Biobased Nanomaterials─The Role of Interfacial Interactions for Advanced Materials.

This review presents recent advances regarding biomass-based nanomaterials, focusing on their surface interactions. Plant biomass-based nanoparticles, like nanocellulose and lignin from industry side streams, hold great potential for the development of lightweight, functional, biodegradable, or recyclable material solutions for a sustainable circular bioeconomy. However, to obtain optimal properties of the nanoparticles and materials made thereof, it is crucial to control the interactions both during particle production and in applications. Herein we focus on the current understanding of these interactions. Solvent interactions during particle formation and production, as well as interactions with water, polymers, cells and other components in applications, are addressed. We concentrate on cellulose and lignin nanomaterials and their combination. We demonstrate how the surface chemistry of the nanomaterials affects these interactions and how excellent performance is only achieved when the interactions are controlled. We furthermore introduce suitable methods for probing interactions with nanomaterials, describe their advantages and challenges, and introduce some less commonly used methods and discuss their possible applications to gain a deeper understanding of the interfacial chemistry of biobased nanomaterials. Finally, some gaps in current understanding and interesting emerging research lines are identified.

Reduced Tiara-like Palladium Complex for Suzuki Cross-Coupling Reactions.

The design of highly active and structurally well-defined catalysts has become a crucial issue for heterogeneous catalysed reactions while reducing the amount of catalyst employed. Beside conventional synthetic routes, the employment of polynuclear transition metal complexes as catalysts or catalyst precursors has progressively intercepted a growing interest. These well-defined species promise to deliver catalytic systems where a strict control on the nuclearity allows to improve the catalytic performance while reducing the active phase loading. This study describes the development of a highly active and reusable palladium-based catalyst on alumina (Pd8 /Al2 O3 ) for Suzuki cross-coupling reactions. An octanuclear tiara-like palladium complex was selected as active phase precursor to give isolated Pd-clusters of ca. 1 nm in size on Al2 O3 . The catalyst was thoroughly characterised by several complementary techniques to assess its structural and chemical nature. The high specific activity of the catalyst has allowed to carry out the cross-coupling reaction in 30 min using only 0.12 mol % of Pd loading under very mild and green reaction conditions. Screening of various substrates and selectivity tests, combined with recycling and benchmarking experiments, have been used to highlight the great potentialities of this new Pd8 /Al2 O3 catalyst.

Single-Element Dual-Interferometer for Precision Inertial Sensing: Sub-Picometer Structural Stability and Performance as a Reference for Laser Frequency Stabilization.

Future GRACE-like geodesy missions could benefit from adopting accelerometer technology akin to that of the LISA Pathfinder, which employed laser interferometric readout at the sub-picometer level in addition to the conventional capacitive sensing, which is at best at the level of 100 pm. Improving accelerometer performance holds great potential to enhance the scientific output of forthcoming missions, carrying invaluable implications for research in climate, water resource management, and disaster risk reduction. To reach sub-picometer displacement sensing precision in the millihertz range, laser interferometers rely on suppression of laser-frequency noise by several orders of magnitude. Many optical frequency stabilization methods are available with varying levels of complexity, size, and performance. In this paper, we describe the performance of a Mach-Zehnder interferometer based on a compact monolithic optic. The setup consists of a commercial fiber injector, a custom-designed pentaprism used to split and recombine the laser beam, and two photoreceivers placed at the complementary output ports of the interferometer. The structural stability of the prism is transferred to the laser frequency via amplification, integration, and feedback of the balanced-detection signal, achieving a fractional frequency instability better than 6 parts in 1013, corresponding to an interferometer pathlength stability better than 1pm/Hz. The prism was designed to host a second interferometer to interrogate the position of a test mass. This optical scheme has been dubbed "single-element dual-interferometer" or SEDI.

Affinity of Keratin Peptides for Cellulose and Lignin: A Fundamental Study toward Advanced Bio-Based Materials.

Keratin is a potential raw material to meet the growing demand for bio-based materials with special properties. Keratin can be obtained from feathers, a by-product from the poultry industry. One approach for keratin valorization is to use the protein to improve the properties of already existing cellulose and lignin-based materials to meet the requirements for replacing fossil-based plastics. To ensure a successful combination of keratin with lignocellulosic building blocks, keratin must have an affinity to these substrates. Hence, we used quartz crystal microbalance with a dissipation monitoring (QCM-D) technique to get a detailed understanding of the adsorption of keratin peptides onto lignocellulosic substrates and how the morphology of the substrate, pH, ionic strength, and keratin properties affected the adsorption. Keratin was fractionated from feathers with a scalable and environmentally friendly deep eutectic solvent process. The keratin fraction used in the adsorption studies consisted of different sized keratin peptides (about 1-4 kDa), which had adopted a random coil conformation as observed by circular dichroism (CD). Measuring keratin adsorption to different lignocellulosic substrates by QCM-D revealed a significant affinity of keratin peptides for lignin, both as smooth films and in the form of nanoparticles but only a weak interaction between cellulose and keratin. Systematic evaluation of the effect of surface, media, and protein properties enabled us to obtain a deeper understanding of the driving force for adsorption. Both the structure and size of the keratin peptides appeared to play an important role in its adsorption. The keratin-lignin combination is an attractive option for advanced material applications. For improved adsorption on cellulose, modifications of either keratin or cellulose would be required.

On-Axis Optical Bench for Laser Ranging Instruments in Future Gravity Missions.

The laser ranging interferometer onboard the Gravity Recovery and Climate Experiment Follow-On mission proved the feasibility of an interferometric sensor for inter-satellite length tracking with sub-nanometer precision, establishing an important milestone for space laser interferometry and the general expectation that future gravity missions will employ heterodyne laser interferometry for satellite-to-satellite ranging. In this paper, we present the design of an on-axis optical bench for next-generation laser ranging which enhances the received optical power and the transmit beam divergence, enabling longer interferometer arms and relaxing the optical power requirement of the laser assembly. All design functionalities and requirements are verified by means of computer simulations. A thermal analysis is carried out to investigate the robustness of the proposed optical bench to the temperature fluctuations found in orbit.

Phenotypic diploidization in plant functional traits uncovered by synthetic neopolyploids in Dianthus broteri.

Whole-genome duplication and post-polyploidization genome downsizing play key roles in the evolution of land plants; however, the impact of genomic diploidization on functional traits still remains poorly understood. Using Dianthus broteri as a model, we compared the ecophysiological behaviour of colchicine-induced neotetraploids (4xNeo) to diploids (2x) and naturally occurring tetraploids (4xNat). Leaf gas-exchange and chlorophyll fluorescence analyses were performed in order to asses to what extent post-polyploidization evolutionary processes have affected 4xNat. Genomic diploidization and phenotypic novelty were evident. Distinct patterns of variation revealed that post-polyploidization processes altered the phenotypic shifts directly mediated by genome doubling. The photosynthetic phenotype was affected in several ways but the main effect was phenotypic diploidization (i.e. 2x and 4xNat were closer to each other than to 4xNeo). Overall, our results show the potential benefits of considering experimentally synthetized versus naturally established polyploids when exploring the role of polyploidization in promoting functional divergence.

Well-Defined Lignin Model Films from Colloidal Lignin Particles.

The transformation of a molecularly complex and irregularly shaped lignin into a nanoscale spherical architecture is anticipated to play a pivotal role in the promotion of lignin valorization. From the standpoint of using colloidal lignin particles (CLPs) as building blocks for a diverse range of applications, it has become essential to study their interactions with soluble compounds of varied origin. However, the lack of model films with well-defined surface properties similar to those of CLPs has hindered fundamental studies using surface-sensitive techniques. Here, we report well-defined and stable thin films prepared from CLPs and demonstrate their suitability for investigation of surface phenomena. Direct adsorption on substrates coated with a cationic anchoring polymer resulted in uniform distribution of CLPs as shown with atomic force microscopy (AFM). Quartz crystal microbalance with dissipation monitoring (QCM-D) experiments revealed higher adsorbed mass of cationic lignin onto the CLP-coated substrate in comparison to the film prepared from dissolved lignin, suggesting preferential adsorption via the carboxylic acid enriched surfaces of CLPs. QCM-D further enabled detection of small changes such as particle swelling or partial dissolution not detectable via bulk methods such as light scattering. The CLP thin films remained stable until pH 8 and displayed only a low degree of swelling. Increasing the pH to 10 led to some instability, but their spherical geometry remained intact until complete dissolution was observed at pH 12. Particles prepared from aqueous acetone or aqueous tetrahydrofuran solution followed similar trends regarding adsorption, pH stability, and wetting, although the particle size affected the magnitude of adsorption. Overall, our results present a practical way to prepare well-defined CLP thin films that will be useful not only for fundamental studies but also as a platform for testing stability and interactions of lignin nanoparticles with materials of technical and biomedical relevance.

Three-Dimensional Printed Cell Culture Model Based on Spherical Colloidal Lignin Particles and Cellulose Nanofibril-Alginate Hydrogel.

Three-dimensional (3D) printing has been an emerging technique to fabricate precise scaffolds for biomedical applications. Cellulose nanofibril (CNF) hydrogels have attracted considerable attention as a material for 3D printing because of their shear-thinning properties. Combining cellulose nanofibril hydrogels with alginate is an effective method to enable cross-linking of the printed scaffolds in the presence of Ca2+ ions. In this work, spherical colloidal lignin particles (CLPs, also known as spherical lignin nanoparticles) were used to prepare CNF-alginate-CLP nanocomposite scaffolds. High-resolution images obtained by atomic force microscopy (AFM) showed that CLPs were homogeneously mixed with the CNF hydrogel. CLPs brought antioxidant properties to the CNF-alginate-CLP scaffolds in a concentration-dependent manner and increased the viscosity of the hydrogels at a low shear rate, which correspondingly provide better shape fidelity and printing resolution to the scaffolds. Interestingly, the CLPs did not affect the viscosity at high shear rates, showing that the shear thinning behavior typical for CNF hydrogels was retained, enabling easy printing. The CNF-alginate-CLP scaffolds demonstrated shape stability after printing, cross-linking, and storage in Dulbecco's phosphate buffer solution (DPBS +) containing Ca2+ and Mg2+ ions, up to 7 days. The 3D-printed scaffolds showed relative rehydration ratio values above 80% after freeze-drying, demonstrating a high water-retaining capability. Cell viability tests using hepatocellular carcinoma cell line HepG2 showed no negative effect of CLPs on cell proliferation. Fluorescence microscopy indicated that HepG2 cells grew not only on the surfaces but also inside the porous scaffolds. Overall, our results demonstrate that nanocomposite CNF-alginate-CLP scaffolds have high potential in soft-tissue engineering and regenerative-medicine applications.

Single-Element Dual-Interferometer for Precision Inertial Sensing.

Tracking moving masses in several degrees of freedom with high precision and large dynamic range is a central aspect in many current and future gravitational physics experiments. Laser interferometers have been established as one of the tools of choice for such measurement schemes. Using sinusoidal phase modulation homodyne interferometry allows a drastic reduction of the complexity of the optical setup, a key limitation of multi-channel interferometry. By shifting the complexity of the setup to the signal processing stage, these methods enable devices with a size and weight not feasible using conventional techniques. In this paper we present the design of a novel sensor topology based on deep frequency modulation interferometry: the self-referenced single-element dual-interferometer (SEDI) inertial sensor, which takes simplification one step further by accommodating two interferometers in one optic. Using a combination of computer models and analytical methods we show that an inertial sensor with sub-picometer precision for frequencies above 10 mHz, in a package of a few cubic inches, seems feasible with our approach. Moreover we show that by combining two of these devices it is possible to reach sub-picometer precision down to 2 mHz. In combination with the given compactness, this makes the SEDI sensor a promising approach for applications in high precision inertial sensing for both next-generation space-based gravity missions employing drag-free control, and ground-based experiments employing inertial isolation systems with optical readout.

Adsorption of Proteins on Colloidal Lignin Particles for Advanced Biomaterials.

Coating of colloidal lignin particles (CLPs), or lignin nanoparticles (LNPs), with proteins was evaluated in order to establish a safe, self-assembly mediated modification technique to tune their surface chemistry. Gelatin and poly- l-lysine formed the most pronounced protein corona on the CLP surface, as determined by dynamic light scattering (DLS) and zeta potential measurements. Spherical morphology of individual protein coated CLPs was confirmed by transmission electron (TEM) and atomic force (AFM) microscopy. A mechanistic adsorption study with several random coiled and globular model proteins was carried out using quartz crystal microbalance with dissipation monitoring (QCM-D). The three-dimensional (3D) protein fold structure and certain amino acid interactions were decisive for the protein adsorption on the lignin surface. The main driving forces for protein adsorption were electrostatic, hydrophobic, and van der Waals interactions, and hydrogen bonding. The relative contributions of these interactions were highly dependent on the ionic strength of the surrounding medium. Capillary electrophoresis (CE) and Fourier transform infrared spectroscopy (FTIR) provided further evidence of the adsorption-enhancing role of specific amino acid residues such as serine and proline. These results have high impact on the utilization of lignin as colloidal particles in biomedicine and biodegradable materials, as the protein corona enables tailoring of the CLP surface chemistry for intended applications.

Adaptation of targeted nanocarriers to changing requirements in antimalarial drug delivery.

The adaptation of existing antimalarial nanocarriers to new Plasmodium stages, drugs, targeting molecules, or encapsulating structures is a strategy that can provide new nanotechnology-based, cost-efficient therapies against malaria. We have explored the modification of different liposome prototypes that had been developed in our group for the targeted delivery of antimalarial drugs to Plasmodium-infected red blood cells (pRBCs). These new models include: (i) immunoliposome-mediated release of new lipid-based antimalarials; (ii) liposomes targeted to pRBCs with covalently linked heparin to reduce anticoagulation risks; (iii) adaptation of heparin to pRBC targeting of chitosan nanoparticles; (iv) use of heparin for the targeting of Plasmodium stages in the mosquito vector; and (v) use of the non-anticoagulant glycosaminoglycan chondroitin 4-sulfate as a heparin surrogate for pRBC targeting. The results presented indicate that the tuning of existing nanovessels to new malaria-related targets is a valid low-cost alternative to the de novo development of targeted nanosystems.

Non-Thermal Plasma Activation of Gold-Based Catalysts for Low-Temperature Water-Gas Shift Catalysis.

Non-thermal plasma activation has been used to enable low-temperature water-gas shift over a Au/CeZrO4 catalyst. The activity obtained was comparable with that attained by heating the catalyst to 180 °C providing an opportunity for the hydrogen production to be obtained under conditions where the thermodynamic limitations are minimal. Using in situ diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS), structural changes associated with the gold nanoparticles in the catalyst have been observed which are not found under thermal activation indicating a weakening of the Au-CO bond and a change in the mechanism of deactivation.

Induced-aggregates in photocatalysis: An unexplored approach to reduce the noble metal co-catalyst content.

Photocatalysis has emerged as a promising and environmentally sustainable solution to produce high-purity hydrogen through ethanol photoreforming. It is commonly accepted that adding co-catalysts, especially noble metals, significantly enhances the catalytic activity of semiconductors. However, the high cost of noble metals such as Pt may limit the real application of this emerging technology. Here we evaluate the possibility of reducing the noble metal loading by creating the appropriate interface between pre-formed semiconductor nanoparticles. Commercial titania (P25) was selected as the semiconductor due to its commercial availability, facilitating the straightforward validation and corroboration of our results. Pt was selected as co-catalyst because one of the most efficient photocatalysts for the ethanol photo-reforming is still based on the use of P25 in combination with Pt. We report that the creation of induced aggregates dramatically improves the total hydrogen produced when very low loadings (≤0.05 wt%) of Pt are used. We have developed a pioneering reactor designed for conducting photoluminescence studies under authentic operational conditions of nanoparticle suspensions in the liquid phase. This approach allows us to obtain the average photoluminescence emission from the P25 agglomerates what it would be impossible to obtain by using standard solid samples holders. Thanks to this equipment, we can conclude that this remarkable improvement of the activity is mainly due to creation of an interface that favors the charge transfer between the particles of the aggregates. According to this, the titania nanoparticles of the agglomerates act as an antenna to collect the photons of the sun-light and produce the photo-excited electrons that will be transferred to the platinum nanoparticles located in the same agglomeration. In contrast, raw P25 with low loadings of Pt would have a high number of titania nanoparticles without platinum, and therefore, inactive. This result would be especially relevant in the case of immobilized photocatalytic systems for real future photocatalytic reactors because the immobilization of the semiconductors would generate similar interactions to the one created by our method. Consequently, the initial semiconductor immobilization followed by the subsequent photo-deposition of the co-catalyst emerges as a promising approach for a substantial reduction of the co-catalyst content.

Insights into spheroids formation in cellulose nanofibrils and Matrigel hydrogels using AFM-based techniques.

The recent FDA decision to eliminate animal testing requirements emphasises the role of cell models, such as spheroids, as regulatory test alternatives for investigations of cellular behaviour, drug responses, and disease modelling. The influence of environment on spheroid formation are incompletely understood, leading to uncertainty in matrix selection for scaffold-based 3D culture. This study uses atomic force microscopy-based techniques to quantify cell adhesion to Matrigel and cellulose nanofibrils (CNF), and cell-cell adhesion forces, and their role in spheroid formation of hepatocellular carcinoma (HepG2) and induced pluripotent stem cells (iPS(IMR90)-4). Results showed different cell behaviour in CNF and Matrigel cultures. Both cell lines formed compact spheroids in CNF but loose cell aggregates in Matrigel. Interestingly, the type of cell adhesion protein, and not the bond strength, appeared to be a key factor in the formation of compact spheroids. The gene expression of E- and N-cadherins, proteins on cell membrane responsible for cell-cell interactions, was increased in CNF culture, leading to formation of compact spheroids while Matrigel culture induced integrin-laminin binding and downregulated E-cadherin expression, resulting in looser cell aggregates. These findings enhance our understanding of cell-biomaterial interactions in 3D cultures and offer insights for improved 3D cell models, culture biomaterials, and applications in drug research.

Exploring Fracture Patterns: Assessing Representation Methods for Bone Fracture Simulation.

Fracture pattern acquisition and representation in human bones play a crucial role in medical simulation, diagnostics, and treatment planning. This article presents a comprehensive review of methodologies employed in acquiring and representing bone fracture patterns. Several techniques, including segmentation algorithms, curvature analysis, and deep learning-based approaches, are reviewed to determine their effectiveness in accurately identifying fracture zones. Additionally, diverse methods for representing fracture patterns are evaluated. The challenges inherent in detecting accurate fracture zones from medical images, the complexities arising from multifragmentary fractures, and the need to automate fracture reduction processes are elucidated. A detailed analysis of the suitability of each representation method for specific medical applications, such as simulation systems, surgical interventions, and educational purposes, is provided. The study explores insights from a broad spectrum of research articles, encompassing diverse methodologies and perspectives. This review elucidates potential directions for future research and contributes to advancements in comprehending the acquisition and representation of fracture patterns in human bone.

Study of the electrocatalytic activity of cerium oxide and gold-studded cerium oxide nanoparticles using a Sonogel-Carbon material as supporting electrode: electroanalytical study in apple juice for babies.

The present work reports a study of the electrocatalytic activity of CeO2 nanoparticles and gold sononanoparticles (AuSNPs)/CeO2 nanocomposite, deposited on the surface of a Sonogel-Carbon (SNGC) matrix used as supporting electrode and the application of the sensing devices built with them to the determination of ascorbic acid (AA) used as a benchmark analyte. Cyclic voltammetry (CV) and differential pulse voltammetry (DPV) were used to investigate the electrocatalytic behavior of CeO2- and AuSNPs/CeO2-modified SNGC electrodes, utilizing different concentrations of CeO2 nanoparticles and different AuSNPs:CeO2 w/w ratios. The best detection and quantification limits, obtained for CeO2 (10.0 mg·mL(-1))- and AuSNPs/CeO2 (3.25% w/w)-modified SNGC electrodes, were 1.59 × 10(-6) and 5.32 × 10(-6) M, and 2.93 × 10(-6) and 9.77 × 10(-6) M, respectively, with reproducibility values of 5.78% and 6.24%, respectively, for a linear concentration range from 1.5 µM to 4.0 mM of AA. The electrochemical devices were tested for the determination of AA in commercial apple juice for babies. The results were compared with those obtained by applying high performance liquid chromatography (HPLC) as a reference method. Recovery errors below 5% were obtained in most cases, with standard deviations lower than 3% for all the modified SNGC electrodes. Bare, CeO2- and AuSNPs/CeO2-modified SNGC electrodes were structurally characterized using scanning electron microscopy (SEM) and energy dispersive X-ray spectroscopy (EDS). AuSNPs and AuSNPs/CeO2 nanocomposite were characterized by UV-vis spectroscopy and X-ray diffraction (XRD), and information about their size distribution and shape was obtained by transmission electron microscopy (TEM). The advantages of employing CeO2 nanoparticles and AuSNPs/CeO2 nanocomposite in SNGC supporting material are also described. This research suggests that the modified electrode can be a very promising voltammetric sensor for the determination of electroactive species of interest in real samples.

Tuning the water interactions of cellulose nanofibril hydrogels using willow bark extract.

Cellulose nanofibrils (CNFs) are increasingly used as precursors for foams, films and composites, where water interactions are of great importance. In this study, we used willow bark extract (WBE), an underrated natural source of bioactive phenolic compounds, as a plant-based modifier for CNF hydrogels, without compromising their mechanical properties. We found that the introduction of WBE into both native, mechanically fibrillated CNFs and TEMPO-oxidized CNFs increased considerably the storage modulus of the hydrogels and reduced their swelling ratio in water up to 5-7 times. A detailed chemical analysis revealed that WBE is composed of several phenolic compounds in addition to potassium salts. Whereas the salt ions reduced the repulsion between fibrils and created denser CNF networks, the phenolic compounds - which adsorbed readily on the cellulose surfaces - played an important role in assisting the flowability of the hydrogels at high shear strains by reducing the flocculation tendency, often observed in pure and salt-containing CNFs, and contributed to the structural integrity of the CNF network in aqueous environment. Surprisingly, the willow bark extract exhibited hemolysis activity, which highlights the importance of more thorough investigations of biocompatibility of natural materials. WBE shows great potential for managing the water interactions of CNF-based products.

Qualitative Analysis of Conversational Chatbots to Alleviate Loneliness in Older Adults as a Strategy for Emotional Health.

This article presents an exploration of conversational chatbots designed to alleviate loneliness among older adults. In addition to technical evaluation, it delves into effective communication between these systems and this demographic group, considering linguistic nuances, communicative preferences, and specific emotional needs. The intrinsic importance of chatbots as innovative solutions in combating loneliness is highlighted, emphasizing their ability to be understanding and empathetic allies, contributing to emotional well-being and socialization. The article explores how improved emotional well-being can positively impact the health and quality of life of older adults. The methodology, rooted in triangulation between a literature review and qualitative research through interviews and focus groups with older adults, provides a comprehensive insight into the findings. Ethical, technical, and design considerations such as privacy, autonomy, technology adaptation, and usability are also addressed. The article concludes with practical recommendations for developing user-friendly interfaces that encourage the active participation of older adults in chatbots. This holistic approach not only analyzes the technical effectiveness of chatbots in mitigating loneliness in older adults but delves into human, ethical, and practical aspects, enriching the understanding and implementation of these agents for social and emotional support.

Formic Acid Decomposition Using Palladium-Zinc Preformed Colloidal Nanoparticles Supported on Carbon Nanofibre in Batch and Continuous Flow Reactors: Experimental and Computational Fluid Dynamics Modelling Studies.

The need to replace conventional fuels with renewable sources is a great challenge for the science community. H2 is a promising alternative due to its high energy density and availability. H2 generation from formic acid (FA) decomposition occurred in a batch and a packed-bed flow reactor, in mild conditions, using a 2% Pd6Zn4/HHT (high heated treated) catalyst synthesised via the sol-immobilisation method. Experimental and theoretical studies took place, and the results showed that in the batch system, the conversion was enhanced with increasing reaction temperature, while in the continuous flow system, the conversion was found to decrease due to the deactivation of the catalyst resulting from the generation of the poisoning CO. Computational fluid dynamics (CFD) studies were developed to predict the conversion profiles, which demonstrated great validation with the experimental results. The model can accurately predict the decomposition of FA as well as the deactivation that occurs in the continuous flow system. Of significance was the performance of the packed-bed flow reactor, which showed improved FA conversion in comparison to the batch reactor, potentially leading to the utilisation of continuous flow systems for future fuel cell applications for on-site H2 production.

Toward Corneal Limbus In Vitro Model: Regulation of hPSC-LSC Phenotype by Matrix Stiffness and Topography During Cell Differentiation Process.

A functional limbal epithelial stem cells (LSC) niche is a vital element in the regular renewal of the corneal epithelium by LSCs and maintenance of good vision. However, little is known about its unique structure and mechanical properties on LSC regulation, creating a significant gap in development of LSC-based therapies. Herein, the effect of mechanical and architectural elements of the niche on human pluripotent derived LSCs (hPSC-LSC) phenotype and growth is investigated in vitro. Specifically, three formulations of polyacrylamide gels with different controlled stiffnesses are used for culture and characterization of hPSC-LSCs from different stages of differentiation. In addition, limbal mimicking topography in polydimethylsiloxane is utilized for culturing hPSC-LSCs at early time point of differentiation. For comparison, the expression of selected key proteins of the corneal cells is analyzed in their native environment through whole mount staining of human donor corneas. The results suggest that mechanical response and substrate preference of the cells is highly dependent on their developmental stage. In addition, data indicate that cells may carry possible mechanical memory from previous culture matrix, both highlighting the importance of mechanical design of a functional in vitro limbus model.