Nitric Oxide Delivery System Research Articles

Ideation and assessment of a nitric oxide delivery system for spontaneously breathing subjects

BackgroundThere is an increasing interest in safely delivering high dose of inhaled nitric oxide (NO) as an antimicrobial and antiviral therapeutics for spontaneously breathing patients. A novel NO delivery system is described. MethodsWe developed a gas delivery system that utilizes standard respiratory circuit connectors, a reservoir bag, and a scavenging chamber containing calcium hydroxide. The performance of the system was tested using a mechanical lung, assessing the NO concentration delivered at varying inspiratory flows. Safety was assessed in vitro and in vivo by measuring nitrogen dioxide (NO2) levels in the delivered NO gas. Lastly, we measured the inspired and expired NO and NO2 of this system in 5 healthy subjects during a 15-min administration of high dose NO (160 parts-per-million, ppm) using our delivery system. ResultsThe system demonstrated stable delivery of prescribed NO levels at various inspiratory flow rates (0–50 L/min). The reservoir bag and a high flow of entering air minimized the oscillation of NO concentrations during inspiration on average 4.6 ppm for each 10 L/min increment in lung inspiratory flow.The calcium hydroxide scavenger reduced the inhaled NO2 concentration on average 0.9 ppm (95% CI -1.58, −0.22; p = .01). We performed 49 NO administrations of 160 ppm in 5 subjects. The average concentration of inspired NO was 164.8±10.74 ppm, with inspired NO2 levels of 0.7±0.13 ppm. The subjects did not experience any adverse events; transcutaneous methemoglobin concentrations increased from 1.05±0.58 to 2.26±0.47%. ConclusionsThe system we developed to administer high-dose NO for inhalation is easy to build, reliable, was well tolerated in healthy subjects.

Drug Delivery System for Refractory Cancer Therapy via an Endogenous Albumin Transport System.

A unique phenomenon in solid tumors, the enhanced permeability and retention (EPR) effect is now well known in the development of macromolecular anticancer therapy. However, cancers with low vascular permeability have posed a challenge for these EPR based therapeutic systems. An intrinsic vascular modulator, such as nitric oxide (NO), could augment the endogenous EPR effect. However, the most important aim has been to construct an effective NO delivery system for cancer. Since it is well known that human serum albumin is one of the most important endogenous NO transport proteins in human circulation, for more than a decade we have demonstrated that S-nitrosated human serum albumin dimer (SNO-HSA-Dimer) becomes an enhancer of the EPR effect. Here, we summarize the enhanced effect of SNO-HSA-Dimer on the anticancer effect of macromolecular anticancer drugs such as PEGylated liposomal doxorubicin (Doxil®). In C26-bearing mice with highly permeable vasculature, SNO-HSA-Dimer is able to increase more 3-fold the tumor accumulation of these anticancer drugs, thereby tripling their anticancer effects. Interestingly, the tumor accumulation of Doxil® in B16-bearing mice, which are characterized by a low permeable vasculature, increased more than 6-fold in the presence of SNO-HSA-Dimer, and the improved accumulation of Doxil® led to both increased survival and decreased tumor volume. These results strongly suggest that the more cancer is refractory, the more the SNO-HSA-Dimer could enhance the EPR effect via an endogenous albumin transport (EAT) system. Accordingly, we conclude that the EAT system is promising as a drug delivery system (DDS) strategy for refractory cancer therapy.

Management of hospitalized patients with pulmonary arterial hypertension and COVID-19 infection.

Management of hospitalized patients with pulmonary arterial hypertension and COVID-19 infection.

Nanoclay-Polyamine Composite Hydrogel for Topical Delivery of Nitric Oxide Gas via Innate Gelation Characteristics of Laponite.

Because nitric oxide (NO) gas is an endogenously produced signaling molecule related to numerous physiological functions, manystudies have been conducted to develop NO delivery systems for potential biomedical applications. However, NO is a reactive radical gas molecule that has a very short life-time and readily transforms into nitrogen oxide species via reaction with oxygen species. Therefore, it is necessary to develop an NO delivery carrier that allows local release of the NO gas at the site of application. In this study, Laponite (LP) nanoclay was used to fabricate an NO delivery carrier through the formation of Laponite-polyamine (LP-PAn) composites. The Laponite clay and pentaethylenehexamine (PEHA) formed a macromolecular structure by electrostatic interaction and the nitric oxide donor, N-diazeniumdiolate (NONOates), was synthesized into the LP-PAn composite. We investigated the conformation of the LP-PAn composite structure and the NO donor formation by ζ potential, X-ray diffraction, and UV-vis and Fourier transform infrared (FT-IR) spectroscopies and also by analyzing the NO release profile. Additionally, we confirmed the applicability in biomedical applications via a cell viability and in vitro endothelial cell tube formation assay.

NO-Releasing Nanoparticles Ameliorate Detrusor Overactivity in Transgenic Sickle Cell Mice via Restored NO/ROCK Signaling.

Sickle cell disease (SCD) is associated with overactive bladder (OAB). Detrusor overactivity, a component of OAB, is present in an SCD mouse, but the molecular mechanisms for this condition are not well-defined. We hypothesize that nitric oxide (NO)/ ras homolog gene family (Rho) A/Rho-associated kinase (ROCK) dysregulation is a mechanism for detrusor overactivity and that NO-releasing nanoparticles (NO-nps), a novel NO delivery system, may serve to treat this condition. Male adult SCD transgenic, combined endothelial NO synthases (eNOSs) and neuronal NOS (nNOS) gene-deficient (dNOS-/-), and wild-type (WT) mice were used. Empty nanoparticle or NO-np was injected into the bladder, followed by cystometric studies. The expression levels of phosphorylated eNOS (Ser-1177), protein kinase B (Akt) (Ser-473), nNOS (Ser-1412), and myosin phosphatase target subunit 1 (MYPT1) (Thr-696) were assessed in the bladder. SCD and dNOS-/- mice had a greater (P < 0.05) number of voiding and nonvoiding contractions compared with WT mice, and they were normalized by NO-np treatment. eNOS (Ser-1177) and AKT (Ser-473) phosphorylation were decreased (P < 0.05) in the bladder of SCD compared with WT mice and reversed by NO-np. Phosphorylated MYPT1, a marker of the RhoA/ROCK pathway, was increased (P < 0.05) in the bladder of SCD mice compared with WT and reversed by NO-np. nNOS phosphorylation on positive (Ser-1412) regulatory site was decreased (P < 0.05) in the bladder of SCD mice compared with WT and was not affected by NO-np. NO-nps did not affect any of the measured parameters in WT mice. In conclusion, dysregulation of NO and RhoA/ROCK pathways is associated with detrusor overactivity in SCD mice; NO-np reverses these molecular derangements in the bladder and decreases detrusor overactivity. SIGNIFICANCE STATEMENT: Voiding abnormalities commonly affect patients with sickle cell disease (SCD) but are problematic to treat. Clarification of the science for this condition in an animal model of SCD may lead to improved interventions for it. Our findings suggest that novel topical delivery of a vasorelaxant agent nitric oxide into the bladder of these mice corrects overactive bladder by improving deranged bladder physiology regulatory signaling.

Tuning Cellular Biological Functions Through the Controlled Release of NO from a Porous Ti-MOF.

Materials for the controlled release of nitric oxide (NO) are of interest for therapeutic applications. However, to date, many suffer from toxicity and stability issues, as well as poor performance. Herein, we propose a new NO adsorption/release mechanism through the formation of nitrites on the skeleton of a titanium-based metal-organic framework (MOF) that we named MIP-177, featuring a suitable set of properties for such an application: (i) high NO storage capacity (3 μmol mg-1solid ), (ii) excellent biocompatibility at therapeutic relevant concentrations (no cytotoxicity at 90 μg mL-1 for wound healing) due to its high stability in biological media (<9 % degradation in 72 hours) and (iii) slow NO release in biological media (≈2 hours for 90 % release). The prospective application of MIP-177 is demonstrated through NO-driven control of mitochondrial respiration in cells and stimulation of cell migration, paving the way for the design of new NO delivery systems for wound healing therapy.

Tuning Cellular Biological Functions Through the Controlled Release of NO from a Porous Ti‐MOF

AbstractMaterials for the controlled release of nitric oxide (NO) are of interest for therapeutic applications. However, to date, many suffer from toxicity and stability issues, as well as poor performance. Herein, we propose a new NO adsorption/release mechanism through the formation of nitrites on the skeleton of a titanium‐based metal–organic framework (MOF) that we named MIP‐177, featuring a suitable set of properties for such an application: (i) high NO storage capacity (3 μmol mg−1solid), (ii) excellent biocompatibility at therapeutic relevant concentrations (no cytotoxicity at 90 μg mL−1 for wound healing) due to its high stability in biological media (&lt;9 % degradation in 72 hours) and (iii) slow NO release in biological media (≈2 hours for 90 % release). The prospective application of MIP‐177 is demonstrated through NO‐driven control of mitochondrial respiration in cells and stimulation of cell migration, paving the way for the design of new NO delivery systems for wound healing therapy.

Potential Protective Effect of Nitric Oxide-Releasing Nanofibers in Hypoxia/Reoxygenation-Induced Cardiomyocyte Injury.

Nitric oxide (NO) is involved in several physiological processes including vasodilation, angiogenesis, immune response, and wound healing, as well as preventing ischemia/reperfusion injury in many organs such as the heart, liver, lungs, and kidneys. Recently, various NO delivery systems such as nanoparticles, nanorods, and nanofibers have been widely studied as potential therapeutic agents. In particular, NO-releasing nanofibers have been attracting much attention for various medicinal applications including regenerative medicine, wound dressings, and coatings for implantable medical devices, due to their flexible and open architectures. In this study, we prepared biocompatible NO-releasing nanofibers by electrospinning using mixed solutions of polymers and methylaminopropyltrimethoxysilane (MAP3), which was modified with N-diazeniumdiolate as an NO donor. In addition, we evaluated their protective effects on hypoxia/reoxygenation (HR) injury in H9c2 cells. The total NO amount released from the resulting MAP3 nanofibers was 1.26 µmol ·mg-1. From the cytotoxicity evaluation of various weights of NO-releasing nanofibers (0 to 2 mg), we selected 1 mg NO-releasing nanofibers for the subsequent experiments. Pre-treatment with NO-releasing nanofibers before hypoxia induction could provide a cytoprotective effect against HR-induced injury in H9c2 cells. The nanofibers could also effectively inhibit the generation of hydrogen peroxide, which was one major contributor to oxidative damage, as well as 8-hydroxyl-2-deoxyguanosine level as an indicator of oxidative DNA damage. In addition, pre-treatment with NO-releasing nano-fibers in a wound model showed wound healing effects similar to those of normal cells. As a result, N-diazeniumdiolate-modified MAP3 nanofibers might protect H9c2 cells from DNA damage by inhibiting the generation of oxidative stress in HR injury. Therefore, we expect that NO-releasing nanofibers could be utilized as a therapeutic strategy for protecting cardiomyocytes from HR injury.

Progress in the development of nitric oxide-releasing biomaterials and their biomedical applications

Nitric oxide (NO) is an essential signaling molecule that participates in various important physiological and pathological processes, such as angiogenesis, neuronal transmission, immunomodulation, and tumor growth. The multifunctional roles of NO in biological processes have substantially increased interest in research regarding effective strategies in delivering exogenous NO for biomedical applications. The development of various NO prodrugs, such as organic nitrates, diazeniumdiolates, S -nitrosothiols, furoxan nitrogen oxides, and metal-nitrosyls, continue to undergo innovative modifications to improve NO payload and to prolong NO release. However, the clinical application of these low-molecular-weight NO donors is limited due to problems associated with burst release and untargeted delivery. The tunable administration of NO by biomaterial-based carriers offers an attractive and efficient strategy to achieve a controlled and targeted delivery of NO to specialized tissues or organs. In this review, we summarize the progress made in the development of NO donors and NO delivery systems and highlight potential biomedical applications aiming to inspire new perspectives in the field of biomaterial-based NO delivery systems.

Developing regulatory property of gelatin-tannic acid multilayer films for coating-based nitric oxide gas delivery system

To utilize potentials of nitric oxide (NO) gas in anti-bacterial, anticancer, wound healing applications, numerous studies have been conducted to develop a NO delivery system in the past few decades. Even though a coating method and film types are essential to apply in biomedical device coating from previous NO delivery systems, release control from the coating system is still challenging. In this study, we introduced a multilayered polymeric coating system to overcome the uncontrollable NO release kinetics of film systems. We used biocompatible gelatin and tannic acid to construct a rough, porous structured film based on the layer-by-layer self-assembly method. The multilayered polymeric structure facilitated the controlled amount of NO release from (Gel/TA)n film and showed burst release in early period owing to their large surface area from the rough, porous structure. We synthesized the proton-responsive NO donor, N-diazeniumdiolate (NONOates), into the (Gel/TA)n film through a chemical reaction under high pressure NO gas. NO release profile was analyzed by a real-time NO analysis machine (NOA 280i). Then, the NO-releasing (Gel/TA)n film was tested its toxicity against human dermal fibroblast cells and bactericidal effects against Staphylococcus aureus.

PEI/NONOates-doped PLGA nanoparticles for eradicating methicillin-resistant Staphylococcus aureus biofilm in diabetic wounds via binding to the biofilm matrix

Wounds infected with methicillin-resistant Staphylococcus aureus (MRSA) biofilm represent a high risk in patients with diabetes. Nitric oxide (NO) has shown promise in dispersing biofilm and wound healing. For an effective treatment of MRSA biofilm-infected wounds, however, NO needs to be supplied to the biofilm matrix in a sustainable manner due to a short half-life and limited diffusion distance of NO. In this study, polyethylenimine/diazeniumdiolate (PEI/NONOate)-doped PLGA nanoparticles (PLGA-PEI/NO NPs) with an ability to bind to the biofilm matrix are developed to facilitate the NO delivery to MRSA biofilm-infected wound. In simulated wound fluid, PLGA-PEI/NO NPs show an extended NO release over 4 days. PLGA-PEI/NO NPs firmly bind to the MRSA biofilm matrix, resulting in a greatly enhanced anti-biofilm activity. Moreover, PLGA-PEI/NO NPs accelerate healing of MRSA biofilm-infected wounds in diabetic mice along with complete biofilm dispersal and reduced bacterial burden. These results suggest that the biofilm-binding NO-releasing NPs represent a promising NO delivery system for the treatments of biofilm-infected chronic wounds.

Targeted delivery of nitric oxide via a ‘bump-and-hole’-based enzyme–prodrug pair

The spatiotemporal generation of nitric oxide (NO), a versatile endogenous messenger, is precisely controlled. Despite its therapeutic potential for a wide range of diseases, NO-based therapies are limited clinically due to a lack of effective strategies for precisely delivering NO to a specific site. In the present study, we developed a novel NO delivery system via modification of an enzyme-prodrug pair of galactosidase-galactosyl-NONOate using a 'bump-and-hole' strategy. Precise delivery to targeted tissues was clearly demonstrated by an in vivo near-infrared imaging assay. The therapeutic potential was evaluated in both rat hindlimb ischemia and mouse acute kidney injury models. Targeted delivery of NO clearly enhanced its therapeutic efficacy in tissue repair and function recovery and abolished side effects due to the systemic release of NO. The developed protocol holds broad applicability in the targeted delivery of important gaseous signaling molecules and offers a potent tool for the investigation of relevant molecular mechanisms.

Biofunctionalization of biomaterials for nitric oxide delivery: potential applications in regenerative medicine

Biofunctionalization of biomaterials for nitric oxide delivery: potential applications in regenerative medicine Kai Jen Tsai,1,* Anna Rammou,1,* Chuanyu Gao,1 Achala de Mel1,2 1UCL Division of Surgery and Interventional Sciences, University College London, London, UK; 2School of Chemistry, University of Nottingham, Nottingham, UK *These authors contributed equally to this work Introduction: Mimicking physiological functions of nitric oxide (NO) has applications in regenerative medicine. However, few NO delivery systems have progressed to clinical trials owing to limitations in delivery. Materials and methods: A novel NO delivery system was explored by integrating S-nitro-N-acetylpenicillamine-functionalized long-chain aliphatic hydrocarbons (LCAHs) into a polyurethane-based polymer. Results and discussion: Contact angle analysis determined the novel delivery system to be significantly more hydrophobic than control. Chemilluminscence showed a four-phase NO release profile of the delivery system with more stable and prolong NO release than control. Conclusion: LCAHs can optimize the duration and rate of NO delivery and present a viable option for use in surgical implants and biomedical applications. Keywords: nitric oxide, biofunctionalization, materials, aliphatic hydrocarbons

Polymer nitric oxide donors potentiate the treatment of experimental solid tumours by increasing drug accumulation in the tumour tissue

The delivery of nitric oxide (NO) specifically to solid tumours was explored in this study as a strategy to augment the passive accumulation of nanomedicines in tumours induced by the Enhanced Permeability and Retention (EPR) effect. An increase in accumulation was achieved by the binding of the chemical precursor of NO, based on an organic nitrate, to a water-soluble synthetic polymer drug carrier. Four structurally different N-(2-hydroxypropyl)methacrylamide (HPMA)-based polymer NO donors were synthesized. Depending on their chemical structure, two of these donors were hydrolytically stable, while two rapidly released the parent nitrate under acidic conditions, mimicking the intracellular environment. The polymer NO donors were shown to overcome the drawbacks related to low-molecular-weight NO releasing compounds, namely systemic toxicity, lack of site specificity, and fast blood clearance. The NO donors showed intracellular NO release upon incubation with tumour cells. In vivo, they potentiated the EPR effect, resulting in an increased accumulation of polymer-bound cytotoxic drug doxorubicin (Dox) in EL4 T-cell lymphoma inoculated in mice. This led to a better therapeutic outcome in the treatment of lymphoma with the high-molecular-weight polymer conjugates carrying Dox but not in the treatment with the free Dox. The localized augmentation of the EPR effect via the tumour-specific NO delivery system can be viewed as a promising strategy to potentiate polymer-based tumour therapy without increasing systemic toxicity.

Combination of nitric oxide and drug delivery systems: tools for overcoming drug resistance in chemotherapy

Chemotherapeutic drugs have made significant contributions to anticancer therapy, along with other therapeutic methods including surgery and radiotherapy over the past century. However, multidrug resistance (MDR) of cancer cells has remained as a significant obstacle in the achievement of efficient chemotherapy. Recently, there has been increasing evidence for the potential function of nitric oxide (NO) to overcome MDR. NO is an endogenous and biocompatible molecule, contrasting with other potentially toxic chemosensitizing agents that reverse MDR effects, which has raised expectations in the development of efficient therapeutics with low side effects. In particular, nanoparticle-based drug delivery systems not only facilitate the delivery of multiple therapeutic agents, but also help bypass MDR pathways, which are conducive for the efficient delivery of NO and anticancer drugs, simultaneously. Therefore, this review will discuss the mechanism of NO in overcoming MDR and recent progress of combined NO and drug delivery systems.

Conjugating S-nitrosothiols with fluorescent nanofibers for the controlled release and real-time detection of nitric oxide

In this work, a novel hybrid fibrous vehicle was developed for the release and real-time detection of nitric oxide (NO). The vehicle consisted of poly(L-lactide)/chitosan (PLLACS), cadmium tellurium quantum dots (CdTe QDs), and conjugating S-nitrosothiols. CdTe QDs were placed on the surface of PLLACS fibers as NO fluorescent probes through coaxial electrospinning; in this method, the PLLACS was used as filament material. Water-dispersible CdTe QDs were synthesized using mercaptoacetic acid as a stabilizer. The amino groups of chitosan served as reactive sites where chitosanbased S-nitrosothiols were formed. The fibrous PLLACS-QDs-NO composite was found to release NO under UV. The released NO could be detected in real time on the basis of the fluorescence quenching mechanism of QDs. These PLLACSQDs-NO nanofibers provide insights into novel designs of biocompatible NO delivery systems.

血清アルブミンを基盤とした一酸化窒素デリバリーシステムの創製と癌および虚血性疾患治療への応用

血清アルブミンを基盤とした一酸化窒素デリバリーシステムの創製と癌および虚血性疾患治療への応用

Development of a Sustained-release System for Nitric Oxide Delivery using Alginate/Chitosan Nanoparticles

Free radical nitric oxide (NO) has been known to interact with various physiological processes, such as wound repair processes and control of vascular tone. However, NO is an unstable molecule and the development of NO delivery systems that enhance its stability has also been studied. In this work, alginate/chitosan nanoparticles have been studied as a drug delivery system of the S- nitrosoglutathione (GSNO) as NO donor. For this, glutathione, GSH, the GSNO precursor, was encapsulated in alginate/chitosan nanoparticles. The presence of GSNO was confirmed by UV spectra at 336 nm. Alginate/chitosan nanoparticles with negative and posi- tive surface charges were obtained by increasing the chitosan amount. The encapsulation efficiency (EE) relied on the nanoparticle zeta potential, obtaining 80% of EE for positive particles. The NO release from GSNO showed that polymeric nanoparticles lead to the stabi- lization of GSNO decomposition, at physiological temperature. Moreover, this system did not exhibit cytotoxicity for fibroblast V79 cells up to the maximum concentration tested (18 μmolL -1 ). These results showed that alginate/chitosan nanoparticles are interesting particles to encapsulate NO donors for biomedical applications where NO might have a therapeutic effect.

Development of a Sustained-release System for Nitric Oxide Delivery using Alginate/Chitosan Nanoparticles

Free radical nitric oxide (NO) has been known to interact with various physiological processes, such as wound repair processes and control of vascular tone. However, NO is an unstable molecule and the development of NO delivery systems that enhance its stability has also been studied. In this work, alginate/chitosan nanoparticles have been studied as a drug delivery system of the Snitrosoglutathione (GSNO) as NO donor. For this, glutathione, GSH, the GSNO precursor, was encapsulated in alginate/chitosan nanoparticles. The presence of GSNO was confirmed by UV spectra at 336 nm. Alginate/chitosan nanoparticles with negative and positive surface charges were obtained by increasing the chitosan amount. The encapsulation efficiency (EE) relied on the nanoparticle zeta potential, obtaining 80% of EE for positive particles. The NO release from GSNO showed that polymeric nanoparticles lead to the stabilization of GSNO decomposition, at physiological temperature. Moreover, this system did not exhibit cytotoxicity for fibroblast V79 cells up to the maximum concentration tested (18 μmolL-1). These results showed that alginate/chitosan nanoparticles are interesting particles to encapsulate NO donors for biomedical applications where NO might have a therapeutic effect. Keywords: Polymeric nanoparticles, alginate, chitosan, nitric oxide, glutathione, cytotoxicity, s-nitrosoglutathione, pharmacology

An observational prospective study of topical acidified nitrite for killing methicillin-resistant Staphylococcus aureus (MRSA) in contaminated wounds

BackgroundEndogenous nitric oxide (NO) kills bacteria and other organisms as part of the innate immune response. When nitrite is exposed to low pH, NO is generated and has been used as an NO delivery system to treat skin infections. We demonstrated eradication of MRSA carriage from wounds using a topical formulation of citric acid (4.5%) and sodium nitrite (3%) creams co-applied for 5 days to 15 wounds in an observational prospective pilot study of 8 patients.FindingsFollowing treatment with topical citric acid and sodium nitrite, 9 of 15 wounds (60%) and 3 of 8 patients (37%) were cleared of infection. MRSA isolates from these patients were all sensitive to acidified nitrite in vitro compared to methicillin-sensitive S. aureus and a reference strain of MRSA.ConclusionsNitric oxide and acidified nitrite offer a novel therapy for control of MRSA in wounds. Wounds that were not cleared of infection may have been re-contaminated or the bioavailability of acidified nitrite impaired by local factors in the tissue.