Inducing chemotactic and haptotactic cues in microfluidic devices for three-dimensional in vitro assays

Microfluidic devices allow for the production of physiologically relevant cellular microenvironments by including biomimetic hydrogels and generating controlled chemical gradients. During transport, the biomolecules interact in distinct ways with the fibrillar networks: as purely diffusive factors in the soluble fluid or bound to the matrix proteins. These two main mechanisms may regulate distinct cell responses in order to guide their directional migration: caused by the substrate-bound chemoattractant gradient (haptotaxis) or by the gradient established within the soluble fluid (chemotaxis). In this work 3D diffusion experiments, in combination with ELISA assays, are performed using microfluidic platforms in order to quantify the distribution of PDGF-BB and TGF-β₁ across collagen and fibrin gels. Furthermore, to gain a deeper understanding of the fundamental processes, the experiments are reproduced by computer simulations based on a reaction-diffusion transport model. This model yields an accurate prediction of the experimental results, confirming that diffusion and binding phenomena are established within the microdevice.     

Hierarchical structural complexity in atomically precise nanocluster frameworks

The supramolecular chemistry of nanoclusters is a flourishing area of nano-research; however, the controllable assembly of cluster nano-building blocks in different arrays remains challenging. In this work, we report the hierarchical structural complexity of atomically precise nanoclusters in micrometric linear chains (1D array), grid networks (2D array) and superstructures (3D array). In the crystal lattice, the Ag₂₉(SSR)₁₂(PPh₃)₄ nanoclusters can be viewed as unassembled cluster dots (Ag₂₉–0D). In the presence of Cs⁺ cations, the Ag₂₉(SSR)₁₂ nano-building blocks are selectively assembled into distinct arrays with different oxygen-carrying solvent molecules―Cs@Ag₂₉(SSR)₁₂(DMF)_x as 1D linear chains (Ag₂₉–1D), Cs@Ag₂₉(SSR)₁₂(NMP)_x as 2D grid networks (Ag₂₉–2D), and Cs@Ag₂₉(SSR)₁₂(TMS)_x as 3D superstructures (Ag₂₉–3D). Such self-assemblies of these Ag₂₉(SSR)₁₂ units have not only been observed in their crystalline state, but also in their amorphous state. Due to the diverse surface structures and crystalline packing modes, these Ag₂₉-based assemblies manifest distinguishable optical absorptions and emissions in both solutions and crystallized films. Furthermore, the surface areas of the nanocluster crystals are evaluated, the maximum value of which occurs when the cluster nano-building blocks are assembled into 2D arrays (i.e. Ag₂₉–2D). Overall, this work presents an exciting example of the hierarchical assembly of atomically precise nanoclusters by simply controlling the adsorbed molecules on the cluster surface.     

市场化进程与上市公司R&D投资： 基于产权特征视角

摘要：基于市场化进程驱动企业R&D投资的理论阐释,本文实证考察了市场化程度对不同产权企业R&D投资的影响。研究结果表明：（1）整体而言,市场化程度不仅与企业R&D投资强度正相关,而且市场化程度与行业竞争的共同作用对R&D投资的影响程度更为显著;（2）央企和民企的R&D投资对市场化程度的敏感性较高,分别为地方国资委所控企业的26倍和23倍,为地方企业集团公司的2倍和18倍;（3）与地方国资委直接控制的公司相比,相同的市场化程度对企业集团R&D投资的促进效果更为明显;（4）只有在市场化进程达到一定程度后,R&D投资的价值效应才可能因市场环境的实质性改善而逐步彰显;而地方政府基于短期利益的干预,很可能削弱了市场化进程对R&D投资价值的边际贡献。上述研究结论为进一步完善企业R&D投资的市场驱动机制和产权约束条件提供了理论和经验依据。     

Effects of foreign acquisitions on R&D activity: Evidence from firm-level data for France

This paper investigates the causal effect of foreign acquisitions on the research and development (R&D) activities of domestic target firms over the period 1994–2004. Using accounting data on French innovative manufacturing firms, we implement appropriate difference-in-difference estimation techniques associated with a matching propensity score procedure. We find that the acquisitions of French firms by foreign companies boost R&D spending. There is a simultaneous rise in the external and in-house R&D expenditures of French acquired firms. R&D is more contracted out to local research providers, in particular to local public laboratories and universities. The increase in internal R&D spending benefits to development, but also basic and applied research. Finally, the growth of the R&D budget is not only financed by internal resources but also receives financing from external partners, especially parent companies. Thus, our results call into question the idea that foreign takeovers hamper the R&D development of target firms and are detrimental to the national innovation system of the host country. First, acquisitions appear to bring efficiency gains enough to counterbalance the various costs of integration and market power effects, pushing firms to invest more in R&D. Acquisitions do not seem to incite managers to take shorter term and more financial driven decisions at the expense of R&D. Second, when motivated by technology sourcing and overseas R&D development, acquisitions might be used to access the specific know-how of target firms and to tap into the knowledge of the national innovation system. Then, purchaser firms could be more likely to develop the innovative capability of target firms and to strengthen their linkages with local partners than to reduce them and relocate R&D abroad.     

Evaluation of 3D-printed molds for fabrication of non-planar microchannels

Replica obtained from micromolds patterned by simple photolithography has features with uniform heights, and attainable microchannels are thus quasi-two-dimensional. Recent progress in three-dimensional (3D) printing has enabled facile desktop fabrication of molds to replicate microchannels with varying heights. We investigated the replica obtained from four common techniques of 3D printing—fused deposition modeling, selective laser sintering, photo-polymer inkjet printing (PJ), and stereolithography (SL)—for the suitability to form microchannels in terms of the surface roughness inherent to the mechanism of 3D printing. There have been limited quantitative studies that focused on the surface roughness of a 3D-printed mold with different methods of 3D printing. We discussed that the surface roughness of the molds affected (1) transparency of the replica and (2) delamination pressure of poly(dimethylsiloxane) replica bonded to flat glass substrates. Thereafter, we quantified the accuracy of replication from 3D-printed molds by comparing the dimensions of the replicated parts to the designed dimensions and tested the ability to fabricate closely spaced microchannels. This study suggested that molds printed by PJ and SL printers were suitable for replica molding to fabricate microchannels with varying heights. The insight from this study shall be useful to fabricate 3D microchannels with controlled 3D patterns of flows guided by the geometry of the microchannels.     

Foreign research and development in Swedish multinationals

In 1987, Sweden's 20 largest multinationals together operated some 170 foreign R&D units, employing 8100 people, 23% of their total R&D personnel. About 60% of the units were added after 1980, the majority through acquisitions of foreign companies.Four major motives for the operation of foreign R&D units are identified: (1) support to local production (5% of foreign R&D employment); (2) ‘market proximity’, i.e. adaptation of centrally developed products (and processes) to local market conditions (32%); (3) exploitation of foreign R&D results and resources (8%); (4) ‘political factors’, i.e. environmental factors more or less directly influenced by government action (34%). The remaining 20% of employment seemed to be motivated by a combination of factors.The primary aim of the first three types of foreign R&D units is to enhance a company's competitive position in ways not possible from a domestic location. Hence, they should be little cause of concern for Swedish policy makers.     

Microscale flow propulsion through bioinspired and magnetically actuated artificial cilia

Recent advances in microscale flow propulsion through bioinspired artificial cilia provide a promising alternative for lab-on-a-chip applications. However, the ability of actuating artificial cilia to achieve a time-dependent local flow control with high accuracy together with the elegance of full integration into the biocompatible microfluidic platforms remains remote. Driven by this motive, the current work has constructed a series of artificial cilia inside a microchannel to facilitate the time-dependent flow propulsion through artificial cilia actuation with high-speed (>40 Hz) circular beating behavior. The generated flow was quantified using micro-particle image velocimetry and particle tracking with instantaneous net flow velocity of up to 10¹μm/s. Induced flow patterns caused by the tilted conical motion of artificial cilia constitutes efficient fluid propulsion at microscale. This flow phenomenon was further measured and illustrated by examining the induced flow behavior across the depth of the microchannel to provide a global view of the underlying flow propulsion mechanism. The presented analytic paradigms and substantial flow evidence present novel insights into the area of flow manipulation at microscale.     

Dynamic radial positioning of a hydrodynamically focused particle stream enabled by a three-dimensional microfluidic nozzle

The ability to confine flows and focus particle streams has become an integral component of the design of microfluidic systems for the analysis of a wide range of samples. Presented here is the implementation of a 3D microfluidic nozzle capable of both focusing particles as well as dynamically positioning those particles in selected flow lamina within the downstream analysis channel. Through the independent adjustment of the three sheath inlet flows, the nozzle controlled the size of a focused stream for 6, 10, and 15 μm polystyrene microparticles. Additional flow adjustment allowed the nozzle to dynamically position the focused particle stream to a specific area within the downstream channel. This unique ability provides additional capability and sample flexibility to the system. In order to gain insight into the fluidic behavior of the system, experimental conditions and results were duplicated within 4.75 μm using a COMSOL Multiphysics^® model to elucidate the structure, direction, proportion, and fate of fluid lamina throughout the nozzle region. The COMSOL Multiphysics model showed that the position and distribution of particles upon entering the nozzle have negligible influence over its focusing ability, extending the experimental results into a wider range of particle sizes and system flow rates. These results are promising for the application of this design to allow for a relatively simple, fast, fully fluidically controlled nozzle for selective particle focusing and positioning for further particle analysis and sorting.     

Hierarchically porous monoliths based on low-valence transition metal (Cu,Co, Mn) oxides: gelation and phase separation

Hierarchically porous monoliths based on copper (Cu), cobalt (Co) and manganese (Mn) oxides with three-dimensionally (3D) interconnected macropores and open nanopores were prepared using metal bromides as precursors via a sol–gel process accompanied by phase separation. The difficulty of gelation for low-valence metal cation was overcome by introducing a highly electronegative Br atom near to the metal atom to control the rates of hydrolysis and polycondensation. The 3D interconnected macropores were obtained using appropriate polymers to induce phase separation. The domain sizes of macropores and skeletons can be controlled by reaction parameters such as concentration and/or average molecular weight of polymers, and the amount of hydrochloric acid. The crystalline metal oxide monoliths with their 3D interconnected macroporous structure preserved were obtained after heat treatment in air.     

On-chip three-dimensional tumor spheroid formation and pump-less perfusion culture using gravity-driven cell aggregation and balanced droplet dispensing

This paper presents a spheroid chip in which three-dimensional (3D) tumor spheroids are not only formed by gravity-driven cell aggregation but also cultured at the perfusion rates controlled by balanced droplet dispensing without fluidic pumps. The previous spheroid chips require additional off-chip processes of spheroid formation and extraction as well as bulky components of fluidic pumps. However, the present spheroid chip, where autonomous medium droplet dispensers are integrated on a well array, achieves the on-chip 3D tumor spheroid formation and perfusion culture using simple structure without bulky fluidic pumps. In the experimental study, we demonstrated that the spheroid chip successfully forms 3D tumor spheroids in the wide diameter range of 220 μm–3.2 mm (uniformity > 90%) using H358, H23, and A549 non-small cell lung cancer cells. At the pump-less perfusion culture (Q = 0.1–0.3 μl/min) of spheroids, the number of H358 cells in the spheroid increased up to 50% from the static culture (Q = 0 μl/min) and the viability of the cultured cells also increased about 10%. Therefore, we experimentally verified that the perfusion environment created by the spheroid chip offers a favourable condition to the spheroids with high increase rate and viability. The present chip achieves on-chip 3D tumor spheroid formation and pump-less perfusion culture with simple structure, thereby exhibiting potential for use in integrated in-vivo-like cell culture systems.     

Rational design of high-quality 2D/3D perovskite heterostructure crystals for record-performance polarization-sensitive photodetection

Polarization-sensitive photodetection is central to optics applications and has been successfully demonstrated in photodetectors of two-dimensional (2D) materials, such as layered hybrid perovskites; however, achieving high polarization sensitivity in such a photodetector remains extremely challenging. Here, for the first time, we demonstrate a high-performance polarization-sensitive photodetector using single-crystalline 2D/3D perovskite heterostructure, namely, (4-AMP)(MA)₂Pb₃Br₁₀/MAPbBr₃ (MA = methylammonium; 4-AMP = 4-(aminomethyl)piperidinium), which exhibits ultrahigh polarization sensitivity up to 17.6 under self-driven mode. To our knowledge, such a high polarization selectivity has surpassed all of the reported perovskite-based devices, and is comparable to, or even better than, the traditional inorganic heterostructure-based photodetectors. Further studies reveal that the built-in electric field formed at the junction can spatially separate the photogenerated electrons and holes, reducing their recombination rate and thus enhancing the performance for polarization-sensitive photodetection. This work provides a new source of polarization-sensitive materials and insights into designing novel optoelectronic devices.     

Advances in three-dimensional rapid prototyping of microfluidic devices for biological applications

The capability of 3D printing technologies for direct production of complex 3D structures in a single step has recently attracted an ever increasing interest within the field of microfluidics. Recently, ultrafast lasers have also allowed developing new methods for production of internal microfluidic channels within the bulk of glass and polymer materials by direct internal 3D laser writing. This review critically summarizes the latest advances in the production of microfluidic 3D structures by using 3D printing technologies and direct internal 3D laser writing fabrication methods. Current applications of these rapid prototyped microfluidic platforms in biology will be also discussed. These include imaging of cells and living organisms, electrochemical detection of viruses and neurotransmitters, and studies in drug transport and induced-release of adenosine triphosphate from erythrocytes.     

Laser-driven programmable non-contact transfer printing of objects onto arbitrary receivers via an active elastomeric microstructured stamp

Transfer printing, as an important assembly technique, has attracted much attention due to its valuable merits to develop novel forms of electronics such as stretchable inorganic electronics requiring the heterogeneous integration of inorganic materials with soft elastomers. Here, we report on a laser-driven programmable non-contact transfer printing technique via a simple yet robust design of active elastomeric microstructured stamp that features cavities filled with air and embedded under the contacting surface, a micro-patterned surface membrane that encapsulates the air cavities and a metal layer on the inner-cavity surfaces serving as the laser-absorbing layer. The micro-patterned surface membrane can be inflated dynamically to control the interfacial adhesion, which can be switched from strong state to weak state by more than three orders of magnitude by local laser heating of the air in the cavity with a temperature increase below 100°C. Theoretical and experimental studies reveal the fundamental aspects of the design and fabrication of the active elastomeric microstructured stamp and the operation of non-contact transfer printing. Demonstrations in the programmable transfer printing of micro-scale silicon platelets and micro-scale LED chips onto various challenging receivers illustrate the extraordinary capabilities for deterministic assembly that are difficult to address by existing printing schemes, thereby creating engineering opportunities in areas requiring the heterogeneous integration of diverse materials such as curvilinear electronics and MicroLED displays.     

The Role of 25-Hydroxy Vitamin D Deficiency in Iron Deficient Children of North India

Extensive data from animal and human studies indicate a role of vitamin D in erythropoiesis. Iron and vitamin D deficiencies are implicated with adverse health effects in children even if they are asymptomatic. The potential relationship between the two remains poorly understood. A cross-sectional study was performed in the period from 1st May 2012 through 30th April 2013 and subjects were classified into vitamin D deficiency (VDD), vitamin D insufficiency (VDI) and vitamin D sufficiency (VDS) groups according to their 25(OH) D levels. A total of 263 children were included in the analysis. Anaemia was present in 66 % of 25(OH) D deficient subjects compared with 35 % in vitamin D sufficient individuals (p < 0.0001). The association of breast feeding and development of VDD was also significant (p < 0.05). Serum levels of 25(OH) D were found lower in female sex and if the analysis was performed in the winter/spring season. Physicians should therefore assess vitamin D levels in all anaemic children and ensure adequate supplementation to prevent deficiencies.     

Collagen-based brain microvasculature model in vitro using three-dimensional printed template

We present an engineered three-dimensional (3D) in vitro brain microvasculature system embedded within the bulk of a collagen matrix. To create a hydrogel template for the functional brain microvascular structure, we fabricated an array of microchannels made of collagen I using microneedles and a 3D printed frame. By culturing mouse brain endothelial cells (bEnd.3) on the luminal surface of cylindrical collagen microchannels, we reconstructed an array of brain microvasculature in vitro with circular cross-sections. We characterized the barrier function of our brain microvasculature by measuring transendothelial permeability of 40 kDa fluorescein isothiocyanate-dextran (Stoke''s radius of ∼4.5 nm), based on an analytical model. The transendothelial permeability decreased significantly over 3 weeks of culture. We also present the disruption of the barrier function with a hyperosmotic mannitol as well as a subsequent recovery over 4 days. Our brain microvasculature model in vitro, consisting of system-in-hydrogel combined with the widely emerging 3D printing technique, can serve as a useful tool not only for fundamental studies associated with blood-brain barrier in physiological and pathological settings but also for pharmaceutical applications.     

A hybrid microfluidic platform for cell-based assays via diffusive and convective trans-membrane perfusion

We present a novel 3D hybrid assembly of a polymer microfluidic chip with polycarbonate track-etched membrane (PCTEM) enabling membrane-supported cell culture. Two chip designs have been developed to establish either diffusive or convective reagent delivery using the integrated PCTEM. While it is well suited to a range of cell-based assays, we specifically employ this platform for the screening of a common antitumor chemotoxic agent (mitomycin C – MMC) on the HL60 myeloid leukemia cell line. The toxic activity of MMC is based on the generation of severe DNA damage in the cells. Using either mode of operation, the HL60 cells were cultured on-chip before, during, and after exposure to MMC at concentrations ranging from 0 to 50 μM. Cell viability was analysed off-chip by the trypan blue dye exclusion assay. The results of the on-chip viability assay were found to be consistent with those obtained off-chip and indicated ca. 40% cell survival at MMC concentration of 50 μM. The catalogue of capabilities of the here described cell assay platform comprises of (i) the culturing of cells either under shear-free conditions or under induced through-membrane flows, (ii) the tight time control of the reagent exposure, (iii) the straightforward assembly of devices, (iv) the flexibility on the choice of the membrane, and, prospectively, (v) the amenability for large-scale parallelization.     

MNCs’ offshore R&D networks in host country's regional innovation system: The case of Taiwan-based firms in China

China has become a hot spot of R&D internationalization and a growing number of Taiwan-based firms have indeed set up R&D units in China. Taking into account China's substantial regional variations in economic development, innovation capacity, and knowledge productivity, such notions as regional innovation system (RIS) and local innovative milieu may become more relevant to the study on relationships between China and its inward R&D internationalization. Therefore, the key issue for this paper is what locational advantages of an RIS within a host country affect the network linkages and networking strategy of multinational corporations’ (MNCs’) offshore R&D units. The paper aims to enrich the current understanding of R&D internationalization in several ways. First, the paper attempts to examine the R&D networking underlying R&D internationalization by Taiwan-based firms in China, with particular reference to the sub-national level inside China. Second, the paper tries to establish a link between the literature of R&D internationalization and that of RIS, with a modified version of Dunning's eclectic paradigm. Efforts are made to map the relationship between foreign subsidiaries’ local R&D networks and their host RISs inside China. Third, the paper takes advantage of a government databank to adopt a quantitative approach, the Seemingly Unrelated Bivariate Probit Regression model, with foreign subsidiaries as the unit of analysis, to highlight the role played by some aspects of the RIS in determining the local R&D networking of Taiwanese subsidiaries in China. Our evident shows that MNCs’ offshore R&D units that purse home-based technology exploitation strategy, the mainstream strategy regarding the developing host country, tend to be located in a host region with a strong knowledge application and exploitation subsystem, while an RIS with a strong knowledge generation and diffusion subsystem, within such a developing country as China, may induce MNCs’ local R&D units to pursue home-base technology augmenting strategy. On balance, not only the location choice but also the local R&D linkages of MNCs’ offshore subsidiaries are related to appropriate fits between the RIS and the subsidiaries’ innovation network inside the host country.     

The local government market as a stimulus to industrial innovation

Supply and demand conditions that affect technological change in four functional areas of local government service delivery are described and analyzed. A consistent picture of small, fragmented markets, ‘lumpy’ demand, or uncertain markets is found, with demand shaped by a heterogeneous body of local procurement traditions and practices, low municipal technical capabilities, and risk-averse behavior by city officials and engineering consultants. Industries involved share a number of characteristics that bode poorly for industrial investment in innovation: low profit margins and uncertain financial futures. Firms that have the resources to invest in R&D do not tend to focus exclusively on municipal markets. The findings suggest that public policies directed toward improving the municipal market for innovative products by strengthening the cities' technological infrastructure will be more effective than policies directed toward the supply side such as increased R&D support.     

Three dimensional passivated-electrode insulator-based dielectrophoresis

In this study, a 3D passivated-electrode, insulator-based dielectrophoresis microchip (3D πDEP) is presented. This technology combines the benefits of electrode-based DEP, insulator-based DEP, and three dimensional insulating features with the goal of improving trapping efficiency of biological species at low applied signals and fostering wide frequency range operation of the microfluidic device. The 3D πDEP chips were fabricated by making 3D structures in silicon using reactive ion etching. The reusable electrodes are deposited on second glass substrate and then aligned to the microfluidic channel to capacitively couple the electric signal through a 100 μm glass slide. The 3D insulating structures generate high electric field gradients, which ultimately increases the DEP force. To demonstrate the capabilities of 3D πDEP, Staphylococcus aureus was trapped from water samples under varied electrical environments. Trapping efficiencies of 100% were obtained at flow rates as high as 350 μl/h and 70% at flow rates as high as 750 μl/h. Additionally, for live bacteria samples, 100% trapping was demonstrated over a wide frequency range from 50 to 400 kHz with an amplitude applied signal of 200 Vpp. 20% trapping of bacteria was observed at applied voltages as low as 50 Vpp. We demonstrate selective trapping of live and dead bacteria at frequencies ranging from 30 to 60 kHz at 400 Vpp with over 90% of the live bacteria trapped while most of the dead bacteria escape.     

The ex ante assessment of knowledge spillovers: Government R&D policy, economic incentives and private firm behavior

This paper investigates the role of government R&D subsidy programs in stimulating knowledge spillovers. R&D subsidies are an effective public policy instrument when knowledge spillovers exist yet ex ante it is difficult to identify projects that have the greatest potential to increase innovation and economic growth. This paper derives a set of project and firm attributes that the literature finds generate knowledge spillovers and uses data on project proposals to estimate the degree to which a government R&D program conforms. We find that projects that were awarded R&D subsidies were more likely to have attributes such as participation in new research joint ventures and connections to universities and other firms. Following the post-award activities of firm, we find that receipt of a government R&D subsidy increased the funding from other sources when compared to firms that were not awarded funding.