Fabrication of Biomaterials and Biostructures Based On Microfluidic Manipulation.

Biofabrication technologies are of importance for the construction of organ models and functional tissue replacements. Microfluidic manipulation, a promising biofabrication technique with micro-scale resolution, can not only help to realize the fabrication of specific microsized structures but also build biomimetic microenvironments for biofabricated tissues. Therefore, microfluidic manipulation has attracted attention from researchers in the manipulation of particles and cells, biochemical analysis, tissue engineering, disease diagnostics, and drug discovery. Herein, biofabrication based on microfluidic manipulation technology is reviewed. The application of microfluidic manipulation technology in the manufacturing of biomaterials and biostructures with different dimensions and the control of the microenvironment is summarized. Finally, current challenges are discussed and a prospect of microfluidic manipulation technology is given. The authors hope this review can provide an overview of microfluidic manipulation technologies used in biofabrication and thus steer the current efforts in this field.

Mussel Inspired Trigger-Detachable Adhesive Hydrogel.

Adhesion to many kinds of surfaces, including biological tissues, is important in many fields but has been proved to be extremely challenging. Furthermore, peeling from strong adhesion is needed in many conditions, but is sometimes painful. Herein, a mussel inspired hydrogel is developed to achieve both strong adhesion and trigger-detachment. The former is actualized by electrostatic interactions, covalent bonds, and physical interpenetration, while the latter is triggered, on-demand, through combining a thixotropic supramolecular network and polymer double network. The results of the experiments show that the hydrogel can adhere to various material surfaces and tissues. Moreover, triggered by shear force, non-covalent interactions of the supramolecular network are destroyed. This adhesion can be peeled easily. The possible mechanism involved is discussed and proved. This work will bring new insight into electronic engineering and tissue repair like skin care for premature infants and burn victims.

Microfluidics for Biosynthesizing: from Droplets and Vesicles to Artificial Cells.

Fabrication of artificial biomimetic materials has attracted abundant attention. As one of the subcategories of biomimetic materials, artificial cells are highly significant for multiple disciplines and their synthesis has been intensively pursued. In order to manufacture robust "alive" artificial cells with high throughput, easy operation, and precise control, flexible microfluidic techniques are widely utilized. Herein, recent advances in microfluidic-based methods for the synthesis of droplets, vesicles, and artificial cells are summarized. First, the advances of droplet fabrication and manipulation on the T-junction, flow-focusing, and coflowing microfluidic devices are discussed. Then, the formation of unicompartmental and multicompartmental vesicles based on microfluidics are summarized. Furthermore, the engineering of droplet-based and vesicle-based artificial cells by microfluidics is also reviewed. Moreover, the artificial cells applied for imitating cell behavior and acting as bioreactors for synthetic biology are highlighted. Finally, the current challenges and future trends in microfluidic-based artificial cells are discussed. This review should be helpful for researchers in the fields of microfluidics, biomaterial fabrication, and synthetic biology.

Engineering of Hydrogel Materials with Perfusable Microchannels for Building Vascularized Tissues.

Vascular systems are responsible for various physiological and pathological processes related to all organs in vivo, and the survival of engineered tissues for enough nutrient supply in vitro. Thus, biomimetic vascularization is highly needed for constructing both a biomimetic organ model and a reliable engineered tissue. However, many challenges remain in constructing vascularized tissues, requiring the combination of suitable biomaterials and engineering techniques. In this review, the advantages of hydrogels on building engineered vascularized tissues are discussed and recent engineering techniques for building perfusable microchannels in hydrogels are summarized, including micromolding, 3D printing, and microfluidic spinning. Furthermore, the applications of these perfusable hydrogels in manufacturing organ-on-a-chip devices and transplantable engineered tissues are highlighted. Finally, current challenges in recapitulating the complexity of native vascular systems are discussed and future development of vascularized tissues is prospected.

A porous graphene sorbent coated with titanium(IV)-functionalized polydopamine for selective lab-in-syringe extraction of phosphoproteins and phosphopeptides.

A novel polydopamine coated three-dimensional porous graphene aerogel sorbent carrying immobilized titanium(IV) ions (denoted as Ti4+@PDA@GA) was fabricated without using an organic solvent. The material is shown to be a viable carbon foam type of monolithic sorbent for selective lab-in-syringe enrichment of phosphoproteins and phosphopeptides. The phosphoproteins can be separated from a sample by aspiration and then bind to the sorbent. The analytes then can be dispensed within 5 min. The weight percent of titanium in the monolith typically is 14%, and the absorption capacities for the model proteins ß-casein and κ-casein are 1300 and 1345 mg g-1, respectively. The absorption capacities for nonphosphoproteins are much smaller, typically 160 mg g-1 for ß-lactoglobulin, 125 mg g-1 for bovine serum, and 4.8 mg g-1 for lysozyme. The results demonstrate that the selectivity for phosphoproteins was excellent on multiple biological samples including standard protein mixtures, spiked human blood serum, and drinking milk. The selective enrichment of phosphopeptides also makes the method a promising tool in phosphoproteomics. Graphical abstract Schematic of a polydopamine coated three-dimensional porous graphene aerogel for immobilization of titanium(IV) ions. The material served as a monolithic sorbent for selective enrichment of phosphopeptides and phosphoproteins from biological samples. The enrichment process can be carried out conveniently using a lab-in-syringe way.

Engineered Microfibers for Tissue Engineering.

Hydrogel microfibers are hydrogel materials engineered into fiber structures. Techniques such as wet spinning, microfluidic spinning, and 3D bioprinting are often used to prepare microfibers due to their ability to precisely control the size, morphology, and structure of the microfibers. Microfibers with different structural morphologies have different functions; they provide a flow-through culture environment for cells to improve viability, and can also be used to induce the differentiation of cells such as skeletal muscle and cardiac muscle cells to eventually form functional organs in vitro through special morphologies. This Review introduces recent advances in microfluidics, 3D bioprinting, and wet spinning in the preparation of microfibers, focusing on the materials and fabrication methods. The applications of microfibers in tissue engineering are highlighted by summarizing their contributions in engineering biomimetic blood vessels, vascularized tissues, bone, heart, pancreas, kidney, liver, and fat. Furthermore, applications of engineered fibers in tissue repair and drug screening are also discussed.

Ultra-Small High-Entropy Alloy Nanoparticles: Efficient Nanozyme for Enhancing Tumor Photothermal Therapy.

High-entropy alloys nanoparticles (HEANPs) are receiving extensive attention due to their broad compositional tunability and unlimited potential in bioapplication. However, developing new methods to prepare ultra-small high-entropy alloy nanoparticles (US-HEANPs) faces severe challenges owing to their intrinsic thermodynamic instability. Furthermore, there are few reports on studying the effect of HEANPs in tumor therapy. Herein, the fabricated PtPdRuRhIr US-HEANPs act as bifunctional nanoplatforms for the highly efficient treatment of tumors. The US-HEANPs are engineered by the universal metal-ligand cross-linking strategy. This simple and scalable strategy is based on the aldol condensation of organometallics to form the target US-HEANPs. The synthesized US-HEANPs exhibit excellent peroxidase-like (POD-like) activity and can catalyze the endogenous hydrogen peroxide to produce highly toxic hydroxyl radicals. Furthermore, the US-HEANPs possess a high photothermal conversion effect for converting 808 nm near-infrared light into heat energy. In vivo and in vitro experiments demonstrated that under the synergistic effect of POD-like activity and photothermal action, the US-HEANPs can effectively ablate cancer cells and treat tumors. It is believed that this work not only provides a new perspective for the fabrication of HEANPs, but also opens the high-entropy nanozymes research direction and their biomedical application.

Characterization of alkaloids in radix Sophora tonkinensis by UPLC-Q-TOF-MS/MS and its application in the comparison of two different habitats.

Sophora tonkinensis is widely used as traditional Chinese medicine for treating the swelling of the gums and tongue and mouth sores due to flame stomach fire. It is mainly origin from Guangxi, Sichuan provinces of China. Alkaloids are considered as the major bioactive components. A method was established for identifying alkaloids in S. tonkinensis root by UPLC-Q-TOF-MS/MS and was applied in characterizing alkaloids in S. tonkinensis root of two different habitats. Consequently, twenty-four alkaloids including six new compounds were identified in S. tonkinensis root. Additionally, the difference of alkaloids in S. tonkinensis from Guozhou, Sichuan province was investigated. In the present study, we firstly characterize total alkaloids in S. tonkinensis root by UPLC-Q-TOF-MS/MS and firstly established the characteristic fragmentation pathway of alkaloids with hydroxy in S. tonkinensis root.

Comprehensive two-dimensional manipulations of picoliter microfluidic droplets sampled from nanoliter samples.

A facile method is presented for achieving comprehensive two-dimensional droplet manipulations in closed microstructures consisting of microwell arrays and a straight microchannel. In this method, picoliter/nanoliter droplets with controllable sizes and numbers are sampled from nanoliter samples/reagents with almost 100% efficiency. Droplet motions are precisely controlled in the ±X-direction and ±Y-direction by managing hydrostatic pressure and magnetic repulsion, respectively. As a demonstration, a fluorescein-labeled droplet and a deionized droplet are successively generated and trapped in adjacent microwells. Then their positions are quickly exchanged without cross-contamination and fusion is implemented on-demand. After operations, hydrophobic ferrofluid can be completely replaced by mineral oil and droplets still remain in microwells safely. A typical fluorescence intensity-based assay is demonstrated: droplet arrays containing copper ion are diluted disproportionately first and then detected by addition of droplet arrays containing Calcein. With the ability of comprehensive two-dimensional droplet manipulations, this method could be used in various miniaturized biochemical analyses including requirements of multistep procedures and in situ monitoring.

Self-Polymerized Dopamine-Decorated Au NPs and Coordinated with Fe-MOF as a Dual Binding Sites and Dual Signal-Amplifying Electrochemical Aptasensor for the Detection of CEA.

Fabrication of functional electrochemical biosensor is a hot topic; however, precise and sensitive cancer detection in early clinical diagnosis is still a great challenge. Continuous efforts have been devoted to explore functional materials for this issue. In this work, we developed a dual binding sites and dual signal-amplifying electrochemical aptasensor of self-polymerized dopamine-decorated Au and coordinated with Fe-MOF (Au@PDA@Fe-MOF) for the detection of carcinoembryonic antigen (CEA). Remarkably, Au@PDA@Fe-MOF features high sensitivity, multiple active sites, good biocompatibility, and excellent selectivity, which is attributed to abundant -COOH in porous Fe-MOF and unsaturated Fe3+ sites on the surface of Fe-MOF as the active binding sites grafting more NH2-functionalized CEA-specific aptamer and redox PDA and Fe-MOF accelerating the movement of electrons for dual signal amplifying. Meanwhile, the electrochemical aptasensor shows favorable repeatability with 1.82% relative standard deviation (RSD) under five independent aptasensors and strong stability with only 3.3% degradation after 12 days of storage. In addition, the aptasensor has wide CEA detection range from 1 fg mL-1 to 1 µg mL-1 with a low detection limit of 0.33 fg mL-1 (S/N = 3). Furthermore, the aptasensor is feasible for accurate and quantitative detection of CEA in serum samples with RSD below 2.32%. The satisfying results demonstrate promising applications of the CEA aptasensor in practical sample analysis and lay an important foundation for other biomarker detection in early clinical diagnosis.

Designed Fabrication of Polymer-Mediated MOF-Derived Magnetic Hollow Carbon Nanocages for Specific Isolation of Bovine Hemoglobin.

It is highly required to develop well-designed separation materials for the specific isolation of certain proteins in proteomic research. Herein, the new type of metal-organic framework (MOF)-derived polymer-mediated magnetic hollow nanocages was fabricated via stress-induced orientation contraction, which was further applied for specific enrichment of proteins. The core-shell nanocomposites comprised of polymer-mediated ZIF-67 cores and polydopamine (PDA) shells, after annealing, generated magnetic hollow carbon nanocages with hierarchical pores and structures. Particularly, the magnetic carbonized PDA@F127/ZIF-67 hollow nanocages exhibited a remarkable adsorption capacity toward bovine hemoglobin (BHB) up to 834.3 mg g-1, which was significantly greater than that of the directed carbonized ZIF-67 nanoparticles. The results also exhibited the notable specificity of the obtained nanocages on complex biosamples, including intact mixed proteins and fetal calf serum. The hierarchically hollow porous structure greatly improves the specific surface area and reduces the mass transfer resistance, leading to enhanced high adsorption for target protein BHB. This novel method will be promising for the applications in purification and enrichment of biomacromolecules for complex biosamples, which successfully solve the problem of low adsorption efficiency and tedious separating process of the previous MOF-derived materials.

Nitrite-Responsive Hydrogel: Smart Drug Release Depending on the Severity of the Nitric Oxide-Related Disease.

Nitric oxide (NO) is known as one of the most important biomarkers of many diseases. However, the development of NO-triggered drug releasing platforms is challenging due to the low concentration and short lifetime of NO in vivo. In this work, a novel nitrite (NO2-)-responsive hydrogel (DHPL-GEL), which can be used for smart drug release depending on the severity of the NO-related disease, is demonstrated. A dihydropyridine cross-linking agent is designed to construct DHPL-GEL to enable the responsive degradation of the hydrogel triggered by NO2-. On-demand release of the drug loaded in DHPL-GEL was observed under the stimulation of various concentrations of NO2- at the physiological level both in vitro and in vivo. In the inflammatory arthritis rat model, the DHPL-GEL drug delivery system showed a better therapeutic effect and less side effects than the traditional therapy and nonresponsive hydrogel drug delivery system, demonstrating the promising application of the NO2--responsive hydrogel for the treatment of NO-related diseases.

Double-Network Hydrogel with Tunable Mechanical Performance and Biocompatibility for the Fabrication of Stem Cells-Encapsulated Fibers and 3D Assemble.

Fabrication of cell-encapsulated fibers could greatly contribute to tissue engineering and regenerative medicine. However, existing methods suffered from not only unavoidability of cell damaging conditions and/or sophisticated equipment, but also unavailability of proper materials to satisfy both mechanical and biological expectations. In this work, a simple method is proposed to prepare cell-encapsulated fibers with tunable mechanical strength and stretching behavior as well as diameter and microstructure. The hydrogel fibers are made from optimal combination of alginate and poly(N-iso-propylacrylamide)-poly(ethylene glycol), characteristics of double-network hydrogel, with enough stiffness and flexibility to create a variety of three dimensional structures like parallel helical and different knots without crack. Furthermore, such hydrogel fibers exhibit better compatibility as indicated by the viability, proliferation and expression of pluripotency markers of embryonic stem cells encapsulated after 4-day culture. The double-network hydrogel possesses specific quick responses to either of alginate lyase, EDTA or lower environmental temperature which facilitate the optional degradation of fibers or fibrous assemblies to release the cells encapsulated for subsequent assay or treatment.

Controlling gas/liquid exchange using microfluidics for real-time monitoring of flagellar length in living Chlamydomonas at the single-cell level.

Chlamydomonas reinhardtii is widely used for studying cilia/flagella, organelles important for human health and disease. In situ monitoring of flagellar assembly/disassembly kinetics in single living cells has been difficult with conventional methods because of time-consuming media exchange and the requirement of whole cell fixation. Here, we develop a PDMS/glass hybrid microfluidic device for real-time tracking of flagellar length in single living cells of Chlamydomonas. Media exchange is precisely controlled by sequential gas-liquid plugs and complete medium replacement occurs within seconds. Rapid medium exchange allows the capture of transient flagellar dynamics. We show that Chlamydomonas cells respond to acidic medium exchange and deflagellate. However, the two flagella may shed asynchronously. After subsequent medium exchange, cells regenerate full-length flagella. Cells are also induced to shorten their flagella after being exposed to extracellular stimuli. The long-term kinetics of flagellar regeneration and disassembly for the whole cell population on the chip are comparable to those from conventional methods; however, individual cells display non-uniform response kinetics. We also find that flagellar growth rate is dependent on flagellar length. This device provides a potential platform to continuously monitor molecular activities associated with changes in flagellar length and to capture transient molecular changes upon flagellar loss, and initiation of flagellar assembly/disassembly.

Metabonomic study on the cumulative cardiotoxicity of a pirarubicin liposome powder.

Pirarubicin (THP) is an anthracycline frequently used in the chemotherapy against acute leukemia, malignant lymphoma and several solid tumors. However, its clinical use is severely limited by the development of a progressive dose-dependent cardiomyopathy that results in irreversible congestive heart failure. To provide a strategy for constraining or minimizing the cumulative cardiotoxicity of THP, a pirarubicin liposome powder (L-THP) was appropriately prepared, and the cumulative cardiotoxicity of L-THP and free THP (F-THP) were investigated on Sprague-Dawley rats after 3 successive doses. Urinary samples for metabonomic study, serum samples for biochemical assay, and heart samples for histopathology test were collected. As a result, the metabonomics-based findings such as PLS-DA plotting showed minimal metabolic alterations in L-THP as compared to F-THP, and correlated with the changes of serum biochemical assay and cardiac histopathology as measurements of damage to heart tissue. Our results confirm that when encapsulated into liposomes, the cumulative cardiotoxicity of THP can be greatly ameliorated. Lipophilic aglycone metabolites of THP associated with redox cycling are cardiotoxic for the possibility of reactive oxygen species (ROS) formation. Also, metabonomic analysis shows that the successive doses of THP will lead to severe metabolic pathways disturbances in the cell energy production. Further, the preliminary efficacy study of L-THP on lung cancer was evaluated in the approach of in vitro cytotoxicity on A549 cells by high content screening (HCS) analysis, and L-THP was found to exhibit better therapeutic index against lung cancer than THP.