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Alzheimer's disease is a severe neurodegenerative disorder, and the discovery of biomarkers is crucial for early diagnosis. While the analysis of biomarkers in cerebrospinal fluid is well accepted, there are currently no blood biomarkers available. Our research focuses on identifying novel plasma biomarkers for Alzheimer's disease. To achieve this, we employed a technique that involves coupling human plasma to mouse organotypic brain slices via microcontact prints. After culturing for two weeks, we assessed Iba1-immunopositive microglia on these microcontact prints. We hypothesized that plasma from Alzheimer's patients contains factors that affect microglial migration. Our data indicated that plasma from Alzheimer's patients significantly inhibited the migration of round Iba1-immunoreactive microglia (13 ± 3, n = 24, p = 0.01) compared to healthy controls (50 ± 16, n = 23). Based on these findings, we selected the most promising plasma samples and conducted mass spectrometry using a differential approach, and we identified four potential biomarkers: mannose-binding protein C, macrophage receptor MARCO, complement factor H-related protein-3, and C-reactive protein. Our method represents a novel and innovative approach to translate research findings from mouse models to human applications. [ABSTRACT FROM AUTHOR]

Soft lithography, in particular microcontact printing (µCP), represents a well‐established and widespread class of lithographic patterning techniques. It is based on a directed deposition of molecules or colloidal particles by a transfer process with a micro‐structured stamp. A critical aspect of µCP is the adhesion cascade that enables the directed transfer of the objects. Here, the interfacial properties of a µCP‐stamp are tuned electrochemically to modify the adhesion cascade. During the printing process, the µCP‐stamp is submerged in an electrolyte solution and acted as a working electrode whose surface properties depended on the externally applied potential, thus enabling in situ rapid switching of its adhesion properties. As a proof of principle, defined particle patterns are selectively removed from a monolayer of colloidal particles. The adhesion at the particle/solid interface and the transfer mechanisms are determined by using the colloidal probe technique based on atomic force microscopy (AFM). In this case, a single particle is brought into contact with an electrode with the same surface chemistry as the µCP‐stamp. Hence, it became possible to determine the electrochemical potential ranges suitable to establish an adhesion cascade. [ABSTRACT FROM AUTHOR]

Introduction: In native heart tissue, functions of cardiac fibroblasts (CFs) include synthesis, remodeling, and degradation of the extracellular matrix (ECM) as well as secreting factors that regulate cardiomyocyte (CM) function. The influence of direct co-culture and CF-derived ECM on CM mechanical function are not fully understood. Methods: Here we use an engineered culture platform that provides control over ECM geometry and substrate stiffness to evaluate the influence of iPSC-CFs, and the ECM they produce, on the mechanical function of iPSC-CMs. Mechanical analysis was performed using digital image correlation to quantify maximum contractile strain, spontaneous contraction rate, and full-field organization of the contractions. Results: When cultured alone, iPSC-CFs produce and remodel the ECM into fibers following the underlying 15° chevron patterned ECM. The substrates were decellularized and confirmed to have highly aligned fibers that covered a large fraction of the pattern area before reseeding with iPSC-CMs, alone or in co-culture with iPSC-CFs. When seeded on decellularized ECM, larger maximum contractile strains were observed in the co-culture condition compared to the CM Only condition. No significant difference was found in contractile strain between the Matrigel and decellularized ECM conditions; however, the spontaneous contraction rate was lower in the decellularized ECM condition. A methodology for quantifying alignment of cell contraction across the entire field of view was developed based on trajectories approximating the cell displacements during contraction. Trajectory alignment was unaltered by changes in culture or ECM conditions. Conclusions: These combined observations highlight the important role CFs play in vivo and the need for models that enable a quantitative approach to examine interactions between the CFs and CMs, as well as the interactions of these cells with the ECM. [ABSTRACT FROM AUTHOR]

Abstract Soft lithography, in particular microcontact printing (µCP), represents a well‐established and widespread class of lithographic patterning techniques. It is based on a directed deposition of molecules or colloidal particles by a transfer process with a micro‐structured stamp. A critical aspect of µCP is the adhesion cascade that enables the directed transfer of the objects. Here, the interfacial properties of a µCP‐stamp are tuned electrochemically to modify the adhesion cascade. During the printing process, the µCP‐stamp is submerged in an electrolyte solution and acted as a working electrode whose surface properties depended on the externally applied potential, thus enabling in situ rapid switching of its adhesion properties. As a proof of principle, defined particle patterns are selectively removed from a monolayer of colloidal particles. The adhesion at the particle/solid interface and the transfer mechanisms are determined by using the colloidal probe technique based on atomic force microscopy (AFM). In this case, a single particle is brought into contact with an electrode with the same surface chemistry as the µCP‐stamp. Hence, it became possible to determine the electrochemical potential ranges suitable to establish an adhesion cascade.

Alzheimer’s disease is a severe neurodegenerative disorder, and the discovery of biomarkers is crucial for early diagnosis. While the analysis of biomarkers in cerebrospinal fluid is well accepted, there are currently no blood biomarkers available. Our research focuses on identifying novel plasma biomarkers for Alzheimer’s disease. To achieve this, we employed a technique that involves coupling human plasma to mouse organotypic brain slices via microcontact prints. After culturing for two weeks, we assessed Iba1-immunopositive microglia on these microcontact prints. We hypothesized that plasma from Alzheimer’s patients contains factors that affect microglial migration. Our data indicated that plasma from Alzheimer’s patients significantly inhibited the migration of round Iba1-immunoreactive microglia (13 ± 3, n = 24, p = 0.01) compared to healthy controls (50 ± 16, n = 23). Based on these findings, we selected the most promising plasma samples and conducted mass spectrometry using a differential approach, and we identified four potential biomarkers: mannose-binding protein C, macrophage receptor MARCO, complement factor H-related protein-3, and C-reactive protein. Our method represents a novel and innovative approach to translate research findings from mouse models to human applications.

Background: Alzheimer’s disease is characterized by extracellular beta-amyloid plaques, intraneuronal tau neurofibrillary tangles and excessive neurodegeneration. The mechanisms of neuron degeneration and the potential of these neurons to form new nerve fibers for compensation remain elusive. The present study aimed to evaluate the impact of beta-amyloid and tau on new formations of nerve fibers from mouse organotypic brain slices connected to collagen-based microcontact prints. Methods: Organotypic brain slices of postnatal day 8–10 wild-type mice were connected to established collagen-based microcontact prints loaded with polyornithine to enhance nerve fiber outgrowth. Human beta-amyloid(42) or P301S mutated aggregated tau was co-loaded to the prints. Nerve fibers were immunohistochemically stained with neurofilament antibodies. The physiological activity of outgrown neurites was tested with neurotracer MiniRuby, voltage-sensitive dye FluoVolt, and calcium-sensitive dye Rhod-4. Results: Immunohistochemical staining revealed newly formed nerve fibers extending along the prints derived from the brain slices. While collagen-only microcontact prints stimulated nerve fiber growth, those loaded with polyornithine significantly enhanced nerve fiber outgrowth. Beta-amyloid(42) significantly increased the neurofilament-positive nerve fibers, while tau had only a weak effect. MiniRuby crystals, retrogradely transported along these newly formed nerve fibers, reached the hippocampus, while FluoVolt and Rhod-4 monitored electrical activity in newly formed nerve fibers. Conclusions: Our data provide evidence that intact nerve fibers can form along collagen-based microcontact prints from mouse brain slices. The Alzheimer’s peptide beta-amyloid(42) stimulates this growth, hinting at a neuroprotective function when physiologically active. This “brain-on-chip” model may offer a platform for screening bioactive factors or testing drug effects on nerve fiber growth.

Upstream exposure of platelets to activating proteins primes platelets for increased downstream adhesion, though the mechanics of platelet translocation before permanently arresting are not well understood. To investigate platelet translocation on platelet-binding proteins, primed platelets transient contacts with immobilized proteins were recorded and analyzed. Using a microfluidic channel, representative of a vascular graft, platelet-activating proteins were covalently attached to the upstream priming, center, and downstream capture positions. Image sequences of platelet interactions with the center protein were captured as platelet-rich plasma (PRP) was perfused through the channel. There was an increase in both platelet pause events and net platelet adhesion on von Willebrand factor, collagen, or fibrinogen following upstream exposure to the same protein. Upstream priming also caused a decrease in average platelet velocity. The duration of transient platelet arrests on the protein-coated surface and the distance that platelets travel between pause events depended on the protein with which they were interacting. The most significant increase in platelet pause events frequency and decrease in average velocity occurred on immobilized von Willebrand factor, compared to the control with no upstream priming. These results demonstrate that platelet priming increases downstream platelet-protein interactions prior to permanent adhesion.

Integrin-dependent adhesion to the extracellular matrix (ECM) mediates mechanosensing and signaling in response to altered microenvironmental conditions. In order to provide tissue- and organ-specific cues, the ECM is composed of many different proteins that temper the mechanical properties and provide the necessary structural diversity. Despite most human tissues being soft, the prevailing view from predominantly in vitro studies is that increased stiffness triggers effective cell spreading and activation of mechanosensitive signaling pathways. To address the functional coupling of ECM composition and matrix rigidity on compliant substrates, we developed a matrix spot array system to screen cell phenotypes against different ECM mixtures on defined substrate stiffnesses at high resolution. We applied this system to both cancer and normal cells and surprisingly identified ECM mixtures that support stiffness-insensitive cell spreading on soft substrates. Employing the motor-clutch model to simulate cell adhesion on biochemically distinct soft substrates, with varying numbers of available ECM–integrin–cytoskeleton (clutch) connections, we identified conditions in which spreading would be supported on soft matrices. Combining simulations and experiments, we show that cell spreading on soft is supported by increased clutch engagement on specific ECM mixtures and even augmented by the partial inhibition of actomyosin contractility. Thus, “stiff-like” spreading on soft is determined by a balance of a cell’s contractile and adhesive machinery. This provides a fundamental perspective for in vitro mechanobiology studies, identifying a mechanism through which cells spread, function, and signal effectively on soft substrates [ABSTRACT FROM AUTHOR]

In recent years, innovative cell-based biosensing systems have been developed, showing impact in healthcare and life science research. Now, there is a need to design mass-production processes to enable their commercialization and reach society. However, current protocols for their fabrication employ materials that are not optimal for industrial production, and their preparation requires several chemical coating steps, resulting in cumbersome protocols. We have developed a simplified two-step method for generating controlled cell patterns on PMMA, a durable and transparent material frequently employed in the mass manufacturing of microfluidic devices. It involves air plasma and microcontact printing. This approach allows the formation of well-defined cell arrays on PMMA without the need for blocking agents to define the patterns. Patterns of various adherent cell types in dozens of individual cell cultures, allowing the regulation of cell–material and cell–cell interactions, were developed. These cell patterns were integrated into a microfluidic device, and their viability for more than 20 h under controlled flow conditions was demonstrated. This work demonstrated the potential to adapt polymeric cytophobic materials to simple fabrication protocols of cell-based microsystems, leveraging the possibilities for commercialization. [ABSTRACT FROM AUTHOR]

Microcontact printing (CP) is widely used to guide neurons to form 2D networks for neuroscience research. However, it is still difficult to establish 3D neuronal cultures on the CP substrate even though 3D neuronal structures are able to recapitulate critical aspects of native tissue. Here, we demonstrate that the reduced cell-substrate adhesion caused by the CP substrate could conveniently facilitate the aggregate formation of large-scale 3D neuron cluster networks. Furthermore, based on the quantitative analysis of the calcium activity of the resulting cluster networks, the effect of cell seeding density and local restriction of the CP substrate on network dynamics was investigated in detail. The results revealed that cell aggregation degree, rather than cell number, could take on the main role of the generation of synchronized network-wide calcium oscillation (network bursts) in the 3D neuron cluster networks. This finding may provide new insights for easy and cell-saving construction of in vitro 3D pathological models of epilepsy, and into deciphering the onset and evolution of network bursts in developmental nerve systems. [ABSTRACT FROM AUTHOR]

Antibody arrays have great implications in many biomedical settings. However, commonly used patterning methods have difficulties in generating antibody arrays with both high resolution and multiplexity, limiting their applications. Here, a convenient and versatile technique for the patterning of multiple antibodies with resolution down to 20 µm is reported using micropillar‐focused droplet printing and microcontact printing. Droplets of antibody solutions are first printed and stably confined on the micropillars of a stamp, and then the antibodies absorbed on the micropillars are contact‐printed to the target substrate, generating antibody patterns faithfully replicating the micropillar array. The effect of different parameters on the patterning results is investigated, including hydrophobicity of the stamps, override time of the droplet printing, incubation time, and the diameters of the capillary tips and micropillars. To demonstrate the utility of the method, multiplex arrays of anti‐EpCAM and anti‐CD68 antibodies is generated to capture breast cancer cells and macrophages, respectively, on the same substrate, and successful capturing of individual cell types and enrichment among the cells are achieved. It is envision that this method would serve as a versatile and useful protein patterning tool for biomedical applications. [ABSTRACT FROM AUTHOR]

Axis formation and related spatial patterning are initiated by symmetry breaking during development. A geometrically confined culture of human pluripotent stem cells (hPSCs) mimics symmetry breaking and cell patterning. Using this, polarized spinal cord organoids (pSCOs) with a self‐organized dorsoventral (DV) organization are generated. The application of caudalization signals promoted regionalized cell differentiation along the radial axis and protrusion morphogenesis in confined hPSC colonies. These detached colonies grew into extended spinal cord‐like organoids, which established self‐ordered DV patterning along the long axis through the spontaneous expression of polarized DV patterning morphogens. The proportions of dorsal/ventral domains in the pSCOs can be controlled by the changes in the initial size of micropatterns, which altered the ratio of center‐edge cells in 2D. In mature pSCOs, highly synchronized neural activity is separately detected in the dorsal and ventral side, indicating functional as well as structural patterning established in the organoids. This study provides a simple and precisely controllable method to generate spatially ordered organoids for the understanding of the biological principles of cell patterning and axis formation during neural development. [ABSTRACT FROM AUTHOR]

Endothelial cells (ECs) elongate in the direction of blood flow, are stiffer, and are considered atheroprotective in areas of the vasculature where flow‐induced shear stress is high and unidirectional and are softer, atherogenic, and polygonal in areas experiencing oscillatory multidirectional flow. To understand the precise roles of EC mechanics and morphology in the uptake of therapeutic nanoparticles (NPs) by atherogenic endothelium, human aortic ECs are induced to adopt prescribed shapes and areas imposed by microcontact patterned adhesive islands. NP uptake per cell increases with increasing spreading area and decreases with increasing cell aspect ratio at constant cell spreading area. Biomechanical analysis shows that elongated cells exhibit higher cellular stress and stiffer membranes than cells with low aspect ratios, indicating a strong correlation between morphology, mechanical phenotype, and NP uptake. Further, ECs elongated by high laminar shear endocytosed NPs to a far lesser extent than those that are nonelongated in the chaotic, lower shear areas when cocultured in the same chamber. Results indicate that conditions leading to atherogenesis, such as low, chaotic shear‐induction of EC polygonal morphology may be used to increase the uptake of therapeutic NPs as a preventative measure against atherosclerosis. [ABSTRACT FROM AUTHOR]

Mechanical stress patterns emerging from collective cell behavior have been shown to play critical roles in morphogenesis, tissue repair, and cancer metastasis. In our previous work, we constrained valvular interstitial cell (VIC) monolayers on circular protein islands to study emergent behavior in a controlled manner and demonstrated that the general patterns of cell alignment, size, and apoptosis correlate with predicted mechanical stress fields if radially increasing stiffness or contractility are used in the computational models. However, these radially symmetric models did not predict the existence of local regions of dense aligned cells observed in seemingly random locations of individual aggregates. The goal of this study is to determine how the heterogeneities in cell behavior emerge over time and diverge from the predicted collective cell behavior. Cell-cell interactions in circular multicellular aggregates of VICs were studied with time-lapse imaging ranging from hours to days, and migration, proliferation, and traction stresses were measured. Our results indicate that elongated cells create strong local alignment within preconfluent cell populations on the microcontact printed protein islands. These cells influence the alignment of additional cells to create dense, locally aligned bands of cells which disrupt the predicted global behavior. Cells are highly elongated at the endpoints of the bands yet have decreased spread area in the middle and reduced mobility. Although traction stresses at the endpoints of bands are enhanced, even to the point of detaching aggregates from the culture surface, the cells in dense bands exhibit reduced proliferation, less nuclear YAP, and increased apoptotic rates indicating a low stress environment. These findings suggest that strong local cell-cell interactions between primary fibroblastic cells can disrupt the global collective cellular behavior leading to substantial heterogeneity of cell behaviors in constrained monolayers. This local emergent behavior within aggregated fibroblasts may play an important role in development and disease of connective tissues. Mechanical stress patterns emerging from collective cell behavior play critical roles in morphogenesis, tissue repair, and cancer metastasis. Much has been learned of these collective behaviors by utilizing microcontact printing to constrain cell monolayers (aggregates) into specific shapes. Here we utilize these tools along with long-term video microscopy tracking of individual aggregates to determine how heterogeneous collective behaviors unique to primary fibroblastic cells emerge over time and diverge from computed stress fields. We find that dense multicellular bands form from local collective behavior and disrupt the global collective behavior resulting in heterogeneous patterns of migration, traction stresses, proliferation, and apoptosis. This local emergent behavior within aggregated fibroblasts may play an important role in development and disease of connective tissues. [Display omitted] [ABSTRACT FROM AUTHOR]

Measuring the traction forces produced by cells provides insight into their behavior and physiological function. Here, we developed a technique (dubbed 'black dots') that microcontact prints a fluorescent micropattern onto a flexible substrate to measure cellular traction forces without constraining cell shape or needing to detach the cells. To demonstrate our technique, we assessed human platelets, which can generate a large range of forces within a population. We find platelets that exert more force have more spread area, are more circular, and have more uniformly distributed F-actin filaments. As a result of the high yield of data obtainable by this technique, we were able to evaluate multivariate mixed effects models with interaction terms and conduct a clustering analysis to identify clusters within our data. These statistical techniques demonstrated a complex relationship between spread area, circularity, F-actin dispersion, and platelet force, including cooperative effects that significantly associate with platelet traction forces. Cells produce contractile forces during division, migration, or wound healing. Measuring cellular forces provides insight into their health, behavior, and function. We developed a technique that calculates cellular forces by seeding cells onto a pattern and quantifying how much each cell displaces the pattern. This technique is capable of measuring hundreds of cells without needing to detach them. Using this technique to evaluate human platelets, we find that platelets exerting more force tend to have more spread area, are more circular in shape, and have more uniformly distributed cytoskeletal filaments. Due to our high yield of data, we were able to apply statistical techniques that revealed combinatorial effects between these factors. [Display omitted] [ABSTRACT FROM AUTHOR]

Advances in organic materials manufacturing have enabled the creation of electronic devices using solution‐processing techniques by employing soluble materials with high conductivity grade. In this exploratory study, the use of micro‐contact for poly(3,4‐ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) polymer ink deposition as high‐quality structured electrodes for organic field‐effect transistors (OFETs) in top‐contact geometry is demonstrated. The optimized OFET's solution‐processed fabrication is a promising strategy to be realized in the simple, cost‐effective roll‐to‐roll manufacturing processes. The electrical performance of the fabricated devices is comparable to transistors with gold electrodes prepared via vacuum deposition, and even exceeding the values of the charge carriers' mobilities and featuring lower contact resistance (Rc), due to lower charge‐carrier injection barrier for carbon‐based organic electrodes. An addition of multi‐walled carbon nanotubes to the PEDOT:PSS decreases Rc even further, changing the work function for better energy alignment with semiconductor materials. [ABSTRACT FROM AUTHOR]

Abstract Endothelial cells (ECs) elongate in the direction of blood flow, are stiffer, and are considered atheroprotective in areas of the vasculature where flow‐induced shear stress is high and unidirectional and are softer, atherogenic, and polygonal in areas experiencing oscillatory multidirectional flow. To understand the precise roles of EC mechanics and morphology in the uptake of therapeutic nanoparticles (NPs) by atherogenic endothelium, human aortic ECs are induced to adopt prescribed shapes and areas imposed by microcontact patterned adhesive islands. NP uptake per cell increases with increasing spreading area and decreases with increasing cell aspect ratio at constant cell spreading area. Biomechanical analysis shows that elongated cells exhibit higher cellular stress and stiffer membranes than cells with low aspect ratios, indicating a strong correlation between morphology, mechanical phenotype, and NP uptake. Further, ECs elongated by high laminar shear endocytosed NPs to a far lesser extent than those that are nonelongated in the chaotic, lower shear areas when cocultured in the same chamber. Results indicate that conditions leading to atherogenesis, such as low, chaotic shear‐induction of EC polygonal morphology may be used to increase the uptake of therapeutic NPs as a preventative measure against atherosclerosis.

Abstract Advances in organic materials manufacturing have enabled the creation of electronic devices using solution‐processing techniques by employing soluble materials with high conductivity grade. In this exploratory study, the use of micro‐contact for poly(3,4‐ethylenedioxythiophene) polystyrene sulfonate (PEDOT:PSS) polymer ink deposition as high‐quality structured electrodes for organic field‐effect transistors (OFETs) in top‐contact geometry is demonstrated. The optimized OFET's solution‐processed fabrication is a promising strategy to be realized in the simple, cost‐effective roll‐to‐roll manufacturing processes. The electrical performance of the fabricated devices is comparable to transistors with gold electrodes prepared via vacuum deposition, and even exceeding the values of the charge carriers’ mobilities and featuring lower contact resistance (Rc), due to lower charge‐carrier injection barrier for carbon‐based organic electrodes. An addition of multi‐walled carbon nanotubes to the PEDOT:PSS decreases Rc even further, changing the work function for better energy alignment with semiconductor materials.

This paper focuses on creating one-dimensional diffractive grooved structures of antigen proteins on glass substrates for the label-free detection of antibodies to dairy allergens. In particular, the fabrication of protein structures is carried out by combining microcontact printing with physisorption, imines coupling, and thiol-ene click chemistry. The work first sets up these patterning methods and discusses and compares the main aspects involved in them (structure, biolayer thickness, functionality, stability). Homogeneous periodic submicron structures of proteins are created and characterized by diffractive measurements, AFM, FESEM, and fluorescence scanning. Then, this patterning method is applied to proteins involved in cow milk allergy, and the resulting structures are implemented as optical transducers to sense specific immunoglobulins G. In particular, gratings of bovine serum albumin, casein, and β-lactoglobulin are created and assessed, reaching limits of detection in the range of 30–45 ng·mL−1 of unlabeled antibodies by diffractive biosensing. [ABSTRACT FROM AUTHOR]

In recent years, innovative cell-based biosensing systems have been developed, showing impact in healthcare and life science research. Now, there is a need to design mass-production processes to enable their commercialization and reach society. However, current protocols for their fabrication employ materials that are not optimal for industrial production, and their preparation requires several chemical coating steps, resulting in cumbersome protocols. We have developed a simplified two-step method for generating controlled cell patterns on PMMA, a durable and transparent material frequently employed in the mass manufacturing of microfluidic devices. It involves air plasma and microcontact printing. This approach allows the formation of well-defined cell arrays on PMMA without the need for blocking agents to define the patterns. Patterns of various adherent cell types in dozens of individual cell cultures, allowing the regulation of cell–material and cell–cell interactions, were developed. These cell patterns were integrated into a microfluidic device, and their viability for more than 20 h under controlled flow conditions was demonstrated. This work demonstrated the potential to adapt polymeric cytophobic materials to simple fabrication protocols of cell-based microsystems, leveraging the possibilities for commercialization.

Eph‐ephrin signaling acts as spatial cue to define the tissue boundaries, the axonal growth, or the organization of compartmentalized tissues in vertebrates. By the regulation of tension, adhesion, and repulsion, intermingling of cells expressing the membrane‐tethered ligand and cells expressing the membrane‐tethered receptor is prevented. Despite being surface‐bound, most of the studies addressing Eph‐ephrin interactions use soluble ligands, which lack the spatial component needed to study tissue patterning. Here, it is shown that spatial patterns of ephrin ligands can modulate the organization of the different compartments in organoid‐derived intestinal epithelial monolayers. A modified version of the microcontact printing technique is used to create spatial cues of ephrin ligand on basement membrane surrogates. It is shown that both ligand concentration and cellular density can impact the strength of the repulsive effect triggered by Eph‐ephrin signaling. Finally, it is demonstrated that by using micropatterned ephrin spatial cues one can modify the orientation of intestinal crypts and align them in the direction set by the pattern. The approach presented here is believed to be an excellent tool to study exogenous signal with relevant spatial distribution in vivo. [ABSTRACT FROM AUTHOR]

Alzheimer's disease (AD) is a neurodegenerative disease characterized by memory loss and progressive deterioration of cognitive functions. Being able to identify reliable biomarkers in easily available body fluids such as blood plasma is vital for the disease. To achieve this, we used a technique that applied human plasma to organotypic brain slice culture via microcontact printing. After a 2-week culture period, we performed immunolabeling for neurofilament and myelin oligodendrocyte glycoprotein (MOG) to visualize newly formed nerve fibers and oligodendrocytes. There was no significant change in the number of new nerve fibers in the AD plasma group compared to the healthy control group, while the length of the produced fibers significantly decreased. A significant increase in the number of MOG+ dots around these new fibers was detected in the patient group. According to our hypothesis, there are factors in the plasma of AD patients that affect the growth of new nerve fibers, which also affect the oligodendrocytes. Based on these findings, we selected the most promising plasma samples and conducted mass spectrometry using a differential approach and we identified three putative biomarkers: aldehyde-dehydrogenase 1A1, alpha-synuclein and protein S100-A4. Our method represents a novel and innovative approach for translating research findings from mouse models to human applications.

Microcontact printing (CP) is widely used to guide neurons to form 2D networks for neuroscience research. However, it is still difficult to establish 3D neuronal cultures on the CP substrate even though 3D neuronal structures are able to recapitulate critical aspects of native tissue. Here, we demonstrate that the reduced cell-substrate adhesion caused by the CP substrate could conveniently facilitate the aggregate formation of large-scale 3D neuron cluster networks. Furthermore, based on the quantitative analysis of the calcium activity of the resulting cluster networks, the effect of cell seeding density and local restriction of the CP substrate on network dynamics was investigated in detail. The results revealed that cell aggregation degree, rather than cell number, could take on the main role of the generation of synchronized network-wide calcium oscillation (network bursts) in the 3D neuron cluster networks. This finding may provide new insights for easy and cell-saving construction of in vitro 3D pathological models of epilepsy, and into deciphering the onset and evolution of network bursts in developmental nerve systems.

Chemical etching of silicon assisted by a number of catalysts is attracting increasing attention in the fabrication of silicon micro–nanostructures. In practical applications, metal-free catalysts, including carbon materials, have been focused on as alternative materials for the assisted etching of silicon. Although this anisotropic etching process is suitable for the fabrication of silicon micro–nanostructures due to its simplicity and cost effectiveness, a number of challenges remain, such as the formation of a porous layer and peeling of the catalyst by the gases produced during the etching process. We herein report vapor-phase etching assisted by graphene oxide and its mechanism in terms of reaction kinetics. By optimizing etching conditions, graphene oxide enhances the etching reaction in the vapor phase without the formation of a porous layer. We also demonstrated the formation of micrometer-sized pores in the desired areas by combining the microcontact printing of graphene oxide with silicon etching in the vapor phase. [ABSTRACT FROM AUTHOR]

Molecular surface gradients can constitute electric field landscapes and serve to control local cell adhesion and migration. Cellular responses to electric field landscapes may allow the discovery of routes to improve osseointegration of implants. Flat molecule aggregate landscapes of amine- or carboxyl-teminated dendrimers, amine-containing protein and polyelectrolytes were prepared on glass to provide lateral electric field gradients through their differing zeta potentials compared to the glass substrate. The local as well as the mesoscopic morphological responses of adhered osteoblasts (MG-63) with respect to the stripes were studied by means of Scanning Ion Conductance Microscopy (SICM) and Fluorescence Microscopy, in situ. A distinct spindle shape oriented parallel to the surface pattern as well as a preferential adhesion of the cells on the glass site have been observed at a stripe and spacing width of 20 μ m. Excessive ruffling is observed at the spindle poles, where the cells extend. To explain this effect of material preference and electro-deformation, we put forward a retraction mechanism, a localized form of double-sided cathodic taxis. [ABSTRACT FROM AUTHOR]

Ischemic stroke is a severe insult in the brain causing cell death, inflammation, and activation of microglia. Microglia are the immune cells of the brain and play a role in any inflammatory process during neurodegeneration. Microglia are round ameboid and migrate to the lesion site, where they differentiate into ramified forms and activated phagocytic microglia. On the other hand, microglia can also release growth factors to repair degeneration. The aim of the present study is to explore the migratory capacity of microglia after ischemic insults. Organotypic brain slices of the mouse cortex (300 μm) were prepared. In order to study migration, the slices were connected to collagen-loaded microcontact prints (with or without monocyte chemoattractant protein-1, MCP-1) on the membranes. Slices were stimulated with lipopolysaccharide (LPS) for maximal microglial activation. Ischemic insults were simulated with oxygen-glucose deprivation (OGD) and acidosis (pH 6.5) for 3 days. After 3 weeks in culture, slices were fixed and immunohistochemically stained for the microglial markers Iba1, CD11b and macrophage-like antigen. Our data show that Iba1+ microglia migrated along the microcontact prints, differentiate and phagocyte 1.0 μm fluorescent microbeads. LPS significantly enhanced the number of round ameboid migrating microglia, while OGD and acidosis enhanced the number of ramified activated microglia. The effect was not visible on slices without any μCP and was most potent in μCP with MCP-1. We conclude that OGD and acidosis activate ramification and exhibit a similar mechanism, while LPS only activates round ameboid microglia. Collagen-loaded microcontact prints connected to mouse brain slices are a potent method to study activation and migration of microglia ex vivo. [ABSTRACT FROM AUTHOR]

[Display omitted] Waxy hydrocarbons diffuse freely in polydimethylsiloxane (PDMS), and this capability can be leveraged to generate inexpensive surface micropatterns that modify adhesion and wetting. Patterns are created by placing a waxy Parafilm sheet on the back of a PDMS stamp containing microscale surface features. When heated, the paraffin liquefies and diffuses through the stamp, creating a thin liquid layer on the micropatterned stamp surface; when placed in contact with a target surface, the layer solidifies and is retained on the target when the stamp is removed. Micropatterns were generated on different materials and surface topographies; pattern geometry was evaluated using optical profilometry and changes in wetting were evaluated using contact angle goniometry. Diffusion of paraffin through PDMS was evaluated using XPS. Wax micropatterns have submicron lateral resolution and thickness ranging from 85 to 380 nm depending on contact time. By using XPS analysis to track paraffin diffusion within the PDMS stamp during this process, we estimate the diffusion coefficient to be 5.3 × 10−7 cm2/s at 65 °C. This means that the paraffin layer at the stamp surface replenishes in less than a second after stamping, so it can be used multiple times without re-inking to deposit complex, multi-layer paraffin patterns. [ABSTRACT FROM AUTHOR]

High-performance biosensors were fabricated by efficiently transferring enzyme onto Pt electrode surfaces using a polydimethylsiloxane (PDMS) stamp. Polypyrrole and Nafion were coated first on the electrode surface to act as permselective films for exclusion of both anionic and cationic electrooxidizable interfering compounds. A chitosan film then was electrochemically deposited to serve as an adhesive layer for enzyme immobilization. Glucose oxidase (GOx) was selected as a model enzyme for construction of a glucose biosensor, and a mixture of GOx and bovine serum albumin was stamped onto the chitosan-coated surface and subsequently crosslinked using glutaraldehyde vapor. For the optimized fabrication process, the biosensor exhibited excellent performance characteristics including a linear range up to 2 mM with sensitivity of 29.4 ± 1.3 μA mM-1 cm-2 and detection limit of 4.3 ± 1.7 μM (S/N = 3) as well as a rapid response time of ~2 s. In comparison to those previously described, this glucose biosensor exhibits an excellent combination of high sensitivity, low detection limit, rapid response time, and good selectivity. Thus, these results support the use of PDMS stamping as an effective enzyme deposition method for electroenzymatic biosensor fabrication, which may prove especially useful for the deposition of enzyme at selected sites on microelectrode array microprobes of the kind used for neuroscience research in vivo.

The vascularization of tissue grafts is critical for maintaining viability of the cells within a transplanted graft. A number of strategies are currently being investigated including very promising microfluidics systems. Here, we explored the potential for generating a vasculature-patterned endothelial cells that could be integrated into distinct layers between sheets of primary cells. Bioinspired from the leaf veins, we generated a reverse mold with a fractal vascular-branching pattern that models the unique spatial arrangement over multiple length scales that precisely mimic branching vasculature. By coating the reverse mold with 50 μg ml-1 of fibronectin and stamping enabled selective adhesion of the human umbilical vein endothelial cells (HUVECs) to the patterned adhesive matrix, we show that a vascular-branching pattern can be transferred by microcontact printing. Moreover, this pattern can be maintained and transferred to a 3D hydrogel matrix and remains stable for up to 4 d. After 4 d, HUVECs can be observed migrating and sprouting into Matrigel. These printed vascular branching patterns, especially after transfer to 3D hydrogels, provide a viable alternative strategy to the prevascularization of complex tissues.

Abstract Surface‐anchored metal‐organic frameworks (surMOFs) integrate nanoporous supramolecular MOF materials directly into architectures for applications such as gas storage, chemical sensing, and energy storage. Layer‐by‐layer solution‐phase deposition of the MOF‐14 components (1,3,5‐tris(4‐carboxyphenyl)benzene and copper (II) dimers, respectively) produces a porous and conformal film on carboxyl‐terminated self‐assembled monolayers (SAMs). In this research, the formation of ultrathin (less than 25 nm) surMOF films on codeposited bicomponent SAMs and microcontact printed SAMs is investigated by atomic force microscopy, ellipsometry, infrared spectroscopy, and contact angle goniometry. SAMs composed of methyl‐terminated alkanethiols assembled on gold substrates inhibit surMOF formation, whereas carboxyl‐terminated alkanethiols promote MOF‐14‐based film growth. To tune the density of carboxyl groups that anchor the film, methyl‐ and carboxyl‐terminated alkanethiols of varying concentrations are codeposited on gold. This systematic study demonstrates how surMOF film formation and morphology are impacted by these SAMs with mixed surface functionalities. Chemical patterning methods for SAMs, such as microcontact printing (μCP), commonly have mixed chemical functionalities within certain regions of the pattern. Insights gained regarding how mixed surface functionalities affect surMOF film formation are applied herein to optimize the μCP method to produce chemically patterned SAMs that selectively direct surMOF assembly to produce high‐quality surMOF film features.

Ischemic stroke is a severe insult in the brain causing cell death, inflammation, and activation of microglia. Microglia are the immune cells of the brain and play a role in any inflammatory process during neurodegeneration. Microglia are round ameboid and migrate to the lesion site, where they differentiate into ramified forms and activated phagocytic microglia. On the other hand, microglia can also release growth factors to repair degeneration. The aim of the present study is to explore the migratory capacity of microglia after ischemic insults. Organotypic brain slices of the mouse cortex (300 μm) were prepared. In order to study migration, the slices were connected to collagen-loaded microcontact prints (with or without monocyte chemoattractant protein-1, MCP-1) on the membranes. Slices were stimulated with lipopolysaccharide (LPS) for maximal microglial activation. Ischemic insults were simulated with oxygen-glucose deprivation (OGD) and acidosis (pH 6.5) for 3 days. After 3 weeks in culture, slices were fixed and immunohistochemically stained for the microglial markers Iba1, CD11b and macrophage-like antigen. Our data show that Iba1+ microglia migrated along the microcontact prints, differentiate and phagocyte 1.0 μm fluorescent microbeads. LPS significantly enhanced the number of round ameboid migrating microglia, while OGD and acidosis enhanced the number of ramified activated microglia. The effect was not visible on slices without any μCP and was most potent in μCP with MCP-1. We conclude that OGD and acidosis activate ramification and exhibit a similar mechanism, while LPS only activates round ameboid microglia. Collagen-loaded microcontact prints connected to mouse brain slices are a potent method to study activation and migration of microglia ex vivo.

Recent trends in 3D cell culturing has placed organotypic tissue models at another level. Now, not only is the microenvironment at the cynosure of this research, but rather, microscopic geometrical parameters are also decisive for mimicking a tissue model. Over the years, technologies such as micromachining, 3D printing, and hydrogels are making the foundation of this field. However, mimicking the topography of a particular tissue-relevant substrate can be achieved relatively simply with so-called template or morphology transfer techniques. Over the last 15 years, in one such research venture, we have been investigating a micro thermoforming technique as a facile tool for generating bioinspired topographies. We call them MatriGrid®s. In this research account, we summarize our learning outcome from this technique in terms of the influence of 3D micro morphologies on different cell cultures that we have tested in our laboratory. An integral part of this research is the evolution of unavoidable aspects such as possible label-free sensing and fluidic automatization. The development in the research field is also documented in this account. [ABSTRACT FROM AUTHOR]

In this work, we have combined the advantages of sequence programmability of DNA nanotechnology and optical birefringence of liquid crystals (LCs). Herein, DNA amphiphiles were adsorbed onto LC droplets. A unique phenomenon of LC droplet aggregation was demonstrated, using DNA-modified LC droplets, through complementary DNA hybridization. Further functionalization of DNA-modified LC droplets with a desired DNA sequence was used to detect a wide range of chemicals and biomolecules, such as Hg2+, thrombin, and enzymes, through LC droplet aggregation and vice versa, which can be seen through the naked eye. These DNA-modified LC droplets can be printed onto a desired patterned surface with temperature-induced responsiveness and reversibility. Overall, our work is the first to report DNA-modified LC droplet, which provides a general detection platform based on the development of DNA aptamers. Additionally, this work inspires the exploration of surface information visualization combined with microcontact printing. [ABSTRACT FROM AUTHOR]

Surface properties are largely responsible for the biological performance of biomedical devices, suggesting the great necessity of surface modification. Micro-contact printing (μCP) is a versatile surface modification technique that is capable of not only producing defined topographical features but also manipulating surface chemical and biological cues through customized inks. Compared to other surface patterning techniques, μCP offers distinct advantages of low cost combined with high reliability and versatility. This review summarizes the principles and characteristics of μCP and presents the latest advances enabled by μCP in the biomedical field, categorized by its applications in constructing cell culture platforms, biosensing platforms, and devices for other biological applications.

Catecholaminergic polymorphic ventricular tachycardia (CPVT) is characterized by an arrhythmogenic mechanism involving disruption of calcium handling. This genetic disease can lead to sudden death in children and young adults during physical or emotional stress. Prior CPVT studies have focused on calcium handling, but mechanical functionality has rarely been investigated in vitro. In this research we combine stem cell-derived cardiomyocytes from a CPVT patient (RyR2-H2464D mutation) and a healthy familial control with an engineered culture platform to evaluate mechanical function of cardiomyocytes. Substrates with Young’s modulus ranging from 10 to 50 kPa were used in conjunction with microcontact printing of ECM proteins into defined patterns for subsequent attachment. Digital Image Correlation (DIC) was used to evaluate collections of contracting cells. The amplitude of contractile strain was utilized as a quantitative indicator of functionality and disease severity. We found statistically significant differences: the maximum contractile strain was consistently higher in patient samples compared to control samples on all substrate stiffnesses. Additionally, the patient cell line had a statistically significantly slower intrinsic contraction rate than the control, which agrees with prior literature. Differences in mechanical strain have not been previously reported, and hypercontractility is not a known characteristic of CPVT. However, functional changes can occur as the disease progresses, thus this observation may not represent behavior observed in adolescent and adult patients. These results add to the limited studies of mechanical function of CPVT CMs reported in literature and identify functional differences that should be further explored.

This paper focuses on creating one-dimensional diffractive grooved structures of antigen proteins on glass substrates for the label-free detection of antibodies to dairy allergens. In particular, the fabrication of protein structures is carried out by combining microcontact printing with physisorption, imines coupling, and thiol-ene click chemistry. The work first sets up these patterning methods and discusses and compares the main aspects involved in them (structure, biolayer thickness, functionality, stability). Homogeneous periodic submicron structures of proteins are created and characterized by diffractive measurements, AFM, FESEM, and fluorescence scanning. Then, this patterning method is applied to proteins involved in cow milk allergy, and the resulting structures are implemented as optical transducers to sense specific immunoglobulins G. In particular, gratings of bovine serum albumin, casein, and β-lactoglobulin are created and assessed, reaching limits of detection in the range of 30–45 ng·mL−1 of unlabeled antibodies by diffractive biosensing.

Background: Alzheimer's disease is characterized by extracellular beta-amyloid plaques, intraneuronal tau neurofibrillary tangles and excessive neurodegeneration. The mechanisms of neuron degeneration and the potential of these neurons to form new nerve fibers for compensation remain elusive. The present study aimed to evaluate the impact of beta-amyloid and tau on new formations of nerve fibers from mouse organotypic brain slices connected to collagen-based microcontact prints., Methods: Organotypic brain slices of postnatal day 8-10 wild-type mice were connected to established collagen-based microcontact prints loaded with polyornithine to enhance nerve fiber outgrowth. Human beta-amyloid(42) or P301S mutated aggregated tau was co-loaded to the prints. Nerve fibers were immunohistochemically stained with neurofilament antibodies. The physiological activity of outgrown neurites was tested with neurotracer MiniRuby, voltage-sensitive dye FluoVolt, and calcium-sensitive dye Rhod-4., Results: Immunohistochemical staining revealed newly formed nerve fibers extending along the prints derived from the brain slices. While collagen-only microcontact prints stimulated nerve fiber growth, those loaded with polyornithine significantly enhanced nerve fiber outgrowth. Beta-amyloid(42) significantly increased the neurofilament-positive nerve fibers, while tau had only a weak effect. MiniRuby crystals, retrogradely transported along these newly formed nerve fibers, reached the hippocampus, while FluoVolt and Rhod-4 monitored electrical activity in newly formed nerve fibers., Conclusions: Our data provide evidence that intact nerve fibers can form along collagen-based microcontact prints from mouse brain slices. The Alzheimer's peptide beta-amyloid(42) stimulates this growth, hinting at a neuroprotective function when physiologically active. This "brain-on-chip" model may offer a platform for screening bioactive factors or testing drug effects on nerve fiber growth., Competing Interests: The authors declare no conflict of interest., (© 2024 The Author(s). Published by IMR Press.)

Parallel lithographic deposition of polymers onto counterpart substrates is a widely applied surface manufacturing operation. However, polymers may only be soluble in organic solvents or are insoluble at all. Solvent evaporation during stamping may trigger hardly controllable capillarity-driven flow processes or phase separation, and polymer solutions may spread on the counterpart substrates. Solvent-free stamping of melts prevents these drawbacks. Here, a stamp design for the deposition of melts is devised, which intrinsically circumvents ink depletion. The stamps' topographically patterned contact surfaces with protruding contact elements contacting the counterpart substrates consist of a nanoporous gold layer with a thickness of a few micrometers. The nanoporous gold layer is attached to a molten polymer layer, which is support for the nanoporous gold layer and ink reservoir at the same time. The nanoporous gold layer in turn stabilizes the topography of the stamps' contact surfaces. As examples, arrays of submicron microdots of polystyrene and poly(vinylidenefluoride-trifluoroethylene) (PVDF-TrFE) are manufactured. The P(VDF-TrFE) microdots are partially crystalline, ferroelectric, and can be locally poled. It is envisioned that the methodology reported here can be automatized and may be extended to functional low-molecular-mass compounds, such as active pharmaceutical ingredients., (© 2024 The Authors. Small published by Wiley‐VCH GmbH.)

Alzheimer's disease is a severe neurodegenerative disorder of the brain, characterized by beta-amyloid plaques, tau pathology, and cell death of cholinergic neurons, resulting in loss of memory. The reasons for the damage of the cholinergic neurons are not clear, but the nerve growth factor (NGF) is the most potent trophic factor to support the survival of these neurons. In the present study we aim to microprint NGF onto semipermeable 0.4 μm pore membranes and couple them with organotypic brain slices of the basal nucleus of Meynert and to characterize neuronal survival and axonal growth. The brain slices were prepared from postnatal day 10 wildtype mice (C57BL6), cultured on membranes for 2–6 weeks, stained, and characterized for choline acetyltransferase (ChAT). The NGF was microcontact printed in 28 lines, each with 35 μm width, 35 μm space between them, and with a length of 8 mm. As NGF alone could not be printed on the membranes, NGF was embedded into collagen hydrogels and the brain slices were placed at the center of the microprints and the cholinergic neurons that survived. The ChAT+ processes were found to grow along with the NGF microcontact prints, but cells also migrated. Within the brain slices, some form of re-organization along the NGF microcontact prints occurred, especially the glial fibrillary acidic protein (GFAP)+ astrocytes. In conclusion, we provided a novel innovative microcontact printing technique on semipermeable membranes which can be coupled with brain slices. Collagen was used as a loading substance and allowed the microcontact printing of nearly any protein of interest. [ABSTRACT FROM AUTHOR]

Alzheimer's disease is a severe neurodegenerative disorder of the brain, characterized by beta-amyloid plaques, tau pathology, and cell death of cholinergic neurons, resulting in loss of memory. The reasons for the damage of the cholinergic neurons are not clear, but the nerve growth factor (NGF) is the most potent trophic factor to support the survival of these neurons. In the present study we aim to microprint NGF onto semipermeable 0.4 μm pore membranes and couple them with organotypic brain slices of the basal nucleus of Meynert and to characterize neuronal survival and axonal growth. The brain slices were prepared from postnatal day 10 wildtype mice (C57BL6), cultured on membranes for 2–6 weeks, stained, and characterized for choline acetyltransferase (ChAT). The NGF was microcontact printed in 28 lines, each with 35 μm width, 35 μm space between them, and with a length of 8 mm. As NGF alone could not be printed on the membranes, NGF was embedded into collagen hydrogels and the brain slices were placed at the center of the microprints and the cholinergic neurons that survived. The ChAT+ processes were found to grow along with the NGF microcontact prints, but cells also migrated. Within the brain slices, some form of re-organization along the NGF microcontact prints occurred, especially the glial fibrillary acidic protein (GFAP)+ astrocytes. In conclusion, we provided a novel innovative microcontact printing technique on semipermeable membranes which can be coupled with brain slices. Collagen was used as a loading substance and allowed the microcontact printing of nearly any protein of interest.

A monolayer of highly motile cells can establish long-range orientational order, which can be explained by hydrodynamic theory of active gels and fluids. However, it is less clear how cell shape changes and rearrangement are governed when the monolayer is in mechanical equilibrium states when cell motility diminishes. In this work, we report that rat embryonic fibroblasts (REF), when confined in circular mesoscale patterns on rigid substrates, can transition from the spindle shapes to more compact morphologies. Cells align radially only at the pattern boundary when they are in the mechanical equilibrium. This radial alignment disappears when cell contractility or cell-cell adhesion is reduced. Unlike monolayers of spindle-like cells such as NIH-3T3 fibroblasts with minimal intercellular interactions or epithelial cells like Madin-Darby canine kidney (MDCK) with strong cortical actin network, confined REF monolayers present an actin gradient with isotropic meshwork, suggesting the existence of a stiffness gradient. In addition, the REF cells tend to condense on soft substrates, a collective cell behavior we refer to as the ‘condensation tendency’. This condensation tendency, together with geometrical confinement, induces tensile prestretch (i.e. an isotropic stretch that causes tissue to contract when released) to the confined monolayer. By developing a Voronoi-cell model, we demonstrate that the combined global tissue prestretch and cell stiffness differential between the inner and boundary cells can sufficiently define the cell radial alignment at the pattern boundary.

Laser-induced graphene (LIG) is a highly promising technology for assembling thin-film electronics with miniaturized surficial patterns, although hard to guarantee both high-quality formation of raw materials and high resolution of feature size. In this study, a combined microcontact printing (μCP) and LIG protocol is creatively utilized to fabricate ultra-thin graphene films with highly ordered micro-stripe arrays. In addition to realize various patterned structures with unique scalable and stackable characteristics, PAA concentration (4–8 wt%) is systematically investigated as a critical parameter to precisely control the feature size of micro-patterns before and after the irradiation process, including line width (before: 5.37–22.44 μm, after: 4.83–22.32 μm) and average height (before: 69.81–187.39 nm, after: 5.72–48.61 nm). By further exploring a defocused multi-lasing strategy, sheet resistance of the graphene array could be optimized dramatically from 634.1 kΩ/sq to 1.9 kΩ/sq, while maintaining low ablation (45.38 % porosity) and high carbon content (84.2 wt%). Lastly, the process is demonstrated to assemble different graphene based electronic devices with miniaturized sizes, including liquid sensors with sensitivity of 2.7 % μL−1 and electrothermal heaters with temperature-ramping rate as high as 24.7 °C/s and working temperature as high as 248 °C. A combined microcontact printing (μCP) and laser-induced graphene (LIG) protocol is creatively developed to assemble ultra-thin graphene film based devices with highly-ordered and stackable micro-stripe-array patterns, tunable feature width (4.83–22.32 μm) and height (5.72–48.61 nm), optimized sheet resistance (1.9 kΩ/sq), and functions of liquid-sensing (sensitivity of 2.7 % μL−1) and joule-heating (working temperature of 248 °C). [Display omitted] [ABSTRACT FROM AUTHOR]

Diamond shows great potentials as an ultimate material candidate for application in microelectromechanical systems (MEMS). In this work, we report microcontact printing (μCP) transfer as a facile technique to fabricate diamond MEMS cantilever arrays. We started from the Deryaguin–Landau–Verwey–Overbeek (DLVO) principles and formulated a colloidal "ink" that contains nanodiamonds (NDs) seeds well dispersed in polymethyl methacrylate/acetone. The NDs seeds were then transfer printed onto a silicon (100) substrate by using a flexible polydimethylsiloxane (PDMS) stamp with predefined pattern. The densely packed NDs pattern (∼109 cm−2) enables facile and selected-area growth of diamond with microwave plasma chemical vapor deposition (MPCVD) technique. A diamond film with minimal graphite contaminants and low residual stresses was obtained after growth at 1118 K for 3 h. The diamond cantilevers were released from the substrate by wet chemical etching using KOH (40 % wt.) and isopropyl alcohol (10 % vol.). An examination of the vibrational kinetics indicates that the diamond film has a high Young's modulus (∼795 GPa) and the cantilevers exhibit consistently distributed resonant frequencies and high quality factors in ambient air. The method opens the door for large-scale, cost-effective production of uniform diamond MEMS devices for sensor and quantum applications. [Display omitted] • Nanodiamonds is well dispersed in polymethyl methacrylate/acetone. • Diamond MEMS cantilevers is fabricated by using microcontact printing. • Diamond MEMS cantilevers exhibit consistently distributed resonant frequencies. [ABSTRACT FROM AUTHOR]

Molecular surface gradients can constitute electric field landscapes and serve to control local cell adhesion and migration. Cellular responses to electric field landscapes may allow the discovery of routes to improve osseointegration of implants. Flat molecule aggregate landscapes of amine- or carboxyl-teminated dendrimers, amine-containing protein and polyelectrolytes were prepared on glass to provide lateral electric field gradients through their differing zeta potentials compared to the glass substrate. The local as well as the mesoscopic morphological responses of adhered osteoblasts (MG-63) with respect to the stripes were studied by means of Scanning Ion Conductance Microscopy (SICM) and Fluorescence Microscopy, in situ. A distinct spindle shape oriented parallel to the surface pattern as well as a preferential adhesion of the cells on the glass site have been observed at a stripe and spacing width of 20 μm. Excessive ruffling is observed at the spindle poles, where the cells extend. To explain this effect of material preference and electro-deformation, we put forward a retraction mechanism, a localized form of double-sided cathodic taxis.