Enzyme-Free Amplification Strategy for Biosensing Using Fe3+-Poly(glutamic acid) Coordination Chemistry.

In this work, we outline a signal amplification strategy using the coordination chemistry between Fe3+ and poly(glutamic acid) (PGA) for biosensing applications. The theoretical calculation based on density functional theory shows that PGA has a much higher binding affinity with Fe3+ than the other metal ions. Guided by this rationale, we prepare a PGA-mediated signal probe through conjugating PGA onto polystyrene (PS) nanoparticles to form a brushlike nanostructure for Fe3+ coordination. This PGA-PS brush (PPB) has a large loading capacity of Fe3+ with a number of 1.92 × 108 Fe atoms per nanoparticle that greatly amplifies the signals for assays in an enzyme-free way. Combined with ferrozine coloration-based readout, this PPB-mediated amplification is further applied for the enzyme-free immunoassay that shows an ultrahigh sensitivity for detection of microcystins-LR (12 pg/mL), a 5-fold enhancement compared with that of traditional enzyme-linked immunosorbent assay (ELISA) (60 pg/mL). In addition, the good stability, rapid response, and long shelf life make this enzyme-free amplification strategy a promising platform for point-of-care biosensing applications.

Double-Enzymes-Mediated Bioluminescent Sensor for Quantitative and Ultrasensitive Point-of-Care Testing.

We report an ultrasensitive, quantitative, and rapid bioluminescent immunosensor (ABS) for point-of-care testing (POCT) of the disease biomarker in clinical samples using double enzymes including alkaline phosphatase (ALP) and luciferase. In the presence of the biomarker, the ALP attached on the surface of immuno-nanocomplex dephosphorylates adenine triphosphate (ATP), subsequently inhibiting the ATP-luciferin-luciferase bioluminescent reaction. The highly sensitive response of ATP (picomolar level) allows for ultrasensitive detection of biomarker via the effective change of the bioluminescence intensity through ALP- and luciferase-catalyzed reactions, which can be quantitatively determined by a portable ATP detector. This ABS fulfills the criteria for POCT that performs sensitive (femtomolar level of biomarkers) and quantitative measurement quickly (less than 1 h) with minimal equipment (portable detector).

Nanocrystalline Cellulose-Assisted Generation of Silver Nanoparticles for Nonenzymatic Glucose Detection and Antibacterial Agent.

Nanocrystalline cellulose (NCC) is a kind of natural biopolymers with merits of large surface area, high specific strength and unique optical properties. This report shows that NCC can serve as the substrate, allowing glucose to reduce Tollen's reagent to produce silver nanoparticles (AgNPs) at room temperature. The generation of AgNPs is affected by the factors such as the concentrations of silver ions, NCC and glucose, as well as the different reaction temperatures. The AgNPs with NCC are applied for the development of a visual, quantitative, nonenzymatic and high-sensitive assay for glucose detection in serum. This assay is also used for monitoring the concentration change of glucose in medium during cell culture. For the antibacterial activity, the minimal inhibitory concentration (MIC) of the generated AgNPs with NCC is much lower than that of commercial AgNPs, attributed to the good dispersion of AgNPs with the presence of NCC. As NCC exhibits unique advantages including green, stable, inexpensive, and abundant, the NCC-based generation of AgNPs may find promising applications in clinical diagnosis, environmental monitoring, and the control of bacteria.

Polydimethylsiloxane Membranes Incorporating Metal-Organic Frameworks for the Sustained Release of Antibacterial Agents.

Cyclodextrin metal-organic frameworks (CD-MOFs) possess great potential in environmental applications due to their high specific surface area and good biocompatibility properties. However, the hydrophilicity of the CD-MOF hinders its ability to maintain a sustained release in water as a carrier. In this study, we prepared a CD-MOF that has codelivery ability for both phytochemicals [caffeic acid (CA)] and silver nanoparticles (Ag NPs) and further incorporated this material (CA@Ag@CD-MOF) into the polydimethylsiloxane (PDMS) matrix to construct a hybrid membrane. This hybrid membrane could effectively maintain the release capacity of the CD-MOF in water, while endowing PDMS with swelling ability in water. The hybrid membrane can achieve a sustained release for up to 48 h in water. In addition, the elastic modulus of the hybrid membrane increases by nearly 100%, and the swelling degree of the hybrid membrane in water increases by 42% compared with that of the pure PDMS membrane, indicating better mechanical properties. The hybrid membrane exhibits excellent antibacterial effects on Escherichia coli O157:H7 (E. coli O157:H7) and Staphylococcus aureus (S. aureus). We expect that this work will be beneficial to the delivery research of the CD-MOF in more environmental scenarios, especially in water treatment.

Horseradish peroxidase-catalyzed formation of polydopamine for ultra-sensitive magnetic relaxation sensing of aflatoxin B1.

Aflatoxin B1 as one of the most toxic mycotoxins poses a major health risk to humans and animals. Highly sensitive detection methods of aflatoxin B1 are urgently required because of its low abundance in biological samples. In this work, we developed a magnetic relaxation sensing strategy using enzyme-catalyzed formation of polydopamine for signal amplification. Horseradish peroxidase can catalyze the reaction to generate polydopamine that assembles magnetic nanoparticles for magnetic relaxation sensing with a high signal-to-noise ratio. Combined with the specific antigen-antibody interaction, this magnetic sensor enables fast and ultra-sensitive detection of aflatoxin B1 by using transverse relaxation time (T2) as a readout. Under optimized conditions, the linear range of this magnetic sensor for detecting aflatoxin B1 is from 10 pg/mL to 10 ng/mL, and the limit of detection is 0.35 pg/mL. This sensor has been challenged for the quantitative analysis of aflatoxin B1 in animal feed samples that is promising for real-world applications.

Broad-Range Magnetic Relaxation Switching Bioassays Using Click Chemistry-Mediated Assembly of Polystyrene Beads and Magnetic Nanoparticles.

Magnetic relaxation switching assays with a broad and tunable detection range can greatly improve current magnetic sensors for biochemical detections, but it remains challenging in terms of the limited detection range and low sensitivity. Herein, we report a methodology that uses click chemistry to assemble different sizes of polystyrene beads and magnetic nanoparticles to prepare versatile magnetic probes for broad-range bioassays with high sensitivity. Small magnetic nanoparticles can be controllably assembled on different sizes of polystyrene beads to form core-satellite structures, acting as broad-range probes that enable the magnetic relaxation switching assays with high sensitivity because different sizes of polystyrene beads can conjugate different numbers of small magnetic nanoparticles. On the basis of click chemistry, we assemble polystyrene beads and magnetic nanoparticles to develop a biosensing technique for analyzing three different antibiotics, with a high sensitivity and a tunable detection range from pg/mL to µg/mL.

A microfluidic tubing method and its application for controlled synthesis of polymeric nanoparticles.

This report describes a straightforward but robust tubing method for connecting polydimethylsiloxane (PDMS) microfluidic devices to external equipment. The interconnection is irreversible and can sustain a pressure of up to 4.5 MPa that is characterized experimentally and theoretically. To demonstrate applications of this high-pressure tubing technique, we fabricate a semicircular microfluidic channel to implement a high-throughput, size-controlled synthesis of poly(lactic-co-glycolic acid) (PLGA) nanoparticles ranging from 55 to 135 nm in diameter. This microfluidic device allows for a total flow rate of 410 mL h(-1), resulting in enhanced convective mixing which can be utilized to precipitate small size nanoparticles with a good dispersion. We expect that this tubing technique would be widely used in microfluidic chips for nanoparticle synthesis, cell manipulation, and potentially nanofluidic applications.

A microfluidic origami chip for synthesis of functionalized polymeric nanoparticles.

This report demonstrates a microfluidic origami chip to synthesize monodisperse, doxorubicin-loaded poly(lactic-co-glycolic acid) nanoparticles with diameters of ~100 nm, a size optimized for cellular uptake and anticancer efficacy, but difficult to achieve with existing approaches. This three-dimensional design in a microchannel may allow for the fabrication of polymeric nanoparticles in this size regime with ease.

Culturing primary human osteoblasts on electrospun poly(lactic-co-glycolic acid) and poly(lactic-co-glycolic acid)/nanohydroxyapatite scaffolds for bone tissue engineering.

In this work, we fabricated polymeric fibrous scaffolds for bone tissue engineering using primary human osteoblasts (HOB) as the model cell. By employing one simple approach, electrospinning, we produced poly(lactic-co-glycolic acid) (PLGA) scaffolds with different topographies including microspheres, beaded fibers, and uniform fibers, as well as the PLGA/nanohydroxyapatite (nano-HA) composite scaffold. The bone-bonding ability of electrospun scaffolds was investigated by using simulated body fluid (SBF) solution, and the nano-HA in PLGA/nano-HA composite scaffold can significantly enhance the formation of the bonelike apatites. Furthermore, we carried out in vitro experiments to test the performance of electrospun scaffolds by utilizing both mouse preosteoblast cell line (MC 3T3 E1) and HOB. Results including cell viability, alkaline phosphatase (ALP) activity, and osteocalcin concentration demonstrated that the PLGA/nano-HA fibers can promote the proliferation of HOB efficiently, indicating that it is a promising scaffold for human bone repair.