Fabrication of photo-induced molecular superoxide radical generator for highly efficient therapy against bacterial wound infection.

The pressing need for highly efficient antibacterial strategies arises from the prevalence of microbial biofilm infections and the emergence of rapidly evolving antibiotic-resistant strains of pathogenic bacteria. Photodynamic therapy represents a highly efficient and compelling antibacterial approach, offering promising prospects for effective control of the development of bacterial resistance. However, the effectiveness of many photosensitizers is limited due to the reduced generation of reactive oxygen species (ROS) in hypoxic microenvironment, which commonly occur in pathological conditions such as inflammatory and bacteria-infected wounds. Herein, we designed and prepared two phenothiazine-derived photosensitizers (NB-1 and NB-2), which can effectively generate superoxide anion radicals (O2●-) through the type I process. Both photosensitizers demonstrate significant efficacy in vitro for the eradication of broad-spectrum bacteria. Moreover, NB-2 possesses distinct advantages including strong membrane binding and strong generation of O2●-, rendering it an exceptionally efficient antibacterial agent against mature biofilms. In addition, laser activated NB-2 could be applied to treat MRSA-infected wound in vivo, which offers new opportunities for potential practical applications.

Multifunctional Smart ZnSe-Nanostructure-Based Fluorescent Aptasensor for the Detection of Ochratoxin A.

Herein, we present a comprehensive investigation of rationally designed zinc selenide (ZnSe) nanostructures to achieve highly negatively charged ZnSe nanostructures. A Microwave-assisted hydrothermal synthesis method was used to synthesize three types of ZnSe nanostructures, i.e., nanorods, µ-spheres and nanoclusters, as characterized by a zeta potential analyzer, X-ray diffraction (XRD), scanning electron microscopy (SEM), Raman spectroscopy and BET, which were labeled as type A, B and C. Three different solvents were used for the synthesis of type A, B and C ZnSe nanostructures, keeping other synthesis conditions such as temperature, pressure and precursors ratio constant. Based on two heating time intervals, 6 and 9 h, types A, B and C were further divided into types A6, A9, B6, B9, C6 and C9. ZnSe nanostructures were further evaluated based on their fluorescent quenching efficiency. The maximum fluorescence quenching effect was exhibited by the ZnSe-B6 type, which can be attributed to its highly negative surface charge that favored its strong interaction with cationic dye Rhodamine B (Rh-B). Further, the optimized ZnSe-B6 was used to fabricate an aptasensor for the detection of a food-based toxin, ochratoxin-A (OTA). The developed aptasensor exhibited a limit of detection of 0.07 ng/L with a wide linear range of 0.1 to 200 ng/L.

Black phosphorus nanosheets/poly(allylamine hydrochloride) based electrochemical immunosensor for the selective detection of human epididymis protein 4.

In this work, an electrochemical immunosensor based on black phosphorus nanosheets (BPNS)/poly(allylamine hydrochloride) (PAH) nanocomposite modified glassy carbon electrode was developed for the detection of ovarian cancer biomarker HE4. PAH has been applied to retain BPNS in its original honeycomb structure and to anchor biomolecules electrostatically on the transducer surface. The as synthesized nanocomposite was characterized by zeta potential analysis, scanning electron microscopy, x-ray photoelectron spectroscopy, transmission electron microscopy, high-resolution transmission electron microscopy. Subsequently, the performance of the electrochemical immunosensor was evaluated through cyclic voltammetry, differential pulse voltammetry and electrochemical impedance spectroscopy. Under the optimal condition, the developed electrochemical immunosensor permitted to detect HE4 with a linear range of 0.1-300 ng ml-1and a detection limit of 0.01 ng ml-1. The developed sensor exhibited good selectivity and specificity to HE4 with negligible interference effect from common biomolecules like bovine serum albumin, lysozyme, protamine, glucose, fructose, hemoglobin and fetal bovine serum. Further, practical application of developed electrochemical immunosensor was demonstrated in spiked human serum which showed satisfactory recovery percentages.

Polypeptide induced perylene probe excimer formation and its application in the noncovalent ratiometric detection of matrix metalloproteinase activity.

Matrix metalloproteinases (MMPs) are important biomarkers for a number of diseases. Thus, the precise determination of MMP activity is of crucial importance. Herein, we report a ratiometric fluorescence method for the sensitive and selective sensing of MMP activity. A number of positively charged MMP substrates (polypeptides) were designed and prepared. These polypeptides could induce aggregation of a negatively charged perylene diimide derivative (PC1). As a result, excimer fluorescence of PC1 was observed. Addition of the corresponding MMP resulted in cleavage of the polypeptide chain and dis-aggregation of PC1, which led to turning on of the PC1 monomer fluorescence. Based on the ratio of the monomer (545 nm, IM) and the excimer (680 nm, IM) fluorescence intensity changes, a ratiometric method I545/I680) was established to detect MMP activity. The enzymatic activity of a number of MMPs (MMP-1, 2, 3, 7, 9 and 13) could be determined with a limit of detection of 4.8, 2.2, 16, 6.0, 1.7 and 5.5 ng mL-1, respectively. Using MMP-2 and MMP-9 as examples, flavonoid herbal extracts as potential inhibitors were studied. It was observed that mangiferin, apigenin, quercetin and isoliquiritigenin had significant inhibiting effects on the enzyme activity. And these herbal extracts also inhibited tumor cell metastasis. Moreover, the developed strategy was also employed to determine the concentration of MMP-9 in human saliva samples. Since the method relies on only noncovalent interactions between the polypeptide and PC1, no covalent labeling of fluorescence dye on the polypeptide substrate is required, and the method is thus simple, broad-spectrum inexpensive and effective. It has the potential to be developed into a clinical test kit.

Controlled synthesis and upconversion luminescence properties of highly uniform and monodisperse ß-NaYF4 :Yb3+ ,Tm3+ nanocrystals.

Ytterbium, thulium (Yb3+ ,Tm3+ ) co-doped ß-NaYF4 upconversion nanocrystals (U-NCs) were synthesized by a convenient hydrothermal method using sodium citrate as a capping agent. X-ray diffraction (XRD) analysis revealed that NCs phase structures were dependent on reaction time. Field-emission scanning electron microscopy (FE-SEM) and transmission electron microscopy (TEM) results showed that the morphology of U-NCs could be controlled by adjusting the concentration of sodium citrate added in the hydrothermal reaction. Moreover, the emission spectra of the U-NCs were investigated to evaluate upconversion efficiency. Strong luminescence intensity of the U-NCs was observed after tuned with optimized concentration of sodium citrate. Furthermore, the U-NCs were incubated with MCF-7 cells at 37°C for 24 h. Under irradiation of a 980 nm laser, the upconversion blue emission of the NCs in MCF-7 cells can be clearly observed through a confocal microscope with an upconversion imaging system and high quality upconversion luminescence images can be acquired. Thus, prepared ß-NaYF4 :Yb3+ ,Tm3+ NCs provide us with highly efficient luminescent probes which can be applied for diverse bio-applications.

Tetraphenylethene probe based fluorescent silica nanoparticles for the selective detection of nitroaromatic explosives.

A simple and sensitive fluorometric method is developed utilizing aggregation-induced emission probe based silica nanoparticles for the detection of nitroaromatic explosives. A positively charged tetraphenylethene based probe (TPE-C2-2+) is doped into silica nanoparticles exploiting electrostatic interactions to produce TPE-SiO2 nanoparticles with a uniform particle size. The TPE-SiO2 nanoparticles exhibit strong fluorescence emission due to the aggregation-induced emission (AIE) effect of the doped TPE probe. The fluorescence emission of TPE-SiO2 offers quantitative and sensitive response to picric acid (PA), 2,4-dinitrotoluene (DNT) and 2,4,6-trinitrotoluene (TNT) which are used as model examples of nitroaromatic compounds. The fluorescence spectroscopy results show that the fluorescence emission of TPE-SiO2 was greatly quenched in the presence of the electron-poor nitroaromatic compounds due to the inner filter effect (IFE) and possibly the contact quenching mechanism. TPE-SiO2 nanoparticles show better sensitivity towards PA and could detect PA down to 0.01 µM with a linear detection range of 0.1-50 µM. The increased chemical stability, efficient high sensitivity and simple synthesis of the TPE-SiO2 nanoparticles demonstrate that they can be used as an excellent fluorescent probe for a wide range of electron-poor compounds, i.e. nitroaromatic compounds. Interference studies show that common interfering species with nitroexplosives such as acids, bases, volatile organic compounds, and salt solutions have a negligible effect during the sensing process.

Identification of milk adulteration by a sensor array based on cationic polymer induced aggregation of a perylene probe.

A novel fluorescence sensor array based on cationic polymer induced self-assembly of a perylene probe is developed. Cationic polymer induced aggregation of the carboxyl modified negatively charged perylene probe, and resulted in large quenching of monomer emission and generation of excimer emission. Upon the addition of negatively charged protein, monomer fluorescence restored with a decrease in excimer fluorescence. Based on these observations, we developed a six-channel sensor array to discriminate five main proteins in milk. In addition, we successfully identified pure milk out of different drinks using the developed sensor array since different drinks contained distinct species and contents of proteins. Furthermore, the sensor array exhibited excellent performance to discriminate milk adulterated by different concentrations of adulterants with 100% accuracy of cross validation. The analysis results also presented excellent linear correlation of adulterants contents and thus the developed sensor array shows great potential for quantitative detection of milk adulteration.

The use of aggregation-induced emission probe doped silica nanoparticles for the immunoassay of human epididymis protein 4.

A sensitive sandwich immunoassay for the sensing of human epididymis protein 4 (HE4) is developed based on aggregation-induced emission probe doped silica nanoparticles. Fluorescent silica nanoparticles (TSiO2) with excellent performance were prepared using a tetraphenylethylene based probe (TPE-C4-4) to produce a controlled aggregation effect, and HE4 was detected using antibody-functionalized fluorescent silica nanoparticles. The fluorescent silica nanoparticles possess good photophysical properties. The TSiO2 NPs were surface modified with carboxyl groups, and the antibody was covalently attached onto the surface of the nanoparticles. In addition, magnetic beads were modified with N-hydroxysulfosuccinimide sodium salt (NHS) on their surface, capable of forming stable peptide bonds with the antibody. TSiO2 NPs modified by an antibody and the magnetic beads modified by an antibody were used for the sandwich immunoassay for HE4 protein. The assay is quite sensitive and selective. The detection limit of HE4 is estimated to be 40 pM. The assay shows promising applications for the early diagnosis of ovarian cancer and the development of ovarian cancer-related drugs.

Development of a disposable electrochemical sensor for detection of cholesterol using differential pulse voltammetry.

In this study, a sensitive and selective electrochemical sensor was fabricated by using a screen printed carbon electrode (SPCE), multi-walled carbon nanotubes (MWCNTs) and ß-cyclodextrin (ß-CD) for detecting cholesterol. MWCNTs were functionalized with benzoic acid moiety by employing diazonium salt chemistry, and, subsequently, a thin film of functionalized CNTs were coated on the surface of SPCE. Afterwards, ß-CD was immobilized on functionalized MWCNTs modified SPCE which acts as a host to recognize guest (cholesterol) molecule specifically. Under the optimal experimental conditions and using differential pulse voltammetry (DPV) as transduction technique the sensor was able to detect cholesterol level ranges from 1 nM to 3 µM, with a detection limit of 0.5 nM. Specificity of the developed sensor towards target analyte (cholesterol) was confirmed in the presence of common interfering species including glucose, uric acid and ascorbic acid. The applicability of proposed sensor was also demonstrated for cholesterol determination in human serum samples with good recovery results (94-96%) and maximum RSD (relative standard deviation) of 4.5%.

Carboxylic group riched graphene oxide based disposable electrochemical immunosensor for cancer biomarker detection.

In this work, we have developed for the first time a carboxylic group riched graphene oxide based disposable electrochemical immunosensor for cancer biomarker detection using methylene blue (MB). The developed immunosensor is highly sensitive for detection of biomarker Mucin1 (MUC1) in human serum samples. Development of this disposable electrochemical immunosensor was premeditated by applying specific monoclonal antibodies against MUC1. In this method, we explored highly conductive surface of carboxylic group (-COOH-) rich graphene oxide (GO) on screen-printed carbon electrodes (SPCE). This modified GO-COOH-SPCE was employed for the detection of MUC1 protein based on the reaction with methylene blue (MB) redox probe using differential pulse voltammetry (DPV) technique. Developed immunosensor exhibited good detection range for MUC1 with excellent linearity (0.1 U/mL- 2 U/mL), with a limit of detection of 0.04 U/mL. Upon potential application of developed biosensor, good recoveries were recorded in the range of 96-96.67% with % R.S.D 4.2. Analytical performance of the developed immunosensor assures the applicability in clinical diagnostic applications.

Chitosan/hydroxyapatite (HA)/hydroxypropylmethyl cellulose (HPMC) spongy scaffolds-synthesis and evaluation as potential alveolar bone substitutes.

Alveolar bone loss is associated with infections and its augmentation is a pre-requisite for the success of dental implants. In present study, we aim to develop and evaluate novel freeze dried doxycycline loaded chitosan (CS)/hydroxyapatite (HA) spongy scaffolds where hydroxypropylmethyl cellulose (HPMC) was added as a crosslinker. Scaffolds displayed compressive strength of 14MPa/cm3 and 0.34 as elastic response. The interconnected pore diameter was 41-273µm, favorably provided the template supporting cells and transport. An overall 10% degradation was seen after 14day's studies at pH 7.4 in PBS. Doxycycline hyclate, a frequently used drug to counter oral infections, demonstrated an initial burst release (6-8h), followed by a sustain release profile for the remaining 64h. CS/HA/HPMC scaffolds were nontoxic and promoted pre-osteoblast cell viability as seen with live/dead calcein staining after 24h where scaffolds with 10% and 25% HPMC by weight of scaffold had more viable cells. Scaffolds with 10%, 20% and 25% HPMC by weight of scaffold showed efficient cellular adhesion as seen in scanning electron microscopy images (day 8) indicating that pre-osteoblast cells were able to adhere well on the surface and into the porous structure via cytoplasmic extensions. Hoechst 33258 nuclear staining at day 2 and 8 indicated cell proliferation which was further supported byMTT assay at day 2, 4 and 8. Although all scaffolds supported pre-osteoblast cell viability, alkaline phosphatase (ALP) staining demonstrated that upon induction, differentiation was pronounced in case of scaffolds with 10% HMPC scaffolds. Conclusively, these materials having all the required mechanical and biological properties are potential candidates for alveolar bone regeneration.

One Step Assembly of Thin Films of Carbon Nanotubes on Screen Printed Interface for Electrochemical Aptasensing of Breast Cancer Biomarker.

Thin films of organic moiety functionalized carbon nanotubes (CNTs) from a very well-dispersed aqueous solution were designed on a screen printed transducer surface through a single step directed assembly methodology. Very high density of CNTs was obtained on the screen printed electrode surface, with the formation of a thin and uniform layer on transducer substrate. Functionalized CNTs were characterized by X-ray diffraction spectroscopy (XRD), Fourier transform infrared spectroscopy (FTIR), thermogravimetric analysis (TGA) and Brunauer-Emmett- Teller (BET) surface area analyzer methodologies, while CNT coated screen printed transducer platform was analyzed by scanning electron microscopy (SEM), atomic force microscopy (AFM), cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The proposed methodology makes use of a minimum amount of CNTs and toxic solvents, and is successfully demonstrated to form thin films over macroscopic areas of screen printed carbon transducer surface. The CNT coated screen printed transducer surface was integrated in the fabrication of electrochemical aptasensors for breast cancer biomarker analysis. This CNT coated platform can be applied to immobilize enzymes, antibodies and DNA in the construction of biosensor for a broad spectrum of applications.

RutheniumII(η6-arene) complexes of thiourea derivatives: synthesis, characterization and urease inhibition.

RuII(arene) complexes have emerged as a versatile class of compounds to design metallodrugs as potential treatment for a wide range of diseases including cancer and malaria. They feature modes of action that involve classic DNA binding like platinum anticancer drugs, may covalent binding to proteins, or multimodal biological activity. Herein, we report the synthesis and urease inhibition activity of RuII(arene) complexes of the general formula [RuII(η6-p-cymene)(L)Cl2] and [RuII(η6-p-cymene)(PPh3)(L)Cl]PF6 with S-donor systems (L) based on heterocyclic thiourea derivatives. The compounds were characterized by 1H-, 13C{1H}- and 31P{1H}-NMR spectroscopy, as well as elemental analysis. The crystal structure of [chlorido(η6-p-cymene)(imidazolidine-2-thione)(triphenylphosphine)ruthenium(II)] hexafluorophosphate 11 was determined by X-ray diffraction analysis. A signal in the range 175-183 ppm in the 13C{1H}-NMR spectrum indicates the presence of a thione rather than a thiolate. This observation was also confirmed in the solid state by X-ray diffraction analysis of 11 which shows a C=S bond length of 1.720 Å. The compounds were tested for urease inhibitory activity and the thiourea-derived ligands exhibited moderate activity, whereas their corresponding Ru(arene) complexes were not active.