Nanomechanical Mapping Research Articles

Triple Function of Amelogenin Peptide-Chitosan Hydrogel for Dentin Repair

Biomimetic strategies like peptide-guided collagen mineralization promise to enhance the effectiveness of dentin remineralization. We recently reported that rationally designed amelogenin-derived peptides P26 and P32 promoted apatite nucleation, mineralized collagen, and showed potential in enamel regrowth and dentin remineralization. To facilitate the clinical application of amelogenin-derived peptides and to uncover their effectiveness in repairing dentin, we have now implemented a chitosan (CS) hydrogel for peptide delivery and have investigated the effects of P26-CS and P32-CS hydrogels on dentin remineralization using 2 in situ experimental models that exhibited different levels of demineralization. The efficacy of the peptide-CS hydrogels in dentin repair was evaluated by characterizing the microstructure, mineral density, mineral phase, and nanomechanical properties of the remineralized samples. The new strategy of atomic force microscopy PeakForce quantitative nanomechanical mapping was used for direct visualization and nanomechanical analysis of repaired dentin lesions across the lesion depth. Results from the 2 models indicated the potential triple functions of peptide-CS hydrogels for dentin repair: building a highly organized protective mineralized layer on dentin, occluding dentinal tubules by peptide-guided in situ mineralization, and promoting biomimetic dentinal collagen remineralization. Importantly, peptides released from the CS hydrogel could diffuse into the dentinal matrix and penetrate the dentinal tubules, leading to both surface and subsurface remineralization and tubule occlusion. Given our previous findings on peptide-CS hydrogels’ potential for remineralizing enamel, we see further promise for hydrogels to treat tooth defects involving multiple hard tissues, as in the case of noncarious cervical lesions.

Deep learning strategy for small dataset from atomic force microscopy mechano-imaging on macrophages phenotypes.

The cytoskeleton is involved during movement, shaping, resilience, and functionality in immune system cells. Biomarkers such as elasticity and adhesion can be promising alternatives to detect the status of cells upon phenotype activation in correlation with functionality. For instance, professional immune cells such as macrophages undergo phenotype functional polarization, and their biomechanical behaviors can be used as indicators for early diagnostics. For this purpose, combining the biomechanical sensitivity of atomic force microscopy (AFM) with the automation and performance of a deep neural network (DNN) is a promising strategy to distinguish and classify different activation states. To resolve the issue of small datasets in AFM-typical experiments, nanomechanical maps were divided into pixels with additional localization data. On such an enlarged dataset, a DNN was trained by multimodal fusion, and the prediction was obtained by voting classification. Without using conventional biomarkers, our algorithm demonstrated high performance in predicting the phenotype of macrophages. Moreover, permutation feature importance was employed to interpret the results and unveil the importance of different biophysical properties and, in turn, correlated this with the local density of the cytoskeleton. While our results were demonstrated on the RAW264.7 model cell line, we expect that our methodology could be opportunely customized and applied to distinguish different cell systems and correlate feature importance with biophysical properties to unveil innovative markers for diagnostics.

Adaptive discrete mapping of dynamic nanomechanical property of soft materials on atomic force microscope

In this paper, an adaptive discrete nanomechanical mapping (A-DNM) technique is proposed for mapping time-varying nanomechanical properties of soft samples using atomic force microscope (AFM). Mapping time-varying nanomechanical properties is needed to investigate samples such as live cells undergoing dynamic evolutions. However, the existing methods based on continuous scanning are either too slow to capture time-elapsing variations of the sample over the entire sampled area, or only capable of mapping a small sample area. Contrarily, we propose to extend a discrete nanomechanical mapping scheme where only a set of chosen discrete locations (points of interests, POIs) are measured, thereby, dramatically increasing the number of measurements acquired at these POIs during the dynamic evolution of the sample occurring in minutes or faster. Specifically, through the proposed A-DNM approach, two challenges are addressed: First, the effect of time-varying sample topography during the nanomechanical evolution is accounted for by updating the estimation of the sample height at each POI during the mapping and compensating for it in the cantilever transition and engagement process. Secondly, an adaptive planning of the mapping sequence is proposed to adjust the distribution of the measurements between the POIs to accomodate the sample dynamic evolution. A POI-dynamics index is introduced to estimate the relative speeds of the nanomechanical evolutions at the POIs based on the measured data, and then online assign more measurements to those POIs of faster nanomechanical evolutions. The proposed approach is demonstrated through an experiment of mapping nanomechanical evolution of a poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) polymer undergoing crystallization process.

Identification of hybrid amyloid strains assembled from amyloid-β and human islet amyloid polypeptide

Cross-fibrillation of amyloid-β (Aβ) peptides and human islet amyloid polypeptides (hIAPP) has revealed a close correlation between Alzheimer’s disease and type 2 diabetes (T2D). Importantly, different amyloid strains are likely to lead to the clinical pathological heterogeneity of degenerative diseases due to toxicity. However, given the complicated cross-interactions between different amyloid peptides, it is still challenging to identify the polymorphism of the hybrid amyloid strains and reveal mechanistic insights into aggregation, but highly anticipated due to their significance. In this study, we investigated the cross-fibrillation of Aβ peptides and different hIAPP species (monomers, oligomers, and fibrils) using combined experimental and simulation approaches. Cross-seeding and propagation of different amyloid peptides monitored by experimental techniques proved that the three species of hIAPP aggregates have successively enhanced Aβ fibrillation, especially for hIAPP fibrils. Moreover, the polymorphism of these morphologically similar hybrid amyloid strains could be distinguished by testing their mechanical properties using quantitative nanomechanical mapping, where the assemblies of Aβ-hIAPP fibrils exhibited the high Young’s modulus. Furthermore, the enhanced internal molecular interactions and β-sheet structural transformation were proved by exploring the conformational ensembles of Aβ-hIAPP heterodimer and Aβ-hIAPP decamer using molecular dynamic simulations. Our findings pave the way for identifying different hybrid amyloid strains by quantitative nanomechanical mapping and molecular dynamic simulations, which is important not only for the precise classification of neurodegenerative disease subtypes but also for future molecular diagnosis and therapeutic treatment of multiple interrelated degenerative diseases.

Nanomechanical behavior of polystyrene/graphene oxide nanocomposites

The in-situ investigation of the nanomechanical features of the polymer graphene nanocomposites has become a challenging and an indispensable task to achieve the required application. Graphene oxide (GO) nanocomposites were prepared at 1.0% weight fraction of GO to reinforce polystyrene (PS) using solution blending approach. The morphology of the resulting nanocomposites was characterized by optical, scanning electron, transmission electron, atomic force, and scattering scanning near-field optical microscopies. These showed a uniform dispersion of graphene oxide nano-sheets in the PS matrix. By adopting Derjaguin–Muller–Toporov (DMT) formula, the nanomechanical properties for the cryogenically fractured surface of the composites were characterized using the traditional atomic force microscopy (AFM), peak-force quantitative nanomechanical mapping (QNM), and tip-force mode functioned with scattering scanning near–field optical microscopy (s-SNOM). Young’s modulus of the PS matrix varied around (1–2) GPa as shown by QNM and s-SNOM similar to what was reported in the literature. However, while putative GO nano-sheets were measured to have a higher elastic modulus than the surrounding matrix in Peak-Force QNM experiments, they were significantly below literature values. By using Tip-Force mode related to s-SNOM, the expected values of Young’s modulus for GO were recovered.

Quantification Characterization of Hierarchical Structure of Polyurethane by Advanced AFM and X-ray Techniques.

Polyurethane (PU) with microphase separation has garnered significant attention due to its highly designable molecular structure and a wide range of adjustable properties. However, there is currently a lack of systematic approaches for quantifying PU's microphase separation. To address this research gap, we utilized an atomic force microscopy (AFM) nanomechanical mapping technique along with Gaussian fitting to recolor and quantitatively analyze the evolution of PU's microphase separation. By varying the ratios of the chain extender to cross-linking agent, we observed the changes in the hydrogen bonding between the soft and hard segments. As the ratio of the chain extender to cross-linking agent decreases, the strength of the hydrogen bonding weakens, resulting in a reduction in the quantity and phase percentage of hard segment (HS) domains. Consequently, the degree of microphase separation between the soft and hard segments decreases, leading to specific alterations in the material's mechanical properties and dynamic viscoelasticity. To further investigate the hierarchical structure of PU, we employed various techniques, such as X-ray analysis, transmission electron microscopy (TEM), and AFM-based infrared spectroscopy (AFM-IR). Our findings reveal a spherulite pattern composed of lamellae within the HS domains, with the cross-linking density gradually increasing from the center to the periphery. Overall, our comprehensive characterization of PU provides valuable insights into its hierarchical structure and establishes a quantitative framework to explore the intricate relationship between the structure and properties.

Operando Nanomechanical Mapping of Silicon Thin Film Electrode during the First Lithiation and Delithiation Investigated By Bimodal Atomic Force Microscopy

Si is emerging as one of the most prospective anode materials for all-solid-state Li-ion batteries due to its extremely high theoretical capacity density of 4200 mAh g-1 and relatively negative potential for lithiation/delithiation. Towards its widespread use, controlling severe volume expansion/contraction during lithiation/delithiation is a challenge because it may cause cracking, fracture, and pulverization that negatively influence the cycle performance and rate capability of the cells [1]. Such mechanical event is induced by the change in the lattice structure due to the formation of LixSi and successive increasing/decreasing of Li content x in LixSi [2].Bimodal atomic force microscopy (AFM) offers quantitative mapping of nanoscale mechanical property, simultaneously with surface topography imaging [3]. Our group previously applied this technique to the cross-section of a composite electrode consist of active materials, binders, conductive additives, and deposited electrolyte solution and successfully demonstrated nanomechanical mapping [4]. Here, we develop an operando bimodal AFM system to track Young’s modulus changes of electrode materials in an all-solid-state battery configuration during the lithiation and delithiation reactions in real-time.A cell in a Cu/Si/LLZT/In/Li configuration was fabricated as follows; a 3 µm-thick Si thin-film and a 100 nm-thick Cu layer were sputter-deposited on one side of Li6.6La3Zr1.6Ta0.4O12 (LLZT) solid electrolyte, an In layer was sputter-deposited on the other side of LLZT, the Cu/Si/LLZT/In sample was milled with an Ar-ion beam to yield a flat and smooth cross-section suitable for AFM analysis, and then a 50 µm-thick Li metal foil was combined with the pre-coated In thin layer. Operando nanomechanical mapping was performed by mounting the cross-section-exposed cell in the custom-made sample holder coupled with a potentiostat for bias applications, in an Ar-filled glove box to prevent any atmospheric influences.Electrochemical lithiation and delithiation were carried out within a potential window of 0.01-1.20 V vs. Li/Li+. Electrochemical characterization indicated that the capacities of the Si electrode for the first lithiation and delithiation are 3300 and 1833 mAh g-1, respectively. These capacities are equal to the formation of Li3.46Si after the first lithiation of the pristine Si and Li1.54Si after the successive delithiation from Li3.46Si. Throughout the lithiation, the averaged Young’s modulus of Si electrode decreased due to the formation of LixSi and increase in the Li content x in LixSi. In the initial stage of lithiation, it drastically decreased because LixSi was formed by the lithiation of pristine Si. Then, it moderately decreased with increasing Li content x in LixSi. Unfortunately, the Young’s modulus maps were only recorded until the formation of Li1.54Si because the successive delithiation was not completed possibly due to the phase transformation of crystalline LixSi to amorphous LixSi [5], which may cause a significant mechanical strain inside the thin film. The measured Young’s modulus values were consistent with those obtained by first principle calculation [6]. Further details are given at the presentation.

Self-assembly of a barnacle cement protein into intertwined amyloid fibres and determination of their adhesive and viscoelastic properties.

The stalked barnacle Pollicipes pollicipes uses a multi-protein cement to adhere to highly varied substrates in marine environments. We investigated the morphology and adhesiveness of a component 19 kDa protein in barnacle cement gland- and seawater-like conditions, using transmission electron microscopy and state-of-the art scanning probe techniques. The protein formed amyloid fibres after 5 days in gland-like but not seawater conditions. After 7-11 days, the fibres self-assembled under gland-like conditions into large intertwined fibrils of up to 10 µm in length and 200 nm in height, with a distinctive twisting of fibrils evident after 11 days. Atomic force microscopy (AFM)-nanodynamic mechanical analysis of the protein in wet conditions determined E' (elasticity), E'' (viscosity) and tan δ values of 2.8 MPa, 1.2 MPa and 0.37, respectively, indicating that the protein is a soft and viscoelastic material, while the adhesiveness of the unassembled protein and assembled fibres, measured using peak force quantitative nanomechanical mapping, was comparable to that of the commercial adhesive Cell-Tak™. The study provides a comprehensive insight into the nanomechanical and viscoelastic properties of the barnacle cement protein and its self-assembled fibres under native-like conditions and may have application in the design of amyloid fibril-based biomaterials or bioadhesives.

Analytical Models for Measuring the Mechanical Properties of Yeast.

The mechanical properties of yeast play an important role in many biological processes, such as cell division and growth, maintenance of internal pressure, and biofilm formation. In addition, the mechanical properties of cells can indicate the degree of damage caused by antifungal drugs, as the mechanical parameters of healthy and damaged cells are different. Over the past decades, atomic force microscopy (AFM) and micromanipulation have become the most widely used methods for evaluating the mechanical characteristics of microorganisms. In this case, the reliability of such an estimate depends on the choice of mathematical model. This review presents various analytical models developed in recent years for studying the mechanical properties of both cells and their individual structures. The main provisions of the applied approaches are described along with their limitations and advantages. Attention is paid to the innovative method of low-invasive nanomechanical mapping with scanning ion-conductance microscopy (SICM), which is currently starting to be successfully used in the discovery of novel drugs acting on the yeast cell wall and plasma membrane.

Correlating Nanomechanical Mapping and Single-Molecule Localization Microscopy for Local Properties in Interpenetrating Network Elastomers

Correlating Nanomechanical Mapping and Single-Molecule Localization Microscopy for Local Properties in Interpenetrating Network Elastomers

Influence of structural disorder on the elastic, frictional, and electrical properties in functionalized polyaniline thin films at the nanoscale investigated by atomic force microscopy

We investigated the influence of structural order on the elastic, frictional, and electrical properties of butylthio-functionalized PANI (PANI-SBu) films by atomic force microscopy (AFM)-based techniques, including PeakForce quantitative nanomechanical mapping, friction force microscopy, and conductive AFM. The PANI-SBu films were prepared by the drop-cast method from the solution of PANI-SBu in N-methyl-2-pyrrolidone that was continuously stirred. The PANI-SBu films were fabricated after different solution stirring times. The shear force during the mechanical stir will disentangle the highly-coiled PANI-SBu polymer chains in the solution. Therefore, the polymer chains in solution cast on the substrates will progressively self-assemble into a more organized structure when solvents evaporate, leading to PANI-SBu films with improved structural order. Our AFM studies discovered that more structurally-ordered PANI-SBu films have substantially larger out-of-plane elastic moduli and charge mobility but smaller kinetic friction coefficients. The denser packing of polymer molecules increases film elasticities and promotes chain-to-chain charge transport. In addition, stiffer PANI-SBu film surfaces are more difficult to deform when sheared by the sliding AFM probe, resulting in less energy dissipation during AFM friction measurements. Thus, smaller kinetic friction coefficients were found. Conversely, more structurally-disordered PANI-SBu films have smaller elasticity and charge mobility but larger kinetic friction coefficients. Our results demonstrate that it is possible to manipulate the elastic, frictional, and electrical properties of PANI-SBu films by controlling their structural order, which can be essential for developing polymer-based composite materials and flexible electronic devices.

Quantitative Elasticity Mapping of Submicron Silica Hollow Particles by PeakForce QNM AFM Mode.

Silica hollow spheres with a diameter of 100-300 nm and a shell thickness of 8±2 nm were synthesized using a self-templating amphiphilic polymeric precursor, i.e., poly(ethylene glycol)-substituted hyperbranched polyethoxysiloxane. Their elastic properties were addressed with a high-frequency AFM indentation method based on the PeakForce QNM (quantitative nanomechanical mapping) mode enabling simultaneous visualization of the surface morphology and high-resolution mapping of the mechanical properties. The factors affecting the accuracy of the mechanical measurements such as a local slope of the particle surface, deformation of the silica hollow particles by a solid substrate, shell thickness variation, and applied force range were analysed. The Young's modulus of the shell material was evaluated as E=26±7 GPa independent of the applied force in the elastic regime of deformations. Beyond the elastic regime, the buckling instability was observed revealing a non-linear force-deformation response with a hysteresis between the loading and unloading force-distance curves and irreversible deformation of the shell at high applied forces. Thus, it was demonstrated that PeakForce QNM mode can be used for quantitative measurements of the elastic properties of submicon-sized silica hollow particles with nano-size shell thickness, as well as for estimation of the buckling behaviour beyond the elastic regime of shell deformations.

Towards artefact-free AFM image presentation and interpretation.

Atomic force microscopy (AFM) is based upon a simple operational principle. However, the presentation and interpretation of AFM images can easily suffer from consequential artefacts that are easily overlooked. Here we discuss results from AFM and its companion variations AFM-IR (AFM combined with infrared spectroscopy) and PF-QNM (an AFM mode called peak-force quantitative nano-mechanical mapping) by imaging 'bee' structures in asphalt binder (bitumen) as examples. We show how common problems manifest themselves and provide solutions, with the intent that authors can present their results clearly and avoid interpreting artefacts as true physical properties, thereby raising the quality of AFM research.

Economical preparation of fluorine‐containing thermoplastic elastomer with good phase miscibility and elasticity based on dynamic vulcanization

AbstractThermoplastic elastomers are a green and sustainable category of high performance polymers and show the great advantage of easy processability relative to conventional rubbers. In this work, a fluorine‐containing thermoplastic elastomer was prepared through dynamic crosslinking of poly(vinylidene fluoride)/poly(ethylene−methyl acrylate−glycidyl methacrylate) (PVDF/EGMA) blends with the aid of amino‐terminated crosslinker (KH602). The reaction kinetics of EGMA with KH602 were evaluated by the DSC Kissinger method with an activation energy (Ea) of 103.04 kJ mol−1. The increased ‘S‐type’ torque curve during dynamic crosslinking indicates the occurrence of chemical crosslinking while Fourier transform infrared spectra further demonstrate the dehydrofluorination of PVDF and the reaction between the amino group of KH602 and the epoxy group in EGMA. The prepared PVDF/EGMA (30/70) thermoplastic vulcanizate shows good reprocessability, mechanical robustness and elasticity. With increasing content of crosslinker, the stress at 300% increases from 2.3 MPa to 3.5 MPa while elongation at break decreases from 1150% to 380%. The PVDF/EGMA blend has a typical ‘sea‐island’ morphology with ca 7 μm dispersed phase while PVDF/EGMA thermoplastic vulcanizates have gradually elongated disperse phases due to improved interfacial miscibility as evidenced by quantitative nanomechanical mapping AFM modulus analysis. Therefore, dynamic vulcanization is an effective and economical approach to integrate thermoplasticity and rubber elasticity into plastic/rubber pairs with good phase miscibility. © 2023 Society of Industrial Chemistry.

Mobile Measurement of a Dynamic Field via Compressed Sensing

In this paper, measuring dynamic signals at points of interest (POIs) using a mobile agent is considered, where the agent is required to repeatedly measure at and transit between the POIs. Dynamic field sensing is needed in areas ranging from nanomechanical mapping of live sample to crop monitoring. Existing work on mobile sensing, however, has been focused on cooperatively tracking one or few known or unknown POIs, whereas the dynamics of the signals is ignored. Challenges arises from capturing and recovering the dynamics at each POI by using data intermittently measured, resulting in temporal-spatial coupling in the mobile sensing. Moreover, trade off between the sensing cost and the performance needs to be addressed. We propose a compressed-sensing based approach to tackle this problem. First, a check-and-removal process based on random permutation and partition of the measurement periods is developed to avoid the temporal-spatial coupling under agent speed constraint. Then a shuffle-and-pair process based on the simulate-annealing is proposed to minimize the transition distance while preserving the performance. It is shown that the distribution of the measurement periods between the POIs converges. The proposed approach is illustrated through a simulation study of measuring the temperature-dependent nanomechanical variations of a polymer sample.

Reactive Blending of Recycled Poly(ethylene terephthalate)/Recycled Polypropylene: Kinetics Modeling of Non-Isothermal Crystallization.

Plastics were developed to change our world for the better. However, plastic pollution has become a serious global environmental crisis. Thermoplastic polyesters and polyolefins are among the most abundant plastic waste. This work presents an in-depth non-isothermal crystallization kinetics analysis of recycled post-consumer poly(ethylene terephthalate) (rPET) and recycled polypropylene (rPP) blends prepared through reactive compounding. The effect of pyromellitic dianhydride (PMDA) on crystallization kinetics and phase morphology of rPET/rPP blends was investigated by differential scanning calorimetry (DSC) and microscopy techniques. DSC results showed that increasing rPP content accelerated rPET crystallization while reducing crystallinity, which indicates the nucleation effect of the rPP phase in blends. Further, it was found that the incorporation of PMDA increased the degree of crystallinity during non-isothermal crystallization, even though the rate of crystallinity decreased slightly due to its restriction effects. The non-isothermal crystallization kinetics was analyzed based on the theoretical models developed by Jeziorny, Ozawa, Mo, and Tobin. The activation energy of the crystallization process derived from Kissinger, Takhor, and Augis-Bennett models was found to increase in rPET/rPP blends with increasing PMDA due to hindered dynamics of the system. Rheological measurements revealed that rPET melt viscosity is remarkably increased in the presence of PMDA and reactive blending with rPP relevant for processing. Moreover, nanomechanical mapping of the rPP phase dispersed in the rPET matrix demonstrated the broadening of the interfacial domains after reactive blending due to the branching effect of PMDA. Findings from this study are essential for the recycling/upcycling thermoplastics through non-isothermal fabrication processes, such as extrusion and injection molding, to mitigate the lack of sorting options.

Development of a red persistent luminescent composite: Electrospun nanofiber polymer coating prevents emission quenching by water

Sulfide persistent luminescent materials are known for their efficient luminescence, although a limitation in application of these materials is their susceptibility to hydrolysis. In this work, the electrospinning technique was used to encapsulate polycrystalline SrS:Eu2+,Sm3+ within PVDF-co-HFP nanofibers, after which thermostimulated luminescence could be observed with the naked eye following treatment with boiling water, confirming that the storage capacity of the powder was maintained after encapsulation. The composite material presented high levels of flexibility, hydrophobicity, and thermal and chemical stability. The structural characteristics of the synthesized SrS:Eu2+,Sm3+ were elucidated using X-ray diffraction (XRD), infrared spectroscopy (FTIR), thermogravimetric analysis (TGA), and differential scanning calorimetry (DSC). The morphologies of the SrS:Eu2+,Sm3+ grains and the electrospun nanofibers were studied by scanning electron microscopy (SEM), transmission electron microscopy (TEM), atomic force microscopy (AFM), X-ray fluorescence (XRF) mapping, and scanning transmission X-ray microscopy (STXM). Mechanical properties including elastic modulus and deformation degree were evaluated by PeakForce quantitative nanomechanical mapping (PF-QNM). The luminescent properties were investigated using photoluminescence spectroscopy (PLS), diffuse reflectance spectroscopy (DRS), and X-ray excited optical luminescence (XEOL), which showed that the photonic functionality of the material in the nanofibers was preserved, even after severe treatment with boiling water.

Scalable Hierarchically Structured Materials from a Multiscale Particle System Enabled by Microscaffolds.

Structural hierarchy is the key to manufacturing multiscale particle-based composite materials. A novel manufacturing method was developed to generate scalable hierarchical structures in concrete. The new method used 3D-printed microscaffolds to interact with the multiscale particle packing in concrete, resulting in a structured lightweight composite material. The size of internal members can vary by more than two orders of magnitude, to adapt to different applications. Based on compression tests and microstructural investigation by optical microscope and quantitative nanomechanical mapping, we found that the new material is 63.93% more efficient in energy absorption capacity compared with traditional lightweight concrete. Our experimental trials also showed that introducing structural hierarchy can reduce the consumption of cementitious material in the system by up to 14% and significantly reduce the use of scaffolds. The method could be applied to a board spectrum of multiscale particle-based materials, such as dental cement and bone implant materials, to improve material performance and efficiency in medical and construction applications.

Application of nanoindentation technique in mechanical characterization of organic matter in shale: Attentive issues, test protocol, and technological prospect

The rise of global shale oil and gas exploration has prompted scholars to pay attention to the mechanics of organic matter (OM) in shale reservoirs. The depth-sensing nanoindentation, a rather mature micromechanical testing technique in materials science, was gradually introduced into the mechanical data acquisition of shale matrix OM in recent years. Although this introduction is commendable, there are still some attentive issues relating to the indentation condition (mainly the reasonable setting of indentation depth parameter) and interpretation of the measured mechanical data, considering the micron-sized and nanoporous nature of OM and its close contact with surrounding minerals in shale composite. This study carried out a series of OM-positioned indentations on a typical shale sample collected from Silurian Longmaxi Formation in South China, in which we specially used the dynamic mechanical analysis that determines the mechanical data as a continuous function of indentation depth. Through it, we evidenced two main indentation effects (i.e., the size and substrate) during indentation of OM, and quantified the depths they appear (at less than 200 nm and greater than 1/20 of OM thickness, respectively). They together limit the indentation depth to a narrow range and also restrict the use of nanoindentation for OM less than 2 μm. Additionally, the porous OM is measured to be in lower mechanical values, suggesting that the porous effect on the indentation response cannot be ignored, especially when making mechanical comparison between different OM particles. Furthermore, we compared nanoindentation with the newly developed AFM-based nanomechanical mapping technique, and proposed to combine the both to achieve a full-scale characterization of shale matrix OM. Overall, this study standardized the indentation conditions to measure the inherent mechanical data of shale matrix OM, which lays the foundation for the forseeable increasing demand for testing and research of OM mechanics in the future.

CD109-Regulated Mechanical Properties Of Endothelial Cells.

CD109 antigen on the endothelial cell surface plays an important role in vascular pathology. The aim of the work was to investigate the effect of the immobilization of CD109 antigen with specific antibodies on nanomechanical properties of human umbilical endothelial cells (HUVECs) using atomic force microscopy in quantitative nanomechanical property mapping mode (PeakForce QNM). Anti-CD109 antibodies induced significant stiffening of the cell surface (Me(LQ; UQ): in 1.45(1.07;2.29) times with respect to control cells for fixed cells and in 4.9(3.6;5.9) times with respect to control cells for living cells, and changes in the spatial distribution of cell surface mechanical properties. The changes in the HUVEC's mechanical properties were accompanied by the activation of the TGF-/Smad2/3 signaling pathway and reorganization of the vimentin and actin cytoskeletal elements. Our findings show that blocking CD109 antigen using anti-CD109 antibodies leads in HUVECs to the processes similar to that occur after cell TGF-β-signaling activation. Therefore, we suggest that CD109 antigen may be involved in regulating the mechanical behavior of endothelial cells. This article is protected by copyright. All rights reserved.