Substrate Compliance Research Articles

Integrin Binding Dynamics Modulate Ligand-Specific Mechanosensing in Mammary Gland Fibroblasts.

SummaryThe link between integrin activity regulation and cellular mechanosensing of tissue rigidity, especially on different extracellular matrix ligands, remains poorly understood. Here, we find that primary mouse mammary gland stromal fibroblasts (MSFs) are able to spread efficiently, generate high forces, and display nuclear YAP on soft collagen-coated substrates, resembling the soft mammary gland tissue. We describe that loss of the integrin inhibitor, SHARPIN, impedes MSF spreading specifically on soft type I collagen but not on fibronectin. Through quantitative experiments and computational modeling, we find that SHARPIN-deficient MSFs display faster force-induced unbinding of adhesions from collagen-coated beads. Faster unbinding, in turn, impairs force transmission in these cells, particularly, at the stiffness optimum observed for wild-type cells. Mechanistically, we link the impaired mechanotransduction of SHARPIN-deficient cells on collagen to reduced levels of collagen-binding integrin α11β1. Thus integrin activity regulation and α11β1 play a role in collagen-specific mechanosensing in MSFs.

Influence of non-hydrodynamic forces on the elastic response of an ultra-thin soft coating under fluid-mediated dynamic loading

The force between two approaching solids in a liquid medium becomes increasingly large with decreasing separation—a phenomenon that prevents contact between the two solids. This growth in force occurs because of the intervening liquid, and studies of such physical systems constitute the classical discipline of lubrication. Furthermore, when the solid(s) are soft, there are quantitative as well as qualitative alterations in the force interaction due to the solids’ deformation. The underlying physics as well as resultant system behavior is even more complex when forces of non-hydrodynamic origin come into play. Two major classes of such forces are the DLVO (Derjaguin–Landau–Verwey–Overbeek) forces and the non-DLVO molecular forces. Studies assessing the coupling of these physical phenomenon are avenues of contemporary research. With this view, we perform an analytical study of oscillatory motion of a rigid sphere over an ultra-thin soft coating with an electrolytic solution filling the gap between them. We delineate the distinctive effects of solvation force as well as substrate compliance. Our key finding is the major augmentation in the force and substrate-deformation characteristics of the system due to solvation force when the confinement reduces to a few nanometers. Consideration of solvation force leads to up to four orders of magnitude and up to three orders of magnitude increment in force and substrate-deformation, respectively. While higher softness leads to higher deformation (as expected), its effect on force and substrate-deformation characteristics exhibits a tendency toward amelioration of the increment due to solvation force.

Enhancement of elastohydrodynamic friction by elastic hysteresis in a periodic structure.

Lubricated contacts are present in many engineering and biological systems involving soft solids. Typical mechanisms considered for controlling the sliding friction in such lubricated conditions involve bulk material compliance, fluid viscosity, viscoelastic response of the material (hysteretic friction), and breaking of the fluid film where dry contact occurs (adhesive friction). In this work we show that a two-phase periodic structure (TPPS), with a varying modulus across the sliding surface, provides significant enhancement of lubricated sliding friction when the system is in the elastohydrodynamic lubrication (EHL) regime. We propose that the enhanced friction is due to extra energy loss during periodic transitions of the sliding indenter between the compliant and stiff regions during which excess energy is dissipated through the fluid layer. This is a form of elastic hysteresis that provides a novel mechanism for friction enhancement in soft solids under lubricated conditions.

Interaction between Compliant Surfaces: How Soft Surfaces Can Reduce Friction.

We describe how a long-range repulsive interaction can surreptitiously modify the effective geometry of approaching compliant surfaces, with significant consequences on friction. We investigated the behavior under shear and compression of mica surfaces coated with poly(N-isopropylacrylamide) pNIPAM-based cationic microgels. We show that local surface deformations as small as a few nanometers must be considered to understand the response of such surfaces under compression and shear, in particular when the range of action of normal and friction forces are significantly different, as is often the case for macromolecular lubrication. Under these conditions, a subtle interplay between normal forces and surface compliance may significantly reduce friction increment by limiting the minimum approach of the surfaces under pressure. We found that stiffening of compressed microgels confined in the region of closest approach make it increasingly difficult to reduce the gap between the mica surfaces, limiting the deformation of microgels distant from the contact apex and their contribution to global friction while increasing the effective contact radius. These findings reveal a simple mechanism for a robust control of lubrication: by properly tuning the stiffness and geometry of the interacting bodies, for an ad hoc long-range interaction, the growth of friction with applied normal load can be significantly hindered. Thus, substrate compliance is as significant as surface interaction in the design of low friction, long life tribological systems.

Substrate Compliance Directs the Osteogenic Lineages of Stem Cells from the Human Apical Papilla via the Processes of Mechanosensing and Mechanotransduction.

It is well recognized that the interaction between stem cells and their physical microenvironment plays a fundamental role in controlling cell behaviors and directing lineage commitment, which eventually determines cell fate. Any change in the physical characteristics of the extracellular matrix in terms of topography, geometry, and stiffness has a strong effect on this interaction. Nevertheless, the precise biomechanism that regulates the responses of stem cells to the biophysical properties of substrates is not fully understood. In this study, we generated a series of polydimethylsiloxane (PDMS) substrates with different stiffness properties and explored the whole process involved in the determination of osteogenic lineage in stem cells from the human apical papilla (hSCAPs) in response to substrate stiffness. We first found that the hSCAPs responded to different substrate stiffnesses by changing their cell morphologies and cytoskeletons (via changes in α-tubulin and β-tubulin in microtubules and F-actin in microfilaments). We then found that the hSCAPs secreted more fibronectin in response to the stiffer substrates. We next found that fibronectin interacted with focal adhesion kinase (FAK) and paxillin in the FA plaques, and moreover, the expressions of FAK and paxillin were enhanced as the substrate stiffness increased. We further found that FAK and paxillin directly interacted with β-catenin. Furthermore, the accumulation of β-catenin in the nuclear region was strengthened as the substrate stiffness increased. We finally detected the changes of Lef-1 and TCF-1 in osteogenic-induced hSCAPs and found that their expressions were enhanced as the substrate stiffness increased. Lef-1 and TCF-1, as the transcriptional factors in the nucleus, potentially bound to the promoter region of Runx2 and might ultimately determine the osteogenic lineage in hSCAPs. These results indicate the important effect of stiffness in the microenvironment on the osteogenic lineage of hSCAPs and increase the understanding of the biomechanisms involved in the molecular signal cascade during mechanosensing, mechanotransduction, and stem cell differentiation, which will be useful in the biological fields of cell-matrix/cell-cell interactions and tissue engineering/regenerative medicine.

Compliant Substrates Disrupt Elastic Energy Storage in Jumping Tree Frogs.

Arboreal frogs navigate complex environments and face diverse mechanical properties within their physical environment. Such frogs may encounter substrates that are damped and absorb energy or are elastic and can store and release energy as the animal pushes off during take-off. When dealing with a compliant substrate, a well-coordinated jump would allow for the recovery of elastic energy stored in the substrate to amplify mechanical power, effectively adding an in-series spring to the hindlimbs. We tested the hypothesis that effective use of compliant substrates requires active changes to muscle activation and limb kinematics to recover energy from the substrate. We designed an actuated force platform, modulated with a real-time feedback controller to vary the stiffness of the substrate. We quantified the kinetics and kinematics of Cuban tree frogs (Osteopilus septentrionalis) jumping off platforms at four different stiffness conditions. In addition, we used electromyography to examine the relationship between muscle activation patterns and substrate compliance during take-off in a knee extensor (m. cruralis) and an ankle extensor (m. plantaris). We find O. septentrionalis do not modulate motor patterns in response to substrate compliance. Although not actively modulated, changes in the rate of limb extension suggest a trade-off between power amplification and energy recovery from the substrate. Our results suggest that compliant substrates disrupt the inertial catch mechanism that allows tree frogs to store elastic energy in the tendon, thereby slowing the rate of limb extension and increasing the duration of take-off. However, the slower rate of limb extension does provide additional time to recover more energy from the substrate. This work serves to broaden our understanding of how the intrinsic mechanical properties of a system may broaden an organism's capacity to maintain performance when facing environmental perturbations.

Rolling adhesion of leukocytes on soft substrates: Does substrate stiffness matter?

Cell rolling on vascular endothelium under hydrodynamic blood flow is critical for realization of many physiological and pathological processes, such as inflammatory response and tumor metastasis. The blood-borne cells are in direct contact with the inner layer of endothelium, formed by a highly compliant layer of endothelial cells. The effect of endothelial stiffness on the adhesion and motion of rolling cells is poorly understood. Inspired by recent in vitro studies, here we implemented a computational method to model the specific adhesion of a rolling cell onto a soft substrate, subjected to a creeping shear flow. The substrate is modeled as an elastic half-space, coated with P- and E-selectin receptors with specific affinity for the complementary ligands located on the moving cell. Of particular importance is to predict the effect of substrate stiffness on cell adhesion and its kinematics and kinetics of motion. Simulation results show that the effect of substrate compliance is minimal when coated with P-selectin. Conversely, the trajectory of rolling cells on E-selectin coated substrates is sensitive to the substrate compliance. This is attributed to the moderation of binding forces applied by the soft substrate which leads to a higher average translational velocity of cells.

Mechanical Durotactic Environment Enhances Specific Glioblastoma Cell Responses.

Background: A hallmark of glioblastoma is represented by their ability to widely disperse throughout the brain parenchyma. The importance of developing new anti-migratory targets is critical to reduce recurrence and improve therapeutic efficacy. Methods: Polydimethylsiloxane substrates, either mechanically uniform or presenting durotactic cues, were fabricated to assess GBM cell morphological and dynamical response with and without pharmacological inhibition of NNMII contractility, of its upstream regulator ROCK and actin polymerization. Results: Glioma cells mechanotactic efficiency varied depending on the rigidity compliance of substrates. Morphologically, glioma cells on highly rigid and soft bulk substrates displayed bigger and elongated aggregates whereas on durotactic substrates the same cells were homogeneously dispersed with a less elongated morphology. The durotactic cues also induced a motility change, cell phenotype dependent, and with cells being more invasive on stiffer substrates. Pharmacological inhibition of myosin or ROCK revealed a rigidity-insensitivity, unlike inhibition of microfilament contraction and polymerization of F-actin, suggesting that alternative signalling is used to respond to durotactic cues. Conclusions: The presence of a distinct mechanical cue is an important factor in cell migration. Together, our results provide support for a durotactic role of glioma cells that acts through actomyosin contractility to regulate the aggressive properties of GBM cells.

Assessment of corneal substrate biomechanics and its effect on epithelial stem cell maintenance and differentiation

Whilst demonstrated extensively in vitro, the control of cell behaviour via modulation of substrate compliance in live tissues has not been accomplished to date. Here we propose that stem cells can be regulated solely through in situ modulation of tissue biomechanics. By first establishing, via high-resolution Brillouin spectro-microscopy, that the outer edge (limbus) of live human corneas has a substantially lower bulk modulus compared to their centre, we then demonstrate that this difference is associated with limbal epithelial stem cell (LESC) residence and YAP-dependent mechanotransduction. This phenotype-through-biomechanics correlation is further explored in vivo using a rabbit alkali burn model. Specifically, we show that treating the burnt surface of the cornea with collagenase effectively restores the tissue’s mechanical properties and its capacity to support LESCs through mechanisms involving YAP suppression. Overall, these findings have extended implications for understanding stem cell niche biomechanics and its impact on tissue regeneration.

Epithelial cells exert differential traction stress in response to substrate stiffness

Epithelial wound healing is essential for maintaining the function and clarity of the cornea. Successful repair after injury involves the coordinated movements of cell sheets over the wounded region. While collective migration has been the focus of studies, the effects that environmental changes have on this form of movement are poorly understood. To examine the role of substrate compliancy on multi-layered epithelial sheet migration, we performed traction force and confocal microscopy to determine differences in traction forces and to examine focal adhesions on synthetic and biological substrates. The leading edges of corneal epithelial sheets undergo retraction or contraction prior to migration, and alterations in the sheet's stiffness are affected by the amount of force exerted by cells at the leading edge. On substrates of 30 kPa, cells exhibited greater and more rapid movement than on substrates of 8 kPa, which are similar to that of the corneal basement membrane. Vinculin and its phosphorylated residue Y1065 were prominent along the basal surface of migrating cells, while Y822 was prominent between neighboring cells along the leading edge. Vinculin localization was diffuse on a substrate where the basement membrane was removed. Furthermore, when cells were cultured on fibronectin-coated acrylamide substrates of 8 and 50 kPa and then wounded, there was an injury-induced phosphorylation of Y1065 and substrate dependent changes in the number and size of vinculin containing focal adhesions. These results demonstrate that changes in substrate stiffness affected traction forces and vinculin dynamics, which potentially could contribute to the delayed healing response associated with certain corneal pathologies.

An in vitro correlation of metastatic capacity and dual mechanostimulation.

Cells are under the influence of multiple forms of mechanical stimulation in vivo. For example, a cell is subjected to mechanical forces from tissue stiffness, shear and tensile stress and transient applied strain. Significant progress has been made in understanding the cellular mechanotransduction mechanisms in response to a single mechanical parameter. However, our knowledge of how a cell responds to multiple mechanical inputs is currently limited. In this study, we have tested the cellular response to the simultaneous application of two mechanical inputs: substrate compliance and transient tugging. Our results suggest that cells within a multicellular spheroid will restrict their response to a single mechanical input at a time and when provided with two mechanical inputs simultaneously, one will dominate. In normal and non-metastatic mammary epithelial cells, we found that they respond to applied stimulation and will override substrate compliance cues in favor of the applied mechanical stimulus. Surprisingly, however, metastatic mammary epithelial cells remain non-responsive to both mechanical cues. Our results suggest that, within our assay system, metastatic progression may involve the down-regulation of multiple mechanotransduction pathways.

Locomotor kinematics of tree squirrels (Sciurus carolinensis) in free‐ranging and laboratory environments: Implications for primate locomotion and evolution

The grasping capabilities and gait kinematics characteristic of primates are often argued to be adaptations for safely moving on small terminal branches. The goal of this study was to identify whether Eastern gray squirrels (Sciurus carolinensis)-arboreal rodents that frequently move and forage on small branches, lack primate-like grasping and gait patterns, and arguably represent extant analogs of a stem primate ancestor-adjust gait kinematics to narrow and nonhorizontal branches. We studied locomotor kinematics of free-ranging and laboratory-housed squirrels moving over various substrates. We used high-speed video to film (a) a population of free-ranging squirrels moving on natural substrates and (b) laboratory-housed squirrels moving on horizontal poles. Substrates were coded as small, medium, or large relative to squirrel trunk diameter, and as inclined, declined, or horizontal. Free-ranging squirrels used more gallops and half-bounds on small- and medium-sized substrates, and more high-impact bounds, with reduced limb-lead durations, on declined substrates. Laboratory squirrels moved at higher speeds than free-ranging squirrels and responded to decreasing diameter by using more gallops and half-bounds, lowering speed, and-controlling for speed-increasing mean duty factor, mean number of supporting limbs, and relative forelimb lead duration. Our inability to detect substantial diameter or orientation-related gait adjustments in the wild may be due to a limited accounting of confounding influences (e.g., substrate compliance). Ultimately, studies assessing stability measures (e.g., center of mass fluctuations and peak vertical force) are required to assess whether primates' enhanced grasping and gait patterns engender performance advantages on narrow or oblique substrates.

Respective Effects of Gelatin-Coated Polydimethylsiloxane (PDMS) Substrates on Self-renewal and Cardiac Differentiation of Induced Pluripotent Stem Cells (iPSCs).

The effects of substrate stiffness on the development of cardiomyocytes have been investigated extensively. Polydimethylsiloxane (PDMS) elastomer is one of biomaterials that are commonly used to explore the effects of substrate compliance on stem cell differentiation. Although the effects of substrate stiffness on cardiac differentiation of pluripotent stem cells, such as embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), have been reported, whether the stiffness of PDMS-based substrates could enhance differentiation of iPSCs toward cardiomyocyte lineage or not remains unknown. In this study, we found that a denser gelatin distribution and a higher gelatin adsorption on the stiffer PDMS. In addition, nanotopographies on PDMS substrates with different stiffness were distinct. iPSCs on the stiffer PDMS substrates showed higher pluripotency marker but lower cardiac gene expressions. In contrast, iPSCs on the softer PDMS substrates revealed lower pluripotency marker but higher cardiac gene expressions. These results indicate that stiffer PDMS substrates with gelatin coating could be used to support iPSC self-renewal and softer PDMS substrates coated with gelatin could be used for enhanced cardiac differentiation of iPSCs.

A user's guide for the quantitative analysis of substrate characteristics and locomotor kinematics in free-ranging primates.

Laboratory studies have yielded important insights into primate locomotor mechanics. Nevertheless, laboratory studies fail to capture the range of ecological and structural variation encountered by free-ranging primates. We present techniques for collecting kinematic data on wild primates using consumer grade high-speed cameras and demonstrate novel methods for quantifying metric variation in arboreal substrates. These methods were developed and applied to our research examining platyrrhine substrate use and locomotion at the Tiputini Biodiversity Station, Ecuador. Modified GoPro cameras equipped with varifocal zoom lenses provided high-resolution footage (1080 p.; 120 fps) suitable for digitizing gait events. We tested two methods for remotely measuring branch diameter: the parallel laser method and the distance meter photogrammetric method. A forestry-grade laser rangefinder was used to quantify substrate angle and a force gauge was used to measure substrate compliance. We also introduce GaitKeeper, a graphical user interface for MATLAB, designed for coding quadrupedal gait. Parallel laser and distance meter methods provided accurate estimations of substrate diameter (percent error: 3.1-4.5%). The laser rangefinder yielded accurate estimations of substrate orientation (mean error = 2.5°). Compliance values varied tremendously among substrates but were largely explained by substrate diameter, substrate length, and distance of measurement point from trunk. On average, larger primates used relatively small substrates and traveled higher in the canopy. Ultimately, these methods will help researchers identify more precisely how primate gait kinematics respond to the complexity of arboreal habitats, furthering our understanding of the adaptive context in which primate quadrupedalism evolved.

A comparative study of defect formation in GaAs nanocrystals selectively grown on nanopatterned and flat Si(001) substrates

Crystal defects present in GaAs nanocrystals ∼15–50 nm in diameter and grown by metal organic vapor phase epitaxy on top of two different nanopatterned Si(001) substrates (nanopillars and nanotips with ∼40–80 nm openings embedded in a SiO2 matrix) and on a planar substrate, have been investigated by means of atomic-resolution aberration-corrected scanning transmission electron microscopy. Conditions of their formation are discussed. The defect analysis of the three GaAs/Si systems reveals a higher defect density in the GaAs crystals grown on nanopillars as compared to those grown on nanotips and the planar substrate, possibly concomitant to the atomic-scale irregularities identified at the patterned Si(001) nanopillars. It is concluded that the misfit strain in the GaAs nanocrystals is fully plastically relaxed while no noticeable substrate compliance effects are observed on any of the studied substrates.

Claw morphometrics in monitor lizards: Variable substrate and habitat use correlate to shape diversity within a predator guild.

Numerous studies investigate morphology in the context of habitat, and lizards have received particular attention. Substrate usage is often reflected in the morphology of characters associated with locomotion, and, as a result, claws have become well‐studied ecomorphological traits linking the two. The Kimberley predator guild of Western Australia consists of 10 sympatric varanid species. The purpose of this study was to quantify claw size and shape in the guild using geometric morphometrics, and determine whether these features correlated with substrate use and habitat. Each species was assigned a Habitat/substrate group based on the substrate their claws interact with in their respective habitat. Claw morphometrics were derived for both wild caught and preserved specimens from museum collections, using a 2D semilandmark analysis. Claw shape significantly separated based on Habitat/substrate group. Varanus gouldii and Varanus panoptes claws were associated with sprinting and extensive digging. Varanus mertensi claws were for shallow excavation. The remaining species’ claws reflected specialization for some form of climbing, and differed based on substrate compliance. Varanus glauerti was best adapted for climbing rough sandstone, whereas Varanus scalaris and Varanus tristis had claws ideal for puncturing wood. Phylogenetic signal also significantly influenced claw shape, with Habitat/substrate group limited to certain clades. Positive size allometry allowed for claws to cope with mass increases, and shape allometry reflected a potential size limit on climbing. Claw morphology may facilitate niche separation within this trophic guild, especially when considered with body size. As these varanids are generalist predators, morphological traits associated with locomotion may be more reliable candidates for detecting niche partitioning than those associated directly with diet.

Motile Dendritic Cells Sense and Respond to Substrate Geometry.

Dendritic cell (DC) migration is required for efficient presentation of antigen to T cells and the initiation of an adaptive immune response. In spite of its importance, many aspects of DC migration have not been characterized. DCs encounter a variety of environments with different stiffness and geometry, but the effect of these parameters on DC migration has not yet been determined. We addressed this question by comparing DC motility on standard migration surfaces (polydimethylsiloxane (PDMS)-coated coverslips) and micropost array detectors (mPADs). These two surfaces differ in both stiffness and geometry. We found that DC migration was affected by substrate type, with significant increases in speed and significant decreases in persistence time on mPADs made of PDMS as compared to spin-coated PDMS coverslips. To determine whether the geometry or compliance of the post arrays was responsible for these changes in DC migration, we quantified DC motility on mPADs of identical geometry but different stiffness. Migration was indistinguishable on these mPADs, suggesting that DCs are responsive to geometry of ligand presentation and not stiffness. Further, by micropatterning ligands on flat PDMS surfaces in similar geometries to the mPAD arrays, we determined that DCs respond to the geometry of printed ligand. Finally, we used a variety of small molecule inhibitors to identify pathways involved in geometry sensing. We saw a significant role for myosin contractility and α5β1 integrin engagement. We also noted significant reorganization of the actin cytoskeleton into dynamic actin rings when DCs were motile on posts. From these experiments, we conclude that DCs are insensitive to substrate compliance in the range tested but respond to changes in geometry via a mechanism that involves integrin function, myosin contractility, and remodeling of the actin cytoskeleton. As a possible explanation, we postulate a consistent role for filopodial extension and contraction as the driver of DC motility.

Effect of Substrate Compliance on Measuring Delamination Properties of Elastic Thin Foil

Through the analysis of a model problem, a thin elastic plate bonded to an elastic foundation, we address several issues related to the miniature bulge test for measuring the energy-release rate associated with the interfacial fracture of a bimaterial system, where one of the constituents is a thin foil. These issues include the effect of the substrate compliance on the interpretation of the energy release rate, interfacial strength, and the identification of the boundary of the deforming bulge or the location of the interfacial crack front. The analysis also suggests a way for measuring the so-called foundation modulus, which characterizes the property of the substrate. An experimental example, a stainless steel thin foil bonded to an aluminum substrate through hot-isostatic-pressing (HIP), is used to illustrate and highlight some of the conclusions of the model analysis.

The influence of substrate roughness, patterning, curvature, and compliance in peeling problems

Biological adhesion, in particular the mechanisms by which animals and plants ‘stick’ to surfaces, has been widely studied in recent years, and some of the structural principles have been successfully applied to bioinspired adhesives. However, modelling of adhesion, such as in single or multiple peeling theories, has in most cases been limited to ideal cases, and due consideration of the role of substrate geometry and mechanical properties has been limited. In this paper, we propose a numerical model to evaluate these effects, including substrate roughness, patterning, curvature, and deformability. The approach is validated by comparing its predictions with classical thin film peeling theoretical results, and is then used to predict the effects of substrate properties. These results can provide deeper insight into experiments, and the developed model can be a useful tool to design and optimize artificial adhesives with tailor-made characteristics.

Atomic force microscopy and nanoindentation investigation of polydimethylsiloxane elastomeric substrate compliancy for various sputtered thin film morphologies.

Crack free electrically continuous metal thin films over soft elastomeric substrates play an integral part in realization of modern day flexible bioelectronics and biosensors. Under nonoptimized deposition conditions, delamination, and/or cracking of the top film as well as the underlying soft substrate hinders optimal performance of these devices. Hence it is very important to understand and control not only the various deposition factors like power, time, or deposition pressure but also investigate the various interfacial physics playing a critical role in assuring thin film adhesion and substrate compliancy. In the present study, various nanomechanical information of the underlying substrate, namely, crack profile, average roughness, Young's modulus, and adhesion force were studied for uncracked and cracked polydimethylsiloxane (PDMS) surfaces along with pristine and conventional plasma treated PDMS samples as control. Quantification of the above parameters were done using three-dimensional surface profiler, scanning electron microscopy, nanoindentation, and atomic force microscopy techniques to elucidate the modulus range, average roughness, and adhesion force. Comparative analysis with control revealed remarkable similarity between increased modulus values, increased surface roughness, and reduced adhesion force accounting for reduced substrate compliancy and resulting in film cracking or buckling which are critical for development of various bioflexible devices. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 725-737, 2018.