A sensitive sandwich-type electrochemical immunosensor using nitrogen-doped graphene/metal-organic framework-derived CuMnCoOx and Au/MXene for the detection of breast cancer biomarker.

In terms of cancer-related deaths among women, breast cancer (BC) is the most common. Clinically, human epidermal growth receptor 2 (HER2) is one of the most commonly used diagnostic biomarkers for facilitating BC cell proliferation and malignant growth. In this study, a disposable gold electrode (DGE) modified with gold nanoparticle-decorated Ti3C2Tx (Au/MXene) was utilized as a sensing platform to immobilize the capturing antibody (Ab1/Au/MXene). Subsequently, nitrogen-doped graphene (NG) with a metal-organic framework (MOF)-derived copper-manganese-cobalt oxide, tagged as NG/CuMnCoOx, was used as a probe to label the detection antibody (Ab2). A sandwich-type immunosensor (NG/CuMnCoOx/Ab2/HER2-ECD /Ab1/Au/MXene/DGE) was developed to quantify HER2-ECD. NG/CuMnCoOx enhances the conductivity, electrocatalytic active sites, and surface area to immobilize Ab2. In addition, Au/MXene facilitates electron transport and captures more Ab1 on its surface. Under optimal conditions, the resultant immunosensor displayed an excellent linear range of 0.0001 to 50.0 ng. mL-1. The detection limit was 0.757 pg·mL-1 with excellent selectivity, appreciable reproducibility, and high stability. Moreover, the applicability for determining HER2-ECD in human serum samples indicates its ability to monitor tumor markers clinically.

Highly sensitive FET sensors for cadmium detection in one drop of human serum with a hand-held device and investigation of the sensing mechanism.

As the heavy metal contamination is becoming worse, monitoring the heavy metal content in water or human body gets more and more important. In this research, a cadmium ion-selective field effect transistor (Cd-ISFET) for rapidly detecting cadmium ions has been developed and the mechanism of the sensor is also investigated in depth. Our Cd-ISFET sensor exhibits high sensitivity beyond the ideal Nernst sensitivity, wide dynamic range, low detection limit (∼10-11M), which is comparable with inductively coupled plasma mass spectrometry, and easy operation enabling people to detect cadmium ion by themselves. From the analysis of electrical measurement results, this Cd-ISFET is preferred to operate at the bias with the maximum transconductance of the FET to enhance the sensor signal. The AC impedance measurement is carried out to directly investigate the mechanism of an ion-selective membrane (ISM). From impedance results, the real part of the total impedance, which is the resistance, was shown to dominate the sensor signal. The potential drop across the ISM is caused by the heavy metal ion in the membrane, which is employed to the gate of the FET via an extended gate electrode. Cadmium ion detection in one drop of human serum with this sensor was demonstrated. This cost-effective and highly sensitive sensor is promising and can be used by anyone and anywhere to prevent people from cadmium poisoning.

Progress in Polymeric Nano-Medicines for Theranostic Cancer Treatment.

Cancer is a life-threatening disease killing millions of people globally. Among various medical treatments, nano-medicines are gaining importance continuously. Many nanocarriers have been developed for treatment, but polymerically-based ones are acquiring importance due to their targeting capabilities, biodegradability, biocompatibility, capacity for drug loading and long blood circulation time. The present article describes progress in polymeric nano-medicines for theranostic cancer treatment, which includes cancer diagnosis and treatment in a single dosage form. The article covers the applications of natural and synthetic polymers in cancer diagnosis and treatment. Efforts were also made to discuss the merits and demerits of such polymers; the status of approved nano-medicines; and future perspectives.

Improving the reproducibility, accuracy, and stability of an electrochemical biosensor platform for point-of-care use.

Electrochemical biosensors possess numerous desirable qualities for target detection, such as portability and ease of use, and are often considered for point-of-care (POC) development. Label-free affinity electrochemical biosensors constructed with semiconductor manufacturing technology (SMT)-produced electrodes and a streptavidin biomediator currently display the highest reproducibility, accuracy, and stability in modern biosensors. However, such biosensors still do not meet POC guidelines regarding these three characteristics. The purpose of this research was to resolve the limitations in reproducibility and accuracy caused by problems with production of the biosensors, with the aim of developing a platform capable of producing devices that exceed POC standards. SMT production settings were optimized and bioreceptor immobilization was improved through the use of a unique linker, producing a biosensor with exceptional reproducibility, impressive accuracy, and high stability. Importantly, the three characteristics of the sensors produced using the proposed platform all meet POC standards set by the Clinical and Laboratory Standards Institute (CLSI). This suggests possible approval of the biosensors for POC development. Furthermore, the detection range of the platform was demonstrated by constructing biosensors capable of detecting common POC targets, including circulating tumor cells (CTCs), DNA/RNA, and curcumin, and the devices were optimized for POC use. Overall, the platform developed in this study shows high potential for production of POC biosensors.

Study of near-infrared light-induced excitation of upconversion nanoparticles as a vector for non-viral DNA delivery.

Clinical requirements have necessitated the development of biomedical nanomaterials that can be implanted into tissues or bodies. Physiological regulation can be achieved in these nanomaterials through external light. The combination of nanomaterials with infrared optics can be termed optogenetics. The low autofluorescence of upconversion nanoparticles (UCNPs) has several applications in the biological field. For optogenetics applications, UCNPs with high fluorescence performance and photostability can solve the penetration depth problem. NaYF4:Yb,Tm nanocrystals with controllable sizes, shapes, and compositions were synthesized using a rapid coprecipitation method in organic solvent. UCNPs using single crystal nanoparticles provide higher chemical stability than those using amorphous phase. However, because UCNPs are usually capped with hydrophobic ligands, it is particularly important to prepare biocompatible UCNPs with specific molecular recognition capabilities. Surface modification and subsequent functionalization are essential for the application of inorganic nanomaterials in the biological environment and are arousing increasing research interest. Due to the high biocompatibility and high loading of materials, mesoporous silica and amine groups were selected as the best candidates. Expression of plasmid DNA in vivo and transfection efficiency were determined by fluorescence microscopy and flow cytometry. The MTT assay was used to evaluate the particle biocompatibility; the results showed that UCNP@mSiO2 has great biocompatibility. Additionally, at neutral pH, the cell surface is negatively charged. Therefore, the surface is functionalized with amino groups and can be electrostatically bound to DNA. Finally, UCNP@mSiO2-NH2 as a vector was applied in live cells by loading DNA; according to the results, DNA-UCNPs were successfully transfected in the primary cells, and NaYF4:Yb,Tm@mSiO2-NH2-DNA were observed to have good transfection efficiency by flow cytometry. It is expected that this work will provide a different method from the traditional adenovirus method and improve the immune response and side effects caused by adenovirus.

The Effects of Different Dynamic Culture Systems on Cell Proliferation and Osteogenic Differentiation in Human Mesenchymal Stem Cells.

The culture environment plays an important role for stem cells' cultivation. Static or dynamic culture preserve differential potentials to affect human mesenchymal stem cells' (hMSCs) proliferation and differentiation. In this study, hMSCs were seeded on fiber disks and cultured in a bidirectional-flow bioreactor or spinner-flask bioreactor with a supplement of osteogenic medium. The hMSCs' proliferation, osteogenic differentiation, and extracellular matrix deposition of mineralization were demonstrated. The results showed that the spinner flask improved cell viability at the first two weeks while the bidirectional-flow reactor increased the cell proliferation of hMSCs through the four-week culture period. Despite the flow reactor having a higher cell number, a lower lactose/glucose ratio was noted, revealing that the bidirectional-flow bioreactor provides better oxygen accessibility to the cultured cells/disk construct. The changes of calcium ions in the medium, the depositions of Ca2+ in the cells/disk constructs, and alkaline phosphate/osteocalcin activities showed the static culture of hMSCs caused cells to mineralize faster than the other two bioreactors but without cell proliferation. Otherwise, cells were distributed uniformly with abundant extracellular matrix productions using the flow reactor. This reveals that the static and dynamic cultivations regulated the osteogenic process differently in hMSCs. The bidirectional-flow bioreactor can be used in the mass production and cultivation of hMSCs for applications in bone regenerative medicine.

Development of lattice-inserted 5-Fluorouracil-hydroxyapatite nanoparticles as a chemotherapeutic delivery system.

Developing an effective vehicle for cancer treatment, hydroxyapatite nanoparticles were fabricated for drug delivery. When 5-Fluorouracil, a major chemoagent, is combined with hydroxyapatite nanocarriers by interclay insertion, the modified hydroxyapatite nanoparticles have superior lysosomal degradation profiles, which could be leveraged as controlled drug release. The decomposition of the hydroxyapatite nanocarriers facilitates the release of 5-Fluorouracil into the cytoplasm causing cell death. Hydroxyapatite nanoparticles with/without 5-Fluorouracil were synthesized and analyzed in this study. Their crystallization properties and chemical composition were examined by X-ray diffraction and Fourier transforms infrared spectroscopy. The 5-Fluorouracil release rate was determined by UV spectroscopy. The biocompatibility of hydroxyapatite-5-Fluorouracil extraction solution was assessed using 3T3 cells via a WST-8 assay. The effect of hydroxyapatite-5-Fluorouracil particles which directly work on the human lung adenocarcinoma (A549) cells was evaluated by a lactate dehydrogenase assay via contact cultivation. A 5-Fluorouracil-absorbed hydroxyapatite particles were also tested. Overall, hydroxyapatite-5-Fluorouracils were prepared using a co-precipitation method wherein 5-Fluorouracil was intercalated in the hydroxyapatite lattice as determined by X-ray diffraction. Energy dispersive scanning examination showed the 5-Fluorouracil content was higher in hydroxyapatite-5-Fluorouracil than in a prepared absorption formulation. With 5-Fluorouracil insertion in the lattice, the widths of the a and c axial constants of the hydroxyapatite crystal increased. The extraction solution of hydroxyapatite-5-Fluorouracil was nontoxic to 3T3 cells, in which 5-Fluorouracil was not released in a neutral phosphate buffer solution. In contrast, at a lower pH value (2.5), 5-Fluorouracil was released by the acidic decomposition of hydroxyapatite. Finally, the results of the lactate dehydrogenase assay revealed that 5-Fluorouracil-hydroxyapatite was highly toxic to A549 cells through direct culture, this phenomenon may result from lysosomal decomposition of particles causing 5-Fluorouracil releasing. The pH-responsive hydroxyapatite-5-Fluorouracil nanoparticles have the potential to be part of a selective drug-delivery system in chemotherapy for cancer treatment.

Fabrication of inorganic hydroxyapatite nanoparticles and organic biomolecules-dual encapsulated alginate microspheres.

Inorganic hydroxyapatite nanoparticles (HANPs) and two kinds of organic biomolecules (i.e., fluorescent dye rhodamine 6G and protein lysozyme) were coencapsulated into alginate microspheres through an air dynamical atomization with optimized operation conditions. The synthesized microspheres have several advantages: HANP provides osteoconductivity and mechanical strength, rhodamine 6G (R6G) and lysozyme act as model drugs, and alginate provides excellent biocompatibility and carboxylate functionality. The results of fluorescent microscopic images indicated the successful dual encapsulation of HANPs and lysozyme inside the alginate microspheres. Furthermore, the results of 3- (4,5-cimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide assay showed that the fabricated alginate microspheres could be uptaken by HepG2 without apparent cytotoxicity. The dual encapsulated alginate microspheres fabricated in this study show great potential in many biomedical applications.

The photothermal effect of silica-carbon hollow sphere-concanavalin A on liver cancer cells.

Hepatocellular carcinoma (HCC) is one of the most common cancers and causes of death by cancer. Concanavalin A (ConA) lectin can specifically bind to the glycoprotein receptors of HCC, which are produced by the aberrant overexpression of liver cancer cells. ConA was used in the current study to conjugate on silica-carbon hollow spheres (SCHSs) and applied in the thermal ablation therapy of liver cancer cell lines under near-infrared (NIR) laser irradiation. We found that the amount of ConA-SCHS complex binding to hepatoma cells was significantly higher than that seen with normal hepatocytes, based on flow cytometric analysis and confocal imaging. Hepatoma cells incubated with ConA-SCHSs were thus more easily killed by the subsequent irradiation with a NIR laser. The results show that the ConA-SCHS complex may enhance the interaction with highly expressed ConA receptors on hepatoma cells, and thus serve as an effective photothermal therapy agent for liver cancer treatment.

Application of carbon nanotubes layered on silicon wafer for the detection of breast cancer marker carbohydrate antigen 15-3 by immuno-polymerase chain reaction.

A highly sensitive detection of breast cancer marker, carbohydrate antigen 15-3 (CA 15-3) by carbon nanotube (CNT) based immuno-polymerase chain reaction was reported. The study was aimed to develop a precise and sensitive method to diagnose breast cancer and its recurrence. The hydrofluoric acid (HF) treated silicon wafer layered with bundled CNT was used as the substrate. The surface was treated with HNO3/H2SO4 to graft carboxyl groups on the tips of CNT. Subsequently, polyoxyethylene bis-amine was grafted to conjugate anti human CA 15-3 antibodies. Water contact angle measurement, scanning electron microscope, Fourier transform infrared spectrometer, Raman spectrometer and sodium dodecyl sulfate polyacrylamide gel electrophoresis were employed to confirm the surface modification. The captured antibodies on the CNT were used to capture the target antigen CA 15-3 and the biotinylated secondary antibodies were subsequently bound with the target antigen. A bi-functional streptavidin was used to link biotinylated DNA to the biotinylated detection antibodies. The biotinylated target DNA was amplified by PCR, and then analyzed by agarose gel electrophoresis. The lower limit of detection of CA 15-3 by the proposed immuno-PCR system was 0.001 U/mL, which is extremely sensitive than the other bioanalytical techniques.

Synthesis of partial stabilized cement-gypsum as new dental retrograde filling material.

The study describes the sol-gel synthesis of a new dental retrograde filling material partial stabilized cement (PSC)-gypsum by adding different weight percentage of gypsum (25% PSC+75% gypsum, 50% PSC+50% gypsum and 75% PSC+25% gypsum) to the PSC. The crystalline phase and hydration products of PSC-gypsum were characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM) analysis. The handling properties such as setting time, viscosity, tensile strength, porosity and pH, were also studied. The XRD and microstructure analysis demonstrated the formation of hydroxyapatite and removal of calcium dihydrate during its immersion in simulated body fluid (SBF) on day 10 for 75% PSC+25% gypsum. The developed PSC-gypsum not only improved the setting time but also greatly reduced the viscosity, which is very essential for endodontic surgery. The cytotoxic and cell proliferation studies indicated that the synthesized material is highly biocompatible. The increased alkaline pH of the PSC-gypsum also had a remarkable antibacterial activity.

A real time detection of the ovarian tumor associated antigen 1 (OVTA 1) in human serum by quartz crystal microbalance immobilized with anti-OVTA 1 polyclonal chicken IgY antibodies.

In this study, we developed a sensitive quartz crystal microbalance (QCM) sensor which employs the anti-ovarian tumor associated antigen 1 (OVTA 1) IgY polyclonal chicken antibodies on the crystal surface for the detection of OVTA 1 in human serum. The anti-OVTA 1 IgY antibodies were anchored on the thiol-activated sensor surface using 1-ethyl-3[3-dimethylaminopropyl] carbodiimide hydrochloride/N-hydroxysuccinimide (EDC-NHS) coupling. The responses for different concentrations of anti-OVTA 1 antibody were also studied and 40µg/mL was favorable for homogenous antibody coverage. The QCM sensor was used to monitor both immobilization and the target binding affinity. The anti-OVTA 1 antibody based QCM has allowed the competent detection of OVTA 1 in a high linear range of 0.5-10µg/mL. The total time needed to complete the detection was as short as 5-6h. The regeneration studies demonstrated that the proposed sensor was reusable up to 9cycles with a slight loss in binding affinity. The detection of OVTA 1 in human serum allows a potential exploitation of the anti-OVTA 1 polyclonal antibody based QCM immunoassay for the screening of OVTA 1 antigen.

Comparison of bioartificial pancreas performance in the bone marrow cavity and intramuscular space.

BACKGROUND AND AIMS: Bone marrow with a widely distributed and well-vascularized microenvironment that is capable of sustaining grafts is a potential site for islet transplantation. The femur bone marrow cavity offers sufficient space that may also receive the implantation of bioartificial pancreas (BAP). METHODS: Mouse insulinoma cells encapsulating in agarose gel were further enclosed in a calcium phosphate cement chamber to create a BAP. BAPs implanted into the femur bone marrow cavity of diabetics were compared with those implanted in the intramuscular space. Blood glucose level and C-peptide were determined perioperatively. RESULTS: The blood glucose level of the diabetics receiving BAPs in the intramuscular space decreased from 413 +/- 24 to 285 +/- 47 mg/dL at 1 day post-surgery. However, the blood glucose level returned to 398 +/- 35 mg/dL with undetectable serum C-peptide at 2 weeks postoperatively that reveals implant failure. The blood glucose level of diabetics receiving BAPs into the femur bone marrow cavity decreased from 422 +/- 32 to 247 +/- 52 mg/dL and maintained in the range of 288 +/- 47 mg/dL during the experimental period with an increase in C-peptide level from 6.1 +/- 2.8 to 104.7 +/- 16.4 pmol/L. CONCLUSIONS: This preliminary study indicates that the effectiveness of BAPs transplanted into the femur bone marrow cavity is superior to that implanted in the intramuscular space, which reveals the bone marrow may be a potential receptor site for the BAP transplantation.