the forehead in comparison to jaw and parotid; these two regions are further characterized by lower creep deformation. Thicker tissue regions display a tendency towards a more compliant and less dissipative response. Comparison of superficial layer thickness and corresponding mechanical measurements suggests that connective tissue density determines the resistance to deformation in suction experiments.

A novel ultra-light suction device for mechanical characterization of skin.

Suction experiments have been extensively applied for skin characterization. In these tests the deformation behavior of superficial tissue layers determines the elevation of the skin surface observed when a predefined negative (suction) pressure history is applied. The ability of such measurements to differentiate between skin conditions is limited by the variability of the elevation response observed in repeated experiments. The scatter was shown to be associated with the force exerted by the observer when holding the instrument against the skin. We have developed a novel suction device and a measurement procedure aiming at a tighter control of mechanical boundary conditions during the experiments. The new device weighs only 3.5 g and thus allows to minimize the force applied on the skin during the test. In this way, it is possible to reliably characterize the mechanical response of skin, also in case of low values of suction pressure and deformation. The influence of the contact force is analyzed through experiments on skin and synthetic materials, and rationalized based on corresponding finite element calculations. A comparative study, involving measurements on four body locations in two subjects by three observers, showed the good performance of the new procedure, specific advantages, and limitations with respect to the Cutometer®, i.e. the suction device most widely applied for skin characterization. As a byproduct of the present investigation, a correction procedure is proposed for the Cutometer measurements, which allows to partially compensate for the influence of the contact force. The characteristics of the new suction method are discussed in view of future applications for diagnostic purposes.

Factors affecting the mechanical behavior of collagen hydrogels for skin tissue engineering.

The effect of the production factors yielding a functional dermal substitute was investigated by means of monotonic and cyclic uniaxial tensile tests, as well as electron microscopy visualizations. The role of (i) plastic compression, (ii) product incubation, and (iii) cell permanence in the collagenous matrix in order to achieve a skin-like behavior were characterized in terms of material and structural stiffness, in-plane kinematics, and cyclic response, as well as pore size and network density. The plastic compression resulted in a denser and stiffer material, while no corresponding change was observed in the behavior of the entire structure. This was related to the progressive reduction in product thickness and amount of excess water, rather than to formation of new crosslinks between fibers. Contrary, irrespective of the presence of human fibroblasts, the product incubation induced both material and structural stiffening, indicating the formation of a denser network. These results were confirmed by similar evolutions in the construct in-plane kinematics and cyclic stress reduction. Finally, comparison of constructs incubated in different culture media indicated a determinant contribution of the biochemical environment, rather than of the seeded cells, to the achieved mechanical properties. The observed features are relevant in terms of mechanical biocompatibility of the implant and might direct future optimizations of the production process in order to rapidly attain the desired mechanical properties.