Directing Cell Delivery to Murine Atherosclerotic Aortic Lesions via Targeting Inflamed Circulatory Interface using Nanocarriers.

Stem cell therapy holds significant potential for many inflammatory diseases and regenerative medicine applications. However, delivery of therapeutic cells to specific disease sites after systemic administration without indiscriminate trafficking to other non-target tissues is a major limitation of current cell therapies. Here, we describe a novel nanocarrier-directed targeted cell delivery system that enables cell surface coating with dendrimer nanocarriers containing adhesion moieties to serve as a global positioning system "GPS" to guide circulating cells to targeted lesions and mediate the anchoring of cells at the inflammation site. By exploiting cell surface ligands/receptors selectively and/or molecular moieties that are highly expressed on activated endothelium in pathologic disease states, nanocarrier-coated cells containing the counterpart binding receptors/ligands can be enabled to specifically traffic to and dock at vasculature within target lesions. We demonstrate the efficacy of the I-domain fragment of LFA-1 ( id LFA-1) complexed to modified nanocarriers to facilitate homing of mesenchymal stem cells (MSCs) to inflamed luminal endothelial cells on which ICAM-1 is highly expressed in a murine model of aortic atherosclerosis. Our method can overcome challenges imposed by the high velocity and dynamic circulatory flow of the aorta to successfully deliver MSCs to atherosclerotic regions and allow for docking of the potentially therapeutic and immunomodulating cells. This targeted cell-delivery platform can be tailored for selective systemic delivery of various types of therapeutic cells to different disease areas.

Impact of Firefighting Aqueous Film-Forming Foams on Human Cell Proliferation and Cellular Mortality.

OBJECTIVES: Evaluate the toxic effects of Aqueous Film-Forming Foams used by firefighters for Class B fire suppression in human-derived kidney cells (HEK-293). METHODS: Three widely used AFFFs were collected from fire departments and were added to HEK-293 cells in various concentrations. Seventy-two hours post-treatment, cellular proliferation and toxicity were examined using commercially available kits. RESULTS: All AFFFs evaluated induced cellular toxicity and significantly decreased cell proliferation, even when cells were treated with concentrations 10-fold lower than the working concentration used for fire suppression. CONCLUSIONS: Despite the reduced usage of PFAS-containing AFFFs in the firefighter work environment, the evaluated AFFFs demonstrated significantly altered cellular proliferation, while also inducing toxicity, indicating the presence of toxic compounds. Both stronger implementation of PFAS-containing AFFFs restrictions and robust evaluation of fluorine-free and next-generation AFFFs are warranted.

Reagentless electrochemical biosensors through incorporation of unnatural amino acids on the protein structure.

Typical protein biosensors employ chemical or genetic labeling of the protein, thus introducing an extraneous molecule to the wild-type parent protein, often changing the overall structure and properties of the protein. While these labeling methods have proven successful in many cases, they also have a series of disadvantages associated with their preparation and function. An alternative route for labeling proteins is the incorporation of unnatural amino acid (UAA) analogues, capable of acting as a label, into the structure of a protein. Such an approach, while changing the local microenvironment, poses less of a burden on the overall structure of the protein. L-DOPA is an analog of phenylalanine and contains a catechol moiety that participates in a quasi-reversible, two-electron redox process, thus making it suitable as an electrochemical label/reporter. The periplasmic glucose/galactose binding protein (GBP) was chosen to demonstrate this detection principle. Upon glucose binding, GBP undergoes a significant conformational change that is manifested as a change in the electrochemistry of L-DOPA. The electroactive GBP was immobilized onto gold nanoparticle-modified, polymerized caffeic acid, screen-printed carbon electrodes (GBP-LDOPA/AuNP/PCA/SPCE) for the purpose of direct measurement of glucose levels and serves as a proof-of-concept of the use of electrochemically-active unnatural amino acids as the label. The resulting reagentless GBP biosensors exhibited a highly selective and sensitive binding affinity for glucose in the micromolar range, laying the foundation for a new biosensing methodology based on global incorporation of an electroactive amino acid into the protein's primary sequence for highly selective electrochemical detection of compounds of interest.

Delivery of therapeutic agents and cells to pancreatic islets: Towards a new era in the treatment of diabetes.

Pancreatic islet cells, and in particular insulin-producing beta cells, are centrally involved in the pathogenesis of diabetes mellitus. These cells are of paramount importance for the endocrine control of glycemia and glucose metabolism. In Type 1 Diabetes, islet beta cells are lost due to an autoimmune attack. In Type 2 Diabetes, beta cells become dysfunctional and insufficient to counterbalance insulin resistance in peripheral tissues. Therapeutic agents have been developed to support the function of islet cells, as well as to inhibit deleterious immune responses and inflammation. Most of these agents have undesired effects due to systemic administration and off-target effects. Typically, only a small fraction of therapeutic agent reaches the desired niche in the pancreas. Because islets and their beta cells are scattered throughout the pancreas, access to the niche is limited. Targeted delivery to pancreatic islets could dramatically improve the therapeutic effect, lower the dose requirements, and lower the side effects of agents administered systemically. Targeted delivery is especially relevant for those therapeutics for which the manufacturing is difficult and costly, such as cells, exosomes, and microvesicles. Along with therapeutic agents, imaging reagents intended to quantify the beta cell mass could benefit from targeted delivery. Several methods have been developed to improve the delivery of agents to pancreatic islets. Intra-arterial administration in the pancreatic artery is a promising surgical approach, but it has inherent risks. Targeted delivery strategies have been developed based on ligands for cell surface molecules specific to islet cells or inflamed vascular endothelial cells. Delivery methods range from nanocarriers and vectors to deliver pharmacological agents to viral and non-viral vectors for the delivery of genetic constructs. Several strategies demonstrated enhanced therapeutic effects in diabetes with lower amounts of therapeutic agents and lower off-target side effects. Microvesicles, exosomes, polymer-based vectors, and nanocarriers are gaining popularity for targeted delivery. Notably, liposomes, lipid-assisted nanocarriers, and cationic polymers can be bioengineered to be immune-evasive, and their advantages to transport cargos into target cells make them appealing for pancreatic islet-targeted delivery. Viral vectors have become prominent tools for targeted gene delivery. In this review, we discuss the latest strategies for targeted delivery of therapeutic agents and imaging reagents to pancreatic islet cells.

Design of Pd-Decorated SrTiO3/BiOBr Heterojunction Materials for Enhanced Visible-Light-Based Photocatalytic Reactivity.

The development of photocatalytic materials that exploit visible light is imperative for their sustainable application in environmental remediation. While a variety of approaches have been attempted, facile routes to achieve such structures remain limited. In this contribution, a direct route for the production of a SrTiO3/BiOBr/Pd heterojunction is presented that employs a low temperature, sustainable production method. The materials were produced in a two-step process wherein BiOBr nanoplates are fabricated in the presence of the SrTiO3 nanospheres, generating a highly integrated composite material. Pd nanoparticle surface decoration was subsequently employed to facilitate and enhance charge separation lifetimes to optimize reactivity. The structures were fully characterized via a suite of approaches to confirm the final material composition and arrangement. Their reactivity was explored for the degradation of both colored and colorless model environmental pollutants, where the SrTiO3/BiOBr/Pd demonstrated significant reactivity using visible light, leading to substrate degradation in <10 min in some cases. The enhanced reactivity was attributed to the significant integration between materials, facilitating electron transfer. Such studies provide key information for the development of new materials with optimized visible-light-driven photocatalytic reactivity for sustainable environmental remediation.

Anion-Selective Electrodes Based On a CH-Hydrogen Bonding Bis-macrocyclic Ionophore with a Clamshell Architecture.

CH-hydrogen bonding provides access to new building blocks for making macrocyclic ionophores with high degrees of preorganization and selective anion recognition. In this study, an anion-binding ionophore in the shape of a clamshell (ClS) was employed that is composed of two cyanostar (CNstar) macrocycles with preorganized cavities linked with a 12-carbon chain. This ionophore allows for anion complexation by CH-hydrogen bonding. The potentiometric performance of membrane-based ion-selective electrodes incorporating this ionophore was evaluated. Different membrane compositions were prepared to determine the optimum concentrations of the ionophore and lipophilic additive in the membrane. The optimized electrode had a slope of -58.2 mV/decade and demonstrated an anti-Hofmeister selectivity pattern toward iodide with a nanomolar detection limit. Electrospray ionization mass spectrometry was employed to study the relative association strengths of ClS with various anions. The observed mass peaks of the ion-ionophore complexes were found to be consistent with the potentiometric selectivity pattern of the corresponding electrodes. Overall, the selectivity of the electrode could be altered by using an ionophore in which the two CNstar macrocycles are linked together with a flexible 12-carbon chain to control the molecularity of the binding event.

Design of a mediator-free, non-enzymatic electrochemical biosensor for glutamate detection.

A mediator-free, non-enzymatic electrochemical biosensor was constructed by covalent immobilization of a genetically engineered periplasmic glutamate binding protein onto gold nanoparticle-modified, screen-printed carbon electrodes (GluBP/AuNP/SPCE) for the purpose of direct measurement of glutamate levels. Glutamate serves as the predominant excitatory neurotransmitter in the central nervous system. As high levels of glutamate are an indicator of many neurologic disorders, there is a need for advancements in glutamate detection technologies. The biosensor was evaluated for glutamate detection by cyclic voltammetry. Binding of glutamate to the immobilized glutamate binding protein results in a conformational change of the latter that alters the microenvironment on the surface of the sensor, which is manifested as a change in signal. Dose-response plots correlating the electrochemical signal to glutamate concentration revealed a detection limit of 0.15 µM with a linear range of 0.1-0.8 µM. Selectivity studies confirmed a strong preferential response of the biosensor for glutamate against common interfering compounds.

Advances in Translational Nanotechnology: Challenges and Opportunities.

The burgeoning field of nanotechnology aims to create and deploy nanoscale structures, devices, and systems with novel, size-dependent properties and functions. The nanotechnology revolution has sparked radically new technologies and strategies across all scientific disciplines, with nanotechnology now applied to virtually every area of research and development in the US and globally. NanoFlorida was founded to create a forum for scientific exchange, promote networking among nanoscientists, encourage collaborative research efforts across institutions, forge strong industry-academia partnerships in nanoscience, and showcase the contributions of students and trainees in nanotechnology fields. The 2019 NanoFlorida International Conference expanded this vision to emphasize national and international participation, with a focus on advances made in translating nanotechnology. This review highlights notable research in the areas of engineering especially in optics, photonics and plasmonics and electronics; biomedical devices, nano-biotechnology, nanotherapeutics including both experimental nanotherapies and nanovaccines; nano-diagnostics and -theranostics; nano-enabled drug discovery platforms; tissue engineering, bioprinting, and environmental nanotechnology, as well as challenges and directions for future research.