Strain Stiffening Research Articles

Versican binds collagen via its G3 domain and regulates the organization and mechanics of collagenous matrices

Type I collagen is the most abundant structural protein in the body and, with other fibrillar collagens, forms the fibrous network of the extracellular matrix. Another group of extracellular matrix polymers, the glycosaminoglycans and glycosaminoglycan-modified proteoglycans, play important roles in regulating collagen behaviors and contribute to the compositional, structural and mechanical complexity of the extracellular matrix. While the binding between collagen and small leucine-rich proteoglycans has been studied in detail, the interactions between collagen and the large bottlebrush proteoglycan versican are not well understood. Here, we report that versican binds collagen directly and regulates collagen structure and mechanics. Versican colocalizes with collagen fibers in vivo and binds to collagen via its C-terminal G3 domain (a non-GAG-modified domain present in all known versican isoforms) in vitro; it promotes the deposition of a highly-aligned collagen-rich matrix by fibroblasts. Versican also shows an unexpected effect on the rheology of collagen gels in vitro, causing decreased stiffness and attenuated shear strain stiffening, and the cleavage of versican in liver results in reduced tissue compression stiffening. Thus, versican is an important collagen binding partner and plays a role in modulating collagen organization and mechanics.

Hemp Protein Isolate-Based Natural Thermal-Reversible Hydrogel as a Novel Wound Dressing Material.

Hydrogels, due to their excellent microstructure and mechanical strength, have become a novel biomaterial in wound dressing. However, plant proteins have never been considered because of their poor original gelling performances and insufficient rheological properties. Here, we reported the fabrication of a plant protein-based thermal-reversible gel using a reverse micelle-extracted hemp protein isolate (HPI). A systematic study was conducted to fully reveal their microstructure, rheological properties, and anti-inflammatory effect to lay a foundation for this newly developed plant protein hydrogel as a potential natural wound dressing. By modulating protein concentration (4% HPI) and temperature (85 °C), a thermal-reversible HPI gel appeared as a filament structure with the major molecular driving force of hydrophobic interactions and hydrogen bonds. By characterizing the rheological properties, lower gel strength and wider linear viscoelastic regime were determined in the thermal-reversible HPI gel compared with a thermal-irreversible HPI gel. Besides, large amplitude oscillatory shear data identified the thermal-reversible gel as a soft gel which demonstrated intracycle strain stiffening and shear thinning behavior. Moreover, the thermal-reversible HPI gel is nontoxic and has benefits in neutrophil growth with injectability and perfect wound coverage. This study opens a novel means to form a natural thermal-reversible hydrogel that can be a new material source for wound dressing.

Electroactive differential growth and delayed instability in accelerated healing tissues

Guided by experiments contrasting electrically accelerated recovery with natural healing, this study formulates a model to investigate the importance of electroactive differential growth and morphological changes in tissue repair. It underscores the clinical potential of leveraging electroactive differential growth for improved healing outcomes. The study reveals that voltage stimulation significantly enhances the healing and growth of biological tissues, accelerating the regeneration process across various growth modalities and steering towards isotropic growth conditions that do not favor any specific growth pathways. Enhancing the electroelastic coupling parameters improves the efficacy of bioelectric devices, initiating contraction and fortification of biological tissues in alignment with the electric field. This process facilitates swift cell migration and proliferation, as well as oriented growth of tissue. In instances of strain stiffening at elevated strains, the extreme critical growth ratio aligns with the predictions of neo-Hookean models. Conversely, for tissues experiencing strain stiffening under moderate to very low strain conditions, the strain stiffening effect substantially delays the onset of electroelastic growth instability, ultimately producing a smooth, hyperelastic surface devoid of any unstable morphologies. Our investigation, grounded in nonlinear electroelastic field and perturbation theories, explores how electric fields influence differential growth and instability in biological tissues. We examine the interactions among dimensionless voltage, internal pressure, electroelastic coupling, radius ratio, and strain stiffening, revealing their effects on promoting growth and delaying instability. This framework offers insights into the mechanisms behind electroactive growth and its instabilities, contributing valuable knowledge to the tissue healing.

Biomechanical and biochemical changes in murine skin during development and aging

Aging leads to biochemical and biomechanical changes in skin, with biological and functional consequences. Despite extensive literature on skin aging, there is a lack of studies which investigate the maturation of the tissue and connect the microscopic changes in the skin to its macroscopic biomechanical behavior as it evolves over time. The present work addresses this knowledge gap using multiscale characterization of skin in a murine model considering newborn, adult and aged mice. Monotonic uniaxial loading, tension relaxation with change of bath, and loading to failure tests were performed on murine skin samples from different age groups, complemented by inflation experiments and atomic force microscopy indentation measurements. In parallel, skin samples were characterized using histological and biochemical techniques to assess tissue morphology, collagen organization, as well as collagen content and cross-linking. We show that 1-week-old skin differs across nearly all measured parameters from adult skin, showing reduced strain stiffening and tensile strength, a thinner dermis, lower collagen content and altered crosslinking patterns. Surprisingly, adult and aged skin were similar across most biomechanical parameters in the physiologic loading range, while aged skin had lower tensile strength and lower stiffening behavior at large force values. This correlates with altered collagen content and cross-links. Based on a computational model, differences in mechanocoupled stimuli in the skin of the different age groups were calculated, pointing to a potential biological significance of the age-induced biomechanical changes in regulating the local biophysical environment of dermal cells. Statement of significanceSkin microstructure and the emerging mechanical properties change with age, leading to biological, functional and health-related consequences. Despite extensive literature on skin aging, only very limited quantitative data are available on microstructural changes and the corresponding macroscopic biomechanical behavior as they evolve over time. This work provides a wide-range multiscale mechanical characterization of skin of newborn, adult and aged mice, and quantifies microstructural correlations in tissue morphology, collagen content, organization and cross-linking. Remarkably, aged skin retained normal hydration and normal biomechanical function in the physiological loading range but showed significantly reduced properties at super-physiological loading. Our data show that age-related microstructural differences have a profound effect not only on tissue-level properties but also on the cell-level biophysical environment.

Hard- and Soft-Coded Strain Stiffening in Metamaterials via Out-of-Plane Buckling Using Highly Entangled Active Hydrogel Elements.

Metamaterials show elaborate mechanical behavior such as strain stiffening, which stems from their unit cell design. However, the stiffening response is typically programmed in the design step and cannot be adapted postmanufacturing. Here, we show hydrogel metamaterials with highly programmable strain-stiffening responses by exploiting the out-of-plane buckling of integrated pH-switchable hydrogel actuators. The stiffening upon reaching a certain extension stems from the initially buckled active hydrogel beams. At low strain, the beams do not contribute to the mechanical response under tension until they straighten with a resulting step-function increase in stiffness. In the hydrogel actuator design, the acrylic acid concentration hard-codes the configuration of the metamaterial and range of possible stiffening onsets, while the pH soft-codes the exact stiffening onset point after fabrication. The utilization of out-of-plane buckling to realize subsequent stiffening without the need to deform the passive structure extends the application of hydrogel actuators in mechanical metamaterials. Our concept of out-of-plane buckled active elements that stiffen only under tension enables strain-stiffening metamaterials with high programmability before and after fabrication.

Asphalt binder characterization using waveform-based viscoelastic measures and time-temperature superposition principle under large strains

In the United States, currently peak-to-peak oscillatory shear stress-strain data is used in the asphalt binder characterization protocols. This practice necessitates the measurement, usage, and reporting of only peak responses for material characterization and specification even though a sinusoidal waveform is applied. However, in this paper it is argued that with the proliferation of modified binding systems, it is critical to capture and examine the complete waveform data when subjected to both small and large strains. This approach will help properly differentiating between materials. In this study, four modified binders were tested at small and large strain levels while recording the complete response waveform. The recorded waveform data was then used to examine the linear and nonlinear viscoelastic characteristics of these binders. The orthogonal stress decomposition technique was used to delineate elastic and viscous contributions in the nonlinear regime. This study examined the time-temperature dependency under both linear and nonlinear regimes. The time-temperature shift factors obtained from linear regime were effectively applied in the nonlinear regime. Further, all the tested binders showed strain stiffening and shear thinning behavior under most of the testing conditions. This study provides useful insights about the material behavior under nonlinear regime and can serve as a benchmark for further investigation of nonlinear viscoelastic behavior of asphalt binders.

A hyperelastic strain energy function for isotropic rubberlike materials

A three parameter novel hyperelastic strain energy function is introduced in this paper for soft and rubber-like materials. The function integrates a non-separable exponential component with a single term Ogden-type polynomial-like function, resulting in an exponential-polynomial based strain energy function. This helps in capturing both small and large deformation (stretch) behaviours of hyperelastic materials. The structure of the model is simple and validated against several experimental datasets including rubbers, hydrogel, and soft tissues. The model is reported to capture key material behaviors, including strain stiffening and various deformation paths. Through comparative studies with well-known models like the Ogden (six parameters) and Yeoh (three parameters), the model’s effectiveness is established. Furthermore, the model successfully addresses pressure-inflation instability in thin spherical balloons. It’s applicability extends to biological materials, as evidenced by its effectiveness in characterizing porcine brain tissue and a monkey’s bladder.

Biopolymer networks packed with microgels combine strain stiffening and shape programmability

Biomaterials that can be reversibly stiffened and shaped could be useful in broad biomedical applications where form-fitting scaffolds are needed. Here we investigate the combination of strong non-linear elasticity in biopolymer networks with the reconfigurability of packed hydrogel particles within a composite biomaterial. By packing microgels into collagen-1 networks and characterizing their linear and non-linear material properties, we empirically determine a scaling relationship that describes the synergistic dependence of the material's linear elastic shear modulus on the concentration of both components. We perform high-strain rheological tests and find that the materials strain stiffen and also exhibit a form of programmability, where no applied stress is required to maintain stiffened states of deformation after large strains are applied. We demonstrate that this non-linear rheological behavior can be used to shape samples that do not spontaneously relax large-scale bends, holding their deformed shapes for days. Detailed analysis of the frequency-dependent rheology reveals an unexpected connection to the rheology of living cells, where models of soft glasses capture their low-frequency behaviors and polymer elasticity models capture their high-frequency behaviors.

Kismet/CHD7/CHD8 affects gut microbiota, mechanics, and the gut-brain axis in Drosophila melanogaster

The gut microbiome affects brain and neuronal development and may contribute to the pathophysiology of neurodevelopmental disorders. However, it is unclear how risk genes associated with such disorders affect gut physiology in a manner that could impact microbial colonization and how the mechanical properties of the gut tissue might play a role in gut-brain bidirectional communication. To address this, we used Drosophila melanogaster with a null mutation in the gene kismet, an ortholog of chromodomain helicase DNA-binding protein (CHD) family members CHD7 and CHD8. In humans, these are risk genes for neurodevelopmental disorders with co-occurring gastrointestinal symptoms. We found that kismet mutant flies have a significant increase in gastrointestinal transit time, indicating the functional homology of kismet with CHD7/CHD8 in vertebrates. Rheological characterization of dissected gut tissue revealed significant changes in the mechanics of kismet mutant gut elasticity, strain stiffening behavior, and tensile strength. Using 16S rRNA metagenomic sequencing, we also found that kismet mutants have reduced diversity and abundance of gut microbiota at every taxonomic level. To investigate the connection between the gut microbiome and behavior, we depleted gut microbiota in kismet mutant and control flies and quantified the flies’ courtship behavior. Depletion of gut microbiota rescued courtship defects of kismet mutant flies, indicating a connection between gut microbiota and behavior. In striking contrast, depletion of the gut microbiome in the control strain reduced courtship activity, demonstrating that antibiotic treatment can have differential impacts on behavior and may depend on the status of microbial dysbiosis in the gut prior to depletion. We propose that Kismet influences multiple gastrointestinal phenotypes that contribute to the gut-microbiome-brain axis to influence behavior. We also suggest that gut tissue mechanics should be considered as an element in the gut-brain communication loop, both influenced by and potentially influencing the gut microbiome and neurodevelopment.

Elasticity of self-organized frustrated disordered spring networks.

There have been some interesting recent advances in understanding the notion of mechanical disorder in structural glasses and the statistical mechanics of these systems' low-energy excitations. Here we contribute to these advances by studying a minimal model for structural glasses' elasticity in which the degree of mechanical disorder-as characterized by recently introduced dimensionless quantifiers-is readily tunable over a very large range. We comprehensively investigate a number of scaling laws observed for various macro, meso and microscopic elastic properties, and rationalize them using scaling arguments. Interestingly, we demonstrate that the model features the universal quartic glassy vibrational density of states as seen in many atomistic and molecular models of structural glasses formed by cooling a melt. The emergence of this universal glassy spectrum highlights the role of self-organization (toward mechanical equilibrium) in its formation, and elucidates why models featuring structural frustration alone do not feature the same universal glassy spectrum. Finally, we discuss relations to existing work in the context of strain stiffening of elastic networks and of low-energy excitations in structural glasses, in addition to future research directions.

Effect of Cross-Link Homogeneity on the High-Strain Behavior of Elastic Polymer Networks.

Cross-link heterogeneity and topological defects have been shown to affect the moduli of polymer networks in the low-strain regime. Probing their role in the high-strain regime, however, has been difficult because of premature network fracture. Here, we address this problem by using a double-network approach to investigate the high-strain behavior of both randomly and regularly cross-linked networks with the same backbone chemistry. Randomly cross-linked poly(n-butyl acrylate) networks with target molecular weights between cross-links of 5-30 kg/mol were synthesized via free-radical polymerization, while regularly cross-linked poly(n-butyl acrylate) networks with molecular weights between cross-links of 7-38 kg/mol were synthesized via cross-linking of tetrafunctional star polymers. Both types of networks were then swollen in a monomer/cross-linker mixture, polymerized to form double networks, and characterized via uniaxial tensile testing. The onset of strain stiffening was found to occur later in regular networks than in random networks with the same modulus but was well-predicted by the target molecular weight between cross-links of each sample. These results indicate that the low- and high-strain behavior of polymer networks result from different molecular-scale features of the material and suggest that controlling network architecture offers new opportunities to both further fundamental understanding of architecture-property relationships and design materials with independently controlled moduli and strain stiffening responses.

Postmortem Digital Image Correlation and Finite Element Modeling Demonstrate Posterior Scleral Deformations during Optic Nerve Adduction Tethering.

Postmortem human eyes were subjected to optic nerve (ON) traction in adduction and elevated intraocular pressure (IOP) to investigate scleral surface deformations. We incrementally adducted 11 eyes (age 74.1 ± 9.3 years, standard deviation) from 26° to 32° under normal IOP, during imaging of the posterior globe, for analysis by three-dimensional digital image correlation (3D-DIC). In the same eyes, we performed uniaxial tensile testing in multiple regions of the sclera, ON, and ON sheath. Based on individual measurements, we analyzed eye-specific finite element models (FEMs) simulating adduction and IOP loading. Analysis of 3D-DIC showed that the nasal sclera up to 1 mm from the sheath border was significantly compressed during adduction. IOP elevation from 15 to 30 mmHg induced strains less than did adduction. Tensile testing demonstrated ON sheath stiffening above 3.4% strain, which was incorporated in FEMs of adduction tethering that was quantitatively consistent with changes in scleral deformation from 3D-DIC. Simulated IOP elevation to 30 mmHg did not induce scleral surface strains outside the ON sheath. ON tethering in incremental adduction from 26° to 32° compressed the nasal and stretched the temporal sclera adjacent to the ON sheath, more so than IOP elevation. The effect of ON tethering is influenced by strain stiffening of the ON sheath.

Synthesis of Bottlebrush Polymers with Spontaneous Self-Assembly for Dielectric Generators.

Cross-linked bottlebrush polymers received significant attention as dielectrics in transducers due to their unique softness and strain stiffening caused by their structure. Despite some progress, there is still a great challenge in increasing their dielectric permittivity beyond 3.5 and cross-linking them to defect-free ultrathin films efficiently under ambient conditions. Here, we report the synthesis of bottlebrush copolymers based on ring-opening metathesis polymerization (ROMP) starting from a 5-norbornene-2-carbonitrile and a norbornene modified with a poly(dimethylsiloxane) (PDMS) chain as a macromonomer. The resulting copolymer was subjected to a postpolymerization modification, whereby the double bonds were used both for functionalization with thiopropionitrile and subsequent cross-linking via a thiol-ene reaction. The solutions of both bottlebrush copolymers formed free-standing elastic films by simple casting. DMA and broadband impedance spectroscopy revealed two glass transition temperatures uncommon for a random copolymer. The self-segregation of the nonpolar PDMS chains and the polynorbornane backbone is responsible for this and is supported by the interfacial polarization observed in broadband impedance spectroscopy and the scattering peaks observed in small-angle X-ray scattering (SAXS). Additionally, the modified bottlebrush copolymer was cross-linked to an elastomer that exhibits increased dielectric permittivity and good mechanical properties with significant strain stiffening, an attractive property of dielectric elastomer generators. It has a relative permittivity of 5.24, strain at break of 290%, elastic modulus at 10% strain of 380 kPa, a breakdown field of 62 V μm-1, and a small actuation of 5% at high electric fields of 48.5 V μm-1. All of these characteristics are attractive for dielectric elastomer generator applications. The current work is a milestone in designing functional elastomers based on bottlebrush polymers for transducer applications.

Micelle-Cross-Linked Hydrogels with Strain Stiffening Properties Regulated by Intramicellar Cross-Linking

Micelle-Cross-Linked Hydrogels with Strain Stiffening Properties Regulated by Intramicellar Cross-Linking

Effect of non-linear strain stiffening in eDAH and unjamming.

In cell clusters, the prominent factors at play encompass contractility-based enhanced tissue surface tension and cell unjamming transition. The former effect pertains to the boundary effect, while the latter constitutes a bulk effect. Both effects share outcomes of inducing significant elongation in cells. This elongation is so substantial that it surpasses the limits of linear elasticity, thereby giving rise to additional effects. To investigate these effects, we employ atomic force microscopy (AFM) to analyze how the mechanical properties of individual cells change under such considerable elongation. Our selection of cell lines includes MCF-10A, chosen for its pronounced demonstration of the extended differential adhesion hypothesis (eDAH), and MDA-MB-436, selected due to its manifestation of cell unjamming behavior. In the AFM analyses, we observe a common trend in both cases: as elongation increases, both cell lines exhibit strain stiffening. Notably, this effect is more prominent in MCF-10A compared to MDA-MB-436. Subsequently, we employ AFM on a dynamic range of 1-200 Hz to probe the mechanical characteristics of cell spheroids, focusing on both surface and bulk mechanics. Our findings align with the results from single cell investigations. Specifically, MCF-10A cells, characterized by strong contractile tissue tension, exhibit the greatest stiffness on their surface. Conversely, MDA-MB-436 cells, which experience significant elongation, showcase their highest stiffness within the bulk region. Consequently, the concept of single cell strain stiffening emerges as a crucial element in understanding the mechanics of multicellular spheroids (MCSs), even in the case of MDA-MB-436 cells, which are comparatively softer in nature.

Controlling ligand density and viscoelasticity in synthetic biomimetic polyisocyanide hydrogels for studying cell behaviours: the key to truly biomimetic hydrogels

Integrin-binding peptide addition and density effects bundling and mechanical properties of polyisocyanide-based hydrogels influencing strain stiffening responsiveness, viscoelasticity, stiffness, matrix architecture and cellular behaviours in 3D.

Parallel governing criteria for non-Newtonian droplet rebound suppression on superhydrophobic surfaces

Non-Newtonian droplets are known to suppress rebound on superhydrophobic (SH) surfaces. Our previous work (Dhar et al., Phys Rev Fluids, 2019, vol 4 [1]) showed that the polymer concentration and impact velocity must exceed a certain threshold to initiate the onset of rebound suppression. Analogous to drag reduction or elastic instabilities, we proposed that rebound suppression occurs only when the Weissenberg number exceeds unity (i.e., Wi >1) at the onset of retraction. In this study, we explore four different types of SH surfaces to examine the universality of the previous observation (the importance of Wi >1 for rebound suppression). We observed that rebound suppression does not occur on all types of SH surfaces. The surfaces where rebound suppression was observed are categorized as ‘Type-I’, while the remaining as ‘Type-II’. The non-uniformity in the rebound suppression phenomena is explained by the role of Cassie to Wenzel transformation (CWT) or impalement. We showed that for ‘Type-I’ surfaces, when the ratio of dynamic pressure (PD) to minimum capillary pressure (PC) exceeds unity (i.e., PD/PC > 1), the rebound is suppressed. The ‘Type-II’ surfaces have characteristic spacing in the order of nanometers, which is three orders of magnitude lower than the microtextured ‘Type-I’ surfaces. Thereby, the minimum capillary pressure is significantly higher for ‘Type-II’ surfaces. The dynamic pressure could not surpass the minimum capillary pressure for the impact velocities and polymer concentrations studied for the ‘Type-II’ surfaces. As PD/PC remained < 1, for ‘Type-II’ surfaces, rebound suppression was not observed. It must be noted that for the same values of PD/PC > 1 values, water droplets didn’t show rebound suppression on ‘Type-I’ surfaces. The non-Newtonian droplets are known to slow down the retraction dynamics due to normal stress, extensional viscosity and higher adsorption at the substrate. We have demonstrated the role of extensional viscosity in this study, particularly how strain stiffening of radial filaments emanating from the droplet aids in dissipating energy, which further aids in arresting drop rebound. Therefore, the existence of two parallel mechanisms: the role of fluid elasticity (manifested through Wi > 1), and the role of CWT or impalement (manifested through PD/PC> 1), are necessary conditions for non-Newtonian drop rebound suppression on superhydrophobic surfaces.

Analysis of the Mullins effect in buckling instability of double-network hydrogel beams under swelling equilibrium

Due to their outstanding physical and mechanical properties, double-network (DN) hydrogels have gained a most attention among various synthetic tough hydrogels. The Mullins effect and buckling instability are two frequent phenomena that may occur simultaneously in slender DN hydrogel structures as high load-bearing candidates. The swelling/deswelling degree of the DN hydrogel affects the coupling phenomena. It is essential to understanding the interplay between these behaviors. In this work, a physically based damage constitutive model for a DN polymer under water-diffusion equilibrium and cyclic loadings is developed. This theory of coupled swelling and load-induced deformations show good capability in capturing the Mullins effect of swollen DN hydrogels and the corresponding swelling ratio. The model parameters are determined by fitting experimental data of a freely swollen DN hydrogel under cyclic compression. Based on the constitutive model and determined parameters, the buckling instability of DN hydrogel beams is studied via the analytical formula of incremental modulus. The stability diagrams of the DN hydrogel beams under virgin loading and reloading are presented. The influences of stress softening, strain stiffening and chemical potential on buckling conditions for compressive stress and slenderness ratio are thoroughly analyzed. It demonstrates that the stress softening is dramatically against the stability of the beam, but the strain-stiffening effect would conversely help widen the stable range. Besides, it is found that a DN gel beam immersed in a sufficient low chemical potential environment has better buckling stability. These theoretical results are valuable in the preparation and structural design of DN hydrogels for repeated use purpose.

Strain-stiffening of chemical bonding enhance strength and fracture toughness of the interface of Fe2B/Fe in situ composite

Fe2B was widely used as the strengthening phase and wear-resistant skeleton in high boron iron-based composite. The weak interfacial bonding of Fe2B/Fe interface is one of the main reasons for the poor mechanical properties of the composite, which motivated us to develop a strain stiffening strategy of interfacial chemical bonds to improve the bond strength and elongation of the interface. The effects of alloying elements Ti, Cr, Mn, Co, Ni, Cu, Y, Nb, Mo, Ta, and W on Fe(001)/Fe2B(001) interface were investigated through first-principles calculations. The interface segregation energy and the work of adhesion showed that Ti, Cr, Mn, Ni, Y, Mo, Ta, and W can segregate at this interface, and the alloying elements Cr, Mn, Ni, Nb, Mo, Ta and W increase the interface bonding strength. The fracture mechanism of the interface resulted from the breaking of the FeFe bond of the bcc-Fe end near the interface. The alloying elements Cr, Mn, and Ni improved the ultimate strength and elongation of bcc-Fe(001)/Fe2B(001) interface. CrB bond displays the obvious strain stiffening in the interface. Therefore, it provides new thought and method for the design of new high boron alloys.

Investigating the phase behaviour of binary suspensions of cellulose nanocrystals and montmorillonite with nonlinear rheology, SAXS and polarized optical microscopy

Using microscopy imaging techniques, X-ray scattering, and nonlinear rheological measurements, we have examined microstructural changes, and phase behavior of cellulose nanocrystals (CNC)/montmorillonite (MMT) composite colloidal systems. Fourier-transform (FT) rheology-stress decomposition methods, Lissajous-Bowditch plots, and other techniques are used to both qualitatively and quantitatively determine the various phases that the suspensions exhibit. In the MAOS (medium amplitude oscillatory shear) region, multiple scaling regions, as well as quadratic scaling regions are obtained. The dependence of the normalized third relative intensity (I3/1) values on angular frequency (ω) also provided new insights into how nonlinear parameters depend on the various CNC/MMT phases. These findings allow us to differentiate various phases exhibited by the composite system, including the isotropic, biphasic, and chiral nematic liquid crystalline (LC) phases. The CNC/MMT nanocomposite system exhibits a variety of liquid crystalline phases, which may contribute to improved mechanical properties for the system, including increased mixture strength, stiffness, and toughness because of the particle alignment in the LC phase. Additionally, the field of optics and photonics may find use for this nanocomposite system in the fabrication of sensors, polarizers, and optical films; energy storage applications where the aligned structures facilitate ion transport; and the formulation of paints and coatings where the strain stiffening and shear thinning behavior of the CNC/MMT composite system are critical.