Bulk Stiffness Research Articles

Structural properties, thermodynamic stability and reaction pathways for solid-state synthesis of Bi2WO6 polymorphs.

This study presents a density functional theory (DFT) investigation into the structural, electronic, and optical properties, thermodynamic stability, phase competition, and reaction pathways of Bi2WO6, a notable compound within the Aurivillius family of oxides. We examine three distinct polymorphs of Bi2WO6: the low-temperature orthorhombic phase (P1), the intermediate-temperature orthorhombic phase (P2), and the high-temperature monoclinic phase (P3). Electronic structure analysis indicates band gaps of 2.339 eV (P1), 2.312 eV (P2), and 2.128 eV (P3), with the valence band primarily composed of O 2p states and the conduction band of Bi 6p and W 5d states. Optical properties, including the dielectric function and absorption spectra, show distinct behaviors for each phase, particularly P3. Elastic and phonon property analyses confirm the mechanical and dynamical stability of all three phases, with the P1 phase exhibiting the highest bulk modulus and stiffness among the polymorphs. Our effective mass calculations suggest that the P3 phase may have better charge carrier mobility compared to the P1 and P2 phases. Structural optimizations reveal marginal differences in total energy among these phases, suggesting their potential coexistence or easy phase transitions under varying conditions. Detailed Gibbs free energy calculations confirm that the P1 phase is the most stable at low temperatures, in agreement with experimental data in the literature. We also construct a chemical reaction network to explore feasible reaction pathways for the solid-state synthesis of Bi2WO6 from Bi2O3 and WO3 precursors, identifying several low-cost reaction pathways, including both direct and multi-step routes involving intermediates such as Bi14WO24 and Bi2W2O9.

Hydrogels and aerogels of cellulose nanofiber derived from barley straw with addition of chitosan using high-intensity ultrasound and supercritical CO2 drying

In this study, cellulose-rich residue (purity of 90.2%) removed from barley straw was used to obtain cellulose nanofiber hydrogels (1–2 wt%) by high-intensity ultrasound treatments at 1200 W for 10–30 min. The highest water retention value (82.9±0.8 g/g) and fibrillation yield (62.12±0.13 wt%) of hydrogel were obtained by 1 and 2 wt% cellulose nanofibers, respectively, after ultrasound treatment at 1200 W for 20 min. Also, chitosan (1–3 wt%) was added to the cellulose nanofiber hydrogels by ultrasound treatment at 1200 W/20 min. The addition of chitosan reduced the water retention value and fibrillation yield. All hydrogels exhibited a typical elastic gel-like behavior. Then, the hydrogels were dried using supercritical CO2 to form aerogels, and their properties (bulk density, porosity, stiffness, functional groups, morphology, and thermal behavior) were determined. The lowest bulk density (0.056±0.011 g/cm3) and the highest porosity (96.50±0.17%) was achieved using 1 wt% cellulose nanofiber. The addition of chitosan at 3 wt% increased aerogel stiffness to 3200.10±8.80 g/mm.

The influence of internal pressure and neuromuscular agents on C. elegans biomechanics: an empirical and multi-compartmental in silico modelling study.

The function of a specific tissue and its biomechanics are interdependent, with pathologies or ageing often being intertwined with structural decline. The biomechanics of Caenorhabditis elegans, a model organism widely used in pharmacological and ageing research, has been established as biomarker for healthy ageing. However, the properties of the constituent tissues, and their contribution to the overall mechanical characteristics of the organism, remain relatively unknown. In this study we investigated the biomechanics of healthy C. elegans cuticle, muscle tissue, and pseudocoelom using a combination of indentation experiments and in silico modelling. We performed stiffness measurements using an atomic force microscope. To approximate the nematode's cylindrical body we used a novel three-compartment nonlinear finite element model, enabling us to analyse of how changes in the elasticity of individual compartments affect the bulk stiffness. We then fine-tuned the parameters of the model to match the simulation force-indentation output to the experimental data. To test the finite element model, we modified distinct compartments experimentally. Our in silico results, in agreement with previous studies, suggest that hyperosmotic shock reduces stiffness by decreasing the internal pressure. Unexpectedly, treatment with the neuromuscular agent aldicarb, traditionally associated with muscle contraction, reduced stiffness by decreasing the internal pressure. Furthermore, our finite element model can offer insights into how drugs, mutations, or processes such as ageing target individual tissues.

Flow affects the structural and mechanical properties of the fibrin network in plasma clots

The fibrin network is one of the main components of thrombi. Altered fibrin network properties are known to influence the development and progression of thrombotic disorders, at least partly through effects on the mechanical stability of fibrin. Most studies investigating the role of fibrin in thrombus properties prepare clots under static conditions, missing the influence of blood flow which is present in vivo. In this study, plasma clots in the presence and absence of flow were prepared inside a Chandler loop. Recitrated plasma from healthy donors were spun at 0 and 30 RPM. The clot structure was characterized using scanning electron microscopy and confocal microscopy and correlated with the stiffness measured by unconfined compression testing. We quantified fibrin fiber density, pore size, and fiber thickness and bulk stiffness at low and high strain values. Clots formed under flow had thinner fibrin fibers, smaller pores, and a denser fibrin network with higher stiffness values compared to clots formed in absence of flow. Our findings indicate that fluid flow is an essential factor to consider when developing physiologically relevant in vitro thrombus models used in researching thrombectomy outcomes or risk of embolization.Graphical

Structural, electronic, optical, and thermoelectric properties of CsPbI3-yBry (y = 0.5, 1.5, 2.5) compounds: First-principle study

BackgroundAmong photovoltaic materials, halide-based perovskites, such as CsPbI3, have been the subject of many research papers due to their suitable and direct bandgap, very good optical properties, and remarkable thermoelectric properties. Enhancing optoelectronic and thermoelectric properties can be achieved by combining halide atoms intelligently. The purpose of this article is to examine the physical properties of Br substitution at different concentrations in CsPbI3 perovskite. MethodsThe structural, electrical, and optical properties of CsPbI3-yBry (y = 0.5, 1.5, 2.5) compounds have been investigated by using the QUANTUM-ESPRESSO/PWSCF computational package which is based on the density functional theory and the pseudo-potential technique. The thermoelectric properties of CsPbI3-yBry perovskites were calculated by solving the Boltzmann transport equations within the constant relaxation time approximation as employed in the BoltzTraP package. Significant findingsThe structural investigation reveals that when the I atom is substituted by Br, the lattice constant decreases, increasing the bulk modulus and stiffness of compounds. The calculated negative formation energy confirmed the thermodynamic stability of CsPbI3-yBry compounds, and the stability improved with an increase in Br concentration. The electronic investigation indicates the semiconductor nature of compounds with a direct band gap, with a slight increase in electronic band gap as Br concentration increases. The noticeable optical absorption coefficient in the visible range of the electromagnetic spectrum, along with high and tunable optical conductivity, confirms the capability of the compounds for use in optoelectronic devices. The positive value of the Seebeck coefficient revealed the p-type semiconducting nature of CsPbI3-yBry perovskites. Lattice thermal conductivity calculation reveals that this quantity is decreased because of the increase in phonon-phonon interaction due to rising temperature. Thermoelectric investigation indicates that a precise mixture of halide I and Br atoms significantly enhances the electrical and thermal conductivity of perovskites. Also, the substitution of halide I atoms by Br can improve the figure of merit (ZT). The maximum value of ZT at room temperature is equal to 0.99, justifying the brilliant thermoelectric properties of the studied compounds.

Bulk stiffness and recovery time – A measure of development of mouse embryos

Objectives: Understanding the onset of embryonic arrest by studying the morphology of the embryos at different stages of their development is a relatively well-understood subject. The qualitative nature of the study makes it difficult to determine and quantify the causes behind the arrest. For successful embryonic development, it is essential to know, non-destructively, and the developmental viability of the embryos at a particular stage. Material and Methods: We investigate the bulk stiffness and recovery time as the viability quantifiers for grading or classifying embryos at the two-cell stage or after the formation of the first cleavage. A simple cantilever beam-based experimental procedure is established to investigate two-cell mouse embryos in two distinct orientations. Results: Bulk stiffness measurement varies with the embryo’s orientation and deformation. A good quality two-cell embryo shows a significant decrease in stiffness ratio with the deformation. Also, the embryo’s recovery time after releasing deformation may indicate a quality-dependent correlation. Conclusion: Our study concludes that the bulk stiffness measurements in the two principal orientations, their ratio, and recovery time correlate with the underlying morphology of two-cell stage mouse embryos.

Generation of functionally distinct T-cell populations by altering the viscoelasticity of their extracellular matrix.

The efficacy of adoptive T-cell therapies largely depends on the generation of T-cell populations that provide rapid effector function and long-term protective immunity. Yet it is becoming clearer that the phenotypes and functions of T cells are inherently linked to their localization in tissues. Here we show that functionally distinct T-cell populations can be generated from T cells that received the same stimulation by altering the viscoelasticity of their surrounding extracellular matrix (ECM). By using a model ECM based on a norbornene-modified collagen type I whose viscoelasticity can be adjusted independently from its bulk stiffness by varying the degree of covalent crosslinking via a bioorthogonal click reaction with tetrazine moieties, we show that ECM viscoelasticity regulates T-cell phenotype and function via the activator-protein-1 signalling pathway, a critical regulator of T-cell activation and fate. Our observations are consistent with the tissue-dependent gene-expression profiles of T cells isolated from mechanically distinct tissues from patients with cancer or fibrosis, and suggest that matrix viscoelasticity could be leveraged when generating T-cell products for therapeutic applications.

Assessing Differential Particle Deformability under Microfluidic Flow Conditions.

Assessing the mechanical behavior of nano- and micron-scale particles with complex shapes is fundamental in drug delivery. Although different techniques are available to quantify the bulk stiffness in static conditions, there is still uncertainty in assessing particle deformability in dynamic conditions. Here, a microfluidic chip is designed, engineered, and validated as a platform to assess the mechanical behavior of fluid-borne particles. Specifically, potassium hydroxide (KOH) wet etching was used to realize a channel incorporating a series of micropillars (filtering modules) with different geometries and openings, acting as microfilters in the direction of the flow. These filtering modules were designed with progressively decreasing openings, ranging in size from about 5 down to 1 μm. Discoidal polymeric nanoconstructs (DPNs), with a diameter of 5.5 μm and a height of 400 nm, were realized with different poly(lactic-co-glycolic acid) (PLGA) and poly(ethylene glycol) (PEG) ratios (PLGA/PEG), namely, 5:1 and 1:0, resulting in soft and rigid particles, respectively. Given the peculiar geometry of DPNs, the channel height was kept to 5 μm to limit particle tumbling or flipping along the flow. After thorough physicochemical and morphological characterization, DPNs were tested within the microfluidic chip to investigate their behavior under flow. As expected, most rigid DPNs were trapped in the first series of pillars, whereas soft DPNs were observed to cross multiple filtering modules and reach the micropillars with the smallest opening (1 μm). This experimental evidence was also supported by computational tools, where DPNs were modeled as a network of springs and beads immersed in a Newtonian fluid using the smoothed particle hydrodynamics (SPH) method. This preliminary study presents a combined experimental-computational framework to quantify, compare, and analyze the characteristics of particles having complex geometrical and mechanical attributes under flow conditions.

NUMERICAL AND EXPERIMENTAL ANALYSIS OF THE CHANGE IN THE MECHANICAL PROPERTIES INDUCED BY DIFFERENT MATERIAL INTERNAL FEATURES

In orthopedic medical devices, differences in elasto-plastic behavior between bone and metallic materials could lead to mechanical issues at the bone-implant interface, such as stress shielding, bone fracture or implant failure. To reduce mismatching-related adverse events between bone and prosthetic mechanical properties, an in-body geometry optimization could be the right approach to reduce prosthetic stiffness. Therefore, this study aims to assess the elastic behavior of four different in-body gap prismatic geometries (quadratic, hexagonal, octagonal, and circular) and how much they reduce bulk stiffness. Uniaxial compression tests were performed on five cubes with a 20[Formula: see text]mm thickness, each containing a different set of internal prismatic gaps. For each design, the elastic response was calculated and compared with a full-volume cube, used as control. All cubes showed a stiffness reduction compared to the control, greater in cubes with quadratic (21%), octagonal (18%), and circular (17%) transversal sections, compared to the hexagonal one (6%). Moreover, finite element models were implemented and tested, showing coherent values obtained through the experimental tests. In addition, a bi-material approach was studied in silico and the results suggested that variable elastic behavior could be obtained by using composite material, providing lower mechanical properties than commonly used commercial prosthetic materials.

Compacted bentonite hydro-mechanical modelling when interaggregate porosity tends to zero

A formulation is proposed for reproducing the hydro-mechanical behaviour of compacted bentonites as interaggregate porosity becomes negligible. In that case, the model assumes the bentonite deformability to be controlled only by the microstructural modelling level in a double porosity approach. Hence, the bulk stiffness of the system is defined by the relationship between the microstructural void ratio and the thermodynamic swelling pressure, requiring no additional parameters to simulate the overall behaviour. An experimental study has been conducted to check the conceptual consistency of the formulation, with favourable results. The quantitative potential of the model has also been satisfactorily verified by numerical simulation of the tests carried out in this study. A displacement-based finite-element model developed for double porosity systems has been modified to run the simulations. The proposed formulation has been implemented using a simple strategy that can be easily replicated for similar numerical models. Both the simulation capabilities and the computational cost efficiency of the proposed formulation confirm its practical interest.

Abstract 4560: Assay-ready tissue-engineered cancer microspheres for high-throughput screening

Abstract For drug discovery, new in vitro cancer models are needed to obtain more translatable study outcomes in a low-cost and high-throughput manner. For this purpose, 3D cancer spheroids have been established as more effective than 2D models. Current commercial techniques, however, rely heavily on self-aggregation of dissociated cells and cannot replicate key features of the native tumor microenvironment, particularly due to a lack of control over extracellular matrix components and heterogeneity in size and aggregate-forming tendencies. Also, current spheroidal techniques are typically limited to one spheroid per well, therefore providing a narrow range of cell numbers per well, disadvantageous for assay development in drug screening. Here, we overcome these challenges by coupling tissue engineering toolsets with microfluidic technologies to create engineered cancer microspheres and sorting desired numbers of microspheres into assay-ready well-plate format. To form the engineered cancer microspheres, MCF7 (non-metastatic) and MDA-MB-231 (metastatic) breast cancer cells were encapsulated within poly(ethylene glycol)-fibrinogen hydrogels using our previously developed microfluidic platform. Highly uniform cancer microspheres (intra and inter-batch coefficient of variation ≤ 5%) with high cell densities (over 20 × 106 cells/ml) were produced rapidly, which is critical for use in drug testing. The microspheres supported the 3D culture of both breast cancer cell lines over at least 14 days in culture. Encapsulated cells displayed cell type-specific differences in morphology, proliferation, metabolic activity, ultrastructure, and overall microsphere size distribution and bulk stiffness. To prepare assay-ready pre-plated microspheres, a COPAS FP flow cytometer was used for its ability to analyze and sort large sample particles such as tumor spheroids and hydrogel cancer microspheres generated in this study. When using a 96-well plate, the sorting rate varied from 2.5 - 6 microspheres per second, depending on the sample concentration. When sorting a desired number of microspheres per well, the accuracy was greater than 95% as verified visually by microscopy. Viability of sorted microspheres was verified 24 hours post-sort. Shipping conditions were established that maintained cell viability for remote use in drug testing. Methods for compound addition by pinning and imaging were tested and optimized. Using these approaches, the microsphere system was shown to be compatible with an automated liquid handling system for administration of drug compounds; MDA-MB-231 microspheres were distributed in 384 well plates and treated with chemotherapeutic drugs. Expected responses were quantitated using CellTiter-Glo® 3D and detected using automated imaging. Overall, our results demonstrate initial applicability for the tissue-engineered cancer microspheres for drug screening. Citation Format: Yuan Tian, Yongwoon Kim, Wen J. Seeto, Shantanu Pradhan, Dmitriy Minond, Rock Pulak, Elizabeth A. Lipke. Assay-ready tissue-engineered cancer microspheres for high-throughput screening. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 4560.

Stereo Coverage and Overall Stiffness of Biomaterial Arrays Underly Parts of Topography Effects on Cell Adhesion.

Surface topography is a biophysical factor affecting cell behaviors, yet the underlying cues are still not clear. Herein, we hypothesized that stereo coverage and overall stiffness of biomaterial arrays on the scale of single cells underly parts of topography effects on cell adhesion. We fabricated a series of microarrays (micropillar, micropit, and microtube) of poly(l-lactic acid) (PLLA) using mold casting based on pre-designed templates. The characteristic sizes of array units were less than that of a single cell, and thus, each cell could sense the micropatterns with varied roughness. With human umbilical vein endothelial cells (HUVECs) as the model cell type, we examined spreading areas and cell viabilities on different surfaces. "Stereo coverage" was defined to quantify the actual cell spreading fraction on a topographic surface. Particularly in the case of high micropillars, cells were confirmed not able to touch the bottom and had to partially hang among the micropillars. Then, in our opinion, a cell sensed the overall stiffness combining the bulk stiffness of the raw material and the stiffness of the culture medium. Spreading area and single cell viability were correlated to coverage and topographic feature of the prepared microarrays in particular with the significantly protruded geometry feather. Cell traction forces exerted on micropillars were also discussed. These findings provide new insights into the surface modifications toward biomedical implants.

Chondrogenic differentiation of human bone marrow MSCs in osteochondral implants under kinematic mechanical load is dependent on the underlying osteo component.

Chondrogenic models utilizing human mesenchymal stromal cells (hMSCs) are often simplistic, with a single cell type and the absence of mechanical stimulation. Considering the articulating joint as an organ it would be beneficial to include more complex stimulation. Within this study we applied clinically relevant kinematic load to biphasic constructs. In each case, the upper layer consisted of fibrin embedded hMSCs retained within an elastomeric polyurethane (PU) scaffold. These were randomly assigned to five base scaffolds, a cell-free fibrin PU base, viable bone, decellularized bone, 3D printed calcium phosphate or clinically used cement. This allowed the study of cross talk between viable bone and chondrogenically differentiating MSCs, while controlling for the change in stiffness of the base material. Data obtained showed that the bulk stiffness of the construct was not the defining factor in the response obtained, with viable and decellularized bone producing similar results to the softer PU base. However, the stiff synthetic materials led to reduced chondrogenesis and increased calcification in the upper MSC seeded layer. This demonstrates that the underlying base material must be considered when driving chondrogenesis of human cells using a clinically relevant loading protocol. It also indicates that the material used for bony reconstruction of osteochondral defects may influence subsequent chondrogenic potential.

Hydrophilic, conductive and flexible piezoresistive PEDOT:PSS 3D sensors with tunable microarchitecture and crosslinked using a PEGylated crosslinker

Poly(3,4-ethylenedioxythiophene):poly(styrene sulfonate) (PEDOT:PSS) is an emerging biomaterial that – due to its stability and conductivity - has been adopted for diverse biomedical technologies, such as biosensors, coatings for electrodes, and substrates for tissue engineering. Currently, some researchers are focusing on the processing of this biomaterial into three-dimensional (3D) scaffolds, either in the form of fibrous structures, porous sponge-like constructs or hydrogels. However, due to its instability in aqueous environments, challenges occur when applying manufacturing techniques to PEDOT:PSS. Moreover, the standardised use of covalent crosslinking via glycidoxypropyltrimethoxysilane (GOPS) negatively impacts the conductive properties of the material. In this work, poly(ethylene glycol) diglycidyl ether (PEGDE) – which was previously demonstrated to stabilise PEDOT:PSS films - has been used as crosslinking reagent to generate 3D electroconductive porous biomaterial sensors via lyophilization. Both isotropic and anisotropic porous scaffolds were obtained, and although macroscopically similar to GOPS-crosslinked samples, these constructs exhibited unique mechanical electrical features. PEGDE crosslinking of 3D PEDOT:PSS sponges enhanced material hydrophilicity, softness, and electrical conductivity. Finally, biphasic electroactive sponges were manufactured via the inclusion of an elastomeric matrix into these constructs to prolong their longevity and achieve varying degrees of bulk stiffness. These were evaluated as piezoresistive sensors and demonstrated a high strain gauge factor with potential for biometric sensing and biomonitoring.

Initial bacterial retention on polydimethylsiloxane of various stiffnesses: The relevance of modulus (mis)match

Initial retention of the bacterium Escherichia coli on model poly(dimethylsiloxane) (PDMS) surfaces was studied as a function of substrate bulk and surface mechanical stiffness values. Our reference PDMS system was designed such that out of the parameters that govern bacterial adhesion only the mechanical stiffness was systematically varied. This was achieved by varying the crosslinking density of PDMS. Following crosslinking, we performed Soxhlet extraction of non-crosslinked, free chains to rule out their effect on bacterial response. Bulk moduli were assessed by dynamic mechanical analysis at 1 rad sec−1 frequency and the values obtained ranged between 0.03 and 1.8 MPa. The increase in crosslink density resulted in increasing surface modulus, as measured by atomic force microscopy, with values ranging between 0.7 and 9 MPa. The number of bacteria retained was then assessed. We observed a decreasing trend with the increase of both bulk and surface mechanical stiffnesses down to a limit corresponding to the Young’s modulus of the bacterial cell surface. For higher values than this threshold, the number of retained bacteria remained constant. We tentatively explain this observation by considering conformal overlay of bacterial and material surfaces.

Gold Nano-Bio-Interaction to Modulate Mechanobiological Responses for Cancer Therapy Applications.

In the present study, we investigate the mechanobiological responses of human lung cancer that may occur through their interactions with two different types of gold nanoparticles: nanostars and nanospheres. Hyperspectral images of nanoparticle-treated cells revealed different spatial distributions of nanoparticles in cells depending on their morphology, with nanospheres being more uniformly distributed in cells than nanostars. Gold nanospheres were also found to be more effective in mechanobiological modulations. They significantly suppressed the migratory ability of cells under different incubation times while lowering the bulk stiffness and adhesion of cells. This in vitro study suggests the potential applications of gold nanoparticles to manage cell migration. Nano-bio-interactions appeared to impact the cytoskeletal organization of cells and consequently alter the mechanical properties of cells, which could influence the cellular functions of cells. According to the results and migratory index model, it is thought that nanoparticle-treated cells experience mechanical changes in their body, which largely reduces their migratory potentials. These findings provide a better understanding of nano-bio-interaction in terms of cell mechanics and highlight the importance of mechanobiological responses in designing gold nanoparticles for cancer therapy.

Cell mediated remodeling of stiffness matched collagen and fibrin scaffolds

Cells are known to continuously remodel their local extracellular matrix (ECM) and in a reciprocal way, they can also respond to mechanical and biochemical properties of their fibrous environment. In this study, we measured how stiffness around dermal fibroblasts (DFs) and human fibrosarcoma HT1080 cells differs with concentration of rat tail type 1 collagen (T1C) and type of ECM. Peri-cellular stiffness was probed in four directions using multi-axes optical tweezers active microrheology (AMR). First, we found that neither cell type significantly altered local stiffness landscape at different concentrations of T1C. Next, rat tail T1C, bovine skin T1C and fibrin cell-free hydrogels were polymerized at concentrations formulated to match median stiffness value. Each of these hydrogels exhibited distinct fiber architecture. Stiffness landscape and fibronectin secretion, but not nuclear/cytoplasmic YAP ratio differed with ECM type. Further, cell response to Y27632 or BB94 treatments, inhibiting cell contractility and activity of matrix metalloproteinases, respectively, was also dependent on ECM type. Given differential effect of tested ECMs on peri-cellular stiffness landscape, treatment effect and cell properties, this study underscores the need for peri-cellular and not bulk stiffness measurements in studies on cellular mechanotransduction.

Abstract 175: Production of cancer tissue-engineered microspheres for high-throughput screening

Abstract There is a need for new in vitro systems that enable pharmaceutical companies to collect more physiologically-relevant information on drug response in a low-cost and high-throughput manner. For this purpose, three-dimensional (3D) spheroidal models have been established as more effective than two-dimensional models. Current commercial techniques, however, rely heavily on self-aggregation of dissociated cells and are unable to replicate key features of the native tumor microenvironment, particularly due to a lack of control over extracellular matrix components and heterogeneity in shape, size, and aggregate forming tendencies. In this study, we overcome these challenges by coupling tissue engineering toolsets with microfluidics technologies to create engineered cancer microspheres. Specifically, we employ biosynthetic hydrogels composed of conjugated poly(ethylene glycol) (PEG) and fibrinogen protein (PEG-Fb) to create engineered breast and colorectal cancer tissue microspheres for 3D culture, tumorigenic characterization, and examination of potential for high-throughput screening (HTS). MCF7 and MDA-MB-231 cell lines were used to create breast cancer microspheres and the HT29 cell line and cells from a stage II patient-derived xenograft (PDX) were encapsulated to produce colorectal cancer (CRC) microspheres. Using our previously developed microfluidic system, highly uniform cancer microspheres (intra-batch coefficient of variation (CV) ≤ 5%, inter-batch CV < 2%) with high cell densities (>20×106 cells/ml) were produced rapidly, which is critical for use in drug testing. Encapsulated cells maintained high viability and displayed cell type-specific differences in morphology, proliferation, metabolic activity, ultrastructure, and overall microsphere size distribution and bulk stiffness. For PDX CRC microspheres, the percentage of human (70%) and CRC (30%) cells was maintained over time and similar to the original PDX tumor, and the mechanical stiffness also exhibited a similar order of magnitude (103 Pa) to the original tumor. The cancer microsphere system was shown to be compatible with an automated liquid handling system for administration of drug compounds; MDA-MB-231 microspheres were distributed in 384 well plates and treated with staurosporine (1 μM) and doxorubicin (10 μM). Expected responses were quantified using CellTiter-Glo® 3D, demonstrating initial applicability to HTS drug discovery. PDX CRC microspheres were treated with Fluorouracil (5FU) (10 to 500 μM) and displayed a decreasing trend in metabolic activity with increasing drug concentration. Providing a more physiologically relevant tumor microenvironment in a high-throughput and low-cost manner, the PF hydrogel-based cancer microspheres could potentially improve the translational success of drug candidates by providing more accurate in vitro prediction of in vivo drug efficacy. Citation Format: Elizabeth A. Lipke, Wen J. Seeto, Yuan Tian, Mohammadjafar Hashemi, Iman Hassani, Benjamin Anbiah, Nicole L. Habbit, Michael W. Greene, Dmitriy Minond, Shantanu Pradhan. Production of cancer tissue-engineered microspheres for high-throughput screening [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2022; 2022 Apr 8-13. Philadelphia (PA): AACR; Cancer Res 2022;82(12_Suppl):Abstract nr 175.

Effect of Graphene Nanofibers on the Morphological, Structural, Thermal, Phase Transitions and Mechanical Characteristics in Metallocene iPP Based Nanocomposites

Several nanocomposites were prepared by extrusion from a commercial metallocene-type isotactic polypropylene (iPP) and different amounts of two types of graphene (G) nanofibers: ones with a high specific surface, named GHS, and the others with a low specific surface, labeled as GLS. The number of graphene layers was found to be around eight for GLS and about five in the GHS. Scanning electron microscopy (SEM) images of the resultant iPP nanocomposites showed a better homogeneity in the dispersion of the GLS nanofibers within the polymeric matrix compared with the distribution observed for the GHS ones. Crystallinity in the nanocomposites turned out to be dependent upon graphene content and upon thermal treatment applied during film preparation, the effect of the nature of the nanofiber being negligible. Graphene exerted a noticeable nucleating effect in the iPP crystallization. Furthermore, thermal stability was enlarged, shifting to higher temperatures, with increasing nanofiber amount. The mechanical response changed significantly with nanofiber type, along with its content, together with the thermal treatment applied to the nanocomposites. Features of nanofiber surface played a key role in the ultimate properties related to superficial and bulk stiffness.

Enhanced Cellular Materials through Multiscale, Variable-Section Inner Designs: Mechanical Attributes and Neural Network Modeling.

In the current work, the mechanical response of multiscale cellular materials with hollow variable-section inner elements is analyzed, combining experimental, numerical and machine learning techniques. At first, the effect of multiscale designs on the macroscale material attributes is quantified as a function of their inner structure. To that scope, analytical, closed-form expressions for the axial and bending inner element-scale stiffness are elaborated. The multiscale metamaterial performance is numerically probed for variable-section, multiscale honeycomb, square and re-entrant star-shaped lattice architectures. It is observed that a substantial normal, bulk and shear specific stiffness increase can be achieved, which differs depending on the upper-scale lattice pattern. Subsequently, extended mechanical datasets are created for the training of machine learning models of the metamaterial performance. Thereupon, neural network (NN) architectures and modeling parameters that can robustly capture the multiscale material response are identified. It is demonstrated that rather low-numerical-cost NN models can assess the complete set of elastic properties with substantial accuracy, providing a direct link between the underlying design parameters and the macroscale metamaterial performance. Moreover, inverse, multi-objective engineering tasks become feasible. It is shown that unified machine-learning-based representation allows for the inverse identification of the inner multiscale structural topology and base material parameters that optimally meet multiple macroscale performance objectives, coupling the NN metamaterial models with genetic algorithm-based optimization schemes.