Nitric oxide generating medical devices and methods

Abstract

An example of a medical device includes a first storage reservoir containing a first formulation, a second storage reservoir containing a second formulation, and a mixing reservoir in selective fluid communication with each of the first storage reservoir and the second storage reservoir. An example of the first formulation in the first storage reservoir includes a copper(I) ligand complex formed from a copper(II) ligand complex and an ascorbate source, and a calcium ion source. The second formulation includes a source of nitrite. A pH of the first formulation ranges from about 5.5 to about 7.5.

Description

NITRIC OXIDE GENERATING MEDICAL DEVICES AND METHODSCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application Serial Number 63 / 665,883, filed June 28, 2024, the content of which is incorporated by reference herein in its entirety.REFERENCE TO SEQUENCE LISTING

[0002] The Sequence Listing submitted herewith is hereby incorporated by reference in its entirety. The name of the file is UMJ243BPCT_2024-379- 02_Sequence_Listing.xml, the size of the file is 5,382 bytes, and the date of creation of the file is June 24, 2025.STATEMENT REGARDING FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0003] This invention was made with government support under HL168099, EB028775, and HL155100 awarded by the National Institutes of Health. The government has certain rights in the invention.BACKGROUND

[0004] Nitric oxide (NO) is an endogenous gas molecule that has been shown to have several important physiological functions, including its unique vasodilating properties, wound healing properties, angiogenesis promoting properties, cancerfighting potency, anti-platelet activity, and anti-microbial / anti-viral activity. In some instances, NO can be used to control infection, prevent biofilm formation, and minimize inflammation and fibrosis.SUMMARY

[0005] Examples of the medical devices and methods set forth herein generate nitric oxide (NO) from the reduction of nitrite, which is mediated via a copper(IIZI) ligandcomplex, the copper(ll) of which first undergoes an ascorbate induced reduction reaction. In these examples, the NO generation is performed in a controlled fashion by i) reducing copper(ll) in the solution phase of an electrolyte using the ascorbate and ii) controlling the availability of a nitrite source with the remaining of the electrolyte components. The formulations used in some of these examples of the medical devices and methods advantageously control the degradation product, oxalate, thus enabling its benefits (i.e. , reducing the reduction of nitrite to nitrous oxide (N2O)) to be maintained and its inhibitory effects (i.e., its ability to inhibit the NO generating reaction) to be minimized. Other examples of the medical devices and methods remove the copper(ll) reduction byproducts before the copper(l) is mixed with the nitrite source. This may provide a more storage stable formulation / electrolyte.BRIEF DESCRIPTION OF THE DRAWINGS

[0006] Features of examples of the present disclosure will become apparent by reference to the following detailed description and drawings, in which like reference numerals correspond to similar, though perhaps not identical, components. For the sake of brevity, reference numerals or features having a previously described function may or may not be described in connection with other drawings in which they appear.

[0007] Fig. 1 is a schematic view of one example of a medical device capable of generating NO using an example of the method disclosed herein;

[0008] Fig. 2 is a schematic view of two other examples of a medical device capable of generating NO using an example of the method disclosed herein;

[0009] Fig. 3 is a schematic view of still two additional examples of a medical device capable of generating NO using an example of the method disclosed herein;

[0010] Fig. 4 is a schematic view of yet another two examples of a medical device capable of generating NO using an example of the method disclosed herein;

[0011] Fig. 5 is a schematic view of a syringe used to generate an NO generating formulation for topical application;

[0012] Fig. 6 is a schematic view of one example of a patch used to generate an NO generating formulation for topical application;

[0013] Fig. 7 A is a perspective view of a catheter inserted into a blood vessel used, where the catheter is to receive the formulations disclosed herein for generating NO;

[0014] Fig. 7B is an exploded view of the catheter of Fig. 7A where the formulations are used as a lock solution;

[0015] Fig. 7C is an exploded view of the catheter of Fig. 7A where the formulations are used to prevent thrombus formation in blood contacting catheters or cannulas;

[0016] Fig. 8 is a schematic view of another example of a patch used to generate an NO generating formulation for topical application;

[0017] Fig. 9 is a graph depicting NO concentration (ppb, Y-axis) versus times (hours, X-axis) for a comparative first formulation;

[0018] Fig. 10 is a bar graph depicting the NO conversion results (percentage, Y-axis) for two comparative first formulations illustrating the interaction between calcium ions and oxalate;

[0019] Fig. 11 A is a graph depicting the NO concentration (ppm, Y-axis) versus time (minutes, X-axis) for five comparative first formulations and one example first formulation;

[0020] Fig. 11 B is a bar graph depicting the NO conversion (percentage, Y-axis) for the five comparative first formulations and the one example first formulation used to generate the data in Fig. 11 A;

[0021] Fig. 12A through Fig. 12D are graphs depicting the control of NO generation by the variation of nitrite (second formulation) infusion into a first formulation of a copper(ll) ligand complex (7 mM) and calcium ascorbate (140 mM) where the rate of NaNO2 was varied from 0-80 pL / min (Fig. 12A); where the correlation between the nitrite infusion rate and the concentration of NO generated is shown in Fig. 12B; where 7.2 mL of 10 mM NaN02was introduced at 15 pL / min (Fig. 12C); and where the average (dashed line) conversion is depicted as a function of nitrite infusion rate (Fig. 12D);

[0022] Fig. 13A through Fig. 13C are graphs respectively depicting the effect of storage on different example formulations including different amounts of calciumascorbate (140 mM = Fig. 13A, 7 mM = Fig. 13B, 35 mM = Fig. 13C), where 250 pL of 10 mM NaN02was infused at 10 pL / minute into each formulation stored over time;

[0023] Fig. 14 is a graph depicting the nitrite conversion to NO (percentage, Y- axis) as a function of calcium ascorbate concentration in the example formulations over six days of storage (electrolyte age in hours, X-axis);

[0024] Fig. 15 is a graph depicting the NO concentration (ppb, Y-axis) versus time (minutes, X-axis) for an example formulation and two comparative example formulations with different reducing agents (2 pL of NaN02was injected to 5 mL of each formulation, where the arrow on the X-axis shows the time of injection), and where the inset shows the details of the lower NO concentrations;

[0025] Fig. 16 includes bright field microscope (BFM) images (top row), fluorescence microscopy (FM) images (middle row), and merged BFM and FM images (bottom row), reproduced in black and white, of a control sample, 3 comparative samples, and 1 sample; and

[0026] Fig. 17A through Fig. 171 are fluorescence microscopy images, reproduced in black and white, of a control sample (Fig. 17A), 7 comparative samples (Figs. 17B-17H), and 1 sample (Fig. 171).DETAILED DESCRIPTION

[0027] Examples of the medical devices and methods set forth herein generate nitric oxide (NO) from the reduction of nitrite, which is mediated via a copper(IIZI) ligand complex, the copper(ll) of which first undergoes an ascorbate induced reduction reaction. These devices and methods control the availability of a limiting reactant (nitrite in one formulation) with the remainder of the electrolyte components (the other formulation), and thus enable control over the NO production.

[0028] Several medical devices are shown in Fig. 1 through Fig. 8. While each of the medical devices is identified by a particular reference numeral, e.g., 10A, 10B, etc., these devices are collectively referred to herein with the reference numeral 10. The medical device 10A shown in Fig. 1A and one example of the medical devices 10B, 10C, 10D shown in Fig. 2 through Fig. 4 are suitable for inhalation therapy.When used for inhalation therapy, these devices 10A, 10B, 10C, 10D eliminate theneed for bulky nitric oxide and nitrogen gas tanks. These devices may alternatively separate the generated nitric oxide and introduce it to a patient’s blood. The treated blood can then be reintroduced into the patient. Another example of the medical devices 10B’, 10C’, 10D’ shown in Fig. 2 through Fig. 4 delivers NO in solution to another medical device, such as a catheter or cannula. The medical devices 10E, 10F, and 10H shown in Fig. 5, Fig. 6, and Fig. 8 are suitable for topical applications and wound dressing. The medical device 10G shown in Fig. 7A is a catheter or cannula, which can be used with the devices 10B’, 10C’, 10D’ shown in Fig. 2 through Fig. 4.

[0029] Some of the medical devices 10 include and / or can be used with first and second formulations respectively including i) a copper(l) ligand complex formed from a copper(ll) ligand complex and an ascorbate source and ii) a source of nitrite. The formulation containing the reduced copper species is also referred to herein as the electrolyte. In this example, the specific components of the first and second formulations and are maintained separately from one another until NO generation is desirable. Other examples use surface-bound copper(ll) ligand complexes, and the ascorbate source and the source of nitrite are maintained separately until NO generation is desirable.

[0030] Referring now to Fig. 1 , the medical device 10A is an NO generator for inhalation therapy. This example device 10A includes a first storage reservoir 14 containing the first formulation 16, which includes: a copper(l) ligand complex formed from a copper(ll) ligand complex and an ascorbate source, and a calcium ion source, wherein a pH of the first formulation ranges from about 5.5 to about 7.5; a second storage reservoir 18 containing a second formulation 20, which includes a source of nitrite; and a mixing reservoir 22 in selective fluid communication with each of the first storage reservoir 14 and the second storage reservoir 18. In this example device 10A, each of the first and second formulations 16, 20 is a liquid, and the mixing reservoir 22 is a reaction chamber including a gas inlet 24 to introduce a purge gas into the reaction chamber; a bubbler 40 in fluid communication with the gas inlet 24 and with the reaction chamber; a gas outlet 26 to remove a gaseous mixture of the purge gas and nitric oxide; and a waste drain 28.

[0031] The first formulation 16 initially includes a copper(ll) ligand complex, an ascorbate source, a calcium ion source, and water, with or without a buffer. The pH of the first formulation 16 is or is adjusted so that it ranges from about 5.5 to about 7.5. This pH range is a suitable pH for the subsequent nitrite reduction reaction.

[0032] The buffer may be added to the first formulation 16 to obtain and maintain the desired pH. It is to be understood that any buffer (combination of salt(s), free acid, and water) may be used that has a pKa that will bring the pH of the formulation / electrolyte to within the reaction pH. As examples, the buffer is selected from the group consisting of phosphate buffered saline, tris(hydroxymethyl)aminomethane, (4-(2-hydroxyethyl)-1 -piperazineethanesulfonic acid), and 2-((morpholino)ethyl)sulfonic acid. When included, the buffer is present in the first formulation 16 at a concentration up to its solubility limit. In an example, the buffer is present at a concentration ranging from about 0.01 M to about 0.5 M.

[0033] The copper(ll) ligand complex is the mediator of the nitrite reduction reaction. In the presence of the ascorbate source, the copper(ll) ions of the ligand complex reduce to copper(l) ions, which function as a catalyst to produce NO from the nitrite that is introduced into the formulation / electrolyte at a controlled dosage.

[0034] Examples of the copper(ll) ligand complex are selected from the group consisting of a Cu(ll)-peptide ligand complex, Cu(ll)-tri(2-pyridylmethyl)amine (CuTPMA), Cu(ll)-tri(2-dimethylamino)ethyl]amine (CuMe6Tren), Cu(ll)-tri(2- pyridylmethyl)phosphine (CuTPMP), Cu(l l)-1 ,4,7-trimethyl-1 ,4-7 -triazacyclononane (Cu(Me3TACN)), Cu(ll)-1 ,4,7-triethyl-1 ,4-7-triazacyclononane (Cu(Et3TACN)), Cu(ll)- 1 , 4,7-tripropy 1-1 ,4-7-triazacyclononane (Cu(Pr3TACN)), Cu( 11)-1 , 4, 7-triisopropyl-1 ,4-7- triazacyclononane (Cu(iPr3TACN)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-ethylate) (Cu(BMPA-Et)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-propanoate) (Cu(BMPA-Pr)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-butylate) (Cu(BMPA-Bu)), Cu(ll)-(N,N-bis-(2- pyridylethyl)amine-N-ethylate) (Cu(BEPA-Et)), Cu(ll)- (N,N-bis-(2-pyridylethyl)amine-N- propanoate) (Cu(BEPA-Pr)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-butylate (Cu(BEPA-Bu)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-methyl-phenolate) (Cu(BMPA-MePhO)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-ethyl-phenolate) (Cu(BMPA-EtPhO)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-propyl-phenolate)(Cu(BMPA-PrPhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-methyl-phenolate) (Cu(BEPA-MePhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-ethyl-phenolate) (Cu(BEPA-EtPhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-propyl-phenolate) (Cu(BEPA-PrPhO)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)ethylate (Cu(PEMA-Et)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)propanoate (Cu(PEMA-Pr)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)butylate (Cu(PEMA-Bu)), Cu(ll)-2-(pyridin-2-yl)-N,N-bis(pyridin-2-ylmethyl)ethan-1 -amine (Cu(PMEA)), Cu(ll)-2,2'-(2-(2-(pyridin-2-yl)ethyl)butane-1 ,4-diyl)dipyridine (Cu(PMAP)), and combinations thereof. When the Cu(ll)-peptide ligand complex is used, a peptide ligand of the Cu(ll)-peptide ligand complex is selected from the group consisting of GKG, GHG, GAG, GDG, GHK, HGH, GGPG (SEQ. ID. NO. 1 ), GPGG (SEQ. ID. NO. 2), RGFTGM (SEQ. ID. NO. 3), PFRMY (SEQ. ID. NO. 4), and DAHK (SEQ. ID. NO. 5). The concentration of copper(ll) ligand complex in the first formulation 16, prior to its reduction, is dependent upon its solubility in water. Any concentration up to its solubility limit may be used. In an example, the copper(ll) ligand complex is present in the first formulation 16 at a concentration ranging from about 6 mM to about 8 mM. In one example, the copper is introduced as copper sulfate separately from the ligand. The concentration of each of these components is within the given range. As a specific example, about 7 mM of the copper(ll) ligand complex is present in the first formulation 16 prior to its reduction, and about 7 mM of the copper(l) ligand complex is present in the formulation after the ascorbate initiated reduction reaction.

[0035] The ascorbate is a reducing agent that reduces the copper(ll) ions of the copper(ll) ligand complex to copper(l) ions, which, in turn reduce nitrite to NO when the nitrite source is introduced. The calcium ions are included in the formulation 16 to mitigate the undesired effects of oxalate, which is a degradation product of ascorbate. In some examples, the ascorbate source and the calcium ion source are different. As examples, the ascorbate source is selected from the group consisting of ascorbic acid, potassium ascorbate, magnesium ascorbate and sodium ascorbate, and the calcium ion source is selected from the group consisting of calcium threonate and calciumchloride. In one example, the calcium threonate is present at a 1 :1 molar ratio with the ascorbate source.

[0036] In these examples, the ascorbate source is present in the first formulation 16 in a concentration up to its solubility, and the calcium ion source is present in the first formulation 16 in a concentration up to one equivalent of the ascorbate concentration. In one example, the concentration of each of the ascorbate source and the calcium ion source ranges from about 5 mM up to 1 M. In other examples, the ascorbate source and the calcium ion source are the same. As an example, the source of both ascorbate and calcium ions is calcium ascorbate. In these examples, the concentration of the source of both ascorbate and calcium ions in the first formulation 16 is up to 1 .2 M. After the copper(ll) reduction reaction, dehydroascorbic acid is present in the formulation 16.

[0037] To form the first formulation 16, the various components are added to the water and mixed until they are dissolved. The pH of the solution is tested, and if outside the range of 5.5 to 7.7, may be adjusted to between 5.5 and 7.5 using a suitable buffering agent as described herein. Alternatively, the pH can be adjusted by adding a source of protons (e.g., hydrochloric acid) or a source of hydroxide ions (e.g., sodium hydroxide), as needed.

[0038] Because the copper(ll) ligand complex and the ascorbate are both present in the first formulation 16 and because the reduction reaction of copper(ll) by ascorbate can take place at room temperature, the generation of copper(l) takes place once the first formulation 16 is formulated. Thus, the first formulation 16 introduced into the storage reservoir 14 includes the copper(ll) ligand complex and the ascorbate as described herein, and as the reduction reaction takes place, the first formulation 16 stored in the storage reservoir 14 contains the copper(l) ligand complex, dehydroascorbic acid, any unreacted components, and any degradation products or byproducts of the copper(ll) reduction reaction (e.g., the practically insoluble calcium salt described herein).

[0039] In the medical device 10A, the first formulation 16 is stored in the first storage reservoir 14. The storage reservoir 14 may be a fluid storage bag, a hardcontainer (e.g., vial, cartridge, or the like), or any other vessel that can hold and release, on demand, the first formulation 16.

[0040] The second formulation 20 includes a source of nitrite in water.

[0041] The source of nitrite is the source of the nitric oxide, as it undergoes the copper(l) induced chemical reduction to generate NO. The source of nitrite is any water soluble inorganic nitrite salt that exhibits storage stability in an aqueous solution and is light insensitive. Some example water soluble, inorganic nitrite salts include nitrite salts of, Na (sodium) or K (potassium). One specific example of the nitrite salt is sodium nitrite (NaNO ). For sustained NO release, these nitrite salts may be combined with an excipient that provides sustained release of the nitrite within the reaction mixture, as described in U.S. Pat. App. Pub. No. 2011 / 0086069, which is incorporated herein by reference in its entirety.

[0042] The concentration of nitrite source in the second formulation 20 is dependent upon its solubility in water. Thus, the nitrite source may be present in the second formulation 20 at a concentration up to its solubility limit. In one example, the nitrite source ranges from about 5 mM to about 15 mM. In another example, the concentration of the nitrite source is 10 mM. The balance of the second formulation 20 is water, either alone or in combination with a second buffer. Any of the examples set forth herein for the first buffer may be used as the second buffer. In an example, the buffer may be added to the second formulation to adjust the pH to within a range of 5.5 to 8.5.

[0043] Because the first and second formulations are ultimately mixed together, it may be desirable to utilize the same buffer in each of the formulations 16, 20.

[0044] In the medical device 10A, the second formulation 20 is stored in the second storage reservoir 18. The storage reservoir 18 may be a fluid storage bag, a hard container, or any other vessel that can hold and release, on demand, the second formulation 20.

[0045] In specific examples of the first and second formulations 16, 20, i) the copper(ll) ligand complex is selected from the group consisting of a Cu(ll)-peptide ligand complex, Cu(ll)-tri(2-pyridylmethyl)amine (CuTPMA), Cu(ll)-tri(2- dimethylamino)ethyl]amine (CuMe6Tren), Cu(ll)-tri(2-pyridylmethyl)phosphine(CuTPMP), Cu(ll)-1 , 4, 7-trimethyl-1 , 4-7 -triazacyclononane (Cu(Me3TACN)), Cu(ll)- 1 , 4, 7-triethy 1-1 ,4-7-triazacyclononane (Cu(Et3TACN)), Cu( 11)-1 , 4, 7-tripropyl-1 ,4-7- triazacyclononane (Cu(Pr3TACN)), Cu(ll)-1 ,4,7-triisopropyl-1 ,4-7-triazacyclononane (Cu(iPr3TACN)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-ethylate) (Cu(BMPA-Et)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-propanoate) (Cu(BMPA-Pr)), Cu(ll)-(N,N-bis- (2-pyridylmethyl)amine-N-butylate) (Cu(BMPA-Bu)), Cu(ll)-(N,N-bis-(2- pyridylethyl)amine-N-ethylate) (Cu(BEPA-Et)), Cu(ll)- (N,N-bis-(2-pyridylethyl)amine-N- propanoate) (Cu(BEPA-Pr)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-butylate (Cu(BEPA-Bu)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-methyl-phenolate) (Cu(BMPA-MePhO)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-ethyl-phenolate) (Cu(BMPA-EtPhO)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-propyl-phenolate) (Cu(BMPA-PrPhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-methyl-phenolate) (Cu(BEPA-MePhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-ethyl-phenolate) (Cu(BEPA-EtPhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-propyl-phenolate) (Cu(BEPA-PrPhO)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)ethylate (Cu(PEMA-Et)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)propanoate (Cu(PEMA-Pr)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)butylate (Cu(PEMA-Bu)), Cu(ll)-2-(pyridin-2-yl)-N,N-bis(pyridin-2-ylmethyl)ethan-1 -amine (Cu(PMEA)), Cu(ll)-2,2'-(2-(2-(pyridin-2-yl)ethyl)butane-1 ,4-diyl)dipyridine (Cu(PMAP)), and combinations thereof; ii) the ascorbate source is ascorbic acid, potassium ascorbate, magnesium ascorbate and sodium ascorbate, and the calcium ion source is selected from the group consisting of calcium threonate and calcium chloride, or calcium ascorbate is both the ascorbate source and the calcium ion source; and iii) the source of nitrite is an inorganic nitrite salt.

[0046] As depicted in Fig. 1 , the medical device 10A further includes a first fluid line 30 connecting the first storage reservoir 14 and the mixing reservoir 22; a valve 32 operatively positioned along the first fluid line 30; a second fluid line 34 connecting the second storage reservoir 18 and the mixing reservoir 22; and a dosing pump 36 operatively position along the second fluid line 34.

[0047] The fluid lines 30, 34 are fluid conduits respectively connecting the reservoirs 14, 18 to the mixing reservoir 22. The fluid lines 30, 34 may be any suitablepolymeric tubing. Each of the fluid lines 30, 34 is set up for one-way flow of the respective formulations 16, 20 into the mixing reservoir 22.

[0048] The valve 32 may be opened as desired to allow a desired amount of the first formulation 16 to flow from the reservoir 14 into the mixing reservoir 22. The valve 32 may be a one-way valve or a flow controller. The valve 32 may be manually operated or automated.

[0049] The dosing pump 36 is a positive displacement pump that injects a predetermined amount of the second formulation 20 into the mixing reservoir 22. The dosing pump 36 is an automatic dispenser that provides an extremely precise flow rate for maximum control of the nitrite source. The dosing pump 36 may be feedback controlled based on measured levels of NO using sensors that are operatively positioned within the mixing reservoir 22 or along the outlet conduit 42.

[0050] The mixing reservoir 22 is a container that receives and mixes the first and second formulations 16, 20. The first formulation 16 is introduced into the mixing reservoir 22 first, and then the controlled injection of the second formulation 20 enables NO generation in a controlled fashion. Within the mixing reservoir 22, the source of nitrite comes into contact with the copper(l) ligand complex generated within the first formulation 16 by the ascorbate initiated reduction of the copper(ll) ligand complex, and thus the copper(l) initiated reduction reaction of the nitrite is initiated to form the nitric oxide. As such, the mixing reservoir 22 is a reaction chamber for the NO generation. The term “mixed formulation” is used herein to refer to the formulation that is generated as a result of mixing the first and second formulations 16, 18.

[0051] Any of the containers set forth herein may be used for the mixing reservoir 22. The mixing reservoir 22 may include a stirring mechanism (e.g., a stir bar), a vibration mechanism, or may be shaken to mix the first and second formulations 16, 20 upon their introduction to the reservoir 22. Alternatively, a mixing mechanism may not be included as the purge gas that is introduced is sufficient to mix the first and second formulations 16, 20.

[0052] In addition to the respective inlets for the formulations 16, 20, the mixing reservoir 22 also includes a purge gas inlet 24, a mixed gas outlet 26, and a waste outlet 28.

[0053] The purge gas inlet 24 is operatively connected to a fluid line 38 that is used to introduce a purse gas into the mixing reservoir 22. The purge gas may also be referred to as a sparge gas or a sweep gas. While not shown, the fluid line 38 may also include a valve for controlling the input of the purse gas. The purge gas may be nitrogen gas (N2), oxygen gas (O2), an oxygen-containing mixed gas (e.g., air), or an oxygen deficient mixed gas. When a substantially pure gas is to be used, the purge gas may be supplied to the inlet 24 from a compressed gas tank. Alternatively, an oxygen scrubber or a nitrogen enriching membrane may be used to generate, respectively, an oxygen deficient mixed gas or a nitrogen rich mixed gas derived from ambient air that contains nitrogen gas, argon gas, carbon dioxide, and potentially small amounts of other non-oxygen gases. The presence of oxygen in the purge gas depends, in part, upon the O2 sensitivity of the ligand chemistry. Moreover, nitrogen or oxygen deficient gas may be more desirable as it does not react with the generated NO, which can lead to a higher concentration of NO in the mixed output stream. Thus, the use of the nitrogen purge gas or any of the oxygen deficient mixed gases may improve the stability in terms of NO generation.

[0054] The mixing reservoir 22 may contain a bubbler 40. In an example, the bubbler 40 is fritted. The bubbler 40 introduces the purge gas into the reaction mixture (i.e. , the mixed formulation) by generating fine bubbles, which are effective for removing the generated NO gas from the liquid phase of the mixed formulation.

[0055] The purge gas that is introduced into the mixing reservoir 22 picks up the nitric oxide that is generated in the mixed formulation. The resulting stream of purge gas and nitric oxide is then transported out of the mixing reservoir 22 through the outlet 26 and into an outlet conduit 42. In each of the examples set forth herein, the outlet conduit 42 may be a tube that has low or no permeability to NO and the other gas(s) in the output stream. The length of the outlet conduit 42 may also be relatively short in order to avoid loss of gas before the stream is delivered to a desirable destination, such as a patient, a separator (e.g., for removing aerosol droplets and excess humidity from the output gas), a nitric oxide extraction device, or an oxygenator.

[0056] In some examples, the output stream may be transported as a result of pressure from the gas source, which may include a regulator to control the flow rate. In other examples, the output stream may be transported as a result of pressure from a vacuum positioned downstream of the outlet 26.

[0057] As shown in Fig 1., the medical device 10A may also include an inspiratory gas conduit 43 that is operatively connected to the outlet conduit 42. This additional gas conduit 43 may be used when the purge gas is nitrogen gas or an oxygen deficient gas, so that an oxygen-containing gas may be introduced into the output stream. The oxygen-containing gas is any ventilation gas that has an oxygen (O2) content ranging from 21 % to 100%. As examples, the oxygen-containing gas may be at least substantially pure oxygen gas O2 or air. In this example, the oxygencontaining gas is delivered from the inspiratory gas conduit 43 into the outlet conduit 42 where it mixes with the output stream of nitric oxide and nitrogen gas or oxygen deficient gas. In an example, the source (not shown) of the oxygen-containing gas may include a regulator to control the flow rate, so that the final mixed gas stream contains from about 20% oxygen to about 99.99% oxygen. Since the oxygencontaining gas is introduced just prior to delivery to a recipient, the time period at which the NO can react with O2 is reduced, and thus the impact on the NO concentration, due to NO2 formation, is minimal or nil.

[0058] The inspiratory gas conduit 43 may be a tube that has low or no permeability to at least the oxygen-containing gas, the nitrogen gas or oxygen deficiency gas, and the nitric oxide. Examples of suitable tubing materials for the inspiratory gas conduit 43 include poly(vinyl chloride) (PVC), polyurethane (PU), polyethylene (PE), fluorinated polymers, etc. These materials may also be used for any of the other conduits, e.g., 30, 34, 38, etc. described herein.

[0059] The medical device shown in Fig. 1 may also include a filter 44 that is operatively positioned in the outlet conduit 42 upstream of the inspiratory gas conduit 43. In one example, the filter 44 is a hydrophobic filter to remove aerosol droplets from the output gas. This example filter 44 is a membrane made up of several NO permeable microporous fibers (e.g., silicone rubber, porous polytetrafluoroethylene (PTFE), polybutadiene, poly(butadiene co-styrene), polycisisoprene, polypropylene(PP), poly(methyl pentene), etc.) or another material that allows the output stream to diffuse through and out of the filter 44 while blocking the aerosol droplets from exiting with the output stream.

[0060] The medical device shown in Fig. 1 may also include another filter 45 that is operatively positioned in the outlet conduit 42 downstream of the inspiratory gas conduit 43. This filter 45 is one that selectively removes contaminants, such as NO2, from the output gas. Suitable NO2 remover filters include ascorbate impregnated silica gel or soda lime.

[0061] As shown in Fig. 1 , the medical device 10A may also include a waste container 46 which can receive the non-gaseous components of the mixed formulation. A valve 48 may be positioned between the mixing reservoir 22 and the waste container 46 so that the mixed formulation and any gases dissolved therein are not prematurely removed from the mixing reservoir 22. As depicted in Fig. 1 , the gas outlet 26 and the waste outlet 28 may be operatively positioned in the upper and lower portions of the mixing reservoir 22, respectively, to utilize the effects of gravity.

[0062] Because the nitrite containing formulation 20 (i.e. , nitrite concentration) and its introduction into the system is controlled, the generation of nitric oxide and its concentration is also controlled. In other words, all of the nitrite that is introduced should react, generating a known amount of NO. Thus, gas sensors may or may not be included in the systems 10 disclosed herein.

[0063] In any of the examples described in reference to Fig. 1 , the NO concentration in the output stream may be feedback controlled. One or more sensors, such as NO sensors and / or NO2 sensors, may be positioned in the mixing reservoir 22, the outlet conduit 42, or in another in another conduit that is split or branched off of the outlet conduit 42. When the inspiratory gas conduit 43 is used, the feedback control sensors may be positioned downstream of the introduction of the oxygencontaining gas to the output stream. When the filter(s) 44, 45 is / are included, the feedback control sensors may be positioned downstream of the f ilter(s) 44, 45.

[0064] It may be desirable to monitor the NO level in order to avoid forming NO2 (nitrogen dioxide, which can be generated from O2 reacting with NO and can be toxic). Example NO sensors include a Shibuki-style sensor, which is based on the oxidationof NO to nitrate (NOs-) at an inner platinum (Pt) electrode position behind a gas permeable membrane, or an amperometric NO sensor. Sensor data (i.e. , the concentration of NO in the output gas stream and / or the concentration of NO2 in the output gas stream) may be used to servo-regulate the second formulation 20 that is added to the mixing chamber 22 in order to achieve an at least substantially constant concentration of NO at the delivery end. The data may also be used to regulate the flow of the inspiratory gas and / or the output gas stream.

[0065] Referring now to Fig. 2, two additional examples of the medical device 10B are depicted. Like the medical device 10A, the medical devices 10B, 10B’ include the first storage reservoir 14 containing the first formulation 16; the second storage reservoir 18 containing the second formulation 20; and a mixing reservoir 22’ in selective fluid communication with each of the first storage reservoir 14 and the second storage reservoir 18. The medical device 10B is suitable for inhalation therapy or for introducing nitric oxide to blood, while the medical device 10B’ is suitable for the delivery of NO from solution.

[0066] In one example of the device 10B, each of the first and second formulations 16, 18 is a liquid; and the medical device 10B further includes a sediment separator 48 in selective fluid communication with, and downstream of, the mixing reservoir 22’; and a liquid / gas contactor 50 in selective fluid communication with, and downstream of, the sediment separator 48 and also in selective fluid communication with the first storage reservoir 14, where the liquid / gas contactor 50 includes a gas inlet 70’ to introduce a purge gas into the liquid / gas contactor 50; and a gas outlet 72’ to remove a gaseous mixture of the purge gas and nitric oxide.

[0067] In another example of the device 10B, each of the first and second formulations 16, 18 is a liquid; and the medical device 10B further includes the sediment separator 48 in selective fluid communication with, and downstream of, the mixing reservoir 22’; and a liquid / liquid contactor 50’ in selective fluid communication with, and downstream of, the sediment separator 48 and also in selective fluid communication with the first storage reservoir 14, where the liquid / liquid contactor 50’ includes a liquid inlet 70’ to introduce a blood into the liquid / gas contactor 50’; and a liquid outlet 72’ to remove a mixture of the blood and nitric oxide.

[0068] It is to be understood that any examples of the first storage reservoir 14, the first formulation 16, the second storage reservoir 18, and the second formulation 20 may be used in the medical device 10B.

[0069] As depicted in Fig. 2, the medical device 10B further includes the first fluid line 30 connecting the first storage reservoir 1 and the mixing reservoir 22’; a circulating pump 52 operatively positioned along the first fluid line 30; the second fluid line 34 connecting the second storage reservoir 18 and the mixing reservoir 22’; the dosing pump 36 operatively position in the second fluid line 34; and a third fluid line 54 connecting the liquid / gas contactor 50 or the liquid / liquid contactor 50’ with the first storage reservoir 14.

[0070] As described in reference to Fig. 1 , fluid lines 30, 34 are fluid conduits respectively connecting the reservoirs 14, 18 to the mixing reservoir 22’, and may be any suitable polymeric tubing. Each of the fluid lines 30, 34 is set up for one-way flow of the respective formulations 16, 20 into the mixing reservoir 22’.

[0071] The circulating pump 52 is positioned in the first fluid line 30 and is used to pump fluids throughout the fluidic circuit that connects the first storage reservoir 14 to the mixing reservoir 22’ to the sediment separator 48 to the liquid / gas contactor 50 or the liquid / liquid contactor 50’ and back to the first storage reservoir 14. The circulating pump 52 may be operated continuously to move the first fluid 16 along this fluidic circuit, and the dosing pump 34 may be operated when it is desirable to generate NO.

[0072] The dosing pump 34 is operated as described in reference to Fig. 1 .

[0073] Similar to the mixing reservoir 22 described in reference to Fig. 1 , the mixing reservoir 22’ is a container that receives and mixes the first and second formulations 16, 20. The reduction reaction to generate NO takes place within the mixing reservoir 22’, and thus the mixing reservoir 22’ is a reaction chamber. Any of the containers set forth herein may be used for the mixing reservoir 22’. In the medical device 10B, the mixing chamber 22’ is a passive mixer because the fluid is circulated through the device 10B. The fluid movement mixes the formulations 16, 20.

[0074] Unlike the mixing reservoir 22, the mixing reservoir 22’ includes a single outlet 26’, through which the mixed formulation and any gas dissolved therein (nitricoxide) are transported through the outlet conduit 42. As mentioned in reference to Fig. 1 , the outlet conduit 42 is at least NO impermeable. In this example, the circulating pump 52 controls the movement of the fluid from the mixing reservoir 22’ to the sediment separator 48.

[0075] The separator 48 has a filter 44’ positioned therein that allows sediment within the mixed formulation to be removed therefrom. When the copper(ll) and ascorbate are reacted within the formulation 16 and the calcium source is present to mitigate the undesired effects of oxalate, calcium salt byproducts may be present. This filter 44’ blocks particulates, such as the calcium salt byproducts that form the sediment, from moving through the separator 48, while allowing the liquid of the mixed formulation and the nitric oxide gas dissolved therein to transport therethrough. The blocked sediment may be transported, for example, to the waste container 46.

[0076] The solution phase of the mixed formulation, which contains dissolved nitric oxide, can then be transported through another fluid line 56 to the I iquid / gas contactor 50.

[0077] As shown in Fig. 2, the liquid / gas contactor 50 or the liquid / liquid contactor 50’ includes a housing 58, which contains a nitric oxide permeable medium 60 separating two spaces 62, 64. The housing 58 may be made of any material that is not permeable to the gases that are introduced thereto (e.g., nitrogen gas, nitric oxide, air, oxygen gas, etc.). In the liquid / gas contactor 50, the space 62 is a liquid space and the space 64 is a gaseous space. In the liquid / liquid contactor 50’, the space 62 is a first liquid space and the space 64 is a second liquid space.

[0078] In the example shown in Fig. 2, the housing 58 includes two inlets 70, 70’ and two outlets 72, 72’. The inlet and outlet 70, 72 lead to and from, respectively, the (first) liquid space 62. The inlet and outlet 70’, 72’ lead to and from, respectively, the gaseous or second liquid space 62. More specifically, the first housing inlet 70 operatively connects the fluid line 56 to the first space 62. As such, the solution phase of the mixed formulation (having NO dissolved therein) is directed from the fluid line 56 into the first space 62. A partial pressure gradient in the liquid / gas contactor 50 or a concentration gradient in the liquid / liquid contactor 50’ across the membrane 60 drivesat least some of the dissolved NO gas out of the solution phase and through the membrane 60 into the gas or liquid in the second space 64.

[0079] The nitric oxide permeable medium 60 blocks the solution phase of the mixed formulation, and this liquid can be redirected to the first storage reservoir 14 via the outlet 72 and the fluid line 54. NO gas that is not driven out of solution and through the membrane 60 remains dissolved in the solution phase of the mixed formulation. Thus, the residual dissolved NO gas is transported with the solution phase along the path that leads from the outlet 72 back to the first storage reservoir 14. The residual dissolved NO gas in the recirculated solution will circulate through the device 10B when the formulation 16 is moved from the storage reservoir 14.

[0080] The nitric oxide permeable medium 60 may be a membrane or a bunch of hollow fibers. The medium 60 may be made up of several NO permeable microporous fibers (e.g., silicone rubber, porous polytetrafluoroethylene (PTFE), polybutadiene, poly(butadiene co-styrene), polycisisoprene, polypropylene (PP), etc.) or another material that allows at least some of the nitric oxide to diffuse through to the second space 64 while blocking the solution.

[0081] As mentioned, the diffused NO gas stream enters the second space 64 of the liquid / gas contactor 50 or the liquid / liquid contactor 50’. In the example shown in Fig. 2, the second space 64 is connected to the second housing inlet 70’ and the second housing outlet 72’.

[0082] In the liquid / gas contactor 50, the second housing inlet 70’ is also operatively connected to a purge gas conduit (shown at 38’) to receive, in the second space 64, any example of the purge gas set forth herein, which mixes with the nitric oxide diffusing through the nitric oxide permeable membrane 60 to form the output gas stream. The purge gas may be delivered to the second space 64 from any suitable gas source. A flow controller is positioned upstream of the outlet 72’ to regulate the flow of the purge gas in and out of the second space 64. The upstream position of the flow controller helps to avoid pressurization within the liquid / gas contactor 50. The flow rate of the purge gas may be continuous or intermittent. As examples, the gas source may be a compressed gas cylinder, a gas pump that delivers ambient air, or any other suitable gas source.

[0083] In the second space 64 of the I iquid / gas contactor 50, diffused NO gas stream mixes with the purge gas to form the output gas stream. The second housing outlet 72’ is operatively connected to another fluid line or conduit 68 that transports the output gas stream to a recipient / patient or other desired destination. The fluid line or conduit 68 may be any suitable polymeric or other tubing that is impermeable to the output gas stream. In an example, the fluid line or conduit 68 also includes a one-way valve so that the output gas stream does not flow back into the liquid / gas contactor 50.

[0084] As shown in Fig. 1 , medical device 10B may also include an inspiratory gas conduit 43 that is operatively connected to the conduit 68. This additional gas conduit 43 may be used when the purge gas is nitrogen gas or an oxygen deficient gas, so that an oxygen-containing gas may be introduced into the output stream. The oxygen-containing gas is any ventilation gas that has an oxygen (O2) content ranging from 21% to 100%. As examples, the oxygen-containing gas may be at least substantially pure oxygen gas O2 or air. In this example, the oxygen-containing gas is delivered from the inspiratory gas conduit 43 into the conduit 68 where it mixes with the output stream of nitric oxide and nitrogen gas or oxygen deficient gas. In an example, the source (not shown) of the oxygen-containing gas may include a regulator to control the flow rate, so that the final mixed gas stream contains from about 20% oxygen to about 99.99% oxygen. The inspiratory gas conduit 43 may be a tube that has low or no permeability to at least the oxygen-containing gas, the nitrogen gas or oxygen deficiency gas, and the nitric oxide. Any of the examples set forth herein may be used.

[0085] The medical device 10B may also include the sensors positioned in the conduit 68 downstream of the inspiratory gas conduit 43 to monitor the content of the final output gas.

[0086] In the liquid / liquid contactor 50’ the second housing inlet 70’ is also operatively connected to an extracorporeal circuit 39 to receive, in the second space 64, blood from a patient. Within the second space, the blood mixes with the nitric oxide diffusing through the nitric oxide permeable membrane 60 to form a treated blood sample. A flow controller may be positioned upstream of the outlet 72’ to regulate the flow of the blood in and out of the second space 64. The upstreamposition of the flow controller helps to avoid pressurization within the liquid / liquid contactor 50’. The flow rate of the blood may be continuous or intermittent.

[0087] In the second space 64 of the liquid / liquid contactor 50’, diffused NO gas stream mixes with the blood to form NO treated blood. The second housing outlet 72’ is operatively connected to another fluid line or conduit 68 that transports the treated blood back to the patient. The fluid line or conduit 68 may be any suitable polymeric or other tubing that is impermeable to the treated blood. In an example, the fluid line or conduit 68 also includes a one-way valve so that the output treated blood does not flow back into the liquid / liquid contactor 50’.

[0088] The medical device 10B including the liquid / liquid contactor 50’ may be used as an extracorporeal circuit to withdraw blood from a patient, treat the blood with nitric oxide, and then deliver the treated blood back to the patient.

[0089] The medical device 10B’ will now be described. As mentioned, the medical device 10B’ shown in Fig. 2 is for delivering nitric oxide that remains dissolved in solution. Thus, unlike the device 10B, this example device 10B’ does not include the liquid / gas contactor 50 or the liquid / liquid contactor 50’, or the fluid line 54, or the circulating pump 52. Rather, this example device 10B’ includes the storage reservoirs 14, 18 (and the respective formulations 16, 20 contained therein); the mixing reservoir 22’; the first fluid line 30 connecting the first storage reservoir 14 and the mixing reservoir 22’; the valve 32 operatively positioned along the first fluid line 30; the second fluid line 34 connecting the second storage reservoir 18 and the mixing reservoir 22’; the dosing pump 36 operatively position along the second fluid line 34; the sediment separator 48 in selective fluid communication with, and downstream of, the mixing reservoir 22’; and an output fluid line 69.

[0090] It is to be understood that any examples of the first storage reservoir 14, the first formulation 16, the second storage reservoir 18, and the second formulation 20 may be used in the medical device 10B’. The formulations 16, 20 may be delivered to the mixing chamber 22’ as described in reference to Fig. 1 (e.g., using the valve 32 and dosing pump 36, respectively), and the mixing reservoir 22’ may include an additional mechanism to stir, vibrate, or otherwise mix the formulations 16, 20.

[0091] In this example device, the mixed formulation (containing dissolved NO) is introduced into the separator 48 as described herein. Particulate material is separated and removed from the solution phase, which is then directed through the output fluid line 69.

[0092] The output fluid line 69 fluidly connects the separator 48 to a catheter or cannular 10G (see Fig. 7A).

[0093] The end of the output fluid line 69 may include a connector (not shown) that attaches to the adapter (hub) 92 of the catheter or cannula 10G (Fig. 7A). In one example, the connector fluidly connects the output fluid line 69 to an inner lumen 84 of the catheter or cannular 10G. As will be described in reference to Fig. 7B, the mixed formulation may be introduced, as a lock solution, to the lumen 84 while the catheter 10G is in place within a patient but is not then-currently in use. In another example, the connector fluidly connects the output fluid line 69 to an outer lumen 88 of a multilumen catheter or cannula 10G. As will be described in reference to Fig. 7C, the mixed formulation may be introduced into the outer lumen 88 (as a static solution) while the catheter or cannula 10G is being used to draw blood from a patient or introduce medications, saline, or the like, to a patient.

[0094] Any of the gas sensors may be positioned in the output fluid line 69.

[0095] Another example of the medical device is similar to device 10B’. This example includes the reservoirs 14, 18 with the formulations 16, 20 stored therein, the fluid lines 30, 34, the valve 32, the dosing pump 36, the mixing reservoir 22’ and the fluid line 42 as described herein. This device does not include the separator 48 or any of the other downstream components shown in Fig. 2. Rather, this device includes a filter (similar to filter 44’) positioned within the mixing reservoir 22’ to remove sediment from the nitric oxide containing solution generated in the mixing reservoir 22’; a liquid outlet (similar to outlet 26’) defined in the mixing reservoir 22’; an extracorporeal circuit (see Fig. 7C) including: an inner lumen 84; an outer lumen 88 surrounding the inner lumen 84; and a nitric oxide permeable wall separating the inner lumen 84 from the outer lumen 88; and a fluid line (e.g., 42) connecting the liquid outlet with the outer lumen 88 of the extracorporeal circuit. In this example, the mixed formulationcontaining NO may be introduced into the outer lumen 88, where the NO will permeate through the nitric oxide permeable wall into a fluid flowing through the inner lumen 84.

[0096] The medical devices 10C, 10C’ shown in Fig. 3 are respectively similar to the medical devices 10B, 10B’ shown in Fig. 2, except that the copper(ll) ligand complex is removed from the formulation 16 and is contained in a reaction chamber 74 that is used to generate the NO after another example of the first formulation 16’ and the second formulation 20 are mixed and introduced thereto.

[0097] In this example, each of the medical devices 10C, 10C’ includes the first storage reservoir 14 containing the other example of the first formulation 16’; the second storage reservoir 18 containing the second formulation 20; a mixing reservoir 22” in selective fluid communication with each of the first storage reservoir 14 and the second storage reservoir 18; and the reaction chamber 74 containing a copper source, the copper source including a resin; and the copper(ll) ligand complex immobilized at a surface of the resin. One example of the device 10C further includes the sediment separator 48 in selective fluid communication with, and downstream of, the reaction chamber 74; and a I iquid / gas contactor 50 in selective fluid communication with, and downstream of, the sediment separator 48 and also in selective fluid communication with the first storage reservoir 14, the liquid / gas contactor 50 including: a gas inlet 70’ to introduce a purge gas into the liquid / gas contactor 50; and a gas outlet 72’ to remove a gaseous mixture of the purge gas and nitric oxide. Another example of the device 10C further includes the sediment separator 48 in selective fluid communication with, and downstream of, the reaction chamber 74; and a liquid / liquid contactor 50’ in selective fluid communication with, and downstream of, the sediment separator 48 and also in selective fluid communication with the first storage reservoir 14, the liquid / liquid contactor 50’ including: a liquid inlet 70’ to introduce blood into the liquid / liquid contactor; and a liquid outlet 72’ to remove a mixture of the blood and nitric oxide.

[0098] It is to be understood that any examples of the first storage reservoir 14, the second storage reservoir 18, and the second formulation 20 may be used in the medical devices 10C, 10C’.

[0099] In the example devices 10C, 10C’, the first formulation 16’ includes water (with or without the first buffer), the ascorbate source, and the calcium ion source. Anyof these components and their respective amounts set forth herein for the first formulation 16 may be used in the formulation 16’. The pH of the first formulation 16’ is also from about 5.5 to about 7.5. This example of the first formulation 16’ is devoid of the copper(ll) ligand complex.

[0100] As depicted in Fig. 3, the medical device 10C further includes the first fluid line 30 connecting the first storage reservoir 14 and the mixing reservoir 22”; the circulating pump 52 operatively positioned along the first fluid line 30; the second fluid line 34 connecting the second storage reservoir 18 and the mixing reservoir 22”; the dosing pump 36 operatively position in the second fluid line 24; and a third fluid line 54 connecting the liquid / gas contactor 50 or the liquid / liquid contactor 50’ with the first storage reservoir 14. Also as depicted in Fig. 3, the medical device 10C’ further includes the first fluid line 30 connecting the first storage reservoir 14 and the mixing reservoir 22”; the valve 32 operatively positioned along the first fluid line 30; the second fluid line 34 connecting the second storage reservoir 18 and the mixing reservoir 22”; and the dosing pump 36 operatively position along the second fluid line 34.

[0101] Similar to the description of Fig. 1 , fluid lines 30, 34 are fluid conduits respectively connecting the reservoirs 14, 18 to the mixing reservoir 22”, and may be any suitable polymeric tubing. Each of the fluid lines 30, 34 is set up for one-way flow of the respective formulations 16’, 20 into the mixing reservoir 22”.

[0102] Similar to the description of Fig. 2 for the device 10B, the circulating pump 52 of the medical device 10C is positioned in the first fluid line 30 and is used to pump fluids throughout the fluidic circuit that connects the first storage reservoir 14 to the mixing reservoir 22” to the sediment separator 48 to the liquid / gas contactor 50 or the liquid / liquid contactor 50’ and back to the first storage reservoir 14. The circulating pump 52 may be operated to continuously move the first fluid 16’ along this fluidic circuit, and the dosing pump 36 may be operated when it is desirable to generate NO.

[0103] For the device 10C’, the valve 32 along the first fluid line 30 is used to selectively enable the first formulation 16’ to flow from the reservoir 14 along the path to the reaction chamber 74. The dosing pump 36 may be operated when it is desirableto generate NO. Another pump may be positioned in the output fluid line 69 to move the fluid through the remainder of the device 10C’ and to the catheter 10G.

[0104] The dosing pump 36 of the devices 10C, 10C’ is operated as described in reference to Fig. 1.

[0105] The mixing reservoir 22” is operated in the same manner as the mixing reservoir 22’ as described in reference to Fig. 2 for the medical device 10B, 10B’, except that, in the medical device 10C, 10C’, it mixes the first formulation 16’ and the second formulation 20. In this example, the formulations 16’, 20 mixed in the mixing reservoir 22” form a “mixed precursor,” which does not yet include the copper(ll) ligand complex. As such, in the medical device 10C, 10C’, the mixing reservoir 22” is not a reaction chamber.

[0106] Like the mixing reservoir 22’, the mixing reservoir 22” includes a single outlet 26”, through which the mixed precursor is transported through another fluid line / conduit 76. This fluid line / conduit 76 may be any polymeric tubing that can transport fluid. As mentioned herein, the circulating pump 52 of the device 10C or another pump of the device 10C’ may control the movement of the fluid(s) from the mixing reservoir 22” to the reaction chamber 74.

[0107] The reaction chamber 74 includes the copper(ll) source, which in conjunction with the mixed precursor, can generate NO. In this example, the copper source includes a resin and any example of the copper(ll) ligand complex attached to the resin. Within the reaction chamber, the copper(ll) ions of the copper(ll) ligand complex are first reduced by the ascorbate in the mixed precursor, and then the nitrite in the mixed precursor is reduced by the generated copper(l) ions of the ligand complex.

[0108] Examples of the resin include those described in U.S. Patent Pub. No. 2009 / 0118819, which is incorporated herein by reference in its entirety. As specific examples, the resin may include polymers or copolymers formed from vinyl monomers, such as N-vinyl pyrrolidone, propylene, styrene, cinnamyl, vinyl chloride, acrylates such as methacrylate, methyl methacrylate, acrylamide and acrylonitrile. A poly(acrylate) includes at least one residue of an acrylate, e.g., where R may be for example, an alkyl such as methyl, H, a halogen, NH2, or CN. “A” may be for example,any substituent, for example, H or an alkyl such as methyl. It is desirable that the ligand of the copper(ll) ligand complex be covalently attached to the resin, and thus the ligand and resin selected should include functional groups that can covalently bond to one another. In one example, succinimide chemistry may be used for immobilizing the ligand to the resin.

[0109] The reaction chamber 74 may be any suitable container that can hold the resin bound copper(ll) ligand complex, that can receive the mixed precursor, and that is inert to the reduction reactions. Any of the containers set forth herein for the mixing reservoir 22, 22’, 22” may be used. In one example, the reaction chamber 74 is in the form of a cartridge, where the mixed precursor is introduced into contact with the resin bound copper(ll) ligand complex that is housed within the cartridge. In one example, the first formulation 16’ is circulated through the device 10C or moved through the device 10C’ continuously, and the injection of the second formulation 20 controls the NO generation.

[0110] In this example devices 10C, 10C’, the outlet conduit 42 operatively connects the reaction chamber 74 and the sediment separator 48. The reaction mixture generated in the reaction chamber 74 is transported through the outlet conduit 42. As mentioned, the outlet conduit 42 is at least NO impermeable. The reaction mixture includes at least water and the nitric oxide gas dissolved therein.

[0111] In the device 10C, the circulating pump 52 controls the movement of the reaction mixture from the reaction chamber 74 to the sediment separator 48. In the device 10C’, the pump positioned in the output fluid line 69 controls the movement of the reaction mixture from the reaction chamber 74 to the sediment separator 48.

[0112] In the example device 10C, the sediment separator 48 and the liquid / gas contactor 50 or the liquid / liquid contactor 50’ may be configured and used as described in reference to Fig. 2, in order to remove sediment from the reaction mixture and to separate the nitric oxide from the liquid of the reaction mixture. The purge gas and inspiratory gas conduit 43 may be used as described for one example of the medical device 10B to generate the output gas, which is transported to the desired destination through the fluid line or conduit 68. Alternatively, the extracorporeal circuit 39 may beused as described for the medical device 10B to generate treated blood, which is transported back to the patient through the fluid line or conduit 68.

[0113] In the example device 10C’, the sediment separator 48 and output fluid line 69 may be configured and used as described in reference to Fig. 2, in order to remove sediment from the reaction mixture and to deliver the solution phase of the reaction mixture (which contains dissolved NO) to an example of the catheter or cannula 10G.

[0114] The medical devices 10D, 10D’ shown in Fig. 4 are respectively similar to the medical devices 10C, 10C’ shown in Fig. 3, except that i) the reaction chamber 74 is replaced with first and second reaction / regeneration chambers 78A, 78B that are in switchable fluid communication with the mixing reservoir 22”, ii) the ascorbate source is removed from the formulation 16’ and is contained in a reductant reservoir 80 that in switchable fluid communication reaction / regeneration chambers 78A, 78B, and iii) the sediment separator 48 is not included.

[0115] In this example, each of the medical devices 10D, 10D’ includes the first storage reservoir 14 containing still another example of the first formulation 16”; the second storage reservoir 18 containing the second formulation 20; a mixing reservoir 22” in selective fluid communication with each of the first storage reservoir 14 and the second storage reservoir 18; the first and second reaction / regeneration chambers 78A, 78B in switchable fluid communication with the mixing reservoir 22”, each of the first and second reaction / regeneration chambers 78A, 78B containing a copper source, the copper source including the resin and the copper(ll) ligand complex immobilized at a surface of the resin; and the reductant reservoir 80 in switchable fluid communication with the first and second reaction / regeneration chambers 78A, 78B, the reductant chamber 81 including the source of ascorbate. The device 10D also includes the I iquid / gas contactor 50 or the liquid / liquid contactor 50’ in switchable fluid communication with, and downstream of, the first and second reaction / regeneration chambers 74A, 74B, and also in selective fluid communication with the first storage reservoir 14.

[0116] It is to be understood that any examples of the first storage reservoir 14, the second storage reservoir 18, and the second formulation 20 may be used in the medical devices 10D, 10D’.

[0117] In these example devices 10D, 10D’, the first formulation 16” includes the first buffer having a pH ranging from about 5.5 to about 7.5. As is discussed in more detail below, the reducing agent (e.g., ascorbate) and its by-product(s) (e.g., oxalate) are removed prior to nitric oxide generation, and thus this example of the formulations 16” does not include the calcium source.

[0118] As depicted in Fig. 4, the medical device 10D further includes the first fluid line 30 connecting the first storage reservoir 14 and the mixing reservoir 22”; the circulating pump 52 operatively positioned along the first fluid line 30; the second fluid line 34 connecting the second storage reservoir 18 and the mixing reservoir 22”; the dosing pump 36 operatively position in the second fluid line 24; and a third fluid line 54 connecting the liquid / gas contactor 50 or the liquid / liquid contactor 50’ with the first storage reservoir 14. Also as depicted in Fig. 4, the medical device 10D’ further includes the first fluid line 30 connecting the first storage reservoir 14 and the mixing reservoir 22”; the valve 32 operatively positioned along the first fluid line 30; the second fluid line 34 connecting the second storage reservoir 18 and the mixing reservoir 22”; and the dosing pump 36 operatively position along the second fluid line 34.

[0119] Similar to the description of Fig. 1 , fluid lines 30, 34 are fluid conduits respectively connecting the reservoirs 14, 18 to the mixing reservoir 22”, and may be any suitable polymeric tubing. Each of the fluid lines 30, 34 is set up for one-way flow of the respective formulations 16”, 20 into the mixing reservoir 22”.

[0120] Similar to the description of Fig. 2 for the medical device 10B, the circulating pump 52 of the medical device 10D is positioned in the first fluid line 30 and is used to pump fluids throughout the fluidic circuit that connects the first storage reservoir 14 to the mixing reservoir 22” to the reaction / regeneration chambers 78A, 78B, to the liquid / gas contactor 50 or the liquid / liquid contactor 50’ and back to the first storage reservoir 14. The circulating pump 52 may be operated continuously to move the first fluid 16” along this fluidic circuit, and the dosing pump 36 may be operatedwhen it is desirable to generate NO. The dosing pump 36 is operated as described in reference to Fig. 1 .

[0121] For the device 10D’, the valve 32 along the first fluid line 30 is used to selectively enable the first formulation 16” to flow along the path to one of the reaction / regeneration chambers 78A, 78B. The dosing pump 36 may be operated as described in reference to Fig. 1 when it is desirable to generate NO. Another pump may be positioned in the output fluid line 69 to move the fluid through the remainder of the device 10D’ and to the catheter 10G.

[0122] The mixing reservoir 22” of the devices 10D, 10D’ is operated in the same manner as the mixing reservoir 22’ as described in reference to Fig. 2, except that it mixes the first formulation 16” and the second formulation 20. Similar to Fig. 3, the mixture of the first formulation 16” and the second formulation 20 is a mixed precursor. In this example, the formulations 16”, 20 mixed in the mixing reservoir 22” do not yet include ascorbate or the copper(ll) ligand complex, and thus the mixing reservoir 22” is not a reaction chamber.

[0123] Like the mixing reservoir 22’, the mixing reservoir 22” includes a single outlet 26”, through which the mixed precursor is transported through another fluid line / conduit 76. This fluid line / conduit 76 may be any polymeric tubing that can transport fluid.

[0124] Positioned between the mixing reservoir 22” and the reaction / regeneration chambers 78A, 78B is a valve 80Athat can switch the flow of the mixed precursor from the mixing reservoir 22” to either the reaction / regeneration chamber 78A or the reaction / regeneration chamber 78B.

[0125] Another valve 80B is positioned between each of reaction / regeneration chambers 78A, 78B and i) the waste container 46 and ii) the downstream components 50 or 50’ or 69. This valve 80B can block flow from one of the reaction / regeneration chambers 78A or 78B while allowing flow from the other of the reaction chambers 78B or 78A, and can switch the flow from the respective chamber 78A or 78B to either the waste container 46 or the device’s downstream component 50 or 50’ or 69. Thus, when the valve 80B is open for flow from the chamber 78A to either the waste container 46 or the device’s downstream component 50 or 50’ or 69, the valve 80Bsimultaneously blocks flow from the other chamber 78B. Similarly, when the valve 80B is open for flow from the chamber 78B to either the waste container 46 or the device’s downstream component 50 or 50’ or 69, the valve 80B simultaneously blocks flow from the other chamber 78A.

[0126] The reaction / regeneration chambers 78A, 78B each include the resin with the copper(ll) ligand complex bound thereto. Any of the resins and copper(ll) ligand complexes described in reference to Fig. 3 may be used. Thus, the reaction / regeneration chambers 78A, 78B may be any suitable container that can hold the copper source (in this example, the resin with the covalently attached copper(ll) ligand), that can receive the mixed precursor, and that is inert to the nitrite reduction reaction. Any of the containers set forth herein for the mixing reservoir 22, 22’, 22” may be used. In one example, the reaction / regeneration chambers 78A, 78B are respective cartridges, where the mixed precursor is introduced into contact with the copper source.

[0127] In order to render copper(ll) in one of the reaction / regeneration chambers, e.g., 78A, active (i.e. , reduced to copper(l) ions), the valve 80A is operated to open the path between the reductant chamber 81 and the reaction / regeneration chamber 78A. This, in conjunction with gravity or a separate pump 83, enables some of the ascorbate contained in the reductant chamber 81 to transport into the reaction / regeneration chamber 78A, where it reacts to reduce the copper(ll) to copper (I). This process activates some of the copper(ll) in the reaction / regeneration chamber 78A by reducing it to copper(l). It is to be understood that an excess of ascorbate relative to the concentration of copper(ll) that is to be reduced is transported to the reaction / regeneration chamber 78A.

[0128] Once the copper in one of the reaction / regeneration chambers 78A is activated, the reaction / regeneration chambers 78A, 78B can be operated in tandem, where the chamber 78A containing the activated copper (the copper(l) ligand complex) is used for NO generation, while the chamber 78B containing the non-activated copper (the copper(ll) ligand complex) is activated.

[0129] During these processes, the valve 80A is adjusted so that the path between the mixing chamber 22” and the reaction / regeneration chamber 78Acontaining activated copper(l) ions is open, and so that the path between the reductant chamber 81 and the reaction / regeneration chamber 78B containing copper(ll) ions is open. The open path between the between the mixing chamber 22” and the reaction / regeneration chamber 78A (containing the activated copper(l) ligand complex) enables the mixed precursor to flow into the reaction / regeneration chamber 78A. The activated copper(l) ions in the chamber 78A reduce the nitrite in the mixed precursor to NO. The open path between the between the reductant chamber 81 and the reaction / regeneration chamber 78B enables the ascorbate to flow into the reaction / regeneration chamber 78B, where the ascorbate reduces the copper(ll) ions to active copper (I) ions. In this example, the reaction / regeneration chamber 78A is the reaction chamber for the generation of NO, while the reaction / regeneration chamber 78B is the regeneration chamber for the activation of the copper(ll) species to the copper(l) species.

[0130] At the outset of these processes, the valve 80B is operated so that the path between this reaction / regeneration chamber 78A and the waste container 46 is opened. This allows unreacted ascorbate and oxalate generated from the activation of the copper in the chamber 78A to be flushed from the reaction / regeneration chamber 78A. As such, sodium ascorbate could be used instead of calcium ascorbate in this example formulation 16”. Thus, the reactant(s) and by-product(s) are removed prior to nitric oxide generation. After a desirable time period (e.g., 5 seconds or less depending upon the size of the reaction chamber 78A) to flush the ascorbate, the valve 80B is switched to open the path from the reaction / regeneration chamber 78A to the contactor 50 or 50’ of the medical device 10D or to the output fluid conduit 69 of the medical device 10D’. While the valve 80B is in an operable position for fluid to flow from the chamber 78A, it blocks flow from the chamber 78B. This enables the ascorbate to remain in the chamber 78B where copper(ll) reduction is taking place.

[0131] While the reaction / regeneration chamber 78A is in use to generate nitric oxide, the copper source in the reaction / regeneration chamber 78B is regenerated or activated with ascorbate. When the copper(l) ions in the reaction / regeneration chamber 78A are depleted, NO generation ceases. During the reduction of nitrite, the copper(l) ligand complex is oxidized to copper(ll) ligand complex, which can besubsequently reduced in another cycle. At this point, the valve 80A is switched. With the valve 80A switch, the mixed precursor is directed through the reaction / regeneration chamber 78B (now containing the active copper(l) ligand complex), and the ascorbate from the reductant chamber 81 is directed to the reaction / regeneration chamber 78B, where at least some of the copper(ll) is activated with the ascorbate. Given the simultaneous operation of the chambers 78A, 78B for respective NO generation and copper(ll) species activation, the devices 10D, 10D’ can be operated continuously without replacement of the depleted reaction / regeneration chambers 78A, 78B and without adding the reducing agent (ascorbate) to the mixed precursor. Alternatively, after a single use or after a suitable number of uses, the reaction / regeneration chambers 78A, 78B could be replaced.

[0132] In the example device 10D, the outlet conduit 42 is branched and connects both of the reaction / regeneration chambers 78A, 78B to the contactor 50 or 50’. In the example device 10D’, the outlet conduit 42 is branched and connects both of the reaction / regeneration chambers 78A, 78B to the output fluid line 69. In each of these examples, the outlet conduit 42 has the valve 80B operatively positioned thereto so that it that can switch the flow so fluid flows from either the reaction / regeneration chamber 78A or the reaction / regeneration chamber 78B. The positioning of the valve 80B also controls where the fluid from the chambers 78A, 78B is directed, i.e. , to the waste container 46 or to the downstream components 50 or 50’ or 69. The reaction mixture generated in the in-use reaction / regeneration chamber 78A or 78B is a solution phase containing dissolved NO, which can be transported through the outlet conduit 42. As mentioned, the outlet conduit 42 is at least NO impermeable.

[0133] In one example of the device 10D, the I iqu id / gas contactor 50, alone or in combination with the inspiratory gas conduit 43, may be configured and used as described in reference to Fig. 2, in order to generate the desired output gas stream. The output gas may then be transported to the desired destination through the fluid line or conduit 68 as described herein. In another example of the device 10D, the liquid / liquid contactor 50’ may be configured and used as described in reference to Fig. 2, in order to generate NO treated blood, which is transported to the patient as described herein.

[0134] In the example device 10D’, the output fluid line 69 may be configured and used as described in reference to Fig. 2, in order to deliver the solution phase of the reaction mixture (which contains dissolved NO) to an example of the catheter or cannula 10G.

[0135] Any of the example devices 10A, 10B, 10C, 10D shown in Fig. 1 through Fig. 4 may include an inhalation unit (not shown). The inhalation unit may be a ventilator, a face mask, a nasal cannula, or some other suitable apparatus for delivering the output gas stream to the airways of the patient. The inhalation unit is positioned downstream, for example, of the filter 44, 44’, 45, the liquid / gas contactor 50, and / or the inspiratory gas conduit 43 (if included).

[0136] Referring now to Fig. 5, another example of the medical device 10E is depicted. Generally, the medical device 10E includes the first storage reservoir 14 containing another example of the first formulation 16”’; the second storage reservoir 18 containing another example of the second formulation 20’; and the mixing reservoir 22’ in selective fluid communication with each of the first storage reservoir 14 and the second storage reservoir 18. The formulations 16’”, 20’ in this device 10E are aqueous-based creams or ointments, and thus can be used in a topical application.

[0137] In one example, the formulation 16’” includes the ascorbate source, the calcium source, and the copper(ll) ligand complex as described herein, and further includes an aqueous carrier; and the formulation 20’ includes the source of nitrite as described herein, and further includes the aqueous carrier. Buffers may be added to these formulations 16’”, 20’.

[0138] As described herein for the formulation 16, the ascorbate source reduces the copper(ll) of the copper(ll) ligand complex within the formulation 16’”, and thus the copper(l) ligand complex is available for reducing the nitrite in the formulation 20’ as soon as the formulations 16’”, 20’ are mixed. Any of the buffers (if included), ascorbate sources, calcium sources, and copper(ll) ligand complexes disclosed herein may be used in this example of the formulation 16’” in any of the concentrations set forth herein forth the formulation 16. Any of the buffers (if included) and nitrite sources may be used in this example of the formulation 20’ in any of the concentrations set forth herein forth the formulation 20.

[0139] In any example of the formulation 16”’ and 20’, the aqueous carrier may be an aqueous based hydrogel, gel, or lotion that can dissolve the other components of the respective formulation 16’”, 20’. Examples of the aqueous based carrier include collagen, gelatin, hyaluronic acid, alginate, chitosan, xanthan gum, pectin, starch, cellulose, or the like.

[0140] In the example shown in Fig. 5, each of the formulations 16’”, 20’ is an aqueous based hydrogel, gel, or lotion, and the first and second storage reservoirs 14, 18 are tubes of syringes (e.g., the tube portion of a syringe or syringe pump). Prior to use, the formulations 16’”, 20’ may be sealed in the respective tubes. When it is desirable to utilize the mixture, the tubes may be opened, and the plunger portion is inserted into the respective tubes and pushed in the direction that will transport the formulations 16’”, 20’ into the mixing chamber 22’.

[0141] In the mixing chamber 22’, the two formulations 16’”, 20’ are combined and the NO generation reaction is initiated.

[0142] The outlet conduit 42 is in fluid communication with the mixing chamber 22’. The mixed formulation (containing NO dissolved therein) is dispensed from the outlet conduit 42.

[0143] Referring now to Fig. 6 and Fig. 8, two other examples of the medical device 10F, 10H are respectively and schematically depicted. Each example device 10F, 10H is a planar patch.

[0144] Generally, the medical device 10F includes the first storage reservoir 14 containing the first formulation 16; the second storage reservoir 18 containing the second formulation 20; and a mixing reservoir 22’” in selective fluid communication with each of the first storage reservoir 14 and the second storage reservoir 18.

[0145] It is to be understood that any examples of the first formulation 16 or 16’” and the second formulation 20 or 20’ may be used in the medical device 10F. The two formulations 16, 20 or 16’”, 20’ should be miscible when combined.

[0146] When the two formulations 16, 20 are to provide an antimicrobial effect with a burst release of NO, the formulation 16 may include 10 mM calcium ascorbate and 7 mM of the copper(ll) ligand complex and the second formulation may include 10 mM of the nitrite source. These same formulations may also provide sustained NOrelease for wound healing. When the two formulations 16, 20 are to provide sustained NO release for wound healing, the formulation 16 may include 10 mM calcium ascorbate and 7 mM of the copper(ll) ligand complex and the second formulation may include 10 mM of a sustained release nitrite source. Sustained release nitrite sources include a mixture of the inorganic nitrite and an excipient, such as an alkanizing agent, a glidant, a lubricant, a bulking agent, a polymer that comprises cellulose, or polyethylene glycol, or any combination thereof, as described in U.S. Pat. App. Pub. No. 2011 / 0086069.

[0147] In this example, the medical device 10F is a patch, and the first and second storage reservoirs 14, 18 are breakable pouches contained within the mixing reservoir 22”’, which is a patch housing. The breakable reservoirs may be plastic pouches (e.g., biaxially-oriented polyethylene terephthalate, such as commercially available MYLAR®) that are filled with the respective formulations 16, 20 or 16’”, 20’. Pressure can be applied to the mixing reservoir 22’” to break the reservoirs 14, 18 and the release the formulations 16, 20 within the mixing reservoir 22’”.

[0148] The mixing reservoir 22’” has the shape of a relatively flat cube, relatively flat rectangular box, or other relatively flat three-dimensional shape desirable for a wound dressing. The mixing reservoir 22’” does have a length, width, and a depth, however, the depth may be relatively small so that the planar patch is flexible and able to conform to the shape of a desired subject (e.g., a limb, appendage, etc. that the planar patch is affixed, adhered, or otherwise secured to).

[0149] The mixing reservoir 22’” may be formed of two materials 84, 86, one, e.g., 84, of which forms the exterior of the planar patch 10F, and the other, e.g., 86, of which forms the interior of the planar patch 10F. When the planar patch 10F is in use, the interior faces the wound and the exterior faces outward.

[0150] The material 84 forms an inset for the reservoirs 14, 18, which can be positioned on top of one another (as shown in Fig. 6) or side by side within the inset. This material 84, and thus this portion of the mixing reservoir 22’”, may be NO impermeable and flexible. This is desirable so that one major surface S2 of the mixing reservoir 22’” does not allow the NO to escape. This major surface S2 will not be in contact with a wound, and thus NO release is not desirable from this surface S2. Anexample of the material 84 that is suitable for forming the inset of the planar patch is polyurethane.

[0151] In the example shown in Fig. 6, the material 86 is secured to the material 84 so that the material 86 covers the reservoirs 14, 18 and forms the other major surface Si of the mixing reservoir 22”’. At least a portion 82 of the material 86, and thus of the mixing reservoir 22’”, will be in contact with the wound that is to receive NO. As such, the material 86 is selected to be NO permeable, so that it can release the NO that is generated within the mixing reservoir 22’” upon the rupture of the reservoirs 14, 18. The material 86 may be a silicone membrane, polyethylene, polyamide, polytetrafluoroethylene (PTFE), polypropylene, polyvinylidene difluoride, etc. The material 86 may be secured to the material 84 using an adhesive or any other securing mechanism that keeps the reservoirs 14, 18 contained between the two materials 84, 86.

[0152] In the example shown in Fig. 6, the portion of the material 86 that surrounds or is at opposed ends of the portion 82 can be configured for affixing the planar patch to a subject. For example, the material 86, and thus the major surface Si of the mixing reservoir 22’”, may include an adhesive at its perimeter or at opposed ends in order to be secured to the desired subject. When the material 86 includes this adhesive, it is to be understood that the portion 82 remains unobstructed so that NO can be released from the portion 82.

[0153] In another example, the material 84 has a perimeter that is wider than the perimeter of the material 86. In this particular example, the extended perimeter of the material 84 can be configured for affixing the planar patch to a subject. For example, the perimeter of the material 84 that extends beyond the material 86 may include an adhesive for securing the patch to the desired subject.

[0154] Fig. 8 illustrates a top, interior view of another example of the planar patch 10H. In this example, the material 84 forms the backing and includes an exposed perimeter for adhesion to the subject. The interior portion of this patch 10H is an absorptive material 88 that is secured to the material 84 and that is capable of receiving the mixed formulation from, for example, the medical device 10E. In this example, the medical device 10H may be used to dispense the mixed formulation ontothe absorptive material 88, and then the medical device 10E may be secured so that the absorptive material 88, and thus the NO generating mixed formulation, is in contact with a wound. Thus, one example of an NO generating kit may include the medical device 10E shown in Fig. 5 and the planar patch 10H shown in Fig. 8.

[0155] Fig. 7A depicts still other examples of the medical device 10G. In one example, the medical device 10G is a catheter or cannula that is used with the medical device 10B’, 10C’ or 10D’ described herein.

[0156] The output fluid line 69 of any of these devices 10B’, 10C’ or 10D’ can be operatively connected to the catheter or cannula 10G to introduce i) an NO generating lock solution into the lumen 84 of the catheter or cannula (Fig. 7B) or ii) an NO generating formulation into the outer lumen 88 of a multi-lumen catheter or cannula (Fig. 7C).

[0157] In Fig. 7B, the NO generating solution (i.e. , mixed formulation or solution phase containing dissolved NO) formed using the medical device 10B’, 10C’ or 10D’ is a catheter or cannula lock solution. The catheter or cannula lock solution is a static solution that can be injected into the inner lumen 84 of the catheter or cannula 10G during a period of non-use of the catheter or cannula 10G when it is inserted into a blood vessel (e.g., artery or vein) or a urinary tract. The catheter lock solution spontaneously and slowly releases NO over a period of many hours to days, e.g., when a sustained release nitrite is used. The NO can permeate through the walls of the catheter or cannula 10G, and thus can help prevent adhesion and growth of a biofilm on both the walls of the inner lumen 84 and the outer surfaces of the catheter or cannula 10G. As such, the catheter lock solution may be an antimicrobial catheter lock solution.

[0158] In Fig. 7C, the NO generating solution (i.e., mixed formulation or solution phase containing dissolved NO) formed using the medical device 10B’, 10C’ or 10D’ is introduced into the outer lumen 88 in order to introduce NO into a fluid contained in the inner lumen 84 and / or to prevent adhesion and growth of a biofilm on the outer surface of the multi-lumen device. In this example, the catheter or cannula 10G is a jacketed catheter or cannula which includes an inner lumen 84; an outer lumen 88 surrounding the inner lumen 84; and a nitric oxide permeable wall separating the inner lumen 84from the outer lumen 88. In this example, the adapter 92 includes a fluid line connecting the output fluid line 86 of the medical device 10B’, 10C’ or 10D’ with the outer lumen 88 of the extracorporeal circuit. The circulation fluid can be injected into the outer lumen 88 of the jacketed catheter or cannula 10G during use (i.e., when it is inserted into a blood vessel). The introduced NO generating solution spontaneously and slowly releases NO over a period of many hours to days. The NO can permeate through the nitric oxide permeable wall of the jacketed catheter or cannula 10G, and thus can help prevent thrombus formation in the blood contacting inner lumen 84.

[0159] It is to be understood that the adapter 92 of the medical device 10G may be configured differently for the examples shown in Fig. 7B and Fig. 7C so that the NO generating solution can be directed to the desired area, i.e., the inner lumen 84 or the outer lumen 88.

[0160] The examples shown in Fig. 7A and 7B or 7A and 7B can be part of a kit including the catheter or cannula 10G and devices 10B’, 10C’, 10D’ described herein.

[0161] With any of the medical devices set forth herein, the method for generating nitric oxide includes introducing a predetermined dosage of the formulation 20, 20’ (containing the source of nitrite) to the formulation 16, 16’, 16”, 16’” having a pH ranging from about 5.5 to about 7.5 and including a copper(l) ligand complex formed from a copper(ll) ligand complex and an ascorbate source, and (in some instances) a calcium ion source, and mixing the two formulations, thereby forming a mixture and generating the nitric oxide in the mixture. In this method, the formulation 20, 20’, including the source of nitrite, is used as the limiting reactant, as opposed to ascorbate, which the present inventors found led to low responsiveness and lingering NO generation (see the Example section).

[0162] In some example methods, the first and second formulations 16, 20 are liquid, and the method further comprises filtering sediment from the mixture; separating at least some of the nitric oxide from the mixture; and sweeping at least some of the nitric oxide with a purge gas. In some examples, the method further includes recycling the mixture after the nitric oxide is swept therefrom (see 10B, 10C, and 10D in Fig. 2 through Fig. 4).

[0163] Some other examples include introducing the mixture into a lumen 84 of a catheter or cannula 10G (see Fig. 7A and Fig. 7B). As examples, the first and second formulations 16, 20 are liquid, and the method further comprises filtering sediment from the mixture; and introducing the mixture into a catheter or cannula 10G. Still other examples include introducing the mixture into an inner lumen 84 of a catheter an outer lumen 88 of a multi-lumen catheter (see Fig. 7A and Fig. 7C).

[0164] In other examples, the first and second formulations16”’, 20’ are liquids or aqueous based formulations, and the method further comprises applying the mixture to a wound (see Fig. 5).

[0165] In still other examples, the formulations 16, 20 are introduced to one another by breaking pouches respectively containing the first formulation 16 and the second formulation 20 so that the first formulation 16 and the second formulation 20 are released into a common area of a patch; and the method further comprises adhering the patch to a wound (see Fig. 6).

[0166] In yet further examples, the first and second formulations 16, 16’, or 16”, 20 are liquids, and the method further comprises: filtering sediment from the mixture; separating at least some of the nitric oxide from the mixture; mixing the at least some of the nitric oxide with blood from a patient to form treated blood; and introducing the treated blood to the patient (Fig. 2, 3, or 4).

[0167] Still another example method includes introducing a predetermined dosage of a first formulation 20 including a source of nitrite to a second formulation 16’ having a pH ranging from about 5.5 to about 7.5 and including an ascorbate source and (in some instances) a calcium ion source; mixing the first formulation 20 and the second formulation 16’, thereby forming a mixture; and introducing the mixture to a reaction chamber 74 containing a copper source, thereby generating nitric oxide within the mixture, wherein the copper source includes: a resin; and a copper(ll) ligand complex immobilized at a surface of the resin. This method is suitable for use with the medical devices 10C, 10C’ shown in Fig. 3.

[0168] As mentioned, some of the medical devices 10A-10H may be included in nitric oxide generating kit. For example, the medical device 10B’, 10C’, or 10D’ may be combined with the medical device 10G in a kit. For another example, the medicaldevice 10B, 1 OC, or 10D may be combined with an inhalation device in a kit. For still another example, the medical device 10E may be combined with the medical device 10H in a kit.

[0169] One example of a kit includes the first storage reservoir 14 containing a first formulation 16, the first formulation 16 including: a copper(ll) ligand complex; an ascorbate source; and a calcium ion source, wherein a pH of the first formulation ranges from about 5.5 to about 7.5; and a second storage reservoir 18 containing a second formulation 20, the second formulation 20 including: a source of nitrite,. Examples of the kit may further include further include a patch 10H including a reservoir (e.g., absorbing material 88) to receive a mixture of the first and second formulations (i.e. , the mixed formulation). When the source of nitrite is a sustained release nitrite component, the kit may further include a multi-lumen catheter 10G including an outer lumen 88 to receive a mixture of the first and second formulations.

[0170] To further illustrate the present disclosure, various examples are given herein. It is to be understood that these examples are provided for illustrative purposes and are not to be construed as limiting the scope of the present disclosure.EXAMPLES

[0171] Expert men tai Example A

[0172] This Experimental Example outlines several comparative examples and examples according to the aspects set forth herein. In each instance, two formulations were used - one (the first formulation) was initially present in the reaction chamber, and the other (the second formulation) was added to initiate nitric oxide generation. Table 1 depicts the composition of the formulations used in each comparative example and example.TABLE 1

[0173] For each of the examples and comparative examples, the first formulations were prepared by mixing the components at room temperature until they were dissolved. The pH was adjusted with sodium hydroxide pellets. For each of the examples and comparative examples (except Comparative Ex. 11 ), the second formulations included the ascorbate salt or the NaNO2 dissolved in the same buffer as the corresponding first formulation.

[0174] For the examples and comparative examples exposed to NO generation testing, the testing was performed at room temperature. The respective second formulation was either injected with a Hamilton syringe into the corresponding first formulation or was infused into the corresponding first formulation through a septum using a Cole Parmer 74900 syringe pump with a disposable plastic syringe via a stainless-steel needle.

[0175] NO generation was measured using a Sievers 280i ozone chemiluminescent NO analyzer (Boulder, CO) connected to a glass reaction cell. The Sievers 280i instrument was used with a 200 mL / min flow restrictor. Measurements were performed using 5 mL of the respective first formulation, which was stirred through vigorous N2 sparging (~150 mL / min) because, as with other similar copper complexes, the reduced copper ligand complex may be oxygen sensitive. Throughout this Experimental Example, concentrations of NO are reported as molar ratios of parts per million (ppm) or parts per billion (ppb) and nitrite conversion efficiencies are reported as the ratio of the amount of generated NO per amount of nitrite added.

[0176] To test the storage stability (shelf life), the first formulation of some of the examples and comparative examples was stored in an airtight amber glass vial at ambient temperature to protect the ascorbate from decomposition by light. After each measurement of NO generation, 5 mL of each first formulation was returned to the amber vial, and all 15 mL of the first formulation was deoxygenated by purging with N2 for 15 minutes to replicate the oxygen-free environment of the NO generation reaction cell for storage.

[0177] In Comparative Ex. 1 , 1 mL of the second formulation (containing 0.28 M ascorbic acid) was infused, at 1 p.L / min, into the reaction chamber containing 5 mL of the corresponding first formulation. The results of the NO generation for ComparativeEx. 1 are shown in Fig. 9. Although NO generation (>50 ppm) was observed, the NO release showed low responsiveness to ascorbic acid infusion, as it took hours for NO to return to baseline after the infusion was stopped. Additionally, upon addition of the second formulation to the first formulation, there was a distinct color change from blue to green, indicating the oxidation change of the copper ligand complex in the mixed formulation. In some of the aspects disclosed herein, NO generation relies upon both the reduction of the copper(ll) ligand complex by the ascorbate and the reduction of nitrite to NO by the reduced copper(l) ligand complex. In Comparative Ex. 1 (where ascorbate is introduced into the first formulation already containing the nitrite), the reduction of nitrite to NO can be rate-limited by either reduction step. By incorporating the ascorbate into the first formulation and switching the component of the second formulation to nitrite, the addition of nitrite becomes the rate limiting step. This enables the copper(ll) reduction to take place in the first formulation before the nitrite is added thereto.

[0178] Comparative Ex. 2 and Comparative Ex. 3 included first formulations that contained sodium ascorbate. For each of the first formulations with these comparative examples, there was a distinct color change from blue to green with the addition of the ascorbate, indicative of the oxidation state change of the copper(ll) ligand complex in the solutions.

[0179] While the results are not reproduced herein, the storage stability tests indicated that the first formulations of both Comparative Ex. 2 and Comparative Ex. 3 had a short shelf life, which was believed to be due to oxalate accumulation. Comparative Ex. 2 (at pH 7.4) did not exhibit a desirable efficiency in converting nitrite to NO, but did have an improved return to baseline relative to Comparative Ex. 1. To stabilize the ascorbate and reduce oxalate formation, the pH of Comparative Ex. 3 was lowered to 5.8. Comparative Ex. 3 (at pH 5.8) did exhibit an improved response time and conversion rate relative to Comparative Ex. 2.

[0180] Oxalate is a degradation product of ascorbate, which can inhibit NO formation. To counteract the undesired effects of oxalate, calcium ions (in the form of calcium ascorbate at concentrations ranging from about 7 mM to about 140 mM as set forth in Table 1 ) were used to remove oxalate by precipitation as a practically insolublecalcium salt. The precipitate was identified as calcium oxalate based on the asymmetric stretching (va), symmetric stretching (vs) oxalate fundamental vibrations assigned to the bands at 1631 and 1321 cm-1,29respectively on the FT-IR ATR spectrum (results not reproduced herein.) Once precipitated out of solution, the solid calcium oxalate was no longer able to interfere with the NO generation. While the data is not reproduced herein, at higher concentrations of the calcium ions (>1 M), it was found that the calcium ions began to outcompete the copper ions for binding with the ligand, and a loss of NO generation was observed. Thus, in the formulations disclosed herein, less then 1 M of the calcium source should be used.

[0181] To illustrate the interaction of the calcium ions with oxalate, the first formulations for Comparative Ex. 4 and Comparative Ex. 5 were prepared with oxalate and with and without calcium ions, respectively, and were tested for NO generation. As shown in Fig. 10, oxalate present at a high concentration and in the absence of calcium ions (Comparative Ex. 5) prevented any nitrite conversion from the bolus injection of 2 pL of 10 mM sodium nitrite, while the presence of calcium ions enabled nitrite conversion from the bolus injection of 2 pL of 10mM sodium nitrite (Comparative Ex. 4).

[0182] The first formulations for Comparative Ex. 6 and Example 7 were each prepared with the copper(ll) ligand complex and 140 mM calcium ascorbate. As shown in Table 1 , the first formulation for Comparative Ex. 6 had the higher pH buffer and the first formulation for Example 7 had the lower pH buffer. The respective second formulations were added. Following mixing, the NO concentration was measured as described herein, and the nitrite conversion efficiency within 10 minutes of mixing was calculated as described herein. The NO concentration (ppb, Fig. 11 A) and nitrite conversion efficiency (%, Fig. 11 B) for Comparative Ex. 6 and Example 7 illustrate that both pH and the presence of the Ca2+ions have an effect on the responsiveness of NO generation to a single injection of nitrite.

[0183] It is known that during acid induced nitrite reduction, ascorbic acid can prevent N2O3 formation in anaerobic conditions and facilitate NO formation. The first formulation for Comparative Ex. 8 was prepared with calcium ascorbate in the lower pH buffer, but without the copper(ll) ligand complex. The second formulation wasadded. Following mixing, the NO concentration was measured as described herein, and the nitrite conversion efficiency within 10 minutes of mixing was calculated as described herein. As shown in Fig. 11 A and Fig. 11 B, there was not any reasonable amount of NO generation detectable without the copper(ll) ligand complex. These results indicate that any NO generation that occurs at pH 5.8 is indeed copper ion mediated.

[0184] Comparative Ex. 9, the first formulation of which included the low pH buffer without any additional components, was used as a control. The second formulation was added. Following mixing, the NO concentration was measured as described herein, and the nitrite conversion efficiency within 10 minutes of mixing was calculated as described herein. As shown in Fig. 11 A and Fig. 11 B, there was not any reasonable amount of NO generation detectable without the copper(ll) ligand complex and the ascorbate.

[0185] The first formulation of Comparative Ex. 10 was prepared with the copper(ll) ligand complex in the lower pH buffer, but without the calcium ascorbate. The second formulation was added. Following mixing, the NO concentration was measured as described herein, and the nitrite conversion efficiency within 10 minutes of mixing was calculated as described herein. As shown in Fig. 11 A and Fig. 11 B, there was not any reasonable amount of NO generation detectable without the calcium ascorbate.

[0186] Additionally, to ensure that complexation of the copper ion with the ligand was necessary to generate NO, Comparative Ex. 11 was prepared. The first formulation in Comparative Ex. 11 was a solution of CuSO4 and calcium ascorbate (without the ligand). The uncoordinated copper ions were immediately reacted with the ascorbate and reduced to Cu( I), which precipitated out of the solution as insoluble cuprous hydroxide in the form of a yellow solid. These results confirmed the need for the ligand to maintain the solubility and thus the catalytic activity of the copper ions. In spite of the precipitation, the second formulation was added. Following mixing, the NO concentration was measured as described herein, and the nitrite conversion efficiency within 10 minutes of mixing was calculated as described herein. As shown in Fig. 11 A and Fig. 11 B, the yellow precipitate has distinctly lower nitrite-reducing activity togenerate NO compared to Example 7. Thus, these results confirmed that the ligand is needed to maintain the solubility and the rapid and responsive catalytic activity of the copper ions.

[0187] The formulations for Example 7 were used in additional tests to demonstrate their responsiveness.

[0188] For some of the additional testing, the first formulation was prepared in accordance with Example 7 as shown in Table 1 with 140 mM of the calcium ascorbate. The rate of second formulation (nitrite) infusion to the first formulation was varied as shown in Fig. 12A. After a change in the infusion rate, the new NO concentration steady state was reached within a couple of minutes, regardless of whether the infusion rate was being increased or decreased. This rapid response rate would allow for tighter feedback control in obtaining the desired NO concentration levels, even in situations where that required amount is variable. As shown in Fig. 12B, the amount of NO generated was proportional with the flow rate of nitrite infused. As shown in Fig. 12C, a controlled and stable amount of NO was generated over several hours of NaNO2 infusion (7.2 mL of 10 mM NaNO2 at 15 pL / minute). In Fig. 12C, there was an observable increase in the amount of noise over time. The source of the noise was identified as an artifact of the reaction cell geometry and correlated with the increasing liquid level over time (data not shown). Additionally, the slight upward slope that was observed may have been the result of first formulation aging. Fig. 12D depicts the average (dashed line) conversion as a function of the nitrite infusion rate.

[0189] Three additional first formulations were prepared in accordance with Example 7 as shown in Table 1 . These formulations were prepared as shown for Example 7 in Table 1 , with 7 mM, 35 mM, and 140 mM calcium ascorbate, respectively. These first formulations were stored as described herein, and the effects of storage on NO generation were tested at no storage (0 hours), after 24 hours of storage, after 48 hours of storage, after 72 hours of storage, and after 144 hours of storage. After the noted storage periods, some of the first formulations were mixed with the second formulation of Example 7, and the NO concentration was measured. The results are shown in Fig. 13A (for NO generation with the first formulationcontaining 140 mM calcium ascorbate), Fig. 13B (for NO generation with the first formulation containing 7 mM calcium ascorbate), and Fig. 13C (for NO generation with the first formulation containing 35 mM calcium ascorbate). Comparing Fig. 13A, Fig. 13B, and Fig. 13C, the first formulation with 140 mM calcium ascorbate was the most consistent in terms of the level of NO generated over storage time. The 7 mM and 35 mM calcium ascorbate concentrations produced more inconsistent levels of NO as storage time progressed. For the first formulation with 140 mM calcium ascorbate, the plateau of NO generation was reached faster after starting the nitrite infusion and also reached the baseline faster after nitrite was stopped, demonstrating consistent responsiveness. The conversion of nitrite to NO was calculated as described herein, and is shown in Fig. 14. As depicted, the conversion efficiency ranged between 60% to 90%, depending on the age of the first formulation and the Ca2+concentration. Based on the results shown in Fig. 13A through Fig. 13C, it may be desirable to store the first formulation for at least one day (24 hours) and up to 6 days (144 hours) in the methods disclosed herein. The results in Fig. 13B and Fig. 13C indicate that use of the first formulation of Example 7 with 7 mM or 35 mM calcium ascorbate may be desirable between 24 hours and 72 hours of its preparation.

[0190] While the data is not reproduced herein, the addition of calcium ions (100 mL 192 mM CaCl2) to a formulation without calcium ions (e.g., 7 mM MeaTACN, 7 mM copper sulfate (CuSO4), 1 M ascorbate, MES, pH 5.8) after extensive storage (e.g., 9 months) can generate a first formulation that has the ability to generate NO when combined with the second formulation (containing nitrite).

[0191] Overall, Experimental Example A supports the effect of calcium ions with equimolar amounts of the copper(ll) ligand and ascorbate. However, there is still an unaccounted mass balance deficit of nitrite. It was demonstrated that the NO generation is precisely controlled by the nitrite infusion (Fig. 12A) when there is an excess of reducing agent present, without the buildup of nitrite in the mixed formulation or subsequent sluggishness. Therefore, the imperfect conversion ratio cannot be explained by an inadequate amount of copper(ll) ligand present. Nitrite reductase enzymes with copper active site are capable of reducing nitrite a step further to N2O. Therefore, the copper(ll) ligand may also be capable of this further reduction. Theproposed mechanism for this reduction involves the formation of a nitrite-Cu complex, followed by dehydration to form a Cu-NO+complex that can then either produce NO, or react with an additional NO or nitrite molecule to form N2O. Oxalate can form complexes with copper. If oxalate has a higher binding affinity for the copper centers than the nitrosyl but lower than nitrite, this could also explain why there was an increase in nitrite conversion at small concentrations of oxalate. The presence of N2O in the generated gas was confirmed using FTIR (data not shown), that may account, at least in part, for the mass balance deficit of NO formation from nitrite reduction.

[0192] Other reducing agents were tested. Comparative Ex. 12, Comparative Ex. 13, and Example 14 were performed with the formulations shown in Table 1 . The NO generation results are shown in Fig. 15. As depicted, neither glutathione nor cysteine were capable of efficient, responsive NO generation from this system. This may be due to the formation of copper-thiolate clusters, which stabilize copper in the Cu(l) state. These clusters may also result in steric hindrance, preventing nitrite from reacting with the Cu(l).

[0193] Experimental Example B

[0194] In this Experimental Example, B16 fibroblast cells and L929 fibroblast cells were pre-treated with DAF-FM Diacetate, a nitric oxide probe that forms a fluorescent molecule when it reacts with nitric oxide. The respective sets of pretreated cells were then exposed to different treatments to examine nitric oxide generation on the different cell types. Some treatments used comparative example compositions and other treatments used example compositions according to the aspects set forth herein. Table 2 depicts the cell sample ID and the composition of the treatment formulations for each comparative example and example.TABLE 2

[0195] Each of the pre-treated cell samples was exposed to the respective treatment outlined in Table 2. For the treatments with two formulations, the fluorescent microscopy formulations were mixed and deposited on the cells.

[0196] Control Sample 1 through Sample 5 were treated, and exposed to bright field microscopy (BFM) and fluorescent microscopy (FM) after 1 hour of treatment. The top row of Fig. 16 depicts the BFM images, the middle row of Fig. 16 depicts the FM images (reproduced in black and white), and the bottom row of Fig. 16 depicts the converged BFM and FM microscopy images (reproduced in black and white). Sample 5 demonstrated an increased NO level compared to Control sample 1 through Comparative Sample 4, as evidenced by the higher level of fluorescence.

[0197] Control Sample 6 through Sample 14 were treated, and exposed to bright field microscopy after 1 hour of treatment. Fig. 17A through Fig. 171 depict the BFM images of Control Sample 6 through Sample 14, respectively. Sample 14 demonstrated an increased NO level compared to each of Control Sample 6 (Fig. 17A), Comparative Sample 7 (Fig. 17B), Comparative Sample 8 (Fig. 17C), Comparative Sample 9 (Fig. 17D), Comparative Sample 10 (Fig. 17E), Comparative Sample 11 (Fig 17F), Comparative Sample 12 (Fig. 17G), and Comparative Sample 13 (Fig. 171), as evidenced by the higher level of fluorescence depicted in Fig. 171.

[0198] Reference throughout the specification to “one example”, “another example”, “an example”, and so forth, means that a particular element (e.g., feature, structure, and / or characteristic) described in connection with the example is included in at least one example described herein, and may or may not be present in other examples. In addition, it is to be understood that the described elements for any example may be combined in any suitable manner in the various examples unless the context clearly dictates otherwise.

[0199] It is to be understood that the ranges provided herein include the stated range and any value or sub-range within the stated range. For example, a pH ranging from about 5.5 to about 7.5 should be interpreted to include not only the explicitly recited limits of from about 5.5 to about 7.5, but also to include individual pH values (e.g., 5.5, 5.7 Daltons, etc.), and sub-ranges of the pH (from about 5.6 to about 5.8,from about 5.5 to about 6.5, etc.). Furthermore, when “about” is utilized to describe a value, this is meant to encompass minor variations (up to + / - 10%) from the stated value.

[0200] In describing and claiming the examples disclosed herein, the singular forms “a”, “an”, and “the” include plural referents unless the context clearly dictates otherwise.

[0201] While several examples have been described in detail, it is to be understood that the disclosed examples may be modified. Therefore, the foregoing description is to be considered non-limiting.

Claims

What is claimed is:

1. A medical device, comprising: a first storage reservoir containing a first formulation, the first formulation including: a copper(l) ligand complex formed from a copper(ll) ligand complex and an ascorbate source; and a calcium ion source, wherein a pH of the first formulation ranges from about 5.5 to about 7.5; a second storage reservoir containing a second formulation, the second formulation including a source of nitrite; and a mixing reservoir in selective fluid communication with each of the first storage reservoir and the second storage reservoir.

2. The medical device as defined in claim 1 , wherein: each of the first and second formulations is a liquid; and the mixing reservoir is a reaction chamber including: a gas inlet to introduce a purge gas into the reaction chamber; a bubbler in fluid communication with the gas inlet and with the reaction chamber; a gas outlet to remove a gaseous mixture of the purge gas and nitric oxide; and a waste drain.

3. The medical device as defined in claim 2, further comprising: a first fluid line connecting the first storage reservoir and the mixing reservoir; a valve operatively positioned along the first fluid line; a second fluid line connecting the second storage reservoir and the mixing reservoir; and a dosing pump operatively positioned along the second fluid line.

4. The medical device as defined in claim 1 , wherein: each of the first and second formulations is a liquid; and the medical device further comprises: a sediment separator in selective fluid communication with, and downstream of, the mixing reservoir; and a liquid / gas contactor in selective fluid communication with, and downstream of, the sediment separator and also in selective fluid communication with the first storage reservoir, the liquid / gas contactor including: a gas inlet to introduce a purge gas into the liquid / gas contactor; and a gas outlet to remove a gaseous mixture of the purge gas and nitric oxide.

5. The medical device as defined in claim 4, further comprising: a first fluid line connecting the first storage reservoir and the mixing reservoir; a circulating pump operatively positioned along the first fluid line; a second fluid line connecting the second storage reservoir and the mixing reservoir; a dosing pump operatively positioned in the second fluid line; and a third fluid line connecting the liquid / gas contactor with the first storage reservoir.

6. The medical device as defined in claim 1 , wherein: each of the first and second formulations is a liquid; and the medical device further comprises: a sediment separator in selective fluid communication with, and downstream of, the mixing reservoir; and a liquid / liquid contactor in selective fluid communication with, and downstream of, the sediment separator and also in selective fluidcommunication with the first storage reservoir, the liquid / liquid contactor including: a liquid inlet to introduce blood into the liquid / liquid contactor; and a liquid outlet to remove a mixture of the blood and nitric oxide.

7. The medical device as defined in claim 6, further comprising: a first fluid line connecting the first storage reservoir and the mixing reservoir; a circulating pump operatively positioned along the first fluid line; a second fluid line connecting the second storage reservoir and the mixing reservoir; a dosing pump operatively positioned in the second fluid line; and a third fluid line connecting the liquid / liquid contactor with the first storage reservoir.

8. The medical device as defined in claim 1 , wherein: each of the first and second formulations is an aqueous based hydrogel, gel, or lotion; the first and second storage reservoirs are tubes of syringes.

9. The medical device as defined in claim 1 , wherein: the medical device is a patch; and the first and second storage reservoirs are breakable pouches contained within the mixing reservoir.

10. The medical device as defined in claim 1 , further comprising: a filter positioned within the mixing reservoir to remove sediment from a nitric oxide containing solution generated in the mixing reservoir; a liquid outlet defined in the mixing reservoir; an extracorporeal circuit including: an inner lumen; an outer lumen surrounding the inner lumen; anda nitric oxide permeable wall separating the inner lumen from the outer lumen; and a fluid line connecting the liquid outlet with the outer lumen of the extracorporeal circuit.

11. The medical device as defined in one of claims 1 -10, wherein: i) the copper(ll) ligand complex is selected from the group consisting of a Cu(ll)- peptide ligand complex, Cu(ll)-tri(2-pyridylmethyl)amine (CuTPMA), Cu(ll)-tri(2- dimethylamino)ethyl]amine (CuMe6Tren), Cu(ll)-tri(2-pyridylmethyl)phosphine (CuTPMP), Cu(ll)-1 , 4, 7-trimethyl-1 , 4-7 -triazacyclononane (Cu(Me3TACN)), Cu(ll)-1 , 4, 7-triethy 1-1 ,4-7-triazacyclononane (Cu(Et3TACN)), Cu( 11)-1 ,4,7-tripropyl-1 ,4-7- triazacyclononane (Cu(Pr3TACN)), Cu(ll)-1 ,4,7-triisopropyl-1 ,4-7-triazacyclononane (Cu(iPr3TACN)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-ethylate) (Cu(BMPA-Et)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-propanoate) (Cu(BMPA-Pr)), Cu(ll)-(N,N-bis- (2-pyridylmethyl)amine-N-butylate) (Cu(BMPA-Bu)), Cu(ll)-(N,N-bis-(2- pyridylethyl)amine-N-ethylate) (Cu(BEPA-Et)), Cu(ll)- (N,N-bis-(2-pyridylethyl)amine-N- propanoate) (Cu(BEPA-Pr)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-butylate (Cu(BEPA-Bu)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-methyl-phenolate) (Cu(BMPA-MePhO)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-ethyl-phenolate) (Cu(BMPA-EtPhO)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-propyl-phenolate) (Cu(BMPA-PrPhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-methyl-phenolate) (Cu(BEPA-MePhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-ethyl-phenolate) (Cu(BEPA-EtPhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-propyl-phenolate) (Cu(BEPA-PrPhO)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)ethylate (Cu(PEMA-Et)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)propanoate (Cu(PEMA-Pr)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)butylate (Cu(PEMA-Bu)), Cu(ll)-2-(pyridin-2-yl)-N,N-bis(pyridin-2-ylmethyl)ethan-1 -amine (Cu(PMEA)), Cu(ll)-2,2'-(2-(2-(pyridin-2-yl)ethyl)butane-1 ,4-diyl)dipyridine (Cu(PMAP)), and combinations thereof; ii) the ascorbate source is selected from the group consisting of ascorbic acid, potassium ascorbate, magnesium ascorbate, and sodium ascorbate, and the calciumion source is selected from the group consisting of calcium threonate and calcium chloride, or calcium ascorbate is both the ascorbate source and the calcium ion source; and iii) the source of nitrite is an inorganic nitrite salt.

12. The medical device as defined in claim 11 , wherein the copper(ll) ligand complex is the Cu(ll)-peptide ligand complex, and the Cu(ll)-peptide ligand complex includes a peptide ligand selected from the group consisting of GKG, GHG, GAG, GDG, GHK, HGH, GGPG, GPGG, RGFTGM, PFRMY, and DAHK.

13. The medical device as defined in claim 11 , wherein the calcium ion source is the calcium threonate, and wherein the calcium threonate is present at a 1 :1 molar ratio with the ascorbate source.

14. A medical device, comprising: a first storage reservoir containing a first formulation, the first formulation including: an ascorbate source; and a calcium ion source, wherein a pH of the first formulation ranges from about 5.5 to about 7.5; a second storage reservoir containing a second formulation, the second formulation including: a source of nitrite; a mixing reservoir in selective fluid communication with each of the first storage reservoir and the second storage reservoir; and a reaction chamber containing a copper source, the copper source including: a resin; and a copper(ll) ligand complex immobilized at a surface of the resin.

15. The medical device as defined in claim 14, wherein: each of the first and second formulations is a liquid; andthe medical device further comprises: a sediment separator in selective fluid communication with, and downstream of, the reaction chamber; and a liquid / gas contactor in selective fluid communication with, and downstream of, the sediment separator and also in selective fluid communication with the first storage reservoir, the liquid / gas contactor including: a gas inlet to introduce a purge gas into the liquid / gas contactor; and a gas outlet to remove a gaseous mixture of the purge gas and nitric oxide.

16. The medical device as defined in claim 15, further comprising: a first fluid line connecting the first storage reservoir and the mixing reservoir; a circulating pump operatively positioned along the first fluid line; a second fluid line connecting the second storage reservoir and the mixing reservoir; a dosing pump operatively positioned in the second fluid line; and a third fluid line connecting the liquid / gas contactor with the first storage reservoir.

17. The medical device as defined in claim 14, wherein: each of the first and second formulations is a liquid; and the medical device further comprises: a sediment separator in selective fluid communication with, and downstream of, the mixing reservoir; and a liquid / liquid contactor in selective fluid communication with, and downstream of, the sediment separator and also in selective fluid communication with the first storage reservoir, the liquid / liquid contactor including: a liquid inlet to introduce blood into the liquid / liquid contactor; anda liquid outlet to remove a mixture of the blood and nitric oxide.

18. The medical device as defined in claim 17, further comprising: a first fluid line connecting the first storage reservoir and the mixing reservoir; a circulating pump operatively positioned along the first fluid line; a second fluid line connecting the second storage reservoir and the mixing reservoir; a dosing pump operatively positioned in the second fluid line; and a third fluid line connecting the liquid / liquid contactor with the first storage reservoir.

19. The medical device as defined in one of claims 14-18, wherein: i) the copper(ll) ligand complex is selected from the group consisting of a Cu(ll)- peptide ligand complex, Cu(ll)-tri(2-pyridylmethyl)amine (CuTPMA), Cu(ll)-tri(2- dimethylamino)ethyl]amine (CuMe6Tren), Cu(ll)-tri(2-pyridylmethyl)phosphine (CuTPMP), Cu(ll)-1 , 4, 7-trimethyl-1 , 4-7 -triazacyclononane (Cu(Me3TACN)), Cu(ll)- 1 , 4, 7-triethy 1-1 ,4-7-triazacyclononane (Cu(Et3TACN)), Cu( 11)-1 , 4, 7-tripropyl-1 ,4-7- triazacyclononane (Cu(Pr3TACN)), Cu(ll)-1 ,4,7-triisopropyl-1 ,4-7-triazacyclononane (Cu(iPr3TACN)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-ethylate) (Cu(BMPA-Et)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-propanoate) (Cu(BMPA-Pr)), Cu(ll)-(N,N-bis- (2-pyridylmethyl)amine-N-butylate) (Cu(BMPA-Bu)), Cu(ll)-(N,N-bis-(2- pyridylethyl)amine-N-ethylate) (Cu(BEPA-Et)), Cu(ll)- (N,N-bis-(2-pyridylethyl)amine-N- propanoate) (Cu(BEPA-Pr)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-butylate (Cu(BEPA-Bu)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-methyl-phenolate) (Cu(BMPA-MePhO)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-ethyl-phenolate) (Cu(BMPA-EtPhO)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-propyl-phenolate) (Cu(BMPA-PrPhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-methyl-phenolate) (Cu(BEPA-MePhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-ethyl-phenolate) (Cu(BEPA-EtPhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-propyl-phenolate) (Cu(BEPA-PrPhO)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)ethylate (Cu(PEMA-Et)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)propanoate(Cu(PEMA-Pr)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)butylate (Cu(PEMA-Bu)), Cu(ll)-2-(pyridin-2-yl)-N,N-bis(pyridin-2-ylmethyl)ethan-1 -amine (Cu(PMEA)), Cu(ll)-2,2'-(2-(2-(pyridin-2-yl)ethyl)butane-1 ,4-diyl)dipyridine (Cu(PMAP)), and combinations thereof; ii) the ascorbate source is selected from the group consisting of ascorbic acid, potassium ascorbate, magnesium ascorbate, and sodium ascorbate, and the calcium ion source is selected from the group consisting of calcium threonate and calcium chloride, or calcium ascorbate is both the ascorbate source and the calcium ion source; and iii) the source of nitrite is an inorganic nitrite salt.

20. A medical device, comprising: a first storage reservoir containing a first formulation, the first formulation including water and having a pH ranging from about 5.5 to about 7.5; a second storage reservoir containing a second formulation, the second formulation including a source of nitrite; a mixing reservoir in selective fluid communication with each of the first storage reservoir and the second storage reservoir; first and second reaction / regeneration chambers in switchable fluid communication with the mixing reservoir, each of the first and second reaction / regeneration chambers containing a copper source, the copper source including: a resin; and a copper(ll) ligand complex immobilized at a surface of the resin; a reductant reservoir in switchable fluid communication with the first and second reaction / regeneration chambers, the reductant chamber including a source of ascorbate at a pH ranging from about 5.5 to about 7.5.

21. The medical device as defined in claim 20, wherein: each of the first and second formulations is a liquid; andthe medical device further comprises a liquid / gas contactor in switchable fluid communication with, and downstream of, the first and second reaction / regeneration chambers, and also in selective fluid communication with the first storage reservoir, the liquid / gas contactor including: a gas inlet to introduce nitrogen or a purge gas into the liquid / gas contactor; and a gas outlet to remove a gaseous mixture of the purge gas and nitric oxide.

22. The medical device as defined in claim 21 , further comprising: a first fluid line connecting the first storage reservoir and the mixing reservoir; a circulating pump operatively positioned along the first fluid line; a second fluid line connecting the second storage reservoir and the mixing reservoir; a dosing pump operatively positioned in the second fluid line; and a third fluid line connecting the liquid / gas contactor with the first storage reservoir.

23. The medical device as defined in claim 20, wherein: each of the first and second formulations is a liquid; and the medical device further comprises: a sediment separator in selective fluid communication with, and downstream of, the mixing reservoir; and a liquid / liquid contactor in selective fluid communication with, and downstream of, the sediment separator and also in selective fluid communication with the first storage reservoir, the liquid / liquid contactor including: a liquid inlet to introduce blood into the liquid / liquid contactor; and a liquid outlet to remove a mixture of the blood and nitric oxide.

24. The medical device as defined in claim 23, further comprising: a first fluid line connecting the first storage reservoir and the mixing reservoir; a circulating pump operatively positioned along the first fluid line; a second fluid line connecting the second storage reservoir and the mixing reservoir; a dosing pump operatively positioned in the second fluid line; and a third fluid line connecting the liquid / liquid contactor with the first storage reservoir.

25. The medical device as defined in one of claims 20-24, wherein: i) the buffer is selected from the group consisting of phosphate buffered saline, tris(hydroxymethyl)aminomethane, (4-(2-hydroxyethyl)-1 -piperazineethanesulfonic acid), and 2-(N-morpholino)ethanesuifonic acid; ii) the copper(ll) ligand complex is selected from the group consisting of a Cu(ll)-peptide ligand complex, Cu(ll)-tri(2-pyridylmethyl)amine (CuTPMA), Cu(ll)-tri(2- dimethylamino)ethyl]amine (CuMe6Tren), Cu(ll)-tri(2-pyridylmethyl)phosphine (CuTPMP), Cu(ll)-1 , 4, 7-trimethyl-1 , 4-7 -triazacyclononane (Cu(Me3TACN)), Cu(ll)-1 , 4, 7-triethy 1-1 ,4-7-triazacyclononane (Cu(Et3TACN)), Cu( 11)-1 ,4,7-tripropyl-1 ,4-7- triazacyclononane (Cu(Pr3TACN)), Cu(ll)-1 ,4,7-triisopropyl-1 ,4-7-triazacyclononane (Cu(iPr3TACN)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-ethylate) (Cu(BMPA-Et)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-propanoate) (Cu(BMPA-Pr)), Cu(ll)-(N,N-bis- (2-pyridylmethyl)amine-N-butylate) (Cu(BMPA-Bu)), Cu(ll)-(N,N-bis-(2- pyridylethyl)amine-N-ethylate) (Cu(BEPA-Et)), Cu(ll)- (N,N-bis-(2-pyridylethyl)amine-N- propanoate) (Cu(BEPA-Pr)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-butylate (Cu(BEPA-Bu)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-methyl-phenolate) (Cu(BMPA-MePhO)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-ethyl-phenolate) (Cu(BMPA-EtPhO)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-propyl-phenolate) (Cu(BMPA-PrPhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-methyl-phenolate) (Cu(BEPA-MePhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-ethyl-phenolate) (Cu(BEPA-EtPhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-propyl-phenolate)(Cu(BEPA-PrPhO)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)ethylate (Cu(PEMA-Et)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)propanoate (Cu(PEMA-Prj), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)butylate (Cu(PEMA-Bu)), Cu(ll)-2-(pyridin-2-yl)-N,N-bis(pyridin-2-ylmethyl)ethan-1 -amine (Cu(PMEA)), Cu(ll)-2,2'-(2-(2-(pyridin-2-yl)ethyl)butane-1 ,4-diyl)dipyridine (Cu(PMAP)), and combinations thereof; iii) the ascorbate source is selected from the group consisting of ascorbic acid, potassium ascorbate, magnesium ascorbate, and sodium ascorbate, and the calcium ion source is selected from the group consisting of calcium threonate and calcium chloride, or calcium ascorbate is both the ascorbate source and the calcium ion source; and iv) the source of nitrite is an inorganic nitrite salt.

26. A method for generating nitric oxide, comprising: introducing a predetermined dosage of a first formulation including a source of nitrite to a second formulation having a pH ranging from about 5.5 to about 7.5 and including a copper(l) ligand complex formed from a copper(ll) ligand complex and an ascorbate source, and a calcium ion source; and mixing the first formulation and the second formulation, thereby forming a mixture and generating the nitric oxide in the mixture.

27. The method as defined in claim 26, wherein the first and second formulations are liquids, and the method further comprises: filtering sediment from the mixture; separating at least some of the nitric oxide from the mixture; and sweeping the at least some of the nitric oxide with a purge gas.

28. The method as defined in claim 27, further comprising recycling the mixture after the at least some of the nitric oxide is separated therefrom.

29. The method as defined in claim 26 wherein the first and second formulations are liquids, and the method further comprises applying the mixture to a wound.

30. The method as defined in claim 26 wherein: the introducing involves breaking pouches respectively containing the first formulation and the second formulation so that the first formulation and the second formulation are released into a common area of a patch; and the method further comprises adhering the patch to a wound.

31. The method as defined in claim 26, further comprising introducing the mixture into an inner lumen of a catheter or an outer lumen of a multi-lumen catheter.

32. The method as defined in claim 26, wherein the first and second formulations are liquids, and the method further comprises: filtering sediment from the mixture; separating at least some of the nitric oxide from the mixture; mixing the at least some of the nitric oxide with blood from a patient to form treated blood; and introducing the treated blood to the patient.

33. The method as defined in one of claims 26-32, wherein: i) the copper(ll) ligand complex is selected from the group consisting of a Cu(ll)- peptide ligand complex, Cu(ll)-tri(2-pyridylmethyl)amine (CuTPMA), Cu(ll)-tri(2- dimethylamino)ethyl]amine (CuMe6Tren), Cu(ll)-tri(2-pyridylmethyl)phosphine (CuTPMP), Cu(ll)-1 , 4, 7-trimethyl-1 , 4-7 -triazacyclononane (Cu(Me3TACN)), Cu(ll)- 1 , 4, 7-triethy 1-1 ,4-7-triazacyclononane (Cu(Et3TACN)), Cu( 11)-1 ,4,7-tripropyl-1 ,4-7- triazacyclononane (Cu(Pr3TACN)), Cu(ll)-1 ,4,7-triisopropyl-1 ,4-7-triazacyclononane (Cu(iPr3TACN)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-ethylate) (Cu(BMPA-Et)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-propanoate) (Cu(BMPA-Pr)), Cu(ll)-(N,N-bis- (2-pyridylmethyl)amine-N-butylate) (Cu(BMPA-Bu)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-ethylate) (Cu(BEPA-Et)), Cu(ll)- (N,N-bis-(2-pyridylethyl)amine-N- propanoate) (Cu(BEPA-Pr)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-butylate (Cu(BEPA-Bu)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-methyl-phenolate) (Cu(BMPA-MePhO)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-ethyl-phenolate) (Cu(BMPA-EtPhO)), Cu(ll)-(N,N-bis-(2-pyridylmethyl)amine-N-propyl-phenolate) (Cu(BMPA-PrPhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-methyl-phenolate) (Cu(BEPA-MePhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-ethyl-phenolate) (Cu(BEPA-EtPhO)), Cu(ll)-(N,N-bis-(2-pyridylethyl)amine-N-propyl-phenolate) (Cu(BEPA-PrPhO)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)ethylate (Cu(PEMA-Et)), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)propanoate (Cu(PEMA-Prj), Cu(ll)-3-((2-(pyridin-2-yl)ethyl)(pyridin-2-ylmethyl)amino)butylate (Cu(PEMA-Bu)), Cu(l l)-2-(pyridin-2-yl)-N, N-bis(pyridin-2-ylmethyl)ethan-1 -amine (Cu(PMEA)), Cu(ll)-2,2'-(2-(2-(pyridin-2-yl)ethyl)butane-1 ,4-diyl)dipyridine (Cu(PMAP)), and combinations thereof; ii) the ascorbate source is selected from the group consisting of ascorbic acid, potassium ascorbate, magnesium ascorbate, and sodium ascorbate, and the calcium ion source is selected from the group consisting of calcium threonate and calcium chloride, or calcium ascorbate is both the ascorbate source and the calcium ion source; and iii) the source of nitrite is an inorganic nitrite salt.

34. The method as defined in claim 33, wherein the copper(ll) ligand complex is the Cu(ll)-peptide ligand complex, and a peptide ligand of the Cu(ll)-peptide ligand complex is selected from the group consisting of GKG, GHG, GAG, GDG, GHK, HGH, GGPG, GPGG, RGFTGM, PFRMY, and DAHK.

35. The method as defined in claim 33, wherein the calcium ion source is calcium threonate present at a 1 :1 molar ratio with the ascorbate source.

36. A method for generating nitric oxide, comprising: introducing a predetermined dosage of a first formulation including a source of nitrite to a second formulation having a pH ranging from about 5.5 to about 7.5 and including an ascorbate source and a calcium ion source; mixing the first formulation and the second formulation, thereby forming a mixed precursor; and introducing the mixed precursor to a reaction chamber containing a copper source, thereby generating nitric oxide within a solution, wherein the copper source includes: a resin; and a copper(ll) ligand complex immobilized at a surface of the resin.

37. The method as defined in claim 36, further comprising: filtering sediment from the solution; separating at least some of the nitric oxide from the solution; and sweeping the at least some of the nitric oxide with a purge gas.

38. The method as defined in claim 36, further comprising introducing the solution containing the nitric oxide into an inner lumen of a catheter or an outer lumen of a multi-lumen catheter.

39. The method as defined in claim 36, further comprising: filtering sediment from the solution; separating at least some of the nitric oxide from the mixture; mixing the at least some of the nitric oxide with blood from a patient to form treated blood; and introducing the treated blood to the patient.

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