Nitric Oxide: Physiological Functions, Delivery, and Biomedical Applications

Nitric oxide (NO) is a gaseous molecule that has a central role in signaling pathways involved in numerous physiological processes (e.g., vasodilation, neurotransmission, inflammation, apoptosis, and tumor growth). Due to its gaseous form, NO has a short half-life, and its physiology role is concentration dependent, often restricting its function to a target site. Providing NO from an external source is beneficial in promoting cellular functions and treatment of different pathological conditions. Hence, the multifaceted role of NO in physiology and pathology has garnered massive interest in developing strategies to deliver exogenous NO for the treatment of various regenerative and biomedical complexities. NO-releasing platforms or donors capable of delivering NO in a controlled and sustained manner to target tissues or organs have advanced in the past few decades. This review article discusses in detail the generation of NO via the enzymatic functions of NO synthase as well as from NO donors and the multiple biological and pathological processes that NO modulates. The methods for incorporating of NO donors into diverse biomaterials including physical, chemical, or supramolecular techniques are summarized. Then, these NO-releasing platforms are highlighted in terms of advancing treatment strategies for various medical problems

Electrostatic flocking of salt-treated microfibers and nanofiber yarns for regenerative engineering

Electrostatic flocking is a textile technology that employs a Coulombic driving force to launch short fibers from a charging source towards an adhesive-covered substrate, resulting in a dense array of aligned fibers perpendicular to the substrate. However, electrostatic flocking of insulative polymeric fibers remains a challenge due to their insufficient charge accumulation. We report a facile method to flock electrostatically insulative poly(ε-caprolactone) (PCL) microfibers (MFs) and electrospun PCL nanofiber yarns (NFYs) by incorporating NaCl during preflock processing. Both MF and NFY were evaluated for flock functionality, mechanical properties, and biological responses. To demonstrate this platform\u27s diverse applications, standalone flocked NFY and MF scaffolds were synthesized and evaluated as scaffold for cell growth. Employing the same methodology, scaffolds made from poly(glycolide-co-L-lactide) (PGLA) (90:10) MFs were evaluated for their wound healing capacity in a diabetic mouse model. Further, a flock-reinforced polydimethylsiloxane (PDMS) disc was fabricated to create an anisotropic artificial vertebral disc (AVD) replacement potentially used as a treatment for lumbar degenerative disc disease. Overall, a salt-based flocking method is described with MFs and NFYs, with wound healing and AVD repair applications presented

Integration of a Copper-Containing Biohybrid (CuHARS) with Cellulose for Subsequent Degradation and Biomedical Control

We previously described the novel synthesis of a copper high-aspect ratio structure (CuHARS) biohybrid material using cystine. While extremely stable in water, CuHARS is completely (but slowly) degradable in cellular media. Here, integration of the CuHARS into cellulose matrices was carried out to provide added control for CuHARS degradation. Synthesized CuHARS was concentrated by centrifugation and then dried. The weighed mass was re-suspended in water. CuHARS was stable in water for months without degradation. In contrast, 25 μg/mL of the CuHARS in complete cell culture media was completely degraded (slowly) in 18 days under physiological conditions. Stable integration of CuHARS into cellulose matrices was achieved through assembly by mixing cellulose micro- and nano-fibers and CuHARS in an aqueous (pulp mixture) phase, followed by drying. Additional materials were integrated to make the hybrids magnetically susceptible. The cellulose-CuHARS composite films could be transferred, weighed, and cut into usable pieces; they maintained their form after rehydration in water for at least 7 days and were compatible with cell culture studies using brain tumor (glioma) cells. These studies demonstrate utility of a CuHARS-cellulose biohybrid for applied applications including: (1) a platform for biomedical tracking and (2) integration into a 2D/3D matrix using natural products (cellulose).This work was funded with the support of grant awards from the National Science Foundation (NSF #1547693 and NSF#1632891), and the MINECO (Spain) project MAT2015-71117-R. M. Darder acknowledges the joint sponsorship by the Fulbright Scholar Program and the Spanish Ministry of Education

03. Halloysite Animal Hair Coating Optimization for Efficient Anti-lice Protection

We demonstrate enhanced immobilization of halloysite tubule nanoclay onto hair surfaces of different mammals including horses, goats, and capybaras and search for the features that allow stable coating with drugs for anti-parasitic protection. Deposition of hair coverage with a micrometer nanotubes’ layer loaded with drugs allows for sustained delivery. Halloysite clay nanotubes are natural, safe, and biocompatible materials. Halloysite self-assembles on the hair surface during a 3-minute application of its aqueous colloids resulting in 2-3 µm thick coating. I referenced Dr. Lvov’s work for this experiment and investigated the topography of animal hair halloysite coatings using 3D laser confocal scanner and SEM. Enhanced animal hair hydrophobicity resulted in more stable nanoclay layers. After being loaded with permethrin, halloysite provides a slow release over 5-10 days which is promising for long-lasting anti-parasitic protection. Halloysite act as a carrier for anti-lice drug delivery by readily attaching to hair allowing for targeted delivery. Water-insoluble drugs such as permethrin can be encapsulated into the anionic amphiphiles modified clay tube’s lumens followed by its aqueous colloid application on hair. Lice harm animals by causing skin diseases, anemia, and death in some cases. Many anti-parasitic treatments are not effective because are quickly removed from hair. I collected goat lice samples and elaborated the experiments on topical anti-lice nanoclay drug delivery on goat and horse hair. It is currently in progress with comparison: 1) lice culture on hair, 2) lice on hair treated with pure permethrin, and 3) hair coated with halloysite nanotubes loaded with the drug

Cellulose-based biomaterials integrated with copper-cystine hybrid structures as catalysts for nitric oxide generation

10 Pág.Bionanocomposite materials were developed from the assembly of polymer-coated copper-cystine high-aspect ratio structures (CuHARS) and cellulose fibers. The coating of the metal-organic materials with polyallylamine hydrochloride (PAH) allows their covalent linkage to TEMPO-oxidized cellulose by means of EDC/NHS. The resulting materials can be processed as films or macroporous foams by solvent casting and lyophilization, respectively. The films show good mechanical behavior with Young's moduli around 1.5 GPa as well as resistance in water, while the obtained foams show an open network of interconnected macropores with average diameters around 130 μm, depending on the concentration of the initial suspension, and compression modulus values around 450 kPa, similar to other reported freeze-dried nanocellulose-based aerogels. Based on these characteristics, the cellulose/PAH-CuHARS composites are promising for potential biomedical applications as implants or wound dressing materials. They have proved to be effective in the decomposition of low molecular weight S-nitrosothiols (RSNOs), similar to those existing in blood, releasing nitric oxide (NO). This effect is attributed to the presence of copper in the crystalline structure of the CuHARS building unit, which can be gradually released in the presence of redox species like ascorbic acid, typically found in blood. The resulting biomaterials can offer the interesting properties associated with NO, like antimicrobial activity as preliminary tests showed here with Escherichia coli and Staphylococcus epidermidis. In the presence of physiological concentration of RSNOs the amount of generated NO (around 360 nM) is not enough to show bactericidal effect on the studied bacteria, but it could provide other properties inherent to NO even at low concentration in the nM range like anti-inflammatory and anti-thrombotic effects. The cytotoxic effect recorded of the films on rat brain endothelial cells (BMVECs) is least significant and proves them to be friendly enough for further biological studies.This work was supported by the project MAT2015-71117-R (MINECO/FEDER, UE) and by NSF grant award #1547693. M. Darder acknowledges the joint sponsorship by the Fulbright Program and the Spanish Ministry of Education (grant PRX16/00153).Peer reviewe

Morphological Changes in Astrocytes by Self-Oxidation of Dopamine to Polydopamine and Quantification of Dopamine through Multivariate Regression Analysis of Polydopamine Images

Astrocytes, also known as astroglia, are important cells for the structural support of neurons as well as for biochemical balance in the central nervous system (CNS). In this study, the polymerization of dopamine (DA) to polydopamine (PDA) and its effect on astrocytes was investigated. The polymerization of DA, being directly proportional to the DA concentration, raises the prospect of detecting DA concentration from PDA optically using image-processing techniques. It was found here that DA, a naturally occurring neurotransmitter, significantly altered astrocyte cell number, morphology, and metabolism, compared to astrocytes in the absence of DA. Along with these effects on astrocytes, the polymerization of DA to PDA was tracked optically in the same cell culture wells. This polymerization process led to a unique methodology based on multivariate regression analysis that quantified the concentration of DA from optical images of astrocyte cell culture media. Therefore, this developed methodology, combined with conventional imaging equipment, could be used in place of high-end and expensive analytical chemistry instruments, such as spectrophotometry, mass spectrometry, and fluorescence techniques, for quantification of the concentration of DA after polymerization to PDA under in vitro and potentially in vivo conditions

Self-Assembled Metal–Organic Biohybrids (MOBs) Using Copper and Silver for Cell Studies

The novel synthesis of metal-containing biohybrids using self-assembly methods at physiological temperatures (37 °C) was compared for copper and silver using the amino acid dimer cystine. Once assembled, the copper containing biohybrid is a stable, high-aspect ratio structure, which we call CuHARS. Using the same synthesis conditions, but replacing copper with silver, we have synthesized cystine-capped silver nanoparticles (AgCysNPs), which are shown here to form stable colloid solutions in contrast to the CuHARS, which settle out from a 1 mg/mL solution in 90 min. Both the copper and silver biohybrids, as synthesized, demonstrate very low agglomeration which we have applied for the purpose of applications with cell culture methods, namely, for testing as anti-cancer compounds. AgCysNPs (1000 ng/mL) demonstrated significant toxicity (only 6.8% viability) to glioma and neuroblastoma cells in vitro, with concentrations as low as 20 ng/mL causing some toxicity. In contrast, CuHARS required at least 5 μg/mL. For comparative purposes, silver sulfate at 100 ng/mL decreased viability by 52% and copper sulfate at 100 ng/mL only by 19.5% on glioma cells. Using these methods, the novel materials were tested here as metal–organic biohybrids (MOBs), and it is anticipated that the functionalization and dynamics of MOBs may result in building a foundation of new materials for cellular applications, including cell engineering of both normal and diseased cells and tissue constructs

Multiplicity dependence of (anti-)deuteron production in pp collisions at s\sqrt{s} = 7 TeV

In this letter, the production of deuterons and anti-deuterons in pp collisions at $\sqrt{s}$ = 7 TeV is studied as a function of the charged-particle multiplicity density at mid-rapidity with the ALICE detector at the LHC. Production yields are measured at mid-rapidity in five multiplicity classes and as a function of the deuteron transverse momentum ($p_{\rm T}$). The measurements are discussed in the context of hadron–coalescence models. The coalescence parameter B$_2$ , extracted from the measured spectra of (anti-)deuterons and primary (anti-)protons, exhibits no significant $p_{\rm T}$-dependence for $p_{\rm T}$ < 3 GeV/c , in agreement with the expectations of a simple coalescence picture. At fixed transverse momentum per nucleon, the B$_2$ parameter is found to decrease smoothly from low multiplicity pp to Pb–Pb collisions, in qualitative agreement with more elaborate coalescence models. The measured mean transverse momentum of (anti-)deuterons in pp is not reproduced by the Blast-Wave model calculations that simultaneously describe pion, kaon and proton spectra, in contrast to central Pb–Pb collisions. The ratio between the $p_{\rm T}$-integrated yield of deuterons to protons, d/p, is found to increase with the charged-particle multiplicity, as observed in inelastic pp collisions at different centre-of-mass energies. The d/p ratios are reported in a wide range, from the lowest to the highest multiplicity values measured in pp collisions at the LHC

Real-time data processing in the ALICE High Level Trigger at the LHC

International audienceAt the Large Hadron Collider at CERN in Geneva, Switzerland, atomic nuclei are collided at ultra-relativistic energies. Many final-state particles are produced in each collision and their properties are measured by the ALICE detector. The detector signals induced by the produced particles are digitized leading to data rates that are in excess of 48 GB/s. The ALICE High Level Trigger (HLT) system pioneered the use of FPGA- and GPU-based algorithms to reconstruct charged-particle trajectories and reduce the data size in real time. The results of the reconstruction of the collision events, available online, are used for high level data quality and detector-performance monitoring and real-time time-dependent detector calibration. The online data compression techniques developed and used in the ALICE HLT have more than quadrupled the amount of data that can be stored for offline event processing