Rapid integration of screen-printed electrodes into thermoplastic organ-on-a-chip devices for real-time monitoring of trans-endothelial electrical resistance.

Trans-endothelial electrical resistance (TEER) is one of the most widely used indicators to quantify the barrier integrity of endothelial layers. Over the last decade, the integration of TEER sensors into organ-on-a-chip (OOC) platforms has gained increasing interest for its efficient and effective measurement of TEER in OOCs. To date, microfabricated electrodes or direct insertion of wires has been used to integrate TEER sensors into OOCs, with each method having advantages and disadvantages. In this study, we developed a TEER-SPE chip consisting of carbon-based screen-printed electrodes (SPEs) embedded in a poly(methyl methacrylate) (PMMA)-based multi-layered microfluidic device with a porous poly(ethylene terephthalate) membrane in-between. As proof of concept, we demonstrated the successful cultures of hCMEC/D3 cells and the formation of confluent monolayers in the TEER-SPE chip and obtained TEER measurements for 4 days. Additionally, the TEER-SPE chip could detect changes in the barrier integrity due to shear stress or an inflammatory cytokine (i.e., tumor necrosis factor-α). The novel approach enables a low-cost and facile fabrication of carbon-based SPEs on PMMA substrates and the subsequent assembly of PMMA layers for rapid prototyping. Being cost-effective and cleanroom-free, our method lowers the existing logistical and technical barriers presenting itself as another step forward to the broader adoption of OOCs with TEER measurement capability.

Engineered Vasculature for Cancer Research and Regenerative Medicine.

Engineered human tissues created by three-dimensional cell culture of human cells in a hydrogel are becoming emerging model systems for cancer drug discovery and regenerative medicine. Complex functional engineered tissues can also assist in the regeneration, repair, or replacement of human tissues. However, one of the main hurdles for tissue engineering, three-dimensional cell culture, and regenerative medicine is the capability of delivering nutrients and oxygen to cells through the vasculatures. Several studies have investigated different strategies to create a functional vascular system in engineered tissues and organ-on-a-chips. Engineered vasculatures have been used for the studies of angiogenesis, vasculogenesis, as well as drug and cell transports across the endothelium. Moreover, vascular engineering allows the creation of large functional vascular conduits for regenerative medicine purposes. However, there are still many challenges in the creation of vascularized tissue constructs and their biological applications. This review will summarize the latest efforts to create vasculatures and vascularized tissues for cancer research and regenerative medicine.

Skin-interfaced electronics: A promising and intelligent paradigm for personalized healthcare.

Skin-interfaced electronics (skintronics) have received considerable attention due to their thinness, skin-like mechanical softness, excellent conformability, and multifunctional integration. Current advancements in skintronics have enabled health monitoring and digital medicine. Particularly, skintronics offer a personalized platform for early-stage disease diagnosis and treatment. In this comprehensive review, we discuss (1) the state-of-the-art skintronic devices, (2) material selections and platform considerations of future skintronics toward intelligent healthcare, (3) device fabrication and system integrations of skintronics, (4) an overview of the skintronic platform for personalized healthcare applications, including biosensing as well as wound healing, sleep monitoring, the assessment of SARS-CoV-2, and the augmented reality-/virtual reality-enhanced human-machine interfaces, and (5) current challenges and future opportunities of skintronics and their potentials in clinical translation and commercialization. The field of skintronics will not only minimize physical and physiological mismatches with the skin but also shift the paradigm in intelligent and personalized healthcare and offer unprecedented promise to revolutionize conventional medical practices.

Organ-On-A-Chip Models of the Blood-Brain Barrier: Recent Advances and Future Prospects.

The human brain and central nervous system (CNS) present unique challenges in drug development for neurological diseases. One major obstacle is the blood-brain barrier (BBB), which hampers the effective delivery of therapeutic molecules into the brain while protecting it from blood-born neurotoxic substances and maintaining CNS homeostasis. For BBB research, traditional in vitro models rely upon Petri dishes or Transwell systems. However, these static models lack essential microenvironmental factors such as shear stress and proper cell-cell interactions. To this end, organ-on-a-chip (OoC) technology has emerged as a new in vitro modeling approach to better recapitulate the highly dynamic in vivo human brain microenvironment so-called the neural vascular unit (NVU). Such BBB-on-a-chip models have made substantial progress over the last decade, and concurrently there has been increasing interest in modeling various neurological diseases such as Alzheimer's disease and Parkinson's disease using OoC technology. In addition, with recent advances in other scientific technologies, several new opportunities to improve the BBB-on-a-chip platform via multidisciplinary approaches are available. In this review, an overview of the NVU and OoC technology is provided, recent progress and applications of BBB-on-a-chip for personalized medicine and drug discovery are discussed, and current challenges and future directions are delineated.

Understanding the Inhibitory and Antioxidant Effects of Pyrroloquinoline Quinone (PQQ) on Copper(II)-Induced α-Synuclein-119 Aggregation.

Parkinson's disease (PD) is associated with the aggregation and misfolding of a-synuclein (a-syn) protein in dopaminergic neurons. The misfolding process is heavily linked to copper dysregulation in PD. Experimental evidence supports the hypothesis that the co-presence of Cu(II) and α-syn facilitates the aggregation of α-syn, affecting the pathological development of PD. Recent literature has shown that pyrroloquinoline quinone (PQQ) contains strong neuroprotective activity by reducing the reactive oxygen species (ROS) production by α-syn. Despite these known facts, minimal studies have been done on the antioxidant effect of PQQ against ROS formation in the presence of Cu(II) and α-syn-119. Thus, it is of great significance to study the interaction between all three components, PQQ, Cu(II), and α-syn-119. In this proof-of-concept study, a variety of chemical techniques were employed to examine the antioxidant effect of PQQ on ROS that α-syn-119 produced in the presence of Cu(II). Our results showed that PQQ effectively prevented ROS formation in SH-SY5Y human differentiated neuronal cells. Thioflavin T (ThT) fluorescence assay, circular dichroism (CD) spectroscopy, and transmission electron microscopy (TEM) were applied, where PQQ was able to actively prevent fibrillation of α-syn-119 in the presence of Cu(II). This finding was further confirmed using electrochemical impedance spectroscopy (EIS), where the binding of PQQ to the α-syn-119 suppressed the aggregation process on the electrode surface. With these encouraging results, we envisage that PQQ and its derivatives can be a promising candidate for further studies as a multitarget therapeutic agent toward PD therapy.

Lab-on-a-Contact Lens: Recent Advances and Future Opportunities in Diagnostics and Therapeutics.

The eye is one of the most complex organs in the human body, containing rich and critical physiological information (e.g., intraocular pressure, corneal temperature, and pH) as well as a library of metabolite biomarkers (e.g., glucose, proteins, and specific ions). Smart contact lenses (SCLs) can serve as a wearable intelligent ocular prosthetic device capable of noninvasive and continuous monitoring of various essential physical/biochemical parameters and drug loading/delivery for the treatment of ocular diseases. Advances in SCL technologies and the growing public interest in personalized health are accelerating SCL research more than ever before. Here, the current status and potential of SCL development through a comprehensive review from fabrication to applications to commercialization are discussed. First, the material, fabrication, and platform designs of the SCLs for the diagnostic and therapeutic applications are discussed. Then, the latest advances in diagnostic and therapeutic SCLs for clinical translation are reviewed. Later, the established techniques for wearable power transfer and wireless data transmission applied to current SCL devices are summarized. An outlook, future opportunities, and challenges for developing next-generation SCL devices are also provided. With the rise in interest of SCL development, this comprehensive and essential review can serve as a new paradigm for the SCL devices.

Electrochemical flow injection analysis of the interaction between pyrroloquinoline quinone (PQQ) and α-synuclein peptides related to Parkinson's disease.

α-Synuclein (α-syn) is a hallmark protein of Parkinson's disease (PD). The aggregation process of α-syn has been heavily associated with the pathogenesis of PD. With the exponentially growing number of potential therapeutic compounds that can inhibit the aggregation of α-syn, there is now a significant demand for a high-throughput analysis system. Herein, a novel flow injection analysis system with an electrochemical biosensor as the detector was developed to study the interaction of a well-described antioxidant and amyloid inhibitor, pyrroloquinoline quinone (PQQ) with α-synuclein peptides. Screen-printed gold electrodes (SPEs) were modified using heptapeptides from α-syn wild-type (WT) and mutants such as lysine knock-out (ETEE) and E46K. Affinity binding events between these peptides and PQQ were analyzed by electrochemical impedance spectroscopy (EIS) and further confirmed by high-performance liquid chromatography (HPLC), liquid chromatography/mass spectrometry (LC/MS), and nuclear magnetic resonance (NMR) spectroscopy. HPLC and LC/MS results revealed that PQQ formed a stable complex with α-syn. NMR results confirmed that the α-syn-PQQ complex was formed via a Schiff base formation-like process. In addition, results showed that lysine residues influenced the binding event, in which the presence of an extra lysine stabilized the α-syn-PQQ complex, and the absence of a lysine significantly decreased the interaction of α-syn with PQQ. Therefore, we concluded that EIS is a promising technique for the evaluation of the interaction between PQQ-based amyloid inhibitors and α-syn. The electrochemical flow injection analysis assembly provided a rapid and low-cost drug discovery platform for the evaluation of small molecule-protein interactions.

Electrochemical biosensors for biometal-protein interactions in neurodegenerative diseases.

Electrochemical biosensors have been adopted into a wide range of applications in the study of biometal-protein interactions in neurodegenerative diseases. Transition metals such as zinc, copper, and iron that are significant to biological functions have been shown to have strong implications in the progressive neural degeneration in Alzheimer's disease (AD), Parkinson's disease (PD), and prion protein diseases. This review presents a summative examination of the progress made in the design, fabrication, and applications of electrochemical biosensors in recent literature at understanding the metal-protein interactions in neurodegenerative diseases. The focus will be drawn on disease-causing biomarkers such as amyloid-ß (Aß) and tau proteins for AD, α-synuclein (α-syn) for PD, and prion proteins (PrP). Topics such as the use of electrochemical biosensing in monitoring biometal-induced conformational changes, elucidation of complexation motifs, production of reactive oxygen species (ROS) as well as the influence on downstream biomolecular interactions will be discussed. Major results and important concepts presented in these studies will be summarized in the hope to spark inspiration for the next generation of electrochemical sensors.

Polyacrylamide hydrogels doped with different shapes of silver nanoparticles: Antibacterial and mechanical properties.

The incorporation of nanoparticles into a hydrogel matrix enables the development of innovative smart materials with enhanced biophysical properties. In this proof-of-concept study, we encapsulated different shapes (spherical, triangular and rod) of silver nanoparticles (AgNPs) within a hydrogel matrix of polyacrylamide (PAA) and N-methylenebisacrylamide (MBA) (PAA-MBA) to investigate whether these hydrogels exhibited shape-dependent antimicrobial and mechanical properties. We examined the mechanism of adsorption of different shapes of AgNPs using time-of-flight secondary ion mass spectrometry (ToF-SIMS). Results showed that the adsorption of AgNPs was primarily occurring on the surface/outer pores of the PAA-MBA hydrogel and that rod AgNPs demonstrated a relatively slower adsorption within the hydrogel matrix. The mechanical properties of AgNP-doped hydrogels were evaluated using rheology and atomic force microscopy (AFM) quantitative imaging. We observed a higher storage and Young's modulus which proved that the incorporation of the various shapes of AgNPs increased the mechanical properties of the hydrogels with no significant differences between the different shapes. While both spherical and triangular AgNP-doped hydrogels showed strong antimicrobial activity, the hydrogel with the rod AgNPs had a relatively lower antimicrobial activity. Overall, our preliminary results demonstrated that nanocomposite hydrogels were promising materials for applications in the future development of wound dressings.

Graphene Oxide Nanoribbons in Chitosan for Simultaneous Electrochemical Detection of Guanine, Adenine, Thymine and Cytosine.

Herein, graphene oxide nanoribbons (GONRs) were obtained from the oxidative unzipping of multi-walled carbon nanotubes. Covalent coupling reaction of 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDC) and N-hydroxy succinimide (NHS) with amine functional groups (-NH2) of the chitosan natural polymer (CH) was used for entrapping GONRs on the activated glassy carbon electrode (GCE/GONRs-CH). The nanocomposite was characterized by high-resolution transmission electron microscopy (HRTEM), and field-emission scanning electron microscopy (FESEM). In addition, the modification steps were monitored using FTIR. The nanocomposite-modified electrode was used for the simultaneous electrochemical determination of four DNA bases; guanine (G), adenine (A), thymine (T) and cytosine (C). The nanocomposite-modified GCE displayed a strong, stable and continuous four oxidation peaks during electrochemistry detection at potentials 0.63, 0.89, 1.13 and 1.27 V for G, A, T and C, respectively. The calibration curves were linear up to 256, 172, 855 and 342 µM with detection limits of 0.002, 0.023, 1.330 and 0.641 µM for G, A, T and C, respectively. The analytical performance of the GCE/GONRs-CH has been used for the determination of G, A, T and C in real samples and obtained a recovery percentage from 91.1%-104.7%. Our preliminary results demonstrated that GCE/GONRs-CH provided a promising platform to detect all four DNA bases for future studies on DNA damage and mutations.

Electrochemical biosensors for the detection and study of α-synuclein related to Parkinson's disease - A review.

Parkinson's disease (PD) is a long-term degenerative disorder that affects predominately dopaminergic neurons in the substantia nigra, which mainly control movement. Alpha-synuclein (α-syn) is a major constituent of Lewy bodies that are reported to be the most important toxic species in the brain of PD patients. In this critical review, we highlight novel electrochemical biosensors that have been recently developed utilizing aptamers and antibodies in connection with various nanomaterials to study biomarkers related to PD such as α-syn. We also review several research articles that have utilized electrochemical biosensors to study the interaction of α-syn with biometals as well as small molecules such as clioquinol, (-)-epigallocatechin-3-gallate (EGCG) and baicalein. Due to the significant advances in nanomaterials in the past decade, electrochemical biosensors capable of detecting multiple biomarkers in clinically relevant samples in real-time have been achieved. This may facilitate the path towards commercialization of electrochemical biosensors for clinical applications and high-throughput screening of small molecules for structure-activity relationship (SAR) studies.

Novel thiolated-PEG linker molecule for biosensor development on gold surfaces.

The surface modifying linker molecules can directly influence the performance and longevity of biosensors. They must allow the attachment of biological recognition layer on the sensor surface, as well as the protection of the surface from fouling effects. Recent advances in this field identified several key factors that can increase the efficiency, stability and the anti-fouling effect of a layer formed by surface modifying linker molecules. Herein, this work presents a simple synthetic procedure, characterization, and application of a novel thiolated-PEG surface modifying molecule (DSPEG2) that could act as a multi-purpose linker for gold surfaces. The analyses of the molecular spatial distribution of DSPEG2 on gold surfaces were performed using time-of-flight secondary ion mass spectrometry (TOF-SIMS) imaging and X-ray photoelectric spectroscopy (XPS). The immobilization of DSPEG2 on gold surfaces was examined using cyclic voltammetry (CV), electrochemical impedance spectroscopy (EIS) and surface plasmon resonance (SPR). Our preliminary results demonstrated that DSPEG2 is a promising novel linker molecule that can be applied in a wide range of biosensors based on gold surfaces.

Polymorphisms of a Collagen-Like Adhesin Contributes to Legionella pneumophila Adhesion, Biofilm Formation Capacity and Clinical Prevalence.

Legionellosis is a severe respiratory illness caused by the inhalation of aerosolized water droplets contaminated with the opportunistic pathogen Legionella pneumophila. The ability of L. pneumophila to produce biofilms has been associated with its capacity to colonize and persist in human-made water reservoirs and distribution systems, which are the source of legionellosis outbreaks. Nevertheless, the factors that mediate L. pneumophila biofilm formation are largely unknown. In previous studies we reported that the adhesin Legionella collagen-like protein (Lcl), is required for auto-aggregation, attachment to multiple surfaces and the formation of biofilms. Lcl structure contains three distinguishable regions: An N-terminal region with a predicted signal sequence, a central region containing tandem collagen-like repeats (R-domain) and a C-terminal region (C-domain) with no significant homology to other known proteins. Lcl R-domain encodes tandem repeats of the collagenous tripeptide Gly-Xaa-Yaa (GXY), a motif that is key for the molecular organization of mammalian collagen and mediates the binding of collagenous proteins to different cellular and environmental ligands. Interestingly, Lcl is polymorphic in the number of GXY tandem repeats. In this study, we combined diverse biochemical, genetic, and cellular approaches to determine the role of Lcl domains and GXY repeats polymorphisms on the structural and functional properties of Lcl, as well as on bacterial attachment, aggregation and biofilm formation. Our results indicate that the R-domain is key for assembling Lcl collagenous triple-helices and has a more preponderate role over the C-domain in Lcl adhesin binding properties. We show that Lcl molecules oligomerize to form large supramolecular complexes to which both, R and C-domains are required. Furthermore, we found that the number of GXY tandem repeats encoded in Lcl R-domain correlates positively with the binding capabilities of Lcl and with the attachment and biofilm production capacity of L. pneumophila strains. Accordingly, the number of GXY tandem repeats in Lcl influences the clinical prevalence of L. pneumophila strains. Therefore, the number of Lcl tandem repeats could be considered as a potential predictor for virulence in L. pneumophila isolates.

Electrochemical Detection of Interaction between Copper(II) and Peptides Related to Pathological α-Synuclein Mutants.

We present a proof of concept study for electrochemical detection of the metal-binding site of α-synuclein (α-syn). Parkinson's disease (PD) is associated with the aggregation and misfolding of α-syn in dopaminergic neurons. Because copper homeostasis is deregulated in PD, it is of great significance to study the metal-binding site of wild-type α-syn (48-53, VVHGVA) and its pathological mutants (H50Q and G51D). Cyclic voltammetry and electrochemical impedance spectroscopy were used to monitor the formation of a peptide-PEG mixed layer on gold surfaces. Differential pulse voltammetry was used to detect and evaluate the interaction of copper(II) with the peptide layer. X-ray photoelectron spectroscopy was used to characterize the formation and attachment of the peptide layer on gold surfaces. Isothermal titration calorimetry was also utilized to evaluate the binding characteristics of the peptides with copper(II) ions. Our results indicated that the effect of a single amino acid mutation on the peptides drastically influenced their ability to interact with copper(II) ions. These results demonstrated that our electrochemical approach provided a rapid and cost-effective platform to study the strong interaction between α-syn and copper(II), which is implicated as one of the factors inducing structural changes in α-syn toward the progression of PD.

Biosensors for the Detection of Interaction between Legionella pneumophila Collagen-Like Protein and Glycosaminoglycans.

The adhesin Legionella collagen-like (Lcl) protein can bind to extracellular matrix components and mediate the binding of Legionella pneumophila to host cells. In this study, electrochemical impedance spectroscopy (EIS) and surface plasmon resonance (SPR)-based biosensors were employed to characterize these interactions between glycosaminoglycans (GAGs) and the adhesin Lcl protein. Fucoidan displayed a high affinity (KD 18 nM) for Lcl protein. Chondroitin sulfate A and dermatan sulfate differ in the position of a carboxyl group replacing D-glucuronate with D-iduronate. Our results indicated that the presence of D-iduronate in dermatan sulfate strongly hindered its interaction with Lcl. These biophysical studies provided valuable information in our understanding of adhesin-ligand interactions related to Legionella pneumophila infections.