In silico modeling of endocrine organ-on-a-chip systems.

The organ-on-a-chip (OoC) is an artificially reconstructed microphysiological system that is implemented using tissue mimics integrated into miniaturized perfusion devices. OoCs emulate dynamic and physiologically relevant features of the body, which are not available in standard in vitro methods. Furthermore, OoCs provide highly sophisticated multi-organ connectivity and biomechanical cues based on microfluidic platforms. Consequently, they are often considered ideal in vitro systems for mimicking self-regulating biophysical and biochemical networks in vivo where multiple tissues and organs crosstalk through the blood flow, similar to the human endocrine system. Therefore, OoCs have been extensively applied to simulate complex hormone dynamics and endocrine signaling pathways in a mechanistic and fully controlled manner. Mathematical and computational modeling approaches are critical for quantitatively analyzing an OoC and predicting its complex responses. In this review article, recently developed in silico modeling concepts of endocrine OoC systems are summarized, including the mathematical models of tissue-level transport phenomena, microscale fluid dynamics, distant hormone signaling, and heterogeneous cell-cell communication. From this background, whole chip-level analytic approaches in pharmacokinetics and pharmacodynamics will be described with a focus on the spatial and temporal behaviors of absorption, distribution, metabolism, and excretion in endocrine biochips. Finally, quantitative design frameworks for endocrine OoCs are reviewed with respect to support parameter calibration/scaling and enable predictive in vitro-in vivo extrapolations. In particular, we highlight the analytical and numerical modeling strategies of the nonlinear phenomena in endocrine systems on-chip, which are of particular importance in drug screening and environmental health applications.

Mono(2-ethylhexyl) phthalate modulates lipid accumulation and reproductive signaling in Daphnia magna.

Mono(2-ethylhexyl) phthalate (MEHP) is a primary metabolite of di(2-ethylhexyl) phthalate (DEHP), which is widely used in industry as a plasticizer. Both DEHP and MEHP have been identified as endocrine disruptors affecting reproduction systems in natural aquatic environments. However, the effects of MEHP exposure on aquatic invertebrates such as Daphnia magna are still poorly understood. In the present study, lipid alterations caused by MEHP in D. magna were identified by analyzing lipid accumulation and nontarget metabolomics. In addition, reproductive endpoints were investigated. MEHP exposure under any conditions upto 2 mg/L was not associated with mortality of D. magna; yet, the number of lipid droplets and the adult female daphnids reproduction rates increased after 96 h of exposure and 21 days of exposure, respectively. MEHP also enhanced lipid metabolism, as evident from 283 potential lipid metabolites, including glycerolipids, glycerophospholipids, and sphingolipids, identified following 48 h of exposure. The MEHP-treated group exhibited significantly higher ecdysone receptor (EcR) and vitellogenin 2 (Vtg2) expression levels at 6 and 24 h. At 48 h, EcR and Vtg2 expression levels were downregulated in the 1 and 2 mg/L MEHP exposure groups. Our data reveal that the EcR pathway changes over MEHP exposure could be associated with lipid accumulation, owing to increased lipid levels and the subsequent increase in the reproduction of MEHP-exposed D. magna.

Combined Effect of Matrix Topography and Stiffness on Neutrophil Shape and Motility.

The crawling behavior of leukocytes is driven by the cell morphology transition, which is a direct manifestation of molecular motor machinery. The topographical anisotropy and mechanical stiffness of the substrates are the main physical cues that affect leukocytes' shape generation and migratory responses. However, their combined effects on the cell morphology and motility have been poorly understood, particularly for neutrophils, which are the fastest reacting leukocytes against infections and wounds. Here, spatiotemporally correlated physical parameters are shown, which determine the neutrophil shape change during migratory processes, in response to surface topography and elasticity. Guided crawling and shape generation of individual neutrophils, activated by a uniform concentration of a chemoattractant, are analyzed by adopting elasticity-tunable micropatterning and live cell imaging techniques. Whole cell-level image analysis is performed based on a planar geometric quantification of cell shape and motility. The findings show that the pattern anisotropy and elastic modulus of the substrate induce synergic effects on the shape anisotropy, deformability, and polarization/alignment of crawling neutrophils. How the morphology-motility relationship is affected by different surface microstructures and stiffness is demonstrated. These results imply that the neutrophil shape-motility correlations can be utilized for controlling the immune cell functions with predefined physical microenvironments.

Transcriptomic and physiological analysis of endocrine disrupting chemicals Impacts on 3D Zebrafish liver cell culture system.

In recent decades, extensive efforts have focused on developing in vitro platforms mimicking fish livers to better understand the acute or chronic effects of toxicants on lower aquatic vertebrates. Fish liver cell lines have emerged as a promising culture system for these in vitro platforms because they complement the currently limited in vitro tools that mostly consist of mammalian cell lines and adhere to the 3Rs: replacement, reduction, and refinement of living animal tests. However, monolayer cell lines have lower transcriptional and physiological responses upon exposure to toxic chemicals than freshly isolated primary cells. To overcome this challenge, we utilized a three-dimensional (3D) spheroid-based in vitro platform, in which hepatocyte cells had self-organized into spheroid forms via E-cadherin bonds. This platform exhibited augmented transcriptomic and phenotypic regulation of liver cells in comparison to monolayer cells. We examined the organoid platform using the zebrafish liver (ZFL) cell line as a model system. ZFL cells spontaneously clustered into 3D spheroids with long-term viability by optimizing cell seeding density on a non-adherent substrate. Interestingly, 3D ZFL spheroids treated with estrogenic chemicals were activated to synthesize a higher level of vitellogenin (Vtg) than monolayer cells. Whole-transcriptome sequencing analysis confirmed that 3D ZFL spheroids had greater transcriptional regulation of genes related to reproductive toxicological response and liver functions, such as the urea cycle, estrogen receptors, and vitellogenin, compared to monolayer cells. These results may contribute to the engineering of novel 3D in vitro platforms for screening harmful chemicals and improving understanding of the underlying liver toxicity mechanisms at the molecular and cellular levels.

Magnetic microgels and nanogels: Physical mechanisms and biomedical applications.

Soft micro- and nanostructures have been extensively developed for biomedical applications. The main focus has been on multifunctional composite materials that combine the advantages of hydrogels and colloidal particles. Magnetic microgels and nanogels can be realized by hybridizing stimuli-sensitive gels and magnetic nanoparticles. They are of particular interest since they can be controlled in a wide range of biological environments by using magnetic fields. In this review, we elucidate physical principles underlying the design of magnetic microgels and nanogels for biomedical applications. Particularly, this article provides a comprehensive and conceptual overview on the correlative structural design and physical functionality of the magnetic gel systems under the concept of colloidal biodevices. To this end, we begin with an overview of physicochemical mechanisms related to stimuli-responsive hydrogels and transport phenomena and summarize the magnetic properties of inorganic nanoparticles. On the basis of the engineering principles, we categorize and summarize recent advances in magnetic hybrid microgels and nanogels, with emphasis on the biomedical applications of these materials. Potential applications of these hybrid microgels and nanogels in anticancer treatment, protein therapeutics, gene therapy, bioseparation, biocatalysis, and regenerative medicine are highlighted. Finally, current challenges and future opportunities in the design of smart colloidal biodevices are discussed.

Agglomeration structure of superparamagnetic nanoparticles in a nematic liquid crystal medium: Image analysis datasets based on cryo-electron microscopy and polarised optical microscopy techniques.

This dataset shows the agglomerate dimension and structure of oleic acid-coated superparamagnetic nanoparticles (SPIONs), which are dispersed in the nematic fluid of a thermotropic liquid crystal (LC), 4-cyano-4'-pentylbiphenyl (5CB). The analysed datasets were acquired from the raw images of the SPION-5CB mixtures obtained using cryogenic transmission electron microscopy (cryo-TEM) and polarised optical microscopy. The image data were quantitatively analysed to extract statistical information on the sizes of SPIONs and their agglomerates and the inter-particle spacing of the agglomerated SPIONs. This dataset supports the fundamental understanding on how colloidal nanospheres behave in an anisotropic fluid, and has a potential to be used as a part of database for automated design of new hybrid materials.

Quantitative prediction of mixture toxicity of AgNO3 and ZnO nanoparticles on Daphnia magna.

Once metal-based engineered nanoparticles (NPs) are released into the aquatic environment, they are expected to interact with other existing co-contaminants. A knowledge gap exists as to how the interaction of NPs with other co-contaminants occurs. Here we selected ZnO NPs among various NPs, with Ag ion existing as a contaminant in the aquatic environment by Ag NPs widely used. A novel modeling strategy was demonstrated enabling quantitative and predictive evaluation of the aqueous mixture nanotoxicity. Individual and binary mixture toxicity tests of ZnO NPs and silver (as AgNO3) on Daphnia magna were conducted and compared to determine whether the presence of Ag ions affects the toxicity of ZnO NPs. Binary mixture toxicity was evaluated based on the concentration addition (CA) and independent action models. The CA dose-ratio dependent model was found to be the model of best fit for describing the pattern of mixture toxicity. The MIX I and MIX III suspensions (higher ratios of ZnO NPs to AgNO3) showed a synergism, whereas the MIX II suspension (lower ratio of ZnO NPs to AgNO3) showed an antagonism. The synergistic mixture toxicity at higher ratios of ZnO NPs to AgNO3 was caused by either the physiological or metabolic disturbance induced by the excessive ionic Zn or increased transport and accumulation in D. magna via the formation of complex of ionic Ag with ZnO NPs. Therefore, the toxicity level contributed via their aggregation and physicochemical properties and the dissolved ions played a crucial role in the mixture toxicities of the NPs.

Mono-(2-ethylhexyl) phthalate induces oxidative stress and lipid accumulation in zebrafish liver cells.

The purpose of the present study was to examine the antioxidant and oxidative stress changes in zebrafish liver (ZFL) cells in the presence of mono-(2-ethylhexyl) phthalate (MEHP). When reactive oxygen species (ROS) and antioxidant levels were measured by immunoassay, significant differences were observed between MEHP-treated and control cells, while catalase levels did not change in any group. MEHP-treated cells had higher levels of ROS, glutathione peroxidase (GPx), glutathione S-transferase (GST), glutathione, and superoxide dismutase (SOD) than control cells. However, lower levels of lipid peroxidation were observed in MEHP-treated cells compared to control cells. After 24 h of MEHP treatment, ROS, SOD, GPx, and GST activity increased in a dose-dependent manner. Cellular lipid droplet formation and endoplasmic reticulum stress were both induced in the presence of MEHP. These findings demonstrated the potential impacts of the association of MEHP with adverse outcomes in fish liver. Future studies will focus on clarifying the molecular mechanism of phthalate toxicity via oxidative stress and peroxisome proliferator activated receptor as the major mechanistic pathway.

Depletion-driven morphological transitions in hexagonal crystallites of virus rods.

The assembly of nanometer-sized building blocks into complex morphologies is not only of fundamental interest but also plays a key role in material science and nanotechnology. We show that the shape of self-assembled superstructures formed by rod-like viruses can be controlled by tuning the attraction via the depletion interaction between the rods. Using non-adsorbing polymers as a depleting agent, we demonstrate that a hierarchical unidimensional self-organization into crystalline clusters emerges progressively upon increasing depletion attraction and enhanced growth kinetics. We observe a polymorphic change proceeding from two-dimensional (2D) crystalline monolayers at weak depletion to one-dimensional (1D) columnar fibers at strong depletion, via the formation of smectic fibrils at intermediate depletion strength. A simple theory for reversible polymerization enables us to determine the typical bond energy between monomeric units making up the smectic fibrils. We also demonstrate that gentle flow-assistance can be used to template filament-like structures into highly aligned supported films. Our results showcase a generic bottom-up approach for tuning the morphology of crystalline superstructures through modification of the interaction between non-spherical building blocks. This provides a convenient pathway for controlling self-organization, dimensionality and structure-formation of anisotropic nanoparticles for use in nanotechnology and functional materials.

In Silico Modeling Method for Computational Aquatic Toxicology of Endocrine Disruptors: A Software-Based Approach Using QSAR Toolbox.

Computational analyses of toxicological processes enables high-throughput screening of chemical substances and prediction of their endpoints in biological systems. In particular, quantitative structure-activity relationship (QSAR) models have been increasingly applied to assess the environmental effects of a plethora of toxic materials. In recent years, some more highlighted types of toxicants are endocrine disruptors (EDs, which are chemicals that can interfere with any hormone-related metabolism). Because EDs may significantly affect animal development and reproduction, rapidly predicting the adverse effects of EDs using in silico techniques is required. This study presents an in silico method to generate prediction data on the effects of representative EDs in aquatic vertebrates, particularly fish species. The protocol describes an example utilizing the automated workflow of the QSAR Toolbox software developed by the Organization for Economic Co-operation and Development (OECD) to enable acute ecotoxicity predictions of EDs. As a result, the following are determined: (1) calculation of the numerical correlations between the concentration for 50% of lethality (LC50) and octanol-water partition coefficient (Kow), (2) output performances in which the LC50 values determined in experiments are compared to those generated by computations, and (3) the dependence of estrogen receptor binding affinity on the relationship between Kow and LC50.

Colloidal gelatin microgels with tunable elasticity support the viability and differentiation of mesenchymal stem cells under pro-inflammatory conditions.

Despite a promising potential for mesenchymal stem cells (MSCs) in tissue regeneration, a major challenge in MSC-based therapy has been associated with poor cell survival and low levels of cell integration into host tissue following transplantation. The objective of this study was to develop a gelatin-based colloidal microgel platform that enables the encapsulation of viable MSCs as well as confer fine-tuning of mechanical stiffness and low cytotoxicity. In this study, we report a facile method of fabricating gelatin-based microgel spheres for the encapsulation of MSCs using a water-in-oil mini-emulsification method, which is covalently crosslinked by genipin. At a given seeding cell number, there was a positive correlation between the size of the microsphere and the number of encapsulated MSCs. Controlling the crosslinking degree of gelatin matrix enabled a fine-tuning of mechanical stiffness of gel microsphere. MSCs within softer microgel exhibit more spread morphology than the cells in the stiffer matrix, while cells within stiffer matrix become more elongated morphology. Importantly, we show that the colloidal gelatin microgel could support the viability and differentiation of encapsulated MSCs in a pro-inflammatory environment. This study demonstrates the feasibility of using genipin-crosslinked gelatin gel microspheres as an injectable carrier of MSCs for tissue engineering applications, which can be further explored for MSC-based cell therapy for tissue repair. © 2018 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 2753-2761, 2018.

Chirality-controlled crystallization via screw dislocations.

Chirality plays an important role in science from enantiomeric separation in chemistry to chiral plasmonics in nanotechnology. However, the understanding of chirality amplification from chiral building blocks to ordered helical superstructures remains a challenge. Here, we demonstrate that topological defects, such as screw dislocations, can drive the chirality transfer from particle to supramolecular structure level during the crystallization process. By using a model system of chiral particles, which enables direct imaging of single particle incorporation into growing crystals, we show that the crystallization kinetic pathway is the key parameter for monitoring, via the defects, the chirality amplification of the crystalline structures from racemic to predominantly homohelical. We provide an explanation based on the interplay between geometrical frustration, racemization induced by thermal fluctuations, and particle chirality. Our results demonstrate that screw dislocations not only promote the growth, but also control the chiral morphology and therefore the functionality of crystalline states.

Liquid-crystalline nanoarchitectures for tissue engineering.

Hierarchical orders are found throughout all levels of biosystems, from simple biopolymers, subcellular organelles, single cells, and macroscopic tissues to bulky organs. Especially, biological tissues and cells have long been known to exhibit liquid crystal (LC) orders or their structural analogues. Inspired by those native architectures, there has recently been increased interest in research for engineering nanobiomaterials by incorporating LC templates and scaffolds. In this review, we introduce and correlate diverse LC nanoarchitectures with their biological functionalities, in the context of tissue engineering applications. In particular, the tissue-mimicking LC materials with different LC phases and the regenerative potential of hard and soft tissues are summarized. In addition, the multifaceted aspects of LC architectures for developing tissue-engineered products are envisaged. Lastly, a perspective on the opportunities and challenges for applying LC nanoarchitectures in tissue engineering fields is discussed.

Alternating Magnetic Field-Responsive Hybrid Gelatin Microgels for Controlled Drug Release.

Magnetically-responsive nano/micro-engineered biomaterials that enable a tightly controlled, on-demand drug delivery have been developed as new types of smart soft devices for biomedical applications. Although a number of magnetically-responsive drug delivery systems have demonstrated efficacies through either in vitro proof of concept studies or in vivo preclinical applications, their use in clinical settings is still limited by their insufficient biocompatibility or biodegradability. Additionally, many of the existing platforms rely on sophisticated techniques for their fabrications. We recently demonstrated the fabrication of biodegradable, gelatin-based thermo-responsive microgel by physically entrapping poly(N-isopropylacrylamide-co-acrylamide) chains as a minor component within a three-dimensional gelatin network. In this study, we present a facile method to fabricate a biodegradable drug release platform that enables a magneto-thermally triggered drug release. This was achieved by incorporating superparamagnetic iron oxide nanoparticles and thermo-responsive polymers within gelatin-based colloidal microgels, in conjunction with an alternating magnetic field application system.

Coexistence of coil and globule domains within a single confined DNA chain.

The highly charged DNA chain may be either in an extended conformation, the coil, or condensed into a highly dense and ordered structure, the toroid. The transition, also called collapse of the chain, can be triggered in different ways, for example by changing the ionic conditions of the solution. We observe individual DNA molecules one by one, kept separated and confined inside a protein shell (the envelope of a bacterial virus, 80 nm in diameter). For subcritical concentrations of spermine (4+), part of the DNA is condensed and organized in a toroid and the other part of the chain remains uncondensed around. Two states coexist along the same DNA chain. These 'hairy' globules are imaged by cryo-electron microscopy. We describe the global conformation of the chain and the local ordering of DNA segments inside the toroid.

Biodegradable colloidal microgels with tunable thermosensitive volume phase transitions for controllable drug delivery.

In this study, we present gelatin-based thermoresponsive colloidal microgels that enable the controlled release of drugs by volume phase transition. The microgel was fabricated by physically entrapping poly(N-isopropylacrylamide-co-acrylamide) chains as a minor component within three-dimensional gelatin networks crosslinked by genipin. We demonstrate that our gelatin-based thermoresponsive microgel exhibits a tunable deswelling to temperature increase, which positively correlated to the release of bovine serum albumin (BSA) as a function of poly(N-isopropylacrylamide-co-acrylamide) concentration. The microgel was enzymatically degradable by collagenase treatment. The extent of BSA release and biodegradability were tuned by controlling the crosslinking degree of the gelatin matrix. Meeting a great need for design and synthesis of auto-degenerating smart microgels that enable the controlled release of therapeutic proteins in responsive to external stimuli, our gelatin-based microgels that satisfy both thermoresponsivity and biodegradability have a great potential in tissue engineering applications as a soft microdevice element for drug delivery.

Electromechanical method coupling non-invasive skin impedance probing and in vivo subcutaneous liquid microinjection: controlling the diffusion pattern of nanoparticles within living soft tissues.

Transdermal drug delivery is the way to transport drug carriers, such as nanoparticles, across the skin barrier to the dermal and/or subcutaneous layer. In order to control the transdermal drug delivery process, based on the heterogeneous and nonlinear structures of the skin tissues, we developed a novel electromechanical method combining in vivo local skin impedance probing, subcutaneous micro-injection of colloidal nanoparticles, and transcutaneous electrical stimulation. Experiments on the nude mice using in vivo fluorescence imaging exhibited significantly different apparent diffusion patterns of the nanoparticles depending on the skin impedance: Anisotropic and isotropic patterns were observed upon injection into low and high impedance points, respectively. This result implies that the physical complexity in living tissues may cause anisotropic diffusion of drug carriers, and can be used as a parameter for controlling drug delivery process. This method also can be combined with microneedle-based drug release systems, micro-fabricated needle-electrodes, and/or advanced in vivo targeting/imaging technologies using nanoparticles.

Novel threadlike structures on the surfaces of mammalian abdominal organs are loose bundles of fibrous stroma with microchannels embedded with fibroblasts and inflammatory cells.

Novel threadlike structures (NTSs) on the surfaces of mammalian abdominal organs have recently attracted interests regarding their ability to transport fluid, enable cell migration, and possibly facilitate cancer metastasis. Nevertheless, histological studies of NTSs have been sporadic and often have inconsistent interpretations of the NTS internal structure. In this article, we provide a synthetic and consistent view of the NTS internal structure: the NTS is a loose bundle of fibrous stroma that forms interstitial channels and microsinusoids infiltrated with inflammatory cells. The fibroblasts are embedded in the stroma and mostly aligned along the major axis of the NTS. The sinusoids, which are in inconsecutive cross sections, have boundaries more or less delineated by extracellular fibers, partly surrounded by endothelial-like cells, or both. We compare these morphological features to other well-known connective tissues (i.e., trabecular meshwork and lymphatic capillary) and discuss the biomechanical and biological functions of NTSs based on their structural characteristics.

Fine structure of extracellular fibers in primo-nodes and vessels.

To investigate the extracellular matrices in the primo-nodes (PNs) and vessels (PVs) (Bonghan corpuscles and ducts), fine structure of the fibers comprising the extracellular matrices was studied in relation to the diameter and the periodicity to identify the types of the fibers. Electron micrographs of the fibers from the previously published works on the PNs and PVs were analyzed by using MATLAB programming and ImageJ software. The extracellular fibrous components of the PNs and PVs could be classified generally into two groups--thin collagen fibrils and thick non-collagenous wavy bent fibers. The thick bent fibers were fibrin-like. The diameters of the thick bent fibers were 30-200 nm and they were highly bent. This group included organ surface PNs and lymphatic PNs/PVs. The diameters of the collagen fibrils were 30-70 nm. Organ surface, heart, and hypodermis PVs were categorized in this group. The periodicity of bent fibers in an organ surface PN was 16.1 +/- 0.2 (SE) nm, and the periodicity of collagen fibrils in the hypodermis PVs was 37.4 +/- 0.5 (SE) nm. Further investigations are required to reveal the physiological implications of the two types of fibers in the PNs and PVs.

Network of endocardial vessels.

BACKGROUND: Although there have been reports on threadlike structures inside the heart, they have received little attention. We aimed to develop a method for observing such structures and to reveal their ultrastructures. METHODS: An in situ staining method, which uses a series of procedures of 0.2-0.4% trypan blue spraying and washing, was applied to observe threadlike structures on the surfaces of endocardia. The threadlike structures were isolated and observed by using confocal laser scanning microscopy (CLSM) and transmission electron microscopy (TEM). RESULTS: Networks of endocardial vessels (20 µm in thickness) with expansions (40-100 µm in diameter) were visualized; they were movable on the endocardium of the bovine atrium and ventricle. CLSM showed that (1) rod-shaped nuclei were aligned along the longitudinal direction of the endocardial vessel and (2) there were many cells inside the expansion. TEM on the endocardial vessel revealed that (1) there existed multiple lumens (1-7 µm in diameter) and (2) the extracellular matrices mostly consisted of collagen fibers, which were aligned along the longitudinal direction of the endocardial vessel or were locally organized in reticular structures. CONCLUSION: We investigated the endocardial circulatory system in bovine cardiac chambers and its ultrastructures, such as nucleic distributions, microlumens, and collagenous extracellular matrices.