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The response of cells to environmental stimuli, under either physiological or pathological conditions, plays a key role in determining cell fate toward either adaptive survival or controlled death. The efficiency of such a feedback mechanism is closely related to the most challenging human diseases, including cancer. Since cellular responses are implemented through physical forces exerted on intracellular components, more detailed knowledge of force distribution through modern imaging techniques is needed to ensure a mechanistic understanding of these forces. In this work, we mapped these intracellular forces at a whole-cell scale and with submicron resolution to correlate intracellular force distribution to the cytoskeletal structures. Furthermore, we visualized dynamic mechanical responses of the cells adapting to environmental modulations in situ. Such task was achieved by using an informatics-assisted atomic force microscope (AFM) indentation technique where a key step was Markov-chain Monte Carlo optimization to search for both the models used to fit indentation force–displacement curves and probe geometry descriptors. We demonstrated force dynamics within cytoskeleton, as well as nucleoskeleton in living cells which were subjected to mechanical state modulation: myosin motor inhibition, micro-compression stimulation and geometrical confinement manipulation. Our results highlight the alteration in the intracellular prestress to attenuate environmental stimuli; to involve in cellular survival against mechanical signal-initiated death during cancer growth and metastasis; and to initiate cell migration. Quantitative knowledge of internal force distribution in living cells is crucial to understand how cellular machinery responds mechanically to changes in the extracellular environment. Conventional force-sensing methods, which rely on pre-attached sensors, are incapable of providing whole-cell information with high spatial resolution. By machine learning assistance, we could improve AFM indentation's force sensitivity to a level that now enabled force (prestress) to be mapped on whole living cells with submicron resolution. The new capability revealed that cells actively redistribute forces among intracellular components to mitigate negative influence from the environment, and such response mechanisms could be used to understand ailing cellular states like cancer. [ABSTRACT FROM AUTHOR]

Mechanical stimuli are fundamental in the development of organs and tissues, their growth, regeneration or disease. They influence the biochemical signals produced by the cells, and, consequently, the development and spreading of a disease. Moreover, tumour cells are usually characterized by a decrease in the cell mechanical properties that may be directly linked to their metastatic potential. Thus, recently, the experimental and computational study of cell biomechanics is facing a growing interest. Various experimental approaches have been implemented to describe the passive response of cells; however, cell variability and complex experimental procedures may affect the obtained mechanical properties. For this reason, in-silico computational models have been developed through the years, to overcome such limitations, while proposing valuable tools to understand cell mechanical behaviour. This being the case, we propose a combined continuous-tensegrity finite element (FE) model to analyse the mechanical response of a cell and its subcomponents, observing how every part contributes to the overall mechanical behaviour. We modelled both Atomic Force Microscopy (AFM) indentation and micropipette aspiration techniques, as common mechanical tests for cells and elucidated also the role of cell cytoplasm and cytoskeleton in the global cell mechanical response. [ABSTRACT FROM AUTHOR]

The early steps of embryogenesis are controlled exclusively by the quality of oocyte that linked closely to its mechanical properties. The mechanical properties of an oocyte were commonly characterized by assuming it was homogeneous such that the result deviated significantly from the true fact that it was composed of subcellular components. In this work, we accessed and characterized the subcellular components of the oocytes and developed a layered high-fidelity finite element model for describing the viscoelastic responses of an oocyte under loading. The zona pellucida (ZP) and cytoplasm were isolated from an oocyte using an in-house robotic micromanipulation platform and placed on AFM to separately characterizing their mechanical profiling by analyzing the creep behavior with the force clamping technique. The spring and damping parameters of a Kelvin–Voigt model were derived by fitting the creeping curve to the model, which were used to define the shear relaxation modulus and relaxation time of ZP or cytoplasm in the ZP and cytoplasm model. In the micropipette aspiration experiment, the model was accurate sufficiently to deliver the time-varying aspiration depth of the oocytes under the step negative pressure of a micropipette. In the micropipette microinjection experiment, the model accurately described the intracellular strain introduced by the penetration. The developed oocyte FEM model has implications for further investigating the viscoelastic responses of the oocytes under different loading settings. [ABSTRACT FROM AUTHOR]

Atomic force microscopy (AFM) indentation experiments are commonly used to study the mechanical properties of graphene, such as Young's modulus and strength. However, applied AFM indentation forces on suspended graphene beams or ribbons are typically limited to several tens of nanonewtons due to the extreme thinness of graphene and their sensitivity to damage caused by the AFM tip indentation. Herein, graphene ribbons with a Si mass attached to their center position are employed, allowing us to introduce an unprecedented, wide range of AFM indentation forces (0–6800 nN) to graphene ribbons before the graphene ribbons are ruptured. It is found that the Young's modulus of double‐layer graphene ribbons decreases as the applied AFM indentation force is larger than ≈3000 nN, which indicates that the stiffness of double‐layer graphene ribbons remains constant before exposing them to AFM indentation forces larger than ≈3000 nN. [ABSTRACT FROM AUTHOR]

The early steps of embryogenesis are controlled exclusively by the quality of oocyte that linked closely to its mechanical properties. The mechanical properties of an oocyte were commonly characterized by assuming it was homogeneous such that the result deviated significantly from the true fact that it was composed of subcellular components. In this work, we accessed and characterized the subcellular components of the oocytes and developed a layered high-fidelity finite element model for describing the viscoelastic responses of an oocyte under loading. The zona pellucida (ZP) and cytoplasm were isolated from an oocyte using an in-house robotic micromanipulation platform and placed on AFM to separately characterizing their mechanical profiling by analyzing the creep behavior with the force clamping technique. The spring and damping parameters of a Kelvin–Voigt model were derived by fitting the creeping curve to the model, which were used to define the shear relaxation modulus and relaxation time of ZP or cytoplasm in the ZP and cytoplasm model. In the micropipette aspiration experiment, the model was accurate sufficiently to deliver the time-varying aspiration depth of the oocytes under the step negative pressure of a micropipette. In the micropipette microinjection experiment, the model accurately described the intracellular strain introduced by the penetration. The developed oocyte FEM model has implications for further investigating the viscoelastic responses of the oocytes under different loading settings.

Hydrogel substrates with a stiffness gradient have been used as a surrogate of the extracellular matrix (ECM) to investigate how cells respond to the stiffness of their surrounding matrix. Various fabrication methods have been proposed to create a stiffness gradient in the hydrogel substrate, and some of them rely on generating a concentration gradient in a prepolymer solution before photo-polymerization. One easy way to do so is to coalesce two prepolymer solution drops of different stiffness values in a narrow confinement formed by two glass surfaces and then to induce polymerization using ultraviolet (UV) light irradiation, as proposed by Lo et al. [Biophys. J. 2000, 79:144–152]. We have improved their method to enable modulating the obtained stiffness gradient and characterized fabricated polyacrylamide (PAAM) gels. We controlled the coalescence and mixing duration of two prepolymer drops using the lab-built Hele-Shaw cell device and glass surfaces with a superhydrophobic barrier. Limited mixing between the drops created a concentration gradient of the gel ingredient, which was converted to a stiffness gradient by UV-based photo-polymerization. Atomic force microscopy (AFM) indentation showed that the fabricated gels had the stiffness gradient zone at the center and that the width of the zone increased with the mixing duration. [ABSTRACT FROM AUTHOR]

Abstract This paper investigates the elastic properties of bone tissue in the adult mouse femur through Atomic Force Microscopy (AFM) indentation with the goal of understanding its microstructure and underlying mechanics at the nano length scale. Both trabecular and cortical bone types are studied. In particular, we examined the elasticity of cortical bone and individual trabeculae in the longitudinal and transverse directions of the samples. For cortical bone, the elastic modulus in the longitudinal direction was found to be 10–15% higher than that in the transverse direction; for trabecular bone, this difference was 42%. For the trabeculae, this value was found to be in a lower range (0.92 ± 0.22 GPa). As per the transverse elastic modulus, an average of 1.58 ± 0.36 GPa was measured for cortical bone, and 0.55 ± 0.21 GPa for trabecular bone. The anisotropy ratio was within the range of 1.2–1.5 for cortical bone and 1.7–2 for trabecular bone. While the elastic modulus of cortical bone varied along the length of the femur with up to 30% variation, no significant differences were observed within each transverse section. The effect of indentation frequency (1–500 Hz) on the longitudinal elastic moduli was also investigated for cortical and trabecular bone, with results showing a correlation between indentation frequency and elastic modulus. Statement of significance: This study examines the adult mouse femur with a twofold aim: to investigate the anisotropy and inhomogeneity of cortical and trabecular bone tissues and to elucidate their elastic behavior at the nanometer length scale. The elastic moduli of cortical bone and individual trabecula are measured in the longitudinal and transverse cross-sections via AFM indentation at selected locations and in specific directions of the adult mouse femur. The results provide insights into the relationship between mechanical properties and structural morphology of cortical and trabecular bone tissue. [ABSTRACT FROM AUTHOR]

Stiffness of the arterial wall and atherosclerotic plaque components is a determinant of the stress field within plaques, which has been suggested to be an indicator of plaque vulnerability. The diversity and inhomogeneous structure of atherosclerotic lesions complicate the characterization of plaque components. In the present study, stiffness of the arterial wall and atherosclerotic plaque components in human coronary arteries was examined in early and developed atherosclerotic lesions. The force-spectroscopy mode of the atomic force microscope and histological examination were used for determination of elastic moduli at specified locations within samples. Fibrous cap (E = 14.1 ± 3.8 kPa) showed lower stiffness than the fibrous tissue beneath the lipid pool (E = 17.6 ± 3.2 kPa). Calcification zones (E = 96.1 ± 18.8 kPa) and lipid pools (E = 2.7 ± 1.8 kPa) were the stiffest and softest components of atherosclerotic lesions, respectively. The increase of media stiffness (%44.8) and reduction of the elastic modulus of the internal elastic lamina (%28.9) was observed in coronary arteries. Moreover, significant differences were observed between the stiffness of medial layer in diseased parts and free-plaque segments in incomplete plaques of coronary arteries. Our results can be used for better understanding of remodeling mechanisms of the arterial wall with plaque development. Graphical abstract Stiffness alteration of the arterial wall and atherosclerotic plaque components with plaque development in coronary arteries. [ABSTRACT FROM AUTHOR]

Atherosclerosis is a major risk factor for cardiovascular disease. However, mechanisms of interaction of atherosclerotic plaque development and local stiffness of the lamellar structure of the arterial wall are not well established. In the current study, the local Young's modulus of the wall and plaque components were determined for three different groups of healthy, mildly diseased and advanced atherosclerotic human abdominal aortas. Histological staining was performed to highlight the atherosclerotic plaque components and lamellar structure of the aortic media, consisting of concentric layers of elastin and interlamellar zones. The force spectroscopy mode of the atomic force microscopy was utilized to determine Young's moduli of aortic wall lamellae and plaque components at the micron level. The high variability of Young's moduli (E) at different locations of the atherosclerotic plaque such as the fibrous cap (E = 15.5± 2.6 kPa), calcification zone (E = 103.7±19.5 kPa), and lipid pool (E = 3.5±1.2 kPa) were observed. Reduction of elastin lamellae stiffness (18.6%), as well as stiffening of interlamellar zones (50%), were detected in the diseased portion of the medial layer of abdominal aortic wall compared to the healthy artery. Additionally, significant differences in the stiffness of both elastin lamellae and interlamellar zones were observed between the diseased wall and disease-free wall in incomplete plaques. Our results elucidate the alternation of the stiffness of different lamellae in the human abdominal aortic wall with atherosclerotic plaque development and may provide new insight on the remodeling of the aortic wall during the progression of atherosclerosis. [ABSTRACT FROM AUTHOR]

We combine non-contact atomic force microscopy (AFM) imaging and AFM indentation in ultra-high vacuum to quantitatively and reproducibly determine the hardness and deformation mechanisms of Pt(111) and a Pt57.5Cu14.7Ni5.3P22.5 metallic glass with unprecedented spatial resolution. Our results on plastic deformation mechanisms of crystalline Pt(111) are consistent with the discrete mechanisms established for larger scales: Plasticity is mediated by dislocation gliding and no rate dependence is observed. For the metallic glass we have discovered that plastic deformation at the nanometer scale is not discrete but continuous and localized around the indenter, and does not exhibit rate dependence. This contrasts with the observation of serrated, rate-dependent flow of metallic glasses at larger scales. Our results reveal a lower size limit for metallic glasses below which shear transformation mechanisms are not activated by indentation. In the case of metallic glass, we conclude that the energy stored in the stressed volume during nanometer-scale indentation is insufficient to account for the interfacial energy of a shear band in the glassy matrix.

Abstract Characterization of the interphase region in carbon fiber reinforced polymer (CFRP) is challenging because of the length scale involved. The interpretation of measured load-displacement curves using indentation is affected by the lack of analytical solutions that account for the fiber constraint effect. A combination of AFM (Atomic Force Microscopy) based indentation and FE (Finite Element) simulations showed a gradient in the elastic modulus of the interphase evaluated along a radial line from the fiber. 3D FEA (Finite Element Analysis) indicated that fiber constraint effect is significant in the region less than 40 nm away from the fiber. Nonetheless, the apparent rise in elastic modulus due to fiber constraint is limited when compared to the gradient in the elastic modulus of the interphase. Additionally, this technique is used to demonstrate that UV irradiation causes a rapid decrease in the modulus of the region near the fiber due to photocatalytic degradation of carbon fiber but subsequently increases due to high cross-linking. Whereas, the modulus of the matrix at 8 mm away from the fiber decreased by 32% after 24 h of UV irradiation. This indicates that the response of epoxy to UV irradiation is influenced by the proximity to the reinforcement. Graphical Abstract Highlights • An apparent increase in interphase modulus due to fiber constraint is limited to 40 nm away from fiber. • The contribution of fiber constraint is less significant considering gradient in interphase modulus. • Response of epoxy to UV irradiation is influenced by the proximity to the fiber reinforcement. • Photocatalytic degradation of carbon fiber due to UV irradiation impacts interphase modulus. [ABSTRACT FROM AUTHOR]

Sonoporation is applied to enhance the permeability of the cell to bioactive materials by employing the acoustic cavitation of microbubbles. This phenomena would be helpful in molecular biology, delivery of large molecules into the cells and gene therapy. Many methods have been applied to monitor the biological effects and trace of sonoporation on the cells such as scanning/transmission electron microscopy, confocal imaging and flow cytometry. Here, we monitored the effect of sonoporation on the cells using electrochemical method with an integrated three electrode system. Electrochemical responses of stimulated cells, compared to flow cytometry and electron microscopy results, presented different patterns of sonoporation in the cells detectable by cyclic voltammetry. In addition, confocal microscopy from actin stress fibers and young’s modulus measured by AFM revealed the correlation of cell mechanics and amount of induced sonopores in the cells. This method could be applied as a new trend in cellular mechanochemical studies. [ABSTRACT FROM AUTHOR]

Recent studies have indicated that the nanoindentation measured stiffness of carcinoma adherent cells is in general lower than normal cells, thus suggesting that cell stiffness may serve as a bio-marker for carcinoma. However, the proper establishment of such a conclusion would require biophysical understanding of the underlying mechanism of the cell stiffness. In this work, we compared the elastic moduli of the actin cytoskeletons of Hey A8 ovarian carcinoma cells with and without metastasis (HM and NM), as measured by 2D atomic force microscopy (AFM) with low-depth nanoindentation via a rate-jump method. The results indicate clearly that HM cells showed lower actin cytoskeleton stiffness atop of their nucleus position and higher actin cytoskeleton stiffness at their rims, compared to NM cells, suggesting that the local stiffness on the cytoskeleton can reflect actin filament distribution. Immunofluorescence staining and scanning electron microscopy (SEM) also indicated that the difference in stiffness in Hey A8 cells with different metastasis is associated with their F-actin rearrangement. Finite-element modelling (FEM) shows that a migrating cell would have its actin filaments bundled together to form stress fibers, which would exhibit lower indentation stiffness than the less aligned arrangement of filaments in a non-migrating cell. The results here indicate that the actin cytoskeleton stiffness can serve as a reliable marker for grading the metastasis of adherent carcinoma cells due to their cytoskeleton change and potentially predicting the migration direction of the cells. [ABSTRACT FROM AUTHOR]

Hard, scratch-resistant and transparent fluoropolymer-based hybrid coatings are successfully prepared through the sol-gel chemistry and investigated, in the attempt to correlate the chemical, physical and surface properties of these materials with the mechanical properties (i.e. hardness and elastic modulus) measured at the nanoscale by atomic force microscopy (AFM). The organic component of these coatings consists in a hydroxyl-functional fluoropolymer resin, which belongs to the class of chlorotrifluoroethylene-vinylether copolymers and exhibits remarkable properties such as easy handling, great weather resistance, good adhesion and flexibility of coatings. A functionalization of this copolymer is also performed using an isocyanate-functionalized silane in order to assure a covalent cross-linking of organic fluorinated resin with inorganic phases. The combination of the silanized chlorotrifluoroethylene-vinylether copolymer with different sols composed of silica and zirconia is used to obtain high scratch resistance and high durability coatings on polycarbonate. A series of three hybrid coatings with different zirconia/silica molar ratios (0.09/0.48) are developed and analyzed by differential scanning calorimetry (DSC), water contact angle measurements, pencil hardness and adhesion tests. AFM scratch hardness, coating wettability and surface composition measured by Fourier transform infrared spectroscopy (FTIR) are regularly monitored over long-term UV–vis light exposure, to assess the durability of the hybrid coatings. Interestingly, the hybrid fluorinated coatings exhibit an improved scratch resistance and a superior long-term stability when exposed to an accelerated weathering, compared to pristine PC substrates. The mixed silica/zirconia hybrid coatings with a low and intermediate zirconia-to-silica ratio also show excellent mechanical strength, high level of hardness and superior integrity after long-term light exposure. [ABSTRACT FROM AUTHOR]

Hertz contact theory has been widely used for the determination of cell elasticity based on AFM indentation experiments. In light of the adhesive contact between AFM tip and cell, this study applied Johnson–Kendall–Roberts ( JKR ) model to fit the indentation force–displacement ( F – D ) curves reported previously. A MIN6 cell has been modeled as first a sphere and then a flattened cell with different thicknesses. The results have shown that both basic JKR model and “generalized” JKR model can best describe the unloading force–displacement behaviors of the indentation curves. The Young׳s modulus of the cell and the work of adhesion of the cell–indenter interface are obtained. In comparison to the Hertzian contact model, the JKR model provides obviously better fitting to the experimental results, indicating that the adhesion is significant in the cell interaction. [ABSTRACT FROM AUTHOR]

Microscopically homogeneous and thin fluorinated hybrid coatings showing significantly high mechanical and photochemical durability are prepared through the sol-gel chemistry and studied in this work. Their physico-chemical and nanomechanical properties are investigated. In this study, a special focus is given to the effects of organic/inorganic ratio and inorganic precursor type on surface and mechanical properties at the nanoscale. Different coatings containing low and high contents of silica as well as a mixed oxide composed of high amount of titania and low amount of silica are investigated. Nanoscratch and nanoindentation tests are performed by atomic force microscopy (AFM) and employed to assess the nanomechanical behavior of the hybrid coatings at a proper scale. Dynamic hardness results obtained from AFM are found to be in good agreement with technological pencil scratch tests performed at a macroscopic scale. Owing to the high inorganic content and good cross-linking degree achieved, coatings with a high amount of silica show considerably high dynamic hardness (nearly 2 GPa), which remains unchanged even after a long-term UV-vis light exposure. Notwithstanding the comparable inorganic amount, the hardness of mixed oxide coatings is markedly lower than high silica coatings and is subjected to modifications when exposed to UV-vis light in the long term, as also corroborated by x-ray photoelectron spectroscopy (XPS). Young's modulus results obtained by AFM nanoindentations appear to be dependent mostly on the inorganic/organic ratio. Higher moduli are determined for samples with higher inorganic/organic ratio, and no particular effect of the nature of inorganic precursor upon such property is observed. [ABSTRACT FROM AUTHOR]

The properties of polymers near an interface are altered relative to their bulk value due both to chemical interaction and geometric confinement effects. For the past two decades, the dynamics of polymers in confined geometries (thin polymer film or nanocomposites with high-surface area particles) has been studied in detail, allowing progress to be made toward understanding the origin of the dynamic effects near interfaces. Observations of mechanical property enhancements in polymer nanocomposites have been attributed to similar origins. However, the existing measurement methods of these local mechanical properties have resulted in a variety of conflicting results on the change of mechanical properties of confined polymers. Here, an atomic force microscopy (AFM)-based method is demonstrated that directly measures the mechanical properties of polymers adjacent to a substrate with nanometer resolution. This method allows us to consistently observe the gradient in mechanical properties away from a substrate in various materials systems, and paves the way for a unified understanding of thermodynamic and mechanical response of polymers. This gradient is both longer (up to 170 nm) and of higher magnitude (50% increase) than expected from prior results. Through the use of this technique, we will be better able to understand how to design polymer nanocomposites and polymeric structures at the smallest length scale, which affects the fields of structures, electronics, and healthcare. [ABSTRACT FROM AUTHOR]

As ultra-thin films or small-scale structures become widely used in electronics and biology, knowledge concerning their near-surface mechanical properties of the materials is increasingly important. Atomic force microscopy (AFM) is employed to determine near-surface elastic modulus via force-penetration curves acquired during indentation. Samples include polydimethylsiloxane (PDMS), parylene, mica, and single-crystal silicon, and indentations are performed with single-crystal silicon and silicon nitride AFM tips. An analysis algorithm based on the secant modulus method is proposed to extract the true penetration curves from the experimental displacement curves. The penetration data is then analyzed in terms of Hertzian model to estimate the elastic modulus. Three concerns in applying nanoscale AFM indentation to the measurement of the elastic modulus of an ultra-thin material are addressed. First, the effect of the lateral force caused by the inclined angle of the cantilevered probe is investigated theoretically and by numerical simulation. A second concern is local plastic deformation induced by a sharp probe tip. In this case, numerical results show a relatively small effect on the force-penetration curves if the plastic deformation is limited to the central area below the probe tip. The deviation of the elastic-plastic simulation from the elastic estimation depends on the yield strength of the material. Finally, the effect of stiffness matching between the AFM probe and the sample is a key issue that is studied numerically, and appropriate stiffness matching criteria are suggested. [ABSTRACT FROM AUTHOR]

Abstract: Living cell cultures exhibit improved adhesion on polymer surfaces engineered with nano-scale structures as compared to their flat counterparts. During fabrication their polymer-chain structure can be altered, thus affecting their mechanical properties. Here, we demonstrate using atomic-force-microscope nanoindentation that the modulus of nanostructured PDMS is doubled, while that of nanostructured ORMOCER increases by an order of magnitude, when compared to their flat counterparts. [Copyright &y& Elsevier]

Abstract: To investigate the roles of intercellular gap junctions and extracellular ATP diffusion in bone cell calcium signaling propagation in bone tissue, in vitro bone cell networks were constructed by using microcontact printing and self-assembled monolayer technologies. In the network, neighboring cells were interconnected through functional gap junctions. A single cell at the center of the network was mechanically stimulated by using an AFM nanoindenter. Intracellular calcium ([Ca2+]i) responses of the bone cell network were recorded and analyzed. In the untreated groups, calcium propagation from the stimulated cell to neighboring cells was observed in 40% of the tests. No significant difference was observed in this percentage when the intercellular gap junctions were blocked. This number, however, decreased to 10% in the extracellular ATP-pathway-blocked group. When both the gap junction and ATP pathways were blocked, intercellular calcium waves were abolished. When the intracellular calcium store in ER was depleted, the indented cell can generate calcium transients, but no [Ca2+]i signal can be propagated to the neighboring cells. No [Ca2+]i response was detected in the cell network when the extracellular calcium source was removed. These findings identified the biochemical pathways involved in the calcium signaling propagation in bone cell networks. [Copyright &y& Elsevier]

The objective of this study was to compare micro- and macroscale friction coefficients of bovine articular cartilage. Microscale measurements were performed using standard atomic force microscopy (AFM) techniques, using a 5 μm spherical probe tip. Twenty-four cylindrical osteochondral plugs were harvested in pairs from adjacent positions in six fresh bovine humeral heads (4–6 months old), and divided into two groups for AFM and macroscopic friction measurements. AFM measurements of friction were observed to be time-independent, whereas macroscale measurements demonstrated the well-documented time-dependent increase from a minimum to an equilibrium value. The microscale AFM friction coefficient (μAFM, 0.152±0.079) and macroscale equilibrium friction coefficient (μeq, 0.138±0.036) exhibited no statistical differences (p=0.50), while the macroscale minimum friction coefficient (μmin, 0.004±0.001) was significantly smaller than μeq and μAFM (p<0.0001). Variations in articular surface roughness (Rq=462±216 nm) did not correlate significantly with μAFM, μeq or μmin. The effective compressive modulus determined from AFM indentation tests using a Hertz contact analysis was E*=45.8±18.8 kPa. The main finding of this study is that μAFM is more representative of the macroscale equilibrium friction coefficient, which represents the frictional response in the absence of cartilage interstitial fluid pressurization. These results suggest that AFM measurements may be highly suited for exploring the role of boundary lubricants in diarthrodial joint lubrication independently of the confounding effect of fluid pressurization to provide greater insight into articular cartilage lubrication. [Copyright &y& Elsevier]

Follicle development in the ovary must be tightly regulated to ensure cyclical release of oocytes (ovulation). Disruption of this process is a common cause of infertility, for example via polycystic ovary syndrome (PCOS) and premature ovarian insufficiency (POI). Recent ex vivo studies suggest that follicle growth is mechanically regulated, however, crucially, the actual mechanical properties of the follicle microenvironment have remained unknown. Here we use atomic force microscopy (AFM) spherical probe indentation to map and quantify the mechanical microenvironment in the mouse ovary, at high resolution and across the entire width of the intact (bisected) ovarian interior. Averaging over the entire organ, we find the ovary to be a fairly soft tissue comparable to fat or kidney (mean Young's Modulus 3.3±2.5 kPa). This average, however, conceals substantial spatial variations, with the overall range of tissue stiffnesses from c. 0.5–10 kPa, challenging the concept that a single Young's Modulus can effectively summarize this complex organ. Considering the internal architecture of the ovary, we find that stiffness is low at the edge and centre which are dominated by stromal tissue, and highest in an intermediate zone that is dominated by large developmentally-advanced follicles, confirmed by comparison with immunohistology images. These results suggest that large follicles are mechanically dominant structures in the ovary, contrasting with previous expectations that collagen-rich stroma would dominate. Extending our study to the highest resolutions (c. 5 μm) showed substantial mechanical variations within the larger zones, even over very short (sub-100 μm) lengths, and especially within the stiffer regions of the ovary. Taken together, our results provide a new, physiologically accurate, framework for ovarian biomechanics and follicle tissue engineering. [ABSTRACT FROM AUTHOR]

Bone tissue remodels throughout life in response to mechanical loads. Impaired activities of bone cells (osteocytes, osteoblasts and osteoclasts) result in a disruption of the bone remodelling cycle, which eventually leads to bone disorders such as osteoporosis. To develop efficient therapeutic strategies against bone disorders, new tools are needed to unravel the bone remodelling cycle at the molecular level. Here, we developed a microfluidic platform, which should allow understanding the bone remodelling cycle in much more detail and ultimately be used to discover new therapeutic compounds. We focused specifically on studying cell-cell communication between osteocytes and osteoblasts cells via connexin 43-gap junctions. Therefore, a new cell printing method was developed to create living cellular bone cell arrays in a microfluidic channel. Several cell printing designs where osteocytes and osteoblasts heterotypically interacted at localized interfaces were evaluated. Physical contacts between the bone cells were characterized at high resolution by correlative atomic force microscopy (AFM)-fluorescence microscopy. We demonstrated that the platform is compatible with single-cell mechanostimulation by AFM nanoindentation and subsequent fluorescent analysis of the mechanoresponse. As a proof of concept, we showed the functionality of the platform by analysing the induced in vivo -like Ca ++ wave in the printed osteocyte-osteoblast network upon mechanical stimulation by fluid flow shear stress., (© 2022 IOP Publishing Ltd.)

Drosophila egg chamber development requires cellular and molecular mechanisms controlling morphogenesis. Previous research has shown that the mechanical properties of the basement membrane contribute to tissue elongation of the egg chamber. Here, we discuss how indentation with the microindenter of an atomic force microscope can be used to determine an effective stiffness value of a Drosophila egg chamber. We provide information on the preparation of egg chambers prior to the measurement, dish coating, the actual atomic force microscope measurement process, and data analysis. Furthermore, we discuss how to interpret acquired data and which mechanical components are expected to influence measured stiffness values., (© 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.)

In-plane strains are commonly found in two-dimensional (2D) metal halide organic-inorganic perovskites (HOIPs). The in-plane mechanical properties of 2D HOIPs are vital for mitigating the strain-induced stability issues of 2D HOIPs, yet their structure and mechanical property relationship largely remains unknown. Here, we employed atomic force microscope indentation to systematically investigate the in-plane Young's moduli E ∥ of 2D lead halide Ruddlesden-Popper HOIPs with a general formula of (R-NH 3 ) 2 PbX 4 , where the spacer molecules R-NH 3 + are linear alkylammonium cations (C m H 2 m +1 -NH 3 + , m = 4, 6, 8, or 12) and X = I, Br, or Cl. Fixing the spacer molecule to butylammonium, we discovered that the E ∥ of 2D HOIPs generally follows the trend of Pb-X bond strength, different from the tendency found in the out-of-plane moduli E ⊥ , showing more prominent effects of the metal halide inorganic framework on E ∥ than E ⊥ . E ∥ exhibits nonmonotonic dependence on the chain length of the linear alkyl spacer molecules, which would first decrease and plateau but then increase again. This is likely due to the competition of the bond strength and structural distortion in the inorganic layer, the relative fraction of the soft organic spacers, and the interfacial mechanical coupling associated with the interdigitation of the alkyl chains. The mechanical anisotropy of 2D HOIPs, marked by E ∥ / E ⊥ , shows wide tunability based on structural composition, particularly for iodide-based 2D HOIPs. Our results provide valuable insights into the structure-property relationships regarding the mechanical anisotropy and in-plane mechanical behaviors of 2D HOIPs, which can guide the materials design and device optimization to achieve required mechanical performance in 2D HOIP-based applications.