Here, we reported a study on the detection and electrical characterization of both cancer cell line and primary tumor cells. Dielectrophoresis (DEP) and electrical impedance spectroscopy (EIS) were jointly employed to enable the rapid and label-free differentiation of various cancer cells from normal ones. The primary tumor cells that were collected from two colorectal cancer patients, cancer cell lines (SW-403, Jurkat, and THP-1), and healthy peripheral blood mononuclear cells (PBMCs) were trapped first at the level of interdigitated microelectrodes with the help of dielectrophoresis. Correlation of the cells dielectric characteristics that was obtained via electrical impedance spectroscopy (EIS) allowed evident differentiation of the various types of cell. The differentiations were assigned to a “dielectric phenotype” based on their crossover frequencies. Finally, Randles equivalent circuit model was employed for highlighting the differences with regard to a series group of charge transport resistance and constant phase element for cancerous and normal cells.

Protein immobilization using biopolymer scaffolds generally involves undesired protein loss of function due to denaturation, steric hindrance or improper orientation. Moreover, most methods for protein immobilization require expensive reagents and laborious procedures. This work presents the synthesis and proof of concept application of two alginate hydrogels that are able to bind proteins with polyhistidine tags by means of interaction with the crosslinking cations. Nickel (II) and cobalt (II) alginate hydrogels were prepared using a simple ionic gelation method. Hydrogels were characterized by optical microscopy and AFM, and evaluated for potential cytotoxicity. In addition, binding capacity was tested towards proteins with or without HisTAG. Hydrogels had moderate cytotoxicity and were able to exclusively bind polyhistidine-tagged proteins with a binding capacity of approximately 300 µg EGFP (enhanced green fluorescent protein) per 1 mL of hydrogel. A lyophilized hydrogel-protein complex dissolved upon the addition of PBS and allowed the protein release and regain of biological activity. In conclusion, the nickel (II) and cobalt (II) alginate biopolymers provided an excellent platform for the “carry and release” of polyhistidine-tagged proteins.

In this study, ligand-free nanogels (LFNGs) as potential antivenom mimics were developed with the aim of preventing hypersensitivity and other side effects following massive bee attacks. For this purpose, poly (ethylene glycol) diacrylate was chosen as a main synthetic biocompatible matrix to prepare the experimental LFNGs. The overall concept uses inverse mini-emulsion polymerization as the main route to deliver nanogel caps with complementary cavities for phospholipase A2 (PLA2) from bee venom, created artificially with the use of molecular imprinting (MI) technologies. The morphology and the hydrodynamic features of the nanogels were confirmed by transmission electron microscopy (TEM) and dynamic light scattering (DLS) analysis. The following rebinding experiments evidenced the specificity of molecularly imprinted LFNG for PLA2, with rebinding capacities up to 8-fold higher compared to the reference non-imprinted nanogel, while the in vitro binding assays of PLA2 from commercial bee venom indicated that such synthetic nanogels are able to recognize and retain the targeted PLA2 enzyme. The results were finally collaborated with in vitro cell-viability experiments and resulted in a strong belief that such LFNG may actually be used for future therapies against bee envenomation.

This study proposes a feasible approach for the rapid, sensitive, and label-free identification of cancerous cells based on dielectrophoretic (DEP) manipulation and electrical characterization. In this method, the concentration of target cells at the level of customized microelectrodes via DEP is first determined, followed by an electrical impedance evaluation. The study demonstrates the capacity of the methodology to electrically differentiate HT-29 cancer cells from healthy blood cells based on their impedance spectra. Within a higher frequency domain, the electrical impedance of trapped cancer cells was significantly lower compared with the normal ones. In order to evaluate the functionality and reproducibility of the proposed method, the influence of the DEP and EIS (electrical impedance spectroscopy) operating voltages on the electrical characterization of trapped HT-29 cells was analyzed.

Influenza virus infections represent an ongoing public health threat as well as an economic burden. Although seasonal influenza vaccines have been available for some decades, efforts are being made to generate new efficient, flexible, and cost-effective technologies to be transferred into production. Our work describes the development of a model influenza hemagglutinin antigen that is capable of inducing protection against viral challenge in mice. High amounts of the H1 hemagglutinin ectodomain, HA_18–528, were expressed in a bacterial system as insoluble inclusion bodies. Solubilization was followed by a thorough differential scanning fluorimetry (DSF)-guided optimization of refolding, which allows for fast and reliable screening of several refolding conditions, yielding tens of milligrams/L of folded protein. Structural and functional analysis revealed native-like folding as well as the presence of a mix of monomers and oligomers in solution. Mice immunized with HA_18–528 were protected when exposed to influenza A virus as opposed to mice that received full-length denatured protein. Sera of mice immunized with HA_18–528 showed both high titers of antigen-specific IgG1 and IgG2a isotypes as well as viral neutralization activity. These results prove the feasibility of the recombinant bacterial expression system coupled with DSF-guided refolding in providing influenza hemagglutinin for vaccine development.

Objectives: The present study evaluated the efficacy of a preparation based on bacterial lysates of Streptococcus oralis, Aggregatibacter actinomycetemcomitans and Fusobacterium nucleatum in the treatment of experimental periodontitis in rats, compared to metronidazole. Methods: Twenty female Wistar rats were used, divided into three groups: control, bacterial lysates and metronidazole, administered for 10 days by oral lavage/gavage. Periodontitis was induced by ligatures contaminated with bacterial suspensions (10⁹ CFU/mL) for 4 weeks. Lysates were obtained by culturing bacterial strains, centrifugation, washing, heat inactivation, ultrasonication and filtration. The evaluations included biocompatibility on HGF-1 fibroblasts, microbiological stability, clinical parameters, hematological, biochemical and histopathological analyses. Results: The lysates demonstrated the absence of cytotoxicity (cell viability 90–100%) and significant antimicrobial effect at the optimal concentration (2 × 10⁹ CFU/mL equivalent). Both treatments significantly reduced periodontal inflammation, with no statistical differences between them. Systemic immunoinflammatory indices (SII, SIRI, AISI) increased comparably, demonstrating controlled immune mobilization, and ALT was maintained within physiological limits. Histopathological examination revealed a reduction in inflammatory infiltrate, connective tissue reorganization and bone regeneration in both treated groups. Conclusions: Bacterial lysates represent a viable therapeutic alternative with comparable efficacy to metronidazole, favorable safety profile and immunomodulatory potential in the treatment of periodontitis.