Porous Alumina Membrane-Based Electrochemical Biosensor for Protein Biomarker Detection in Chronic Wounds.

A label-free electrochemical detection platform for the sensitive and rapid detection of Flightless I (Flii) protein, a biomarker of wound chronicity, has been developed using nanoporous anodic alumina (NAA) membranes modified with Flii antibody recognition sites. The electrochemical detection is based on the nanochannel blockage experienced upon Flii capture by immobilized antibodies within the nanochannels. This capture impedes the diffusion of redox species [[Fe(CN)6]4-/3-] toward a gold electrode attached at the backside of the modified NAA membrane. Partial blockage causes a decrease in the oxidation current of the redox species at the electrode surface which is used as an analytical signal by the reported biosensor. The resulting biosensing system allows detection of Flii at the levels found in wounds. Two types of assays were tested, sandwich and direct, showing <3 and 2 h analysis time, respectively, a significant reduction in time from the nearly 48 h required for the conventional Western blot assay. Slightly higher sensitivity values were observed for the sandwich-based strategy. With faster analysis, lack of matrix effects, robustness, ease of use and cost-effectiveness, the developed sensing platform has the potential to be translated into a point-of-care (POC) device for chronic wound management and as a simple alternative characterization tool in Flii research.

A time based objective evaluation of the erosive effects of various beverages on enamel and cementum of deciduous and permanent teeth.

BACKGROUND: Erosion of the teeth is a chronic irreversible process leading to loss of surface enamel and even the dentin, in turn causing sensitivity and pain. Increased consumption of carbonated beverages remains a major cause for dental erosion. However, many of the so called safe beverages that are consumed may also have sufficiently low pH to cause dental erosion. One of the parameters to measure the dental erosion is estimation of hardness and surface roughness. Thus, this study aims to evaluate the difference in hardness and surface roughness of enamel and cementum using three beverages namely (carbonated drink, lime soda, lime juice) in deciduous and permanent teeth. MATERIAL AND METHODS: Ten permanent and three deciduous teeth samples each were kept in lime juice, lime soda, carbonated beverage and tap water. The VHN using Vickers hardness tester and Ra value using surface profilometer were assessed at baseline, 1 day and 10 days. RESULTS: At the end of 10 days the decrease in hardness of enamel of permanent teeth was maximum for teeth immersed in carbonated beverage followed by lime soda and lime juice. However, in the deciduous teeth it was observed that the VHN drop was maximum at 1 day in relation to teeth immersed in carbonated beverage followed by lime juice and lime soda. The hardness of cementum decreased significantly at the end of ten days both in deciduous as well as permanent teeth. CONCLUSIONS: The present study shows that many of the most commonly used beverages like lime juice and lime soda have a sufficiently low pH to cause erosion of the enamel surface as well as that of cementum of both deciduous and permanent teeth. Though protective mechanisms do exist in the oral cavity to neutralize the acids present in these beverages, continuous usage of these beverages leads to irreversible damage to the tooth structure. Key words:Dental erosion, hardness, surface roughness, permanent teeth, deciduous teeth.

Magnetic Nanoparticles Enhance Pore Blockage-Based Electrochemical Detection of a Wound Biomarker.

A novel pore blockage-based electrochemical immunosensor based on the combination of 100 nm-magnetic nanoparticles (MNPs), as signal enhancers, and 200 nm-pore diameter nanoporous anodic alumina (NAA) membranes, as sensing platform, is reported. A peptide conjugate mimicking flightless I (Flii), a wound healing biomarker, was chosen as target analyte. The sensing platform consists of an anti-Flii antibody (Ab1)-modified NAA membrane attached onto a gold electrode. Anti-KLH antibody (Ab2)-modified MNPs (MNP-Ab2) were used to selectively capture the Flii peptide conjugate in solution. Sensing was based on pore blockage of the Ab1-modified NAA membrane caused upon specific binding of the MNP-Ab2-analyte complex. The degree of pore blockage, and thus the concentration of the Flii peptide conjugate in the sample, was measured as a reduction in the oxidation current of a redox species ([Fe(CN)6]4-) added in solution. We demonstrated that pore blockage is drastically enhanced by applying an external magnetic field at the membrane backside to facilitate access of the MNP-Ab2-analyte complex into the pores, and thus ensure its availability to bind to the Ab1-modified NAA membrane. Combining the pore blockage-based electrochemical magnetoimmunosensor with an externally applied magnetic field, a limit of detection (LOD) of 0.5 ng/ml of Flii peptide conjugate was achieved, while sensing in the absence of magnetic field could only attain a LOD of 1.2 µg/ml. The developed sensing strategy is envisaged as a powerful solution for the ultra-sensitive detection of an analyte of interest present in a complex matrix.

Microfluidic Cell Microarray Platform for High Throughput Analysis of Particle-Cell Interactions.

With advances in nanotechnology, particles with various size, shape, surface chemistry, and composition can be easily produced. Nano- and microparticles have been extensively explored in many industrial and clinical applications. Ensuring that the particles themselves are not possessing toxic effects to the biological system is of paramount importance. This paper describes a proof of concept method, in which a microfluidic system is used in conjunction with a cell microarray technique aiming to streamline the analysis of particle-cell interaction in a high throughput manner. Polymeric microparticles, with different particle surface functionalities, were first used to investigate the efficiency of particle-cell adhesion under dynamic flow. Silver nanoparticles (AgNPs, 10 nm in diameter) perfused at different concentrations (0 to 20 µg/mL) in parallel streams over the cell microarray exhibited a higher toxicity compared to the static culture in the 96-well-plate format. This developed microfluidic system can be easily scaled up to accommodate a larger number of microchannels for high throughput analysis of the potential toxicity of a wide range of particles in a single experiment.

Advances in Nanoporous Anodic Alumina-Based Biosensors to Detect Biomarkers of Clinical Significance: A Review.

There is a strong and growing demand for compact, portable, rapid, and low-cost devices to detect biomarkers of interest in clinical and point-of-care diagnostics. Such devices aid in early diagnosis of diseases without the need to rely on expensive and time-consuming large instruments in dedicated laboratories. Over the last decade, numerous biosensors have been developed for detection of a wide range of clinical biomarkers including proteins, nucleic acids, growth factors, and bacterial enzymes. Various transduction techniques have been reported based on biosensor technology that deliver substantial advances in analytical performance, including sensitivity, reproducibility, selectivity, and speed for monitoring a wide range of human health conditions. Nanoporous anodic alumina (NAA) has been used extensively for biosensing applications due to its inherent optical and electrochemical properties, ease of fabrication, large surface area, tunable properties, and high stability in aqueous environment. This review focuses on NAA-based biosensing systems for detection of clinically significant biomarkers using various detection techniques with the main focus being on electrochemical and optical transduction methods. The review covers an overview of the importance of biosensors for biomarkers detection, general (surface and structural) properties and fabrication of NAA, and NAA-based biomarker sensing systems.

On-demand Antimicrobial Treatment with Antibiotic-Loaded Porous Silicon Capped with a pH-Responsive Dual Plasma Polymer Barrier.

Chronic wounds are a major socio-economic problem. Bacterial infections in such wounds are a major contributor to lack of wound healing. An early indicator of wound infection is an increase in pH of the wound fluid. Herein, we describe the development of a pH-responsive drug delivery device that can potentially be used for wound decontamination in situ and on-demand in response to an increase in the pH of the wound environment. The device is based on a porous silicon film that provides a reservoir for encapsulation of an antibiotic within the pores. Loaded porous silicon is capped with dual plasma polymer layers of poly(1,7-octadiene) and poly(acrylic acid), which provide a pH-responsive barrier for on-demand release of the antibiotic. We demonstrate that release of the antibiotic is inhibited in aqueous buffer at pH 5, whereas the drug is released in a sustainable manner at pH 8. Importantly, the released drug was bacteriostatic against the Pseudomonas aeruginosa wound pathogen. In the future, incorporation of the delivery device into wound dressings could potentially be utilized for non-invasive decontamination of wounds.