Virus adsorbent systems based on Amazon holocellulose and nanomaterials.

The environment preservation has been an important motivation to find alternative, functional, and biodegradable materials to replace polluting petrochemicals. The production of nonbiodegradable face masks increased the concentration of microplastics in the environment, highlighting the need for sustainable alternatives, such as the use of local by-products to create efficient and eco-friendly filtering materials. Furthermore, the use of smart materials can reduce the risk of contagion and virus transmission, especially in the face of possible mutations. The development of novel materials is necessary to ensure less risk of contagion and virus transmission, as well as to preserve the environment. Taking these factors into account, 16 systems were developed with different combinations of precursor materials (holocellulose, polyaniline [ES-PANI], graphene oxide [GO], silver nanoparticles [AgNPs], and activated carbon [AC]). Adsorption tests of the spike protein showed that the systems containing GO and AC were the most efficient in the adsorption process. Similarly, plate tests conducted using the VSV-IN strain cultured in HepG2 cells showed that the system containing all phases showed the greatest reduction in viral titer method. In agreement, the biocompatibility tests showed that the compounds extracted from the systems showed low cytotoxicity or no significant cytotoxic effect in human fibroblasts. As a result, the adsorption tests of the spike protein, viral titration, and biocompatibility tests showed that systems labeled as I and J were the most efficient. In this context, the present research has significantly contributed to the technological development of antiviral systems, with improved properties and increased adsorption efficiency, reducing the viral titer and contributing efficiently to public health. In this way, these alternative materials could be employed in sensors and devices for filtering and sanitization, thus assisting in mitigating the transmission of viruses and bacteria. RESEARCH HIGHLIGHTS: Sixteen virus adsorbent systems were developed with different combinations of precursor materials (holocellulose, polyaniline (ES-PANI), graphene oxide (GO), silver nanoparticles (AgNPs), and activated carbon (AC)). The system that included all of the nanocomposites holocellulose, PANI, GO, AgNPs, and AC showed the greatest reduction in viral titration. The biocompatibility tests revealed that all systems caused only mild or moderate cytotoxicity toward human fibroblasts.

Nanoarchitectonic E-Tongue of Electrospun Zein/Curcumin Carbon Dots for Detecting Staphylococcus aureusin Milk.

We report a nanoarchitectonic electronic tongue made with flexible electrodes coated with curcumin carbon dots and zein electrospun nanofibers, which could detect Staphylococcus aureus(S. aureus) in milk using electrical impedance spectroscopy. Electronic tongues are based on the global selectivity concept in which the electrical responses of distinct sensing units are combined to provide a unique pattern, which in this case allowed the detection of S. aureus through non-specific interactions. The electronic tongue used here comprised 3 sensors with electrodes coated with zein nanofibers, carbon dots, and carbon dots with zein nanofibers. The capacitance data obtained with the three sensors were processed with a multidimensional projection technique referred to as interactive document mapping (IDMAP) and analyzed using the machine learning-based concept of multidimensional calibration space (MCS). The concentration of S. aureus could be determined with the sensing units, especially with the one containing zein as the limit of detection was 0.83 CFU/mL (CFU stands for colony-forming unit). This high sensitivity is attributed to molecular-level interactions between the protein zein and C-H groups in S. aureus according to polarization-modulated infrared reflection-absorption spectroscopy (PM-IRRAS) data. Using machine learning and IDMAP, we demonstrated the selectivity of the electronic tongue in distinguishing milk samples from mastitis-infected cows from milk collected from healthy cows, and from milk spiked with possible interferents. Calibration of the electronic tongue can also be reached with the MCS concept employing decision tree algorithms, with an 80.1% accuracy in the diagnosis of mastitis. The low-cost electronic tongue presented here may be exploited in diagnosing mastitis at early stages, with tests performed in the farms without requiring specialized laboratories or personnel.

PLGA-PVA-PEG Single Emulsion Method as a Candidate for Aminolevulinic Acid (5-ALA) Encapsulation: Laboratory Scaling Up and Stability Evaluation.

One of the most widely used molecules used for photodynamic therapy (PDT) is 5-aminolevulinic acid (5-ALA), a precursor in the synthesis of tetrapyrroles such as chlorophyll and heme. The 5-ALA skin permeation is considerably reduced due to its hydrophilic characteristics, decreasing its local bioavailability and therapeutic effect. For this reason, five different systems containing polymeric particles of poly [D, L-lactic-co-glycolic acid (PLGA)] were developed to encapsulate 5-ALA based on single and double emulsions methodology. All systems were standardized (according to the volume of reagents and mass of pharmaceutical ingredients) and compared in terms of laboratory scaling up, particle formation and stability over time. UV-VIS spectroscopy revealed that particle absorption/adsorption of 5-ALA was dependent on the method of synthesis. Different size distribution was observed by DLS and NTA techniques, revealing that 5-ALA increased the particle size. The contact angle evaluation showed that the system hydrophobicity was dependent on the surfactant and the 5-ALA contribution. The FTIR results indicated that the type of emulsion influenced the particle formation, as well as allowing PEG functionalization and interaction with 5-ALA. According to the 1H-NMR results, the 5-ALA reduced the T1 values of polyvinyl alcohol (PVA) and PLGA in the double emulsion systems due to the decrease in molecular packing in the hydrophobic region. The results indicated that the system formed by single emulsion containing the combination PVA-PEG presented greater stability with less influence from 5-ALA. This system is a promising candidate to successfully encapsulate 5-ALA and achieve good performance and specificity for in vitro skin cancer treatment.

Low-cost bacterial nanocellulose-based interdigitated biosensor to detect the p53 cancer biomarker.

Low-cost sensors to detect cancer biomarkers with high sensitivity and selectivity are essential for early diagnosis. Herein, an immunosensor was developed to detect the cancer biomarker p53 antigen in MCF7 lysates using electrical impedance spectroscopy. Interdigitated electrodes were screen printed on bacterial nanocellulose substrates, then coated with a matrix of layer-by-layer films of chitosan and chondroitin sulfate onto which a layer of anti-p53 antibodies was adsorbed. The immunosensing performance was optimized with a 3-bilayer matrix, with detection of p53 in MCF7 cell lysates at concentrations between 0.01 and 1000 Ucell. mL-1, and detection limit of 0.16 Ucell mL-1. The effective buildup of the immunosensor on bacterial nanocellulose was confirmed with polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS) and surface energy analysis. In spite of the high sensitivity, full selectivity with distinction of the p53-containing cell lysates and possible interferents required treating the data with a supervised machine learning approach based on decision trees. This allowed the creation of a multidimensional calibration space with 11 dimensions (frequencies used to generate decision tree rules), with which the classification of the p53-containing samples can be explained.

Correction: Sexual pheromone detection using PANI·Ag nanohybrid and PANI/PSS nanocomposite nanosensors.

Correction for 'Sexual pheromone detection using PANI·Ag nanohybrid and PANI/PSS nanocomposite nanosensors' by Janine Martinazzo et al., Anal. Methods, 2021, 13, 3900-3908, DOI: 10.1039/d1ay00987g.

Sexual pheromone detection using PANI·Ag nanohybrid and PANI/PSS nanocomposite nanosensors.

In this study, polyaniline/poly(styrene sulfonate) (PANI/PSS) nanocomposite and polyaniline·silver (PANI·Ag) nanohybrid thin films were obtained in cantilever nanosensors surface. The developed films were characterized in relation to topography, roughness, thickness, height, and structural properties. The topography study revealed that both films have a globular morphology, thickness and height in nanoscale. The gas sensing performance was investigated for sexual pheromone from the neotropical brown stink bug, Euschistus heros (F.). The sensitivities of both nanosensors based on PANI/PSS nanocomposite and PANI·Ag nanohybrid films were similar. The PANI·Ag nanohybrid nanosensor had a limit of detection of less than 3.1 ppq and limit of quantification of 10.05 ppq. The nanosensor layers were analyzed by UV-vis and FTIR showing the incorporation of Ag nanoparticles in the nanohybrid. We found that pheromone compound was adsorbed in sensing layer resulting in a reduction in the resonance frequency. The detection mechanism help us understand the good results of LOD, LOQ, sensitivity, selectivity and repeatability. The presented device has great potential for detection of the sexual pheromone from E. heros.

Controlled molecular architectures in microfluidic immunosensors for detecting Staphylococcus aureus.

Detection of pathogenic microorganisms is essential for food quality control and diagnosis of various diseases, which is currently performed with high-cost, sophisticated methods. In this paper, we report on a low-cost detection method based on impedance spectroscopy to detect Staphylococcus aureus (S. aureus). The immunosensors were made with microfluidic devices made of interdigitated electrodes coated with layer-by-layer (LbL) films of chitosan and chondroitin sulfate, on which a layer of anti-S. aureus antibodies was adsorbed. The limit of detection was 2.83 CFU mL-1 with a limit of quantification of 9.42 CFU mL-1 for immunosensors with 10-bilayer LbL films. This level of sensitivity is sufficient to detect traces of bacteria that cause mastitis in milk, which we have confirmed by distinguishing milk samples containing various concentrations of S. aureus from pure milk and milk contaminated with Escherichia coli (E. coli) and Salmonella. Distinction of these samples was made possible by projecting the electrical impedance data with the interactive document mapping (IDMAP) technique. The high sensitivity and selectivity are attributed to the highly specific interaction with anti-S. aureus antibodies captured with polarization-modulated reflection absorption spectroscopy (PM-IRRAS), with adsorption on the antibodies explained with the Langmuir-Freundlich model. Since these immunosensors are stable for up to 25 days and detection measurements can be made within minutes, the methodology proposed is promising for monitoring S. aureus contamination in the food industry and hospitals, and in detecting bovine mastitis.

Polysaccharide Multilayer Films in Sensors for Detecting Prostate Tumor Cells Based on Hyaluronan-CD44 Interactions.

The increasing need for point-of-care diagnosis has sparked the development of label-free sensing platforms, some of which are based on impedance measurements with biological cells. Here, interdigitated electrodes were functionalized with layer-by-layer (LbL) films of hyaluronan (HA) and chitosan (CHI) to detect prostatic tumor cells (PC3 line). The deposition of LbL films was confirmed with atomic force microscopy and polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS), which featured the vibrational modes of the HA top layer capable of interacting specifically with glycoprotein CD44 receptors overexpressed in tumor cells. Though the CHI/HA LbL films cannot be considered as a traditional biosensor due to their limited selectivity, it was possible to distinguish prostate tumor cells in the range from 50 to 600 cells/µL in in vitro experiments with impedance spectroscopy. This was achieved by treating the impedance data with information visualization methods, which confirmed the distinguishing ability of the films by observing the absence of false positives in a series of control experiments. The CD44-HA interactions may, therefore, be exploited in clinical analyses and point-of-care diagnostics for cancer, particularly if computational methods are used to process the data.

Immunosensors containing solution blow spun fibers of poly(lactic acid) to detect p53 biomarker.

This paper reports on biosensors made with a matrix of polylactic acid (PLA) fibers, which are suitable for immobilization of the anti-p53 active layer for detection of p53 biomarker. The PLA fibers were produced with solution blow spinning, a method that is advantageous for its simplicity and possibility to tune the fiber properties. For the biosensors, the optimized time to deposit the fibers was 60 s, with which detection of p53 could be achieved with the limit of detection of 11 pg/mL using electrical impedance spectroscopy. This sensitivity is also sufficient to detect the p53 biomarker in patient samples, which was confirmed by distinguishing samples from cell lines with distinct p53 concentrations in a plot where the impedance spectra were visualized with the interactive document mapping (IDMAP) technique. The high sensitivity and selectivity of the biosensors may be attributed to the specific interaction between the active layer and p53 modeled with a Langmuir-Freundlich and Freundlich isotherms and inferred from the analysis of the vibrational bands at 1550, 1650 and 1757 cm-1 using polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS). The successful immobilization of the active layer is evidence that the approach based on solution blown spun fibers may be replicated to other types of biosensors.

Detection of the Prostate Cancer Biomarker PCA3 with Electrochemical and Impedance-Based Biosensors.

Diagnosis of prostate cancer via PCA3 biomarker detection is promising to be much more efficient than with the prostatic specific antigens currently used. In this study, we present the first electrochemical and impedance-based biosensors that are capable of detecting PCA3 down to 0.128 nmol/L. The biosensors were made with a layer of PCA3-complementary single-stranded DNA (ssDNA) probe, immobilized on a layer-by-layer (LbL) film of chitosan (CHT) and carbon nanotubes (MWCNT). They are highly selective to PCA3, which was confirmed in impedance measurements and with polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS). Using information visualization methods, we could also distinguish between cell lines expressing the endogenous PCA3 long noncoding RNA (lncRNA) from cells that did not contain detectable levels of this biomarker. Since the methods involved in fabrication the biosensors are potentially low cost, one may hope to deploy PCA3 tests in any laboratory of clinical analyses and even for point-of-care diagnostics.

Microfluidic-Based Genosensor To Detect Human Papillomavirus (HPV16) for Head and Neck Cancer.

High-risk human papillomavirus (HPV) infection, mainly with HPV16 type, has been increasingly considered as an important etiologic factor in head and neck cancers. Detection of HPV16 is therefore crucial for these types of cancer, but clinical tests are not performed routinely in public health systems owing to the high cost and limitations of the existing tests. In this article, we report on a potentially low-cost genosensor capable of detecting low concentrations of HPV16 in buffer samples and distinguishing, with high accuracy, head and neck cancer cell lines according to their HPV16 status. The genosensor consisted of a microfluidic device that had an active layer of a HPV16 capture DNA probe (cpHPV16) deposited onto a layer-by-layer film of chitosan and chondroitin sulfate. Impedance spectroscopy was the principle of detection utilized, leading to a limit of detection of 10.5 pM for complementary ssDNA HPV16 oligos (ssHPV16). The genosensor was also able to distinguish among HPV16+ and HPV16- cell lines, using the multidimensional projection technique interactive document mapping. Hybridization between the ssHPV16 oligos and cpHPV16 probe was confirmed with polarization-modulated infrared reflection-absorption spectroscopy, where PO2 and amide I and amide II bands from adenine and thymine were monitored. The electrical response could be modeled as resulting from an adsorption process represented in a Freundlich model. Because the fabrication procedures of the microfluidic devices and genosensors and the data collection and analysis can be implemented at low cost, the results presented here amount to a demonstration of possible routine screening for HPV infections.

A simple architecture with self-assembled monolayers to build immunosensors for detecting the pancreatic cancer biomarker CA19-9.

The challenge of the early diagnosis of pancreatic cancer in routine clinical practice requires low-cost means of detection, and this may be achieved with immunosensors based on electrical or electrochemical principles. In this paper, we report a potentially low-cost immunosensor built with interdigitated gold electrodes coated with a self-assembled monolayer and a layer of anti-CA19-9 antibodies, which is capable of detecting the pancreatic cancer biomarker CA19-9 using electrical impedance spectroscopy. Due to specific, irreversible adsorption of CA19-9 onto its corresponding antibody, according to data from polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS), the immunosensor is highly sensitive and selective. It could detect CA19-9 in commercial samples with a limit of detection of 0.68 U mL-1, in addition to distinguishing between blood serum samples from patients with different concentrations of CA19-9. Furthermore, by treating the capacitance data with information visualization methods, we were able to verify the selectivity and robustness of the immunosensor with regard to false positives, as the samples containing higher CA19-9 concentrations, including those from tumor cells, could be distinguished from those with possible interferents.

Carbon Nanotube Matrix for Highly Sensitive Biosensors To Detect Pancreatic Cancer Biomarker CA19-9.

Biosensors fabricated with nanomaterials promise faster, cheaper, and more efficient alternatives to traditional, often bulky devices for early cancer diagnosis. In this study, we fabricated a thin film sensing unit on interdigitated gold electrodes combining polyethyleneimine and carbon nanotubes in a layer by layer fashion, onto which antibodies anti-CA19-9 were adsorbed with a supporting layer of N-hydroxysuccinimide and 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide solution. By use of impedance spectroscopy, the pancreatic cancer biomarker CA19-9 was detected in a buffer with limit of detection of 0.35 U/mL. This high sensitivity allowed for distinction between samples of blood serum from patients with distinct probabilities to develop pancreatic cancer. The selectivity of the biosensor was confirmed in subsidiary experiments with HT-29 and SW-620 cell lines and possible interferents, e.g., p53 protein, ascorbic acid, and glucose, where significant changes in capacitance could only be measured with HT-29 that contained the CA19-9 biomarker. Chemisorption of CA19-9 molecules onto the layer of anti-CA19-9 antibodies was the mechanism responsible for sensing while electrostatic interactions drove the adsorption of carbon nanotubes, according to polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS). The adsorption behavior was successfully described by the Langmuir-Freundlich isotherm.

Analysis of Scanning Electron Microscopy Images To Investigate Adsorption Processes Responsible for Detection of Cancer Biomarkers.

Adsorption processes are responsible for detection of cancer biomarkers in biosensors (and immunosensors), which can be captured with various principles of detection. In this study, we used a biosensor made with nanostructured films of polypyrrole and p53 antibodies, and image analysis of scanning electron microscopy data made it possible to correlate morphological changes of the biosensor with the concentration of cells containing the cancer biomarker p53. The selectivity of the biosensor was proven by distinguishing images obtained with exposure of the biosensor to cells containing the biomarker from those acquired with cells that did not contain it. Detection was confirmed with cyclic voltammetry measurements, while the adsorption of the p53 biomarker was probed with polarization-modulated infrared reflection absorption (PM-IRRAS) and a quartz crystal microbalance (QCM). Adsorption is described using the Langmuir-Freundlich model, with saturation taking place at a concentration of 100 Ucells/mL. Taken together, our results point to novel ways to detect biomarkers or any type of analyte for which detection is based on adsorption as is the case of the majority of biosensors.

Adsorption according to the Langmuir-Freundlich model is the detection mechanism of the antigen p53 for early diagnosis of cancer.

Biosensors for early detection of cancer biomarkers normally depend on specific interactions between such biomarkers and immobilized biomolecules in the sensing units. Though these interactions are expected to yield specific, irreversible adsorption, the underlying mechanism appears not to have been studied in detail. In this paper, we show that adsorption explained with the Langmuir-Freundlich model is responsible for detection of the antigen p53 associated with various types of cancers. Irreversible adsorption was proven between anti-p53 antibodies immobilized on the biosensors and the antigen p53, with the adequacy of the Langmuir-Freundlich model being confirmed with three independent experimental methods, viz. polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS), nanogravimetry using a quartz crystal microbalance and electrochemical impedance spectroscopy. The method based on this irreversible adsorption was sufficiently sensitive (limit of detection of 1.4 pg mL(-1)) for early diagnosis of Hodgkin lymphoma, pancreatic and colon carcinomas, and bladder, ovarian and lung cancers, and could distinguish between MCF7 cells containing the antigen p53 from Saos-2 cells that do not contain it.

Supramolecular Control in Nanostructured Film Architectures for Detecting Breast Cancer.

The need for early detection of various diseases, including breast cancer, has motivated research into nanomaterials that can be assembled in organized films which serve as biosensors. Owing to the variety of possible materials and film architectures, procedures are required to design optimized biosensors. In this study, we combine surface-specific methods to monitor the assembly of antibodies on nanostructured films with two distinct architectures. In the first, a layer of the antibody type mouse anti-HER2 (clone tab250) was immobilized on a self-assembled monolayer (SAM) of 11-mercaptoundecanoic acid modified with N-hydroxysuccinimide (NHS) and 1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide (EDC). In the second approach, a SAM of cysteamine was coated with a biotin/spreptavidin bilayer on which a layer of biotinylated antibody type MSx2HUp185/her biotin was adsorbed. The rougher, less passivating coating with cysteamine determined from cyclic voltammetry and scanning electron microscopy led to biosensors that are more sensitive to detect the breast cancer ERBB2 (HER2) biomarker in impedance spectroscopy measurements. This higher distinguishing ability of the cysteamine-containing film architecture was proven with information visualization methods to treat the impedance data. Polarization-modulated infrared reflection absorption spectroscopy (PM-IRRAS) confirmed that biosensing resulted from the antibody-ERBB2 antigen affinity.