Engineering a Point-of-Care Paper-Microfluidic Electrochemical Device Applied to the Multiplexed Quantitative Detection of Biomarkers in Sputum.

Health initiatives worldwide demand affordable point-of-care devices to aid in the reduction of morbidity and mortality rates of high-incidence infectious and noncommunicable diseases. However, the production of robust and reliable easy-to-use diagnostic platforms showing the ability to quantitatively measure several biomarkers in physiological fluids and that could in turn be decentralized to reach any relevant environment remains a challenge. Here, we show the particular combination of paper-microfluidic technology, electrochemical transduction, and magnetic nanoparticle-based immunoassay approaches to produce a unique, compact, and easily deployable multiplex device to simultaneously measure interleukin-8, tumor necrosis factor-α, and myeloperoxidase biomarkers in sputum, developed with the aim of facilitating the timely detection of acute exacerbations of chronic obstructive pulmonary disease. The device incorporates an on-chip electrochemical cell array and a multichannel paper component, engineered to be easily aligned into a polymeric cartridge and exchanged if necessary. Calibration curves at clinically relevant biomarker concentration ranges are produced in buffer and artificial sputum. The analysis of sputum samples of healthy individuals and acutely exacerbated patients produces statistically significant biomarker concentration differences between the two studied groups. The device can be mass-produced at a low cost, being an easily adaptable platform for measuring other disease-related target biomarkers.

Designing antimicrobial biomembranes via clustering amino-modified cellulose nanocrystals on silk fibroin ß-sheets.

The continuous rising of infections caused by multidrug-resistant pathogens is becoming a global healthcare concern. Developing new bio-based materials with unique chemical and structural features that allow efficient interaction with bacteria is thus important for fighting this phenomenon. To address this issue, we report an antimicrobial biomaterial that results from clustering local facial amphiphilicity from amino-modified cellulose on silk fibroin ß-sheets, by simply blending the two components through casting technology. A simple but effective method for creating a membrane that is antibacterial and non-cytotoxic. Amino-modified cellulose nanocrystals (CNC-NH2) were mixed with proteinaceous silk fibroin (SF) which resulted in a material with improved crystallinity, higher ß-sheet content, and exposed amino-groups at its surface features, proven by Fourier transform infrared (FTIR) spectroscopy and X-ray photoelectron spectroscopy (XPS), that does not occur when the components are individually assembled. The resulting material possesses important antibacterial activity inducing >3 CFU log10 reduction of Escherichia coli and Staphylococcus epidermidis, while the pristine membranes show no antibacterial effect. The chemical interactions occurring between SF and CNC-NH2 during casting, exposing the amino moieties at the surface of the material, are proposed as the main reason for this antimicrobial activity. Importantly, the membranes are non-cytotoxic, showing their potential to be used as a new bioinspired material with intrinsic antibacterial activity for biomedical applications. Those may include coatings for medical devices for the control of healthcare-associated infections, with no need for including external antimicrobial agents in the material.

Ionic Liquids as Biocompatible Antibacterial Agents: A Case Study on Structure-Related Bioactivity on Escherichia coli.

The potential of ionic liquids (ILs) to be used as antimicrobial agents for biomedical applications has been hindered by the fact that most of them are cytotoxic toward mammalian cells. Understanding the mechanism of bacterial and mammalian cellular damage of ILs is key to their safety design. In this work, we evaluate the antimicrobial activity and mode of action of several ILs with varying anions and cations toward the clinically relevant Gram-negative Escherichia coli. Langmuir monolayer technique was used to evaluate if the IL's mode of action was related to the bacterial cell membrane interaction for an effective E. coli killing. 1-Decyl-3-methylimidazolium bis(trifluoromethylsulfonyl) imide [DMIM][TFSI] and trihexyltetradecyl phosphonium bis(trifluoromethylsulfonyl) imide [P6,6,6,14][TFSI] were surface-active and induced bacterial cell lysis, through a membrane-disruption phenomenon on bacteria, in a mechanism that was clearly related to the long alkyl chains of the cation. 1-Ethyl-3-methylimidazolium hydrogen sulfate [EMIM][HSO4] was highly antimicrobial toward E. coli and found suitable for biological applications since it was harmless to mammalian cells at most of the tested concentrations. The results suggest that the imidazolium cation of the ILs is mostly responsible not only for their antimicrobial activity but also for their cytotoxicity, and the inclusion of different anions may tailor the ILs' biocompatibility without losing the capacity to kill bacteria, as is the case of [EMIM][HSO4]. Importantly, this IL was found to be highly antimicrobial even when incorporated in a polymeric matrix.

Exploring electroactive microenvironments in polymer-based nanocomposites to sensitize bacterial cells to low-dose embedded silver nanoparticles.

The search for alternative antimicrobial strategies capable of avoiding resistance mechanisms in bacteria are highly needed due to the alarming emergence of antimicrobial resistance. The application of physical stimuli as a mean of sensitizing bacteria for the action of antimicrobials on otherwise resistant bacteria or by allowing the action of low quantity of antimicrobials may be seen as a breakthrough for such purpose. This work proposes the development of antibacterial nanocomposites using the synergy between the electrically active microenvironments, created by a piezoelectric polymer (poly(vinylidene fluoride-co-trifluoroethylene) (PVDF-TrFE)), with green-synthesized silver nanoparticles (AgNPs). The electrical microenvironment is generated via mechanical stimulation of piezoelectric PVDF-TrFE/AgNPs films using a lab-made mechanical bioreactor. The generated material's electrical response further translates to bacterial cells, namely Escherichia coli and Staphylococcus epidermidis which in combination with AgNPs and the specific morphological features of the material induce important antibacterial and antibiofilm activity. Both porous and non-porous PVDF composites have shown antibacterial characteristics when stimulated at a mechanical frequency of 4 Hz being the effect boosted when AgNPs were incorporated in the nanocomposite, reducing in more than 80% the S. epidermidis bacterial growth in planktonic and biofilm form. The electroactive environments sensitize the bacteria allowing the action of a low dose of AgNPs (1.69% (w/w)). Importantly, the material did not compromise the viability of mammalian cells, thus being considered biocompatible. The piezoelectric stimulation of PVDF-based polymeric films may represent a breakthrough in the development of antibacterial coatings for devices used at hospital setting, taking advantage on the use of mechanical stimuli (pressure/touch) to exert antibacterial and antibiofilm activity. STATEMENT OF SIGNIFICANCE: The application of physical methods in alternative to the common chemical ones is seen as a breakthrough for avoiding the emergence of antimicrobial resistance. Antimicrobial strategies that take advantage on the capability of bacteria to sense physical stimuli such as mechanical and electrical cues are scarce. Electroactive nanocomposites comprised of poly(vinylidene fluoride-co-trifluoroethylene (PVDF-TrFE) and green-synthesized silver nanoparticles (AgNPs) were developed to obtain material able to inhibit the colonization of microorganisms. By applying a mechanical stimuli to the nanocomposite, which ultimately mimics movements such as walking or touching, an antimicrobial effect is obtained, resulting from the synergy between the electroactive microenvironments created on the surface of the material and the AgNPs. Such environments sensitize the bacteria to low doses of antimicrobials.

Biodegradable Hydrogels Loaded with Magnetically Responsive Microspheres as 2D and 3D Scaffolds.

Scaffolds play an essential role in the success of tissue engineering approaches. Their intrinsic properties are known to influence cellular processes such as adhesion, proliferation and differentiation. Hydrogel-based matrices are attractive scaffolds due to their high-water content resembling the native extracellular matrix. In addition, polymer-based magnetoelectric materials have demonstrated suitable bioactivity, allowing to provide magnetically and mechanically activated biophysical electrical stimuli capable of improving cellular processes. The present work reports on a responsive scaffold based on poly (L-lactic acid) (PLLA) microspheres and magnetic microsphere nanocomposites composed of PLLA and magnetostrictive cobalt ferrites (CoFe2O4), combined with a hydrogel matrix, which mimics the tissue's hydrated environment and acts as a support matrix. For cell proliferation evaluation, two different cell culture conditions (2D and 3D matrices) and two different strategies, static and dynamic culture, were applied in order to evaluate the influence of extracellular matrix-like confinement and the magnetoelectric/magneto-mechanical effect on cellular behavior. MC3T3-E1 proliferation rate is increased under dynamic conditions, indicating the potential use of hydrogel matrices with remotely stimulated magnetostrictive biomaterials for bone tissue engineering.

Magnetically Activated Electroactive Microenvironments for Skeletal Muscle Tissue Regeneration.

This work reports on magnetoelectric biomaterials suitable for effective proliferation and differentiation of myoblast in a biomimetic microenvironment providing the electromechanical stimuli associated with this tissue in the human body. Magnetoelectric films are obtained by solvent casting through the combination of a piezoelectric polymer, poly(vinylidene fluoride-trifluoro-ethylene), and magnetostrictive particles (CoFe2O4). The nonpoled and poled (with negative and positive surface charge) magnetoelectric composites are used to investigate their influence on C2C12 myoblast adhesion, proliferation, and differentiation. It is demonstrated that the proliferation and differentiation of the cells are enhanced by the application of mechanical and/or electrical stimulation, with higher values of maturation index under mechanoelectrical stimuli. These results show that magnetoelectric cell stimulation is a full potential approach for skeletal muscle tissue engineering applications.

Electroactive Smart Materials: Novel Tools for Tailoring Bacteria Behavior and Fight Antimicrobial Resistance.

Despite being very simple organisms, bacteria possess an outstanding ability to adapt to different environments. Their long evolutionary history, being exposed to vastly different physicochemical surroundings, allowed them to detect and respond to a wide range of signals including biochemical, mechanical, electrical, and magnetic ones. Taking into consideration their adapting mechanisms, it is expected that novel materials able to provide bacteria with specific stimuli in a biomimetic context may tailor their behavior and make them suitable for specific applications in terms of anti-microbial and pro-microbial approaches. This review maintains that electroactive smart materials will be a future approach to be explored in microbiology to obtain novel strategies for fighting the emergence of live threatening antibiotic resistance.

Tailoring Bacteria Response by Piezoelectric Stimulation.

Bacteria are simple organisms with a remarkable capacity for survival by adapting to different environments, which is a result of their long evolutionary history. Taking into consideration these adapting mechanisms, this work now investigates the effect of electrically active microenvironments on bacteria and on how this stimulation may trigger bacteria growth inhibition or proliferation. Electrical microenvironments are generated via stimulation of a piezoelectric polymer with a mechanical cue, thus developing an electrical response and a variation on the surface charge of the polymeric material. Specifically, Gram-positive Staphylococcus epidermidis and Gram-negative Escherichia coli were grown overnight under static and dynamic conditions on piezoelectric poly(vinylidene) fluoride (PVDF) films to further study bacteria behavior under: (i) the effect of the material surface charge in static conditions, (ii) the mechanical effect, and (iii) the piezoelectric effect, the last two performed under dynamic conditions. Bacteria viability in planktonic and biofilm forms was measured, and the microorganism morphology was characterized. Whereas E. coli responds little to any of the stimuli application, S. epidermidis growth can be regulated through the material surface charge and by the applied frequency. Positively charged PVDF induces bacterial growth inhibition in planktonic and adhered cells in static conditions, whereas antifouling properties are obtained when a mechanical or piezoelectric effect at 4 Hz stimuli is applied. By increasing the stimuli to 40 Hz, however, the adhesion of bacteria is promoted. In conclusion, the behavior of certain bacteria species is tailored through the application of piezoelectric materials, which provide sufficient mechanoelectrical stimuli for growth or inhibition of bacteria, allowing for the design of suitable anti- and promicrobial strategies. Such strategies are only found in studies related to mammalian cells, whereas in bacterial cells this type of stimuli are still unknown. Thus, this work provides one of the first insights on the effect of piezoelectric stimuli on bacterial cells.