Detecting Bacteria with Ultralow Concentrations by Enzymatic Cascade Reaction-Amplifying Strategy.

Bacteria can cause infectious diseases even at ultralow concentrations (<1 CFU/mL). It is important to rapidly identify bacterial contamination at ultralow concentrations. Herein, FITC-labeled gelatinase-sensitive nanoparticles (GNPs@FITCs) and NFM@GNP@FITCs are designed and fabricated as ultralow concentration bacteria detection platforms based on an enzymatic cascade reaction-amplifying strategy. Bacterial secretions could trigger the dissociation of GNPs@FITCs to release FITC, with gelatinase used as the model secretion. The detectable signal of ultralow concentration bacteria could be amplified effectively by the gelatinase-triggered cascade reaction. Bacterial concentration was evaluated by the change in fluorescence density. The results showed that the GNPs@FITCs and NFM@GNP@FITCs could be used for identifying bacterial contamination qualitatively, even when the bacterial contamination is lower than 1 CFU/mL. Moreover, the method has better timeliness and convenience, when compared with national standards. As solid films, NFM@GNP@FITCs have better long-term storage stability than GNPs@FITCs. The potential applications of GNPs@FITC and NFM@GNP@FITCs were proved by detecting pathogenic bacteria in food. All the results showed that the method has great potential for screening pathogenic bacterial contamination qualitatively.

Multifunctional Nanoplatform for Single NIR Laser-Regulated Efficient PDT/PTT/Chemotherapy.

The combination of phototherapy and chemotherapy with superior advantages is a promising strategy for cancer therapy. However, combination therapy is generally regulated by two different wavelengths of light or other stimuli, which results in complex operations and inevitable systemic side effects, even affecting therapeutic efficacy. Herein, we design a signal NIR light-regulated nanoplatform via the self-assembly process of reactive oxygen species (ROS)-sensitive prodrug (DTD), human serum albumin (HSA), and IR780 for combined photothermal/photodynamic therapy and chemotherapy. Upon 808 nm laser irradiation, IR780 in nanoparticles generates abundant ROS and a significant photothermal effect to achieve photothermal/photodynamic therapy. Meanwhile, the generating ROS further cleans up the thioketal link to release DOX for chemotherapy. Hence, signal NIR light can effectively control the process of combination therapy. In vivo and in vitro experiment results demonstrate that the multifunctional nanoparticles exhibit excellent antitumor efficacy via the combination of phototherapy and chemotherapy controlled by a signal NIR laser. Overall, the signal NIR light-regulated nanoparticles with combination therapy performance provide a versatile platform for enhancing antitumor efficacy.

A rapid screening platform for antibiotic susceptibility testing based on a simple colorimetric method.

Rapid screening platforms for antibiotic susceptibility testing (AST) are important in inhibiting bacterial resistance in clinical practice. Herein, a rapid screening platform is reported for AST, which is based on nanofiber membrane enrichment bacteria-assisted cell counting Kit-8 (CCK8) colorimetry. The absorbance of CCK8 formazan has a linear relationship with the number of bacteria. The interference of antibiotics in the absorbance of CCK8 formazan could be eliminated by separating planktonic bacteria from the culture medium using nanofiber membranes. The total detection time is 7-9 h, using the new screening platform, which is significantly shorter than that with the traditional method, and the limit of detection of this method is 10 CFU mL-1. The evaluation results of antibiotic susceptibility are identical when using the new screening method and traditional methods. This method meets the definition of "rapid testing" for antibiotic susceptibility by most microbiologists. Furthermore, the new screening platform for antibiotic susceptibility testing ability in vitro was proved using E. coli in urine and blood, and S. aureus in wound fluid as practical samples. All the results showed that the new screening platform is a promising method for rapid antibiotic susceptibility testing in vitro.

3-Aminophenylboronic acid-functionalized molybdenum disulfide quantum dots for fluorescent determination of hypochlorite.

A simple method is reported for hypochlorite determination based on fluorescence 3-aminophenylboronic acid-functionalized molybdenum disulfide quantum dots (B-MoS2 QDs). B-MoS2 QDs with strong fluorescence at 380 nm have been successfully synthesized by the amidation reaction between APBA and hydrothermal MoS2 QDs. Hypochlorite sensing was proposed utilizing the fluorescent quenching effect of 3,3',5,5'-tetramethylbenzidine dihydrochloride (TMB) on B-MoS2 QDs and the fast redox reaction between hypochlorite and TMB. The fluorescent quenching effect of TMB to B-MoS2 QDs was proved to be caused by static dynamic quenching and inner filter effect. A good linear relationship was obtained in the hypochlorite concentration range from 1 to 20 µM, and the limit of detection (LOD) was 36.8 nM. The proposed fluorescent detection assay was simple and fast, taking only 5 min at room temperature. Satisfactory results were obtained in the standard spike recovery tests on tap water and milk samples, which indicate high potential in constructing fluorescent bio-detection assays.

Ascorbic acid detector based on fluorescent molybdenum disulfide quantum dots.

A rapid and facile method is reported for the detection of ascorbic acid using molybdenum disulfide quantum dots (MoS2 QDs) as a fluorescence sensor. Water-soluble and biocompatible MoS2 QDs with the maximum fluorescence emission at 506 nm have been successfully synthesized by hydrothermal method and specific detection for ascorbic acid (AA) was constructed to utilize the modulation of metal ion on the fluorescence of MoS2 QDs and the affinity and specificity between the ligand and the metal ion. The fluorescence of MoS2 QDs was quenched by the irreversible static quenching of Fe3+ through the formation of a MoS2 QDs/Fe3+ complex, while the pre-existence of AA can retain the fluorescence of MoS2 QDs through the redox reaction between AA and Fe3+. Based on this principle, a good linear relationship was obtained in the AA concentration range 1 to 150 µM with a detection limit of 50 nM. The proposed fluorescent sensing strategy was proven to be highly selective, quite simple, and rapid with a requirement of only 5 min at room temperature (RT), which is particularly useful for rapid and easy analysis. Satisfactory results were obtained when applied to AA determination in fruits, beverages, and serum samples as well as AA imaging in living cells, suggesting its great potential in constructing other fluorescence detection and imaging platforms.

Wire templated electrodeposition of vessel-like structured chitosan hydrogel by using a pulsed electrical signal.

Herein, by performing a templated electrodeposition process with an oscillating electrical signal stimulation, a vessel-like structured chitosan hydrogel (diameter about 0.4 mm) was successfully prepared in the absence of salt conditions. Experimental results demonstrated that the hydrogel growth (e.g. the thickness) is linearly correlated with the imposed charge transfer and can be well quantified by using a theoretical moving front model. Morphological observations indicated that the heterogeneous multilayer structure was spatially and temporally controlled by an externally employed electrical signal sequence while the channel structure could be determined by the shaped electrode. Moreover, the oscillating ON-OFF cycles were proved to strongly affect the film structure, leading to a more compact hydrogel coating with a lower water content, higher crystallinity, complex layer architecture and relatively strong mechanical properties that could be easily peeled off as a free-standing hollow tube. Importantly, all the experiments were conducted under mild conditions that allowed additional enhancing materials to be added in to further improve the mechanical and/or biological properties. Thus, this work advances a very promising self-assembly technology for the construction of a multi-functional hydrogel coating and artificial blood vessel regeneration.

Glycomimetic-Conjugated Photosensitizer for Specific Pseudomonas aeruginosa Recognition and Targeted Photodynamic Therapy.

Due to the rapid development of bacterial resistance, there is an urgent need to explore new antibacterial agents to substitute for traditional antibiotic therapy. Photodynamic therapy has been identified as a promising bactericidal method to conquer antibiotic-resistant pathogens. To solve the problem of photosensitizer damage to normal tissues in vivo, we developed a boron-dipyrrolemethene (BODIPY)-based glycosylated photosensitizer for ablating Pseudomonas aeruginosa ( P. aeruginosa). This glycosylated photosensitizer exhibited good water solubility and generated 1O2 rapidly in an aqueous solution under light exposure. The photosensitizer did not cause detectable toxicity to human cells in the dark. Importantly, the photosensitizer was able to selectively attach to P. aeruginosa over normal cells, thus resulting in effective pathogen ablation by reactive oxygen species. Moreover, the photosensitizer inhibited over 90% of the biofilm formation produced by P. aeruginosa. The results indicate that the design of the macromolecular photosensitizer-induced bacterial death and inhibited biofilm formation provide a novel strategy for overcoming bacterial infection.

Hierarchically Three-Dimensional Nanofiber Based Textile with High Conductivity and Biocompatibility As a Microbial Fuel Cell Anode.

Microbial fuel cells (MFCs) encompass complex bioelectrocatalytic reactions that converting chemical energy of organic compounds to electrical energy. Improving the anode configuration is thought to be a critical step for enhancing MFCs performance. In present study, a hierarchically structured textile polypyrrole/poly(vinyl alcohol-co-polyethylene) nanofibers/poly(ethylene terephthalate) (referred to PPy/NFs/PET) is shown to be excellent anode for MFCs. This hierarchical PPy/NFs/PET anode affords an open porous and three-dimensional interconnecting conductive scaffold with larger surface roughness, facilitating microbial colonization and electron transfer from exoelectrogens to the anode. The mediator-less MFC equipped with PPy/NFs/PET anode achieves a remarkable maximum power density of 2420 mW m(-2) with Escherichia coli as the microbial catalyst at the current density of 5500 mA m(-2), which is approximately 17 times higher compared to a reference anode PPy/PET (144 mW m(-2)). Considering the low cost, low weight, facile fabrication, and good winding, this PPy/NFs/PET textile anode promises a great potential for high-performance and cost-effective MFCs in a large scale.

An Acid-Triggered Degradable and Fluorescent Nanoscale Drug Delivery System with Enhanced Cytotoxicity to Cancer Cells.

To reduce side-effects of anticancer drugs, development of nanocarriers with precise biological functions is a critical requirement. In this study, the multifunctional nanoparticles combining imaging and therapy for tumor-targeted delivery of hydrophobic anticancer drugs were prepared via self-assembly of amphiphilic copolymers obtained using RAFT polymerization, specifically, acid-labile ortho ester and galactose. First, boron-dipyrromethene dye-conjugated chain transfer agent provides fluorescent imaging capability for diagnostic application. Second, nanoparticles were stable under physiological conditions but degraded in acidic tumor microenvironment, leading to enhanced anticancer efficacy. Third, the application of biocompatible glycopolymers efficiently increased the target-to-background ratio through carbohydrate-protein interactions. Data from cell viability, cellular internalization, flow cytometry, biodistribution and anticancer efficacy tests showed that the drug-loaded nanoparticles were capable of inhibiting cancer cell proliferation with significantly enhanced capacity. Our newly developed multifunctional nanoparticles may thus facilitate the development of effective drug delivery systems for application in diagnosis and therapy of cancer.

Photopolymerized multifunctional sodium alginate-based hydrogel for antibacterial and coagulation dressings.

Trauma caused by tissue damage in clinical applications has posed a serious threat to public safety. Dressings with a single function cannot meet the needs of wound healing, but multifunctional dressings are difficult to achieve and obtain. To address this issue, this research designed a facile one-pot photo-crosslinking method to prepare multifunctional sodium alginate-based hydrogel dressings for effective wound healing. According to irregular wounds, sodium alginate-based hydrogel dressings can be quickly prepared anytime and anywhere. The structure and physicochemical properties of hydrogels are regulated by modulating the proportion of main components sodium alginate and acrylamide. The results showed the sodium alginate-based composite hydrogel as a candidate multifunctional dressing that exhibits excellent stretchability and compressibility, viscoelasticity, and suitable tissue-like adhesion. In vitro drug release and antibacterial experiments indicated that the hydrogel has effective antibacterial properties against S. aureus and P. aeruginosa. Furthermore, the haemostatic behaviour of the hydrogel was demonstrated using the coagulation activation test, whole blood-clotting test, and blood cell and platelet adhesion experiments. All these results demonstrated that the sodium alginate-based hydrogel had high application potential as a multifunctional medical dressing for wound healing.

Near-IR-Regulated Composite Hydrogel with Real-Time Infection Monitoring and a Combined Antibacterial Effect for Efficient Wound Management.

Chronic wounds induced by bacterial infection have seriously affected the health of people in the world. So, it is meaningful to develop a novel strategy with real-time infection monitoring and excellent antibacterial performance for enhancing wound management. Herein, we constructed a composite hydrogel by loading the pH indicator bromothymol blue (BTB) and gold nanocages containing 2,2'-azobis[2-(2-imidazolin-2-yl)propane] dihydrochloride (Au NCs@AIPH) into a polyacrylamide-co-poly(acrylic anhydride-modified oxidized sodium alginate) (PAM-co-PAOSA) hydrogel. In vitro and in vivo experimental results demonstrated that the composite hydrogel could effectively detect bacteria and diagnose the infection status of a wound in real time by showing visible color changes. In addition, the composite hydrogel containing Au NCs@AIPH possessed an excellent photothermal effect under near-IR (NIR) laser irradiation. The photothermal effect further activated AIPH to generate toxic free radicals to form combined antibacterial therapy for accelerating wound healing. Moreover, the composite hydrogel showed great biocompatibility. Therefore, the multifunctional hydrogel provided a novel wound management strategy for bacterial infection diagnosis and combined therapy in an infected wound.

NIR regulated upconversion nanoparticles@metal-organic framework composite hydrogel dressing with catalase-like performance and enhanced antibacterial efficacy for accelerating wound healing.

Developing a hydrogel dressing with excellent antibacterial efficacy for accelerating wound healing is high desirable in clinical applications. In this work, NIR regulated metal-organic framework composite hydrogel dressing was constructed for enhanced antibacterial efficacy and accelerated wound healing via the compounding between hydrogel and UCNPs@ZrMOF-Pt nanoparticles. The visible light emitted from upconvertion nanoparticles (UCNPs) activated porphyrin based metal-organic framework (MOF) in composite hydrogel to generate 1O2 for photodynamic antibacterial therapy under NIR laser irradiation. Moreover, the UCNPs@ZrMOF-Pt in composite hydrogel with catalase-like performance could effectively convert the high concentration H2O2 in wound to abundant O2, which relieved the hypoxic in infected wound. Thus, the photodynamic antibacterial efficacy was remarkably enhanced, leading to accelerate the wound healing. This work presented a novel strategy for high efficient antibacterial therapy and accelerated wound healing.

An ionic/thermal-responsive agar/alginate wet-spun microfiber-shaped hydrogel combined with grooved/wrinkled surface patterns and multi-functions.

A dual stimuli-responsive wet-spun microfiber-shaped hydrogel is prepared by injecting a hot blend of two stimuli biopolymers alginate (i.e., ionic-responsive) and agar (i.e., temperature-responsive) into a pre-cooling and metal cation containing coagulation bath. Experimental results indicate the fiber microstructure could be manipulated by the extrusion rate and cooling temperature, achieving an anisotropic shrinkage characteristic and novel grooved/wrinkled surface patterns. Importantly, the integration of metal cations (e.g., Ca2+and/or Zn2+) was confirmed to significantly improve the hydrogel mechanical properties (i.e., double networks) and enhanced blue fluorescent intensity as a typical metal-polymer complexation formed within the agar gel matrix. Moreover, the functionality-independent double networks enabled typical pH-shape memory and sustainable antibacterial properties have also been demonstrated. Therefore, combing the facile fabricating approach and multifunctionality, this study would advance the development of stimuli-responsive hydrogel microfiber for complex biomedical systems.

Rapid and ratiometric fluorescent detection of hypochlorite by glutathione functionalized molybdenum disulfide quantum dots.

We proposed a rapid and ratiometric fluorescent detection method for hypochlorite by glutathione functionalized molybdenum disulfide quantum dots (G-MoS2 QDs). The G-MoS2 QDs were obtained through a hydrothermal method and the maximum fluorescence intensity was obtained at 430 nm under excitation of 360 nm. The fluorescence of G-MoS2 QDs at 430 nm can be weakened by curcumin through the inner filter effect, meanwhile the fluorescence of curcumin at 540 nm appeared. Hypochlorite can fast oxidize curcumin and weaken the inner filter effect, thus the fluorescence of curcumin at 540 nm decreased and the fluorescence of G-MoS2 QDs at 430 nm increased. This process takes only 30 s at room temperature. This is the rationale behind our rapid ratiometric fluorescent detection model for hypochlorite. Two linear detection ranges for hypochlorite are obtained with concentration from 1 to 20 µM and 20 to 30 µM, the limit of detection (LOD) was 11.5 nM. The standard spike recovery tests on milk and tap water samples showed satisfactory results, which extended the application of G-MoS2 QDs in the field of ratiometric fluorescence detection assays.

Ionic crosslinking of alginate/carboxymethyl chitosan fluorescent hydrogel for bacterial detection and sterilization.

Herein, a polysaccharide-based fluorescent hydrogel with multi-responsiveness simply implemented by concurrent effects of ionic crosslinking/rehydration processes is presented. Specifically, the alginate and carboxymethyl chitosan are chosen to prepare the interpenetrating polymer matrix while a pair of metal cations has been selectively sequentially integrated to alter hydrogel mechanical and fluorescent properties. Experimental results indicate the hydrogels show tunable fluorescent emission in response to multiple cations and pH conditions, and display a reversible "ON/OFF" fluorescent response to Mn+/ethylenediaminetetraacetic acid. Moreover, this synergistic ionic crosslinking strategy is proved to be highly effective in preparing multifunctional metallohydrogels possessing robust/anisotropic mechanical properties, typical shape memory and cation/pH-responsive fluorescence performance, and a proof-of-application for bacterial detection and sterilization has also been demonstrated. Therefore, we believe this study would provide new insights into multifunctional luminescent hydrogels for advanced biomedical systems.

pH-sensitive alginate hydrogel for synergistic anti-infection.

This work designed a pH-sensitive sodium alginate hydrogel for combating bacterial infection caused by tissue damage. The antibacterial hydrogels were prepared using sodium alginate, citric acid, and vancomycin by one-step in situ method. Vancomycin (Van) was loaded into hydrogels via reversible imine bonds for controlled drug delivery. The morphology, swelling properties, and antibacterial activity of hydrogel were characterized. The hydrogel shown strong water absorbent behavior and pH-dependent performance. The result under weak acid conditions, the drug release rate of van-loaded gel was faster than neutral and alkaline conditions and followed zero-order kinetic release model, and the cumulative release amount could reach 86.7 % over 320 min. The van-loaded gel had highly effective antibacterial activity in a weak acid environment, the combination of citric acid and vancomycin had a synergistic therapeutic effect for acute infection. The drug-loaded hydrogel shows good biocompatibility. Compared with gauze, the drug-loaded hydrogel exhibited good coagulation properties, high platelet adhesion, high fluid absorption capacity, and proper balance of fluid on the wound bed. This work proposed this simple alginate-based drug delivery system has potential applications in the field of clinical treatment of infections.

Gelatinase-responsive photonic crystal membrane for pathogenic bacteria detection and application in vitro health diagnosis.

Selective identification and rapid detection for multiple pathogenic bacteria are important to prevent bacterial infection. Herein, gelatinase-responsive photonic crystal membrane (PCM) is reported as rapid, selective and direct detection platform for multiple pathogenic bacteria. PCMs exhibit angle-independent structural color, and reflection spectra has negligible change after irradiation and storage under different temperature. The ultra-thin response layer makes shorter detection time (30 min) than the reported gelatin-based photonic crystal detection platform (10-12 h). There is obvious positive correlation between reflection spectra change and gelatinase concentration, even if the gelatinase was as low as 4.8 pM. PCM has high stability against multiple interferers, which are beneficial to improve the detection accuracy and reliability. PCM also show "selective" ability to identify typical pathogens from atypical pathogens through specific response to gelatinase, and good selectivity gives PCM direct detection ability without pretreatment. PCM could detect the pathogens bacteria concentration range spanning 7 orders of magnitude from 10 to 107 CFU/mL, and the relative error between gold standard method and the new platform is less than 10%. Furthermore, PCM's in vitro health diagnosis ability was proved by detecting pathogenic bacteria in artificial wound fluid and urine. All the results show that PCM is well promising platform for selective identifying and rapid detecting pathogenic bacteria in vitro diagnosis.

A simple colorimetric method for viable bacteria detection based on cell counting Kit-8.

In this study, Cell Counting Kit-8 (CCK-8) was introduced to detect the concentration of live bacteria for the first time depending on the redox reaction between CCK-8 solution and dehydrogenase. CCK-8 solution can be reduced to form water soluble orange-yellow formazan by the dehydrogenase present in bacterial cells, and the concentration of live bacteria is proportional to the absorbance value of formazan at 450 nm. Based on this principle, Staphylococcus aureus and Escherichia coli were chosen as the model bacteria. The optimal detection conditions were investigated and a good linear relationship was obtained in the concentration range from 2.600 × 102 to 1.160 × 109 CFU mL-1 with a linear equation of Y = 0.06305 log10 X-0.1153 (X in CFU mL-1, R2 = 0.9747) for S. aureus and 9.750 × 102 to 6.000 × 108 CFU mL-1 with a linear equation of Y = 0.06122 log10 X-0.1358 (X in CFU mL-1, R2 = 0.9958) for E. coli. The CCK-8 based viable bacteria detection method can be completed within 2 h with a wide bacterial detection concentration range. Satisfactory results were obtained when applied to an actual sample analysis and there is a good consistency between the proposed CCK-8 based method and the traditional plate counting method. More importantly, this method can realize the one-time detection of a large number of samples with high sensitivity, which suggests its great potential in high-throughput bacterial detection.

Interpenetrating polysaccharide-based hydrogel: A dynamically responsive versatile medium for precisely controlled synthesis of nanometals.

Herein, we reported an interpenetrating polysaccharide-based hydrogel in which carboxymethyl chitosan (CMC) chains were physically dispersed throughout the thermoplastic elastomer gel network has been developed as a versatile platform for precisely controlled synthesis of nanometals. Results indicated the interpenetrated CMC chains could serve as multifunctional fillers for metal ions adsorption and stabilization while the thermally reconfigurable agarose (Agar) gel medium provides three-dimensional semi-solid framework for entrapping and recollecting of the fabricated nanometals. Specifically, the CMC chains were found to strongly coordinate with silver ions as a dynamically responsive metal-biopolymer complex within the bulk gel network as confirmed by the enhanced mechanical properties and regulated shape memory performances. Moreover, by varying CMC concentrations and coupling with a layer-stacking approach, multiple biochemical gradients could be facilely generated for in-situ synthesis of silver nanoparticles, achieving a narrow size of ~7 nm, confined sphere-shape and high concentrations. The monodispersed nanometals are confirmed to be highly active (e.g., considerable catalytic performance), and which could be easily recycled from the bulk gel system via a heating treatment. Thus, this work would provide a generic methodology for the multifunctional metallogel assembly and great possibility for controllable and largescale synthesis of noble nanometals toward biomedical applications.

Nanofiber-reinforced bulk hydrogel: preparation and structural, mechanical, and biological properties.

Alginate-based hydrogels are increasingly being used as biomaterials for tissue engineering, drug carriers, and wound dressing; however, their poor mechanical strength limits their applications. Nanofiber reinforcement is an effective method for increasing the mechanical strength of hydrogels. However, the macro preparation of nanofiber-reinforced hydrogels with a bulk structure is challenging. Herein, we describe the fabrication of nanofiber-reinforced bulk alginate hydrogel composites. The mechanical properties of hydrogels were significantly improved, and the reinforcement law of nanofiber was systematically studied. The maximum tensile stress (0.76 MPa) was obtained with 30% nanofiber content, which was 87% higher than that of pure alginate hydrogel. The compressive stress of the composite hydrogel exhibited "J-curve" behavior with gradually increasing nanofiber content, which indicated that the composited hydrogels were suitable as biomaterials. Furthermore, in 2 h, the hydrogels killed more than 90% of the bacteria that were present, and the bacteriostatic rate reached 100% after 12 h of treatment. More importantly, the sterile environment continued to be maintained, and the composited hydrogel also had satisfactory cytocompatibility and cell adhesion. Compared with pure alginate hydrogel, the roughness of the composited hydrogel surface was increased, which resulted in stronger cell adhesion. Therefore, the composite hydrogel demonstrated improved mechanical and biological properties, and exhibited the potential for clinical application.