Substrate stress relaxation regulates neural stem cell fate commitment.

Adult neural stem cells (NSCs) reside in the dentate gyrus of the hippocampus, and their capacity to generate neurons and glia plays a role in learning and memory. In addition, neurodegenerative diseases are known to be caused by a loss of neurons and glial cells, resulting in a need to better understand stem cell fate commitment processes. We previously showed that NSC fate commitment toward a neuronal or glial lineage is strongly influenced by extracellular matrix stiffness, a property of elastic materials. However, tissues in vivo are not purely elastic and have varying degrees of viscous character. Relatively little is known about how the viscoelastic properties of the substrate impact NSC fate commitment. Here, we introduce a polyacrylamide-based cell culture platform that incorporates mismatched DNA oligonucleotide-based cross-links as well as covalent cross-links. This platform allows for tunable viscous stress relaxation properties via variation in the number of mismatched base pairs. We find that NSCs exhibit increased astrocytic differentiation as the degree of stress relaxation is increased. Furthermore, culturing NSCs on increasingly stress-relaxing substrates impacts cytoskeletal dynamics by decreasing intracellular actin flow rates and stimulating cyclic activation of the mechanosensitive protein RhoA. Additionally, inhibition of motor-clutch model components such as myosin II and focal adhesion kinase partially or completely reverts cells to lineage distributions observed on elastic substrates. Collectively, our results introduce a unique system for controlling matrix stress relaxation properties and offer insight into how NSCs integrate viscoelastic cues to direct fate commitment.

Microscopic dynamics underlying the stress relaxation of arrested soft materials.

Arrested soft materials such as gels and glasses exhibit a slow stress relaxation with a broad distribution of relaxation times in response to linear mechanical perturbations. Although this macroscopic stress relaxation is an essential feature in the application of arrested systems as structural materials, consumer products, foods, and biological materials, the microscopic origins of this relaxation remain poorly understood. Here, we elucidate the microscopic dynamics underlying the stress relaxation of such arrested soft materials under both quiescent and mechanically perturbed conditions through X-ray photon correlation spectroscopy. By studying the dynamics of a model associative gel system that undergoes dynamical arrest in the absence of aging effects, we show that the mean stress relaxation time measured from linear rheometry is directly correlated to the quiescent superdiffusive dynamics of the microscopic clusters, which are governed by a buildup of internal stresses during arrest. We also show that perturbing the system via small mechanical deformations can result in large intermittent fluctuations in the form of avalanches, which give rise to a broad non-Gaussian spectrum of relaxation modes at short times that is observed in stress relaxation measurements. These findings suggest that the linear viscoelastic stress relaxation in arrested soft materials may be governed by nonlinear phenomena involving an interplay of internal stress relaxations and perturbation-induced intermittent avalanches.

Compressible and Elastic Reduced Graphene Oxide Sponge for Stable and Dendrite-Free Lithium Metal Anodes.

Dendritic Li deposition, an unstable solid-electrolyte interphase (SEI), and a nearly infinite relative volume change during cycling are three major obstacles to the practical application of Li metal batteries. Herein, we introduce a compressible and elastic reduced graphene oxide sponge (rGO-S) to simultaneously eliminate Li dendrite growth, stabilize the SEI, and accommodate the volume change. The volume change is contained by compressing and expanding the rGO-S anode, which effectively releases the Li plating-induced stress during cycling. The smooth and dense Li metal is deposited on rGO-S without dendrites, which preserves the SEI, reduces consumption of the electrolyte, and prevents the formation of Li debris. The half-cells employing rGO-S show a steady and high Coulombic efficiency. The Li@rGO-S symmetric cells demonstrate excellent cycling stability over 1200 cycles with a low overpotential. When paired with LiFePO4 (LFP), the Li@rGO-S||LFP full cells exhibit a high specific capacity (150.3 mAh g-1 at 1C), superior rate performance, and good capacity retention.

Integrins play a role in stress relaxation of perivascular adipose tissue.

Perivascular adipose tissue (PVAT) is known for being anti-contractile in healthy tissues. We discovered a new function of PVAT, the ability to stress relax and maintain a tone in response to a stretch. This is of note because stress relaxation has been attributed to smooth muscle, of which PVAT has none that is organized in a functional layer. We test the hypothesis the interactions of integrins with collagen play a role in stress relaxation. Our model is the thoracic aorta of the male Dahl SS rat. The PVAT and aorta were physically separated for most assays. Results from single nuclei RNA sequencing (snRNAseq) experiments, histochemistry and isometric contractility were also used. Masson Trichrome staining made evident the expression of collagen in PVAT. From snRNA seq experiments of the PVAT, mRNA for multiple collagen and integrin isoforms were detected: the α1 and ß1 integrin were most highly expressed. Pharmacological inhibition of integrin/collagen interaction was effected by the specific α1ß1 distintegrin obtustatin or general integrin inhibitor RGD peptide. RGD peptide but not obtustatin increased the stress relaxation. Cell-cell communication inference identified integrins αv and α5, two major RGD motif containing isoforms, as potential signaling partners of collagens. Collectively, these findings validate that stress relaxation can occur in a non-smooth muscle tissue, doing so in part through integrin-collagen interactions that may not include α1ß1 heterodimers. The importance of this lies in considering PVAT as a vascular layer that possesses mechanical functions.

Enhanced microscopic dynamics in mucus gels under a mechanical load in the linear viscoelastic regime.

Mucus is a biological gel covering the surface of several tissues and ensuring key biological functions, including as a protective barrier against dehydration, pathogen penetration, or gastric acids. Mucus biological functioning requires a finely tuned balance between solid-like and fluid-like mechanical response, ensured by reversible bonds between mucins, the glycoproteins that form the gel. In living organisms, mucus is subject to various kinds of mechanical stresses, e.g., due to osmosis, bacterial penetration, coughing, and gastric peristalsis. However, our knowledge of the effects of stress on mucus is still rudimentary and mostly limited to macroscopic rheological measurements, with no insight into the relevant microscopic mechanisms. Here, we run mechanical tests simultaneously to measurements of the microscopic dynamics of pig gastric mucus. Strikingly, we find that a modest shear stress, within the macroscopic rheological linear regime, dramatically enhances mucus reorganization at the microscopic level, as signaled by a transient acceleration of the microscopic dynamics, by up to 2 orders of magnitude. We rationalize these findings by proposing a simple, yet general, model for the dynamics of physical gels under strain and validate its assumptions through numerical simulations of spring networks. These results shed light on the rearrangement dynamics of mucus at the microscopic scale, with potential implications in phenomena ranging from mucus clearance to bacterial and drug penetration.

Characterization of the strain-rate-dependent mechanical response of single cell-cell junctions.

Cell-cell adhesions are often subjected to mechanical strains of different rates and magnitudes in normal tissue function. However, the rate-dependent mechanical behavior of individual cell-cell adhesions has not been fully characterized due to the lack of proper experimental techniques and therefore remains elusive. This is particularly true under large strain conditions, which may potentially lead to cell-cell adhesion dissociation and ultimately tissue fracture. In this study, we designed and fabricated a single-cell adhesion micro tensile tester (SCAµTT) using two-photon polymerization and performed displacement-controlled tensile tests of individual pairs of adherent epithelial cells with a mature cell-cell adhesion. Straining the cytoskeleton-cell adhesion complex system reveals a passive shear-thinning viscoelastic behavior and a rate-dependent active stress-relaxation mechanism mediated by cytoskeleton growth. Under low strain rates, stress relaxation mediated by the cytoskeleton can effectively relax junctional stress buildup and prevent adhesion bond rupture. Cadherin bond dissociation also exhibits rate-dependent strengthening, in which increased strain rate results in elevated stress levels at which cadherin bonds fail. This bond dissociation becomes a synchronized catastrophic event that leads to junction fracture at high strain rates. Even at high strain rates, a single cell-cell junction displays a remarkable tensile strength to sustain a strain as much as 200% before complete junction rupture. Collectively, the platform and the biophysical understandings in this study are expected to build a foundation for the mechanistic investigation of the adaptive viscoelasticity of the cell-cell junction.

Dynamic Exchange in 3D Cell Culture Hydrogels Based on Crosslinking of Cyclic Thiosulfinates.

Dynamic polymer materials are highly valued substrates for 3D cell culture due to their viscoelasticity, a time-dependent mechanical property that can be tuned to resemble the energy dissipation of native tissues. Herein, we report the coupling of a cyclic thiosulfinate, mono-S-oxo-4-methyl asparagusic acid, to a 4-arm PEG-OH to prepare a disulfide-based dynamic covalent hydrogel with the addition of 4-arm PEG-thiol. Ring opening of the cyclic thiosulfinate by nucleophilic substitution results in the rapid formation of a network showing a viscoelastic fluid-like behaviour and relaxation rates modulated by thiol content through thiol-disulfide exchange, whereas its viscoelastic behaviour upon application as a small molecule linear crosslinker is solid-like. Further introduction of 4-arm PEG-vinylsulfone in the network yields a hydrogel with weeks-long cell culture stability, permitting 3D culture of cell types that lack robust proliferation, such as human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs). These cells display native behaviours such as cell elongation and spontaneous beating as a function of the hydrogel's mechanical properties. We demonstrate that the mode of dynamic cyclic thiosulfinate crosslinker presentation within the network can result in different stress relaxation profiles, opening the door to model tissues with disparate mechanics in 3D cell culture.

Mechanism of elastic energy storage of honey bee abdominal muscles under stress relaxation.

Energy storage of passive muscles plays an important part in frequent activities of honey bee abdomens due to the muscle distribution and open circulatory system. However, the elastic energy and mechanical properties of structure in passive muscles remain unclear. In this article, stress relaxation tests on passive muscles from the terga of the honey bee abdomens were performed under different concentrations of blebbistatin and motion parameters. In stress relaxation, the load drop with the rapid and slow stages depending on stretching velocity and stretching length reflects the features of myosin-titin series structure and cross-bridge-actin cyclic connections in muscles. Then a model with 2 parallel modules based on the 2 feature structures in muscles was thus developed. The model described the stress relaxation and stretching of passive muscles from honey bee abdomen well for a good fitting in stress relaxation and verification in loading process. In addition, the stiffness change of cross-bridge under different concentrations of blebbistatin is obtained from the model. We derived the elastic deformation of cross-bridge and the partial derivatives of energy expressions on motion parameters from this model, which accorded the experimental results. This model reveals the mechanism of passive muscles from honey bee abdomens suggesting that the temporary energy storage of cross-bridge in terga muscles under abdomen bending provides potential energy for springback during the periodic abdomen bending of honey bee or other arthropod insects. The finding also provides an experimental and theoretical basis for the novel microstructure and material design of bionic muscle.

Compressive mechanical properties of vitrified porcine menisci are superior to frozen and similar to fresh porcine menisci.

The common practice of freezing meniscal allograft tissue is limited due to the formation of damaging ice crystals. Vitrification, which eliminates the formation of damaging ice crystals, may allow the mechanical properties of meniscal allograft tissue to be maintained during storage and long-term preservation. The primary objective of this study was to investigate the differences between fresh, frozen, and vitrified porcine lateral menisci examining compressive mechanical properties in the axial direction. Unconfined compressive stress-relaxation testing was conducted to quantify the mechanical properties of fresh, frozen and vitrified porcine lateral menisci. The compressive mechanical properties investigated were peak and equilibrium stress, secant, instantaneous and equilibrium modulus, percent stress-relaxation, and relaxation time constants from three-term Prony series. Frozen menisci exhibited inferior compressive mechanical properties in comparison with fresh menisci (significant differences in peak and equilibrium stress, and secant, instantaneous and equilibrium modulus) and vitrified menisci (significant differences in peak stress, and secant and instantaneous modulus). Interestingly, fresh and vitrified menisci exhibited comparable compressive mechanical properties (stress, modulus and relaxation parameters). These findings are significant because (1) vitrification was successful in maintaining mechanical properties at values similar to fresh menisci, (2) compressive mechanical properties of fresh menisci were characterized providing a baseline for future research, and (3) freezing affected mechanical properties confirming that freezing should be used with caution in future investigations of meniscal mechanical properties. Vitrification was superior to freezing for preserving compressive mechanical properties of menisci which is an important advance for vitrification as a preservation option for meniscal allograft transplantation.

Nanotwin orientation on history-dependent stress decay in Cu nanopillar under constant strain.

Cu with nanotwin (NT) possesses great electrical, mechanical, and thermal properties and has potential for electronic applications. Various studies have reported the effect of NT orientation on Cu mechanical properties. However, its effect on Cu stress-relaxation behavior has not been clarified, particularly in nano-scale. In this study, Cu nanopillars with various orientations were examined by a picoindenter under constant strain and observed byin situTEM. The angles between the twin plane and the loading direction in the examined nanopillars were 0°, 60°, to 90°, and a benchmark pillar of single-crystal Cu without NT was examined. The stress drops were respectively 10%, 80%, 4%, and 50%. Owing to the interaction by NT, the dislocation behavior in nanopillars was different from that in bulk or in thin film samples. Especially, the rapid slip path of dislocations to go to the free surface of the nanopillar induced a dislocation-free zone in the 0° nanopillar, which led to work-softening. On the contrary, a high dislocation density was observed in the 90° nanopillar, which was generated by dislocation interaction and obstruction of dislocation slip by twin planes, and it led to work-hardening. The findings reveal the NT orientation in Cu nanopillars affected stress relaxation significantly.

Nanomechanical study of aqueous-processed h-BN reinforced PVA composites.

Hexagonal boron nitride (h-BN) as a filler has significantly improved the mechanical properties of various polymers composites. Among them, polyvinyl alcohol (PVA) is particularly important for its wide range of industrial applications and biocompatibility nature. However, preparing a homogenous composite of h-BN and PVA in water is troublesome as the aqueous processing of h-BN without any additives is challenging. In this context, a pre-processing technique is used to produce an additive-free aqueous dispersion of h-BN. The uniformly dispersed composites are then prepared with different concentrations of h-BN. Free-standing thin films are fabricated using the doctor blade technique, and nanoindentation is employed to understand their deformation behaviour at smaller length scale for better understanding of micro-mechanism involved. Reduced elastic modulus and hardness of 10 wt% h-BN/PVA composite film are enhanced by ∼93% and ∼159%, respectively, compared to pristine PVA. Frequency sweep dynamic mechanical analysis is performed between 1 and 50 Hz, and the elastic properties of composite materials are found to improve significantly upon addition of h-BN nanosheets. Besides, the impact of h-BN incorporation in stress relaxation behaviour and hardness depth profiling are also investigated. The observed improvement in mechanical properties of the composites may be attributed to the uniform distribution of the nanosheets and the strong interfacial interaction between h-BN and PVA, which ensures efficient mechanical stress transfer at the interface.

When Dynamic Diselenide Bonds Meet Dynamic Imine Bonds in Polymeric Materials.

In both natural and artificial functional systems, the cooperation between different dynamic interactions is of vital importance for realizing complicated functions. Dynamic covalent bonds are one kind of relatively stable dynamic interactions and have shown synergistic effect in natural systems such as functional proteins. However, synergistic interactions between different dynamic covalent bonds in polymeric materials are still unclear. Herein, polymeric materials containing diselenide and imine bonds are prepared, and then the synergistic effect between the two dynamic covalent bonds is quantitatively evaluated in typical processes of dynamic materials. The results reveal that dynamic covalent bonds show weak synergistic effect in the degradation process and have strong synergistic effect in stress relaxation process. Therefore, introducing multiple dynamic covalent bonds in polymeric materials can extensively enhance their dynamic properties.

An Effective Approach to Improve the Thermal Conductivity, Strength, and Stress Relaxation of Carbon Nanotubes/Epoxy Composites Based on Vitrimer Chemistry.

An effective method was developed to improve the interfacial interaction between Mutiwalled carbon nanotubes (MWCNTs) and epoxy matrix. The performance of thermal conductivity and strength of the epoxy vitrimer were enhanced by polydopamine (PDA) coating. Polydopamine is a commonly used photothermal agent, which of course, was effective in modifying MWCNTs used in photoresponsive epoxy resin. The surface temperature of the epoxy composite with 3% MWCNTs@PDA fillers added increased from room temperature to 215 °C in 48 s. The metal-catechol coordination interactions formed between the catechol groups of PDA and Zn2+ accelerated the stress relaxation of epoxy vitrimer. Moreover, the shape memory, repairing, and recycling of epoxy vitrimer were investigated. Therefore, dopamine coating is a multifunctional approach to enhance the performance of epoxy vitrimer.

Viscoelastic Liquid Matrix with Faster Bulk Relaxation Time Reinforces the Cell Cycle Arrest Induction of the Breast Cancer Cells via Oxidative Stress.

The reactivating of disseminated dormant breast cancer cells in a soft viscoelastic matrix is mostly correlated with metastasis. Metastasis occurs due to rapid stress relaxation owing to matrix remodeling. Here, we demonstrate the possibility of promoting the permanent cell cycle arrest of breast cancer cells on a viscoelastic liquid substrate. By controlling the molecular weight of the hydrophobic molten polymer, poly(ε-caprolactone-co-D,L-lactide) within 35-63 g/mol, this study highlights that MCF7 cells can sense a 1000 times narrower relaxation time range (80-290 ms) compared to other studies by using a crosslinked hydrogel system. We propose that the rapid bulk relaxation response of the substrate promotes more reactive oxygen species generation in the formed semi-3D multicellular aggregates of breast cancer cells. Our finding sheds light on the potential role of bulk stress relaxation in a viscous-dominant viscoelastic matrix in controlling the cell cycle arrest depth of breast cancer cells.

Gradual Stress-Relaxation of Hydrogel Regulates Cell Spreading.

There is growing evidence that the mechanical properties of extracellular matrices (ECMs), including elasticity and stress-relaxation, greatly influence the function and form of the residing cells. However, the effects of elasticity and stress-relaxation are often correlated, making the study of the effect of stress-relaxation on cellular behaviors difficult. Here, we designed a hybrid network hydrogel with a controllable stress-relaxation gradient and a constant elasticity. The hydrogel is crosslinked by covalent bonds and dynamic peptide-metal ion coordination interactions. The stress-relaxation gradient is controlled by spatially controlling the coordination and covalent crosslinker ratios. The different parts of the hydrogel exhibit distinct stress-relaxation amplitudes but the have same stress-relaxation timescale. Based on this hydrogel, we investigate the influence of hydrogel stress-relaxation on cell spreading. Our results show that the spreading of cells is suppressed at an increasing stress-relaxation amplitude with a fixed elasticity and stress-relaxation timescale. Our study provides a universal route to tune the stress-relaxation of hydrogels without changing their components and elasticity, which may be valuable for systematic investigations of the stress-relaxation gradient in cell cultures and organoid constructions.

Fabrication and Evaluation of Gellan Gum/Hyaluronic Acid Hydrogel for Retinal Tissue Engineering Biomaterial and the Influence of Substrate Stress Relaxation on Retinal Pigment Epithelial Cells.

Cell therapies for age-related macular degeneration (AMD) treatment have been developed by integrating hydrogel-based biomaterials. Until now, cell activity has been observed only in terms of the modulus of the hydrogel. In addition, cell behavior has only been observed in the 2D environment of the hydrogel and the 3D matrix. As time-dependent stress relaxation is considered a significant mechanical cue for the control of cellular activities, it is important to optimize hydrogels for retinal tissue engineering (TE) by applying this viewpoint. Herein, a gellan Gum (GG)/Hyaluronic acid (HA) hydrogel was fabricated using a facile physical crosslinking method. The physicochemical and mechanical properties were controlled by forming a different composition of GG and HA. The characterization was performed by conducting a mass swelling study, a sol fraction study, a weight loss test, a viscosity test, an injection force study, a compression test, and a stress relaxation analysis. The biological activity of the cells encapsulated in 3D constructs was evaluated by conducting a morphological study, a proliferation test, a live/dead analysis, histology, immunofluorescence staining, and a gene expression study to determine the most appropriate material for retinal TE biomaterial. Hydrogels with moderate amounts of HA showed improved physicochemical and mechanical properties suitable for injection into the retina. Moreover, the time-dependent stress relaxation property of the GG/HA hydrogel was enhanced when the appropriate amount of HA was loaded. In addition, the cellular compatibility of the GG/HA hydrogel in in vitro experiments was significantly improved in the fast-relaxing hydrogel. Overall, these results demonstrate the remarkable potential of GG/HA hydrogel as an injectable hydrogel for retinal TE and the importance of the stress relaxation property when designing retinal TE hydrogels. Therefore, we believe that GG/HA hydrogel is a prospective candidate for retinal TE biomaterial.

Effect of Heat Treatment on Residual Stress of Cold Sprayed Nickel-based Superalloys.

Residual stress formation during cold spraying process may result in deteriorative effects on the performance of coating materials. The objective of this investigation is to characterize residual stress built-up in two well-known nickel-based superalloys (Inconel 625 and Inconel 718) deposited using cold spraying technique. To this end, the residual stress was precisely measured using x-ray diffraction method. Here, residual stress in the subsurface regions was only studied because the surface properties may alter during sample preparation. The average residual stress was slightly higher in Inconel 625 compared to the Inconel 718 sample. Heat treatment at 800 °C helped in the reduction of porosities which exerted tensile stress in subsurface regions of both coatings. Stresses with opposite signs could cancel each other and result in reduction of residual stress after heat treatment. However, the recovery of residual stress was higher for Inconel 718 coating. In the next step as-sprayed and heat-treated coating samples were subjected to microindentation test to measure their hardness and study the crack formation in the samples. The as-sprayed Inconel 625 exhibited higher hardness than Inconel 718, but the hardness of Inconel 625 decreased more drastically after heat treatment. While the cracks were formed on both as-sprayed samples around indents, no cracks were found in the heat-treated samples. The results from this study will contribute to better understanding the performance of cold spray deposited superalloys under service conditions and the effect of stress relaxation heat treatment on elimination of residual stress.

Time-varying regularity of changes in biomechanical properties of the corneas after removal of anterior corneal tissue.

BACKGROUND: The corneal biomechanical properties with the prolongation of time after corneal refractive surgery are important for providing a mechanical basis for the occurrence of clinical phenomena such as iatrogenic keratectasia and refractive regression. The aim of this study was to explore the changes of corneal elastic modulus, and stress relaxation properties from the 6-month follow-up observations of rabbits after a removal of anterior corneal tissue in simulation to corneal refractive surgery. METHODS: The anterior corneal tissue, 6 mm in diameter and 30-50% of the original corneal thickness, the left eye of the rabbit was removed, and the right eye was kept as the control. The rabbits were normally raised and nursed for 6 months, during which corneal morphology data, and both of corneal hysteresis (CH) and corneal resistance factor (CRF) were gathered. Uniaxial tensile tests of corneal strips were performed at months 1, 3, and 6 from 7 animals, and corneal collagen fibrils were observed at months 1, 3, and 6 from 1 rabbit, respectively. RESULTS: Compared with the control group, there were statistical differences in the curvature radius at week 2 and month 3, and both CH and CRF at months 1, 2, and 6 in experiment group; there were statistical differences in elastic modulus at 1, 3, and month 6, and stress relaxation degree at month 3 in experiment group. The differences in corneal elastic modulus, stress relaxation degree and the total number of collagen fibrils between experiment and control groups varied gradually with time, and showed significant changes at the 3rd month after the treatment. CONCLUSIONS: Corneas after a removal of anterior corneal tissue undergo dynamic changes in corneal morphology and biomechanical properties. The first 3 months after treatment could be a critical period. The variation of corneal biomechanical properties is worth considering in predicting corneal deformation after a removal of anterior corneal tissue.

Grain boundary sliding and non-constancy strain during stress relaxation of pure Mg.

Stress relaxation during plastic deformation has been reported to improve ductility of metallic materials. In this study, the stress relaxation behaviour in pure magnesium is investigated during interrupted uniaxial tensile tests. During intermittent stopping of the machine for relaxation studies, the total strain is expected to remain constant. However, an anomalous non-constancy in total strain is observed in the present work. The total strain increases with relaxation time. Additional in-situ tensile tests indicate that the non-constant total strain is restricted only in the gauge area of the specimen, indicating a likely shear dominated deformation such as grain boundary sliding (GBS) responsible for the anomalous behaviour. The role of GBS during relaxation is studied using the deformation induced evolution of surface inhomogeneity. Determinations of surface profiling step heights at grain boundaries and inclination of grains were used to quantify the effect of GBS. The estimated activation volume of 4.35 b 3 further confirms the role of slip induced GBS on the deformation. A new stress relaxation model accommodating GBS is proposed and is found to fit the experimental data accurately.

An Improved Large-Scale Stress-Controlled Apparatus for Long-Term Seepage Study of Coarse-Grained Cohesive Soils.

Clay-gravel mixture has been widely used in high embankment dams and understanding its seepage characteristics is critical to dam safety. From the instrumental perspective, the realization of continuous pressurized water supply becomes a key technical challenge, significantly restricting the working conditions replicated in previous seepage apparatuses. To this end, a novel water provision system, relying on parallel-disposed sensor-based pressure devices, was introduced, so that the application of an existing large-scale stress-controlled apparatus can be expanded to long-term seepage tests regarding coarse-grained cohesive soils. Constant-head permeability tests were conducted on original-graded clay-gravel mixtures to investigate their hydraulic properties, incorporating the influence of stress relaxation. Test results show that with 35% gravel content, the clay-gravel mixture is suitable for dam construction as the core material. The stress relaxation holds a marginal effect on the hydraulic conductivity of soil. The functionality of this improved apparatus is verified, especially under long-term seepage conditions.