Container and method for nitric oxide sterilization

Abstract

[0003] Apparatus and methods for sterilizing an object are provided in which the object is sealed in a container that includes a nitric oxide-permeable microbial barrier that allows nitric oxide to penetrate into the container to achieve sterilization conditions within the container. In a typical embodiment, the sealed container is placed within a sealable second container, and nitric oxide is supplied to or generated within the second container. From the sealed second container, nitric oxide passes through the nitric oxide-permeable microbial barrier into the container with the object, providing a sterilizing nitric oxide concentration. Advantageously, the contemplated apparatus and methods enable rapid sterilization at low temperatures without posing a hazard to operators or the environment.

Description

Detailed Description of the Invention

[0001] [CROSS-REFERENCE TO RELATED APPLICATIONS]

[0001] This application claims priority to co-pending U.S. Provisional Patent Application No. 63 / 438,073, filed January 10, 2023, which is incorporated herein by reference.

[0002] [Field of the Invention]

[0002] The field of the invention is apparatus and methods for sterilizing objects in an enclosed environment using nitric oxide as a sterilant, and in particular relates to containers having a nitric oxide-permeable microbial barrier that allows nitric oxide to penetrate into the container to achieve sterile conditions within the container.

[0003] [Background of the invention] The background discussion includes information that may be useful in understanding the present invention. This is not an admission that any of the information provided herein is prior art or relevant to the claimed invention, or that any publication specifically or implicitly cited is prior art.

[0004]

[0004] All publications and patent applications herein are incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually indicated to be incorporated by reference. In the event that the definition or use of a term in the incorporated references is inconsistent or contradicts the definition of a term provided herein, the definition of the term provided herein shall apply and the definition of the term in the reference shall not apply.

[0005] Sterilization of medical devices and equipment is essential for their safe use, and the most common sterilization methods include steam autoclaving and irradiation. However, not all medical devices, instruments, and biological products can withstand such harsh conditions, and other sterilization methods must be used. Among other options, sterilization can be achieved using certain gases. For example, ethylene oxide is the most commonly used sterilization gas for items such as surgical kits, catheters, cardiac implants, stents, and IV sets. Unfortunately, ethylene oxide poses several physical and health hazards that merit special attention. According to the U.S. Department of Labor's Occupational Safety and Health Administration, acute exposure to ethylene oxide gas can result in respiratory irritation and lung damage, headache, nausea, vomiting, diarrhea, shortness of breath, and cyanosis. Chronic exposure has been linked to the development of cancer, reproductive toxicity, mutagenic changes, neurotoxicity, and sensitization. In fact, the Environmental Protection Agency has classified ethylene oxide as a carcinogen.

[0006] To avoid some of the difficulties with ethylene oxide, other sterilization gases can be used. For example, formaldehyde can be produced from formalin and has been used as a sterilization gas at relatively high concentrations (e.g., 8–16 mg / L). While significantly less flammable than ethylene oxide, formaldehyde gas is produced and used at operating temperatures of approximately 70–75°C, preventing its use with heat-sensitive materials or equipment. Alternatively, hydrogen peroxide vapor can be used as a sterilization gas. Hydrogen peroxide vapor is typically produced by vacuum evaporation. Among other advantages, hydrogen peroxide vapor typically has a rapid cycle time (e.g., 30–45 minutes), can be effective at low temperatures (e.g., 20°C), and produces environmentally safe by-products (water, oxygen). Furthermore, hydrogen peroxide vapor generally has reasonable material compatibility and ease of operation. However, hydrogen peroxide vapor can be reactive with selected polymers and is not approved by the FDA for sterilization of medical devices. On the other hand, ozone can be used as a sterilizing gas that is relatively effective even at low temperatures. Unfortunately, ozone is highly chemically unstable and can be reactive with biological agents. Furthermore, production of quantities sufficient for sterilization typically requires specialized equipment.

[0007]

[0007] Furthermore, none of the currently known sterilization systems and methods are readily adaptable for safe use in the field or clinical setting, as currently known systems typically require specialized equipment and / or pose health hazards to operators or the environment. Furthermore, currently known sterilization devices typically, in most instances, require electricity to operate.

[0008]

[0008] Thus, while various devices and methods for sterilizing objects are known in the art, all or nearly all of the various devices and methods for sterilizing objects suffer from several drawbacks, particularly when the sterilant is toxic and / or delivered at high temperatures. Thus, there remains a need for improved devices and methods that allow for sterilization in a safe and efficient manner.

[0009] [Summary of the Invention]

[0009] The subject matter of the present invention is directed to an apparatus and method for sterilizing an object in a safe and conceptually simple manner by sealing the object in a container that includes a nitric oxide-permeable microbial barrier that allows nitric oxide to penetrate into the container to achieve sterilization conditions within the container.

[0010] In one aspect of the present subject matter, the inventors contemplate a kit including a first container configured to receive and sealingly enclose an object, and a second container configured to sealingly enclose the first container. It is further generally contemplated that when the first container includes a nitric oxide-permeable microbial barrier and the second container contains nitric oxide at or above a sterile nitric oxide concentration, the nitric oxide-permeable microbial barrier has sufficient nitric oxide permeability to allow for a sterile nitric oxide concentration in the first container.

[0011] In some embodiments, the first container may be sealed via adhesive, heat, snap-fit, clamp, or threaded seal, and / or it is preferred (but not required) that the first container be flexible. If desired, at least a portion of the first container may be transparent or may include a transparent portion. Furthermore, it will be understood that the second container may be configured to sealingly enclose at least one additional first container. While not limiting the subject matter of the present invention, particularly contemplated objects relate to the medical field and thus may include medical devices, surgical or dental tools, various implants, catheters and / or trocars, IV sets, or biologic products.

[0012]

[0012] In further embodiments, the nitric oxide permeable microbial barrier comprises a porous, breathable film; particularly contemplated porous, breathable films include perforated films, composite films of microporous polymers and inorganic fillers, and nonwoven polymer fibrous webs. For example, the first container may be configured as a pouch comprising a nonwoven polymer fibrous web as the nitric oxide permeable microbial barrier on one side and a transparent PET / LDPE polyester film on the other side. Alternatively, or in addition, the nitric oxide permeable microbial barrier may also comprise a nonporous, breathable film. Among other suitable options, contemplated nonporous, breathable films include polymer composites comprising polyurethane polymers, poly(N-isopropylacrylamide) polymers, side-chain crystalline polymers, and paraffin wax.

[0013] Regardless of the type of nitric oxide-permeable microbial barrier, the nitric oxide-permeable microbial barrier has a microbial resistance of at least 0.2×10 -5 cm 2 / sec, or at least 1.0 x 10 -5 cm 2 It is typically preferred to have an apparent diffusion coefficient of nitric oxide of 1000 ppm / sec. It is further contemplated that the sterilizing nitric oxide concentration will be a steady state concentration of 1 to 50 ppb of nitric oxide.

[0014] If desired, the second container may contain a nitric oxide-releasing source, which may or may not be coupled to or form part of the second container. For example, suitable nitric oxide-releasing sources may include S-nitroso-N-acetyl-D-penicillamine (SNAP), nitrite, S-nitrosothiol, S-nitrosocysteine, S-nitrosoglutathione, diazeniumdiolate compounds, arginine, organic nitrite, or a biological source adapted to produce nitric oxide. As will be readily understood, such nitric oxide-releasing sources may be contained in a separate container, linked to a carrier, or covalently attached to a polymer. It should be understood that, depending on the type of nitric oxide-releasing source, the source may release nitric oxide upon irradiation with light.

[0015]

[0015] In a further embodiment, the first and / or second container may also include a carrier containing a chromophore that undergoes a color change in the presence of nitric oxide; exemplary chromophores include 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid), methyl orange, or thymol blue.

[0016]

[0016] Accordingly, in another aspect of the inventive subject matter, the inventors also contemplate a method of sterilizing an object, comprising placing the object in a first container and then sealing the first container, the first container comprising a nitric oxide-permeable microbial barrier. Such a method further comprises placing the sealed first container in a second container and sealing the second container, thereby enclosing the sealed first container within the second container. In a further step, nitric oxide is then delivered to or generated in the second container up to at least a sterilizing nitric oxide concentration. Most typically, the nitric oxide-permeable microbial barrier has sufficient nitric oxide permeability to allow a sterilizing nitric oxide concentration in the first container when the second container contains nitric oxide at or above the sterilizing nitric oxide concentration. Thus, in a further step of a contemplated method, the object is exposed to a sterilizing nitric oxide concentration in the first container for a time sufficient to sterilize the object.

[0017] In some embodiments, the first and / or second containers are sealed using adhesive seals, heat seals, snap-fit ​​seals, clamp seals, or threaded seals. It is further contemplated that the first and / or second containers are flexible. As noted above, at least a portion of the first container may be transparent, and / or the second container may be configured to sealingly enclose at least one additional first container. While not limiting the subject matter of the present invention, particularly contemplated objects relate to the medical field and thus may include medical devices, surgical or dental tools, various implants, catheters and / or trocars, IV sets, or biologic products.

[0018]

[0018] The same considerations provided above regarding the nitric oxide permeable microbial barrier, the sterilizing nitric oxide concentration, the nitric oxide releasing source, and the carrier containing the chromophore apply and will not be repeated here.

[0019]

[0019] Accordingly, the inventor also contemplates a method for non-contact sterilization of an object, comprising the steps of exposing the object to a sterilizing nitric oxide concentration for a time sufficient to sterilize the object, wherein during the exposing step, (a) the object is sealed in a first container comprising a nitric oxide-permeable microbial barrier, and (b) nitric oxide is delivered to the object across the nitric oxide-permeable microbial barrier.

[0020] Most typically, but not necessarily, the sterilizing nitric oxide concentration is a steady-state concentration of nitric oxide between 1 and 50 ppb, the time is between 20 and 240 minutes, and / or the object is exposed to the sterilizing nitric oxide concentration at a temperature between 20 and 50° C. The same considerations provided above regarding the nitric oxide permeable microbial barrier and sterilizing nitric oxide concentration apply and will not be repeated here.

[0021]

[0021] Furthermore, in the most typical embodiment, during the exposing step, the first container is contained within a second sealed container. As will be appreciated, nitric oxide can then be supplied to the second container from a nitric oxide-releasing source or generated in the second container, as described above. If desired, a color change can be detected in a chromophore (typically linked to a carrier and located within the first container) to confirm the presence of nitric oxide. Sterility of an object is achieved using the above device in the above method and can be confirmed or tested in accordance with ASTM E1766-15.

[0022] Various objects, features, aspects, and advantages of the subject matter of the present invention will become more apparent from the following detailed description of preferred embodiments, taken in conjunction with the accompanying drawings in which like numerals represent like elements. [Brief explanation of the drawings]

[0023] [Figure 1] Photographs of a nitric oxide indicator sealed within a polymer film / Tyvek sealed pouch seen through the pouch's transparent film and the nitric oxide indicator outside the sealed pouch prior to exposure to nitric oxide. [Figure 2] 2 is a photograph of the nitric oxide indicator of FIG. 1 sealed within a polymer film / Tyvek sealed pouch and the nitric oxide indicator of FIG. 1 outside the sealed pouch after exposure to nitric oxide.

[0024] [Detailed explanation]

[0025] The inventors have discovered that objects in a container can be sterilized with nitric oxide if the container contains a permeable microbial barrier that allows nitric oxide to penetrate into the container in an amount sufficient to provide a steady-state concentration of nitric oxide effective to sterilize the object.

[0025]

[0026] Advantageously, the systems and methods contemplated herein enable a simple and effective sterilization process in which objects can be sterilized in a non-contact manner and the objects can be maintained in a closed container that prevents contamination of the objects after sterilization. Furthermore, sterilization of objects using such systems and methods with nitric oxide can be carried out at desirably low temperatures (e.g., 20-50°C) and ambient pressures (e.g., about 1 bar) and can be completed within a relatively short period of time (e.g., 30-180 minutes).

[0026]

[0027] To that end, non-contact sterilization of objects is generally contemplated to occur in a closed container containing a nitric oxide-permeable microbial barrier. Most typically, the nitric oxide-permeable microbial barrier has sufficient nitric oxide permeability to allow a sterilizing nitric oxide concentration in the container when the container is subjected to an environment containing nitric oxide at or above the sterilizing nitric oxide concentration. With respect to the sterilizing nitric oxide concentration, it should be understood that the exact concentration will depend on various factors, including the type and amount of microbial contamination present, the surface and geometry of the object to be sterilized, etc. However, it is generally contemplated that the sterilizing nitric oxide concentration will be a steady-state concentration of 1-50 ppb or higher of nitric oxide.

[0027]

[0028] Therefore, to achieve the desirably short time to achieve a sterile nitric oxide concentration in a closed container, the nitric oxide permeable microbial barrier typically has a microbial concentration of at least 0.01 x 10-5 cm 2 / sec, more typically at least 0.05 x 10 -5 cm 2 / sec, or at least 0.75 x 10 -5 cm 2 / sec, or at least 0.1 x 10 -5 cm 2 / sec, or at least 0.25 x 10 -5 cm 2 / sec, or at least 0.40 x 10 -5 cm 2 / sec, or at least 0.50 x 10 -5 cm 2 / sec, or at least 0.60 x 10 -5 cm 2 / sec, or at least 0.70 x 10 -5 cm 2 / sec, or at least 0.8 x 10 -5 cm 2 / sec, or at least 0.9 x 10 -5 cm 2 / sec, or at least 1.0 x 10 -5 cm 2 / sec, or at least 1.2 x 10 -5 cm 2 / sec, or at least 1.4 x 10 -5 cm 2 / sec, or at least 1.6 x 10 -5 cm 2 / sec, or at least 1.8 x 10 -5 cm 2 / sec, or at least 2.0 x 10 -5 cm 2 / sec, or at least 2.25 x 10 -5 cm 2 / sec, or at least 2.5 x 10 -5 cm 2 / sec, or at least 2.75 x 10 -5 cm 2 / sec, or at least 3.0 x 10 -5 cm 2 The apparent diffusion coefficient of nitric oxide is 0.05 x 10 / sec. Therefore, an adequate nitric oxide permeable microbial barrier is typically -5 cm2 / sec~0.5×10 -5 cm 2 / sec, or 0.5 x 10 -5 cm 2 / sec~1.5×10 -5 cm 2 / sec, or 1.0 x 10 -5 cm 2 / sec~3.0×10 -5 cm 2 / sec has an apparent diffusion coefficient of nitric oxide.

[0028]

[0029] As will be readily appreciated, the types of suitable materials can vary considerably, so long as the material has the above-described permeability to nitric oxide and forms a microbial barrier. Regarding permeability to nitric oxide, numerous test methods are known in the art, and an exemplary test method suitable for use herein can be found in ACS Biomater. Sci. Eng. 2016, 2, 1483-1492, which is incorporated herein by reference. Similarly, regarding microbial barrier properties, numerous test methods are known in the art, and an exemplary test method suitable for use herein is ASTM F2638 (Standard Test Method Using Aerosol Filtration to Determine the Performance of Porous Packaging Materials as Surrogate Microbial Barriers).

[0029]

[0030] Therefore, suitable nitric oxide-permeable microbial barriers for use herein include various porous and non-porous breathable films. For example, porous breathable films typically include (laser or microneedle) perforated films, composite films made from one or more fillers in a microporous polymer, and various nonwoven polymer fiber webs. Similarly, when the nitric oxide-permeable microbial barrier is a non-porous breathable film, suitable films include polymer composites containing various polyurethane polymers, poly(N-isopropylacrylamide) polymers, side-chain crystalline polymers, and paraffin wax. Additional polymers and suitable parameters are described in Trends in Food Science & Technology, Vol. 76, 2018, pp. 15-27, which is incorporated herein by reference.

[0030]

[0031] Viewed from a different perspective, suitable nitric oxide-permeable microbial barriers can be made from a wide variety of natural and synthetic polymers, such as silicone rubber, polyurethane, Tyvek, PVC, EVA, polyester, polycarbonate, thermoplastic polyurethane, polylactic acid, polycaprolactone, cellulose, and copolymers, and combinations thereof. Furthermore, these polymers can be microporous, form porous webs, or be permeable to nitric oxide under elevated temperatures or mechanical stress, etc. Furthermore, it is contemplated that any microbial barrier suitable for use in packaging materials for contents sterilized with ethylene oxide is considered suitable for use herein.

[0031]

[0032] As will be readily appreciated, the specific nature of the objects to be sterilized can vary widely. However, it is specifically contemplated that the objects are for use in medicine and thus typically include various medical devices, surgical or dental tools, implants, catheters, IV sets, wound dressings, sutures, staples, and biologics (which may or may not be associated with implantable or injectable materials). Other non-medical uses include the sterilization of cosmetic formulations, containers, food products, and / or articles used in food preparation, as well as common household items (e.g., toothbrushes and toothbrush heads) and articles used in infant care (e.g., bottles, nipples, pacifiers, etc.).

[0032]

[0033] Thus, the configuration of a container containing an object for sterilization can vary considerably, so long as the container includes the nitric oxide-permeable microbial barrier described above, and so long as such a container can be sealed to hold the object sterile without risk of inadvertent contamination of the object. In some embodiments, such as those for household products, the container can be a relatively rigid container with a screw-top that includes a nitric oxide-permeable microbial barrier. On the other hand, if the object is for medical use, the container may be configured as a single-use pouch comprising a nonwoven polymeric fibrous web as a nitric oxide-permeable microbial barrier on one side and a transparent PET / LDPE polyester film on another side (which may be heat-sealed together or may be hermetically sealed closed after the object is placed in the pouch). Accordingly, in view of the above, it should be understood that the nitric oxide-permeable microbial barrier may form part of the structure of the container (e.g., a wall or side of the pouch) or the nitric oxide-permeable microbial barrier may be connected to a frame or carrier or otherwise secured to at least a portion of the container (e.g., glued, sewn, welded, etc.). Furthermore, it should be noted that the nitric oxide-permeable microbial barrier may be permanently connected to the container (e.g., most single-use embodiments) or the nitric oxide-permeable microbial barrier may be removably connected to the container (e.g., most multi-use embodiments). For example, if the nitric oxide-permeable microbial barrier is removably connected to the container, the nitric oxide-permeable microbial barrier may be reused or replaced with a new nitric oxide-permeable microbial barrier.

[0033]

[0034] The appropriate container volume (after placing the object) is 1 to 10 cm 3 , or 10 to 50 cm 3 , or 50 to 500 cm 3 , or 500cm 3 ~5,000cm 3, and larger. Similarly, contemplated containers may be flexible (deformable using manual force and not retain their shape after application of force) or rigid (not deformable using manual force and retain their shape after application of force), or may include a flexible portion. Furthermore, it is also contemplated that in some embodiments, the container may include a visually transparent portion through which the contents of the container are at least partially visible and / or a color change in a chromogen can be observed. Thus, containers contemplated herein are configured to receive a single object or multiple objects for sterilization.

[0034]

[0035] As will be readily appreciated, suitable manners for sealingly closing a container vary widely. For example, after the object is received in the container, the container may be sealed using an adhesive seal, a heat seal, a snap-fit ​​seal, a clamp seal, a threaded seal, or the like. Thus, the seal of the container may be a permanent seal or a reusable seal that may form an integral part of the container or may be external to the container.

[0035]

[0036] It should be understood that nitric oxide can be provided to an object sealed in a container in a variety of ways, depending on the specific use. Most typically, however, the sealed container with the object is placed in a secondary, external container, after which nitric oxide can be delivered from an external nitric oxide source into the secondary, external container, or a nitric oxide source is placed in the secondary container, and nitric oxide is generated or released from the nitric oxide source after the secondary container is sealed. Therefore, it should be noted that the container with the object does not need to contain any source of nitric oxide or a valve structure for receiving nitric oxide. Instead, nitric oxide is delivered to the object from outside the container through a nitric oxide-permeable microbial barrier.

[0036]

[0037] Among other suitable nitric oxide sources, it is generally contemplated that nitric oxide is provided as a gas (typically pure (e.g., at least 90 mol%, or at least 95 mol%, or at least 98 mol% pure)) from a gas storage such as a compressed gas cylinder, or is generated from the decomposition of a precursor chemical that can be decomposed in situ via heat, light irradiation (photolytic cleavage), pH change, electrochemical reaction, or enzymatic methods to produce nitric oxide as a reaction product. Furthermore, to avoid other undesirable chemical interactions with the objects to be sterilized, it is generally preferred that the nitric oxide source produce nitric oxide as the only reactive species.

[0037]

[0038] Thus, suitable nitric oxide sources include nitric oxide-donating polymers and various polymer-based materials using various nitric oxide moieties. In some embodiments, the nitric oxide donor can be covalently attached to or mixed with the polymer. Furthermore, the nitric oxide donor can also be used in solid, liquid, or gel form. Among other options, particularly contemplated nitric oxide sources include S-nitroso-N-acetyl-D-penicillamine (SNAP), nitrite, S-nitrosocysteine, S-nitrosoglutathione, diazeniumdiolate compounds, arginine (through enzymatic action), and various organic nitrites. Non-limiting examples of suitable S-nitroso-N-acetyl-D-penicillamine and other photolabile S-nitrosothiols covalently attached to polymers are described in U.S. Pat. No. 9,884,943 B2 and WO 2020 / 018488 A1, both of which are incorporated herein by reference.

[0038]

[0039] Other examples of nitric oxide sources include vapor-phase delivery from polymers, acidified nitrites, or nitrates; nitric oxide-donating molecules such as diazeniumdiolates, nitrosothiols, nitrosyl compounds, or other methods of NO generation, such as enzymatic generation of nitric oxide, chemical generation of nitric oxide from ascorbic acid or metal catalysis, electrochemical generation of nitric oxide, photolytic cleavage of a bond to release nitric oxide, direct delivery of nitric oxide gas, etc. Further considerations and compositions suitable for use herein are described in WO 2022 / 164894, which is incorporated herein by reference.

[0039]

[0040] Regardless of the specific source of nitic oxide, it is generally contemplated that the source and / or container is configured to generate at least a sterilizing nitric oxide concentration within the container surrounding the object and within an outer container surrounding the container with the object. In most embodiments, the sterilizing nitric oxide concentration is a steady-state concentration of nitric oxide of 1-500 ppb, or 1-10 ppb, or 10-50 ppb, or 50-250 ppb, or 250 ppb-500 ppb, or higher. Accordingly, suitable sterilizing nitric oxide concentrations are 1 ppb, or at least 5 ppb, or at least 10 ppb, or at least 50 ppb, or at least 100 ppb, or at least 200 ppb, and higher.

[0040]

[0041] Therefore, in view of the above, it should be noted that a secondary (external) container can be configured to enclose only a single internal container that encloses an object, or a secondary (external) container can be configured to enclose only multiple internal containers that each enclose one or more objects for sterilization. Thus, the secondary container can be a room that can be closed to maintain sterilizing nitric oxide concentrations, or an enclosure, chest, or other configured container with a door that allows for hermetic closure. In further contemplated embodiments, the secondary container can be a bag or pouch that receives the internal container with the object, and the external container can be sealed or clamped shut. Thus, the ratio of the internal volume of the external container to the single internal container can be at least 1.5:1, or at least 2:1, or at least 3:1, or at least 5:1, or at least 10:1, or at least 50:1, or at least 100:1, and higher.

[0041]

[0042] It will therefore be appreciated that, viewed from different perspectives, the nature and type of sterilization may dictate, at least to some extent, the configuration of the inner and outer containers. For example, if sterilization is performed in a hospital sterilization facility, the outer container may be a room or a relatively large container (e.g., at least 5 m) enclosing a large number of inner containers (e.g., at least 200). 3 On the other hand, if sterilization is performed in a dental or medical office, the outer container may be a moderately sized enclosure (e.g., 2 m) enclosing a moderate number (e.g., 10-50) of inner containers. 3 Additionally, when sterilization is performed in a clinical setting or in the field, the outer container may be a pouch or bag (e.g., 10-500 cm) that encloses a single inner container. 3 Similarly, if sterilization is performed in a home setting for household items, the outer container may be a box (e.g., having an internal volume of 100-5,000 cm) enclosing a small number of inner containers (e.g., 1-5). 3 (having an internal volume of 1000 .mu.m).

[0042]

[0043] In further contemplated embodiments, the outer container may or may not include one or more elements for controlling one or more environmental parameters that affect, at least to some extent, the period of time required to ensure sterility of the object. For example, the outer container may include insulating materials, heating and / or cooling circuits, an energy source for facilitating the release / cleavage of nitric oxide from the precursor material, and / or may be coupled to a system for increasing pressure in the outer container.

[0043]

[0044] As will be readily understood, sterilization of objects in the inner container typically depends, at least in part, on the concentration, temperature, pressure, and exposure time of nitric oxide in the outer and inner containers. However, it is generally contemplated that, using the systems and methods provided herein, sterilization can be carried out at temperatures of 10-20°C, or 15-25°C, or 15-30°C, or 20-40°C, or 15-45°C, or 15-50°C, or 20-55°C. Accordingly, suitable sterilization temperatures are typically at least 10°C, or at least 15°C, or at least 20°C, or at least 25°C, or at least 30°C, or at least 35°C, or at least 40°C, or at least 45°C, or at least 50°C, but preferably less than 60°C, or less than 55°C, or less than 50°C, or less than 45°C, or less than 40°C. Similarly, it is generally preferred that the pressure of sterilization be at ambient pressure, although elevated pressures such as at least 0.1 barg, or at least 0.2 barg, or at least 0.5 barg, or at least 1.0 barg, or at least 2 barg are expressly contemplated herein. Furthermore, it is noted that the time taken to sterilize the objects in the inner container can be at least 10 minutes, or at least 20 minutes, or at least 40 minutes, or at least 60 minutes, or at least 120 minutes, or at least 180 minutes, or at least 300 minutes, but is preferably less than 240 minutes, or less than 210 minutes, or less than 150 minutes, or less than 90 minutes, or less than 30 minutes.

[0044]

[0045] In some embodiments, while sterilization may follow a specific protocol to ensure sterility, it is also contemplated that a sterility indicator may be included in the inner container (and in some cases, the outer container) to indicate sterility. For example, particularly contemplated sterility indicators include dyes reactive with nitric oxide, particularly nitric oxide-reactive chromophores that change color upon exposure to nitric oxide. For example, suitable chromophores include 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), methyl orange (MeORG), thymol blue (ThBlu), and any reasonable combination thereof. Such chromophores may be linked to the carrier, or the carrier may be impregnated with the chromophore. As will be readily understood, the carrier may further include a layer (which may or may not include a low-adhesion adhesive) covering the chromophore to control the diffusion of nitric oxide to the chromophore and / or underlying layer, allowing the carrier to be fixed to the interior portion of the object or inner container. Among other options, particularly suitable sterility indicating devices and compositions are described in WO 2022 / 164905, which is incorporated herein by reference.

[0045]

[0046] It will be understood, of course, that the sterility of an object is achieved using the above-described apparatus in the above-described manner and can be confirmed or tested in accordance with ASTM E1766-15. Thus, viewed from a different perspective, an object sterilized using the systems, apparatus, and methods provided herein will have a reduction in the number of viable microorganisms by at least one order of magnitude, more typically at least two orders of magnitude, or at least three orders of magnitude, or at least four orders of magnitude, or at least five orders of magnitude, or at least six orders of magnitude, after sterilization compared to before sterilization. Advantageously, and as already noted above, sterilization can be carried out at ambient pressure, at moderate to low temperatures (e.g., 20-24°C), and for a relatively short period of time (e.g., 20-60 minutes). Furthermore, once sterilized, the object can be kept in a container, protected from external microbial contamination. [Example]

[0046]

[0047] To validate the concept of non-contact sterilization of objects using nitric oxide, we tested a commercially available packaging container (Oliver Healthcare Packaging, Grand Rapids, MI 49504) configured as a pouch, with one side made from a nonwoven polymeric fibrous web (Tyvek 1073B) as a nitric oxide-permeable microbial barrier that served as a porous, breathable film. The other side was made from a flexible, transparent, transparent PET / LDPE polyester film (TPF-0501A 48 PET / 200 LDPE) that was heat-sealed to the nonwoven polymeric fibrous web to allow visualization of the contents within the container. To test the permeation of nitric oxide across the nonwoven polymeric fibrous web, we placed a paper strip treated with ABTS (2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)), which served as a chromophore that turned green upon exposure to nitric oxide. The test container was then heat sealed to enclose the test strip, and a second test strip served as an external control. Both the sealed container and the external test strip were then sealed within a secondary plastic bag that contained a PDMS polymer strip that contained S-nitroso-N-acetyl-D-penicillamine (SNAP-PDMS) as a source material for nitric oxide release.

[0047]

[0048] FIG. 1 is a photograph showing an indicator strip sealed in a pouch and a second indicator strip outside the pouch before the pouch and second strip were placed in a secondary bag. As can be seen from FIG. 1, both indicator strips were the same color, indicating no exposure to nitric oxide. The SNAP-PDMS was then irradiated with light to generate nitric oxide in the secondary plastic pouch, and the setup was maintained at room temperature (25°C for 16 hours). As can be clearly seen from FIG. 2, both indicator strips turned dark green, demonstrating exposure to nitric oxide. It should be noted in this context that the first indicator strip remained contained within the sealed packaging, and both strips turned color with the same intensity. This experiment clearly demonstrates that objects in the pouches disclosed herein can be sterilized by non-contact sterilization.

[0048]

[0049] In some embodiments, numbers expressing properties such as amounts and concentrations of ingredients, reaction conditions, and the like, used to describe and claim particular embodiments of the present invention are understood to be modified in some instances by the term "about." When referring to a specified measurable value (parameter, amount, time period, etc.), the terms "about" and "approximately" as used herein are meant to encompass the specified value and variations therefrom, for example, variations of + / - 10% or less, or + / - 5% or less, or + / - 1% or less, or + / - 0.1% or less, to the extent that such variations are appropriate for the disclosed embodiments. Accordingly, the value to which the modifier "about" or "approximately" refers is itself specifically disclosed. Recitation of ranges of values ​​herein is intended merely to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise specified herein, each individual value is incorporated herein as if individually recited herein.

[0049]

[0050] All methods described herein can be performed in any suitable order unless otherwise indicated herein or clearly contradicted by the context. Any and all examples provided with respect to specific embodiments herein, or the use of exemplary language (e.g., "etc."), are intended merely to better clarify the invention and do not otherwise pose limitations on the scope of the claimed invention. No language in the specification should be construed as indicating any non-claimed element essential to the practice of the invention.

[0050]

[0051] As used throughout this description and the claims that follow the description, the meanings of "a," "an," and "the" include plural references unless the context clearly dictates otherwise. Also, as used throughout this description, the meaning of "in" includes "in" and "on," unless the context clearly dictates otherwise. Also, unless the context dictates otherwise, the term "coupled to," as used herein, is intended to include both direct coupling (the two elements that are coupled together touch each other) and indirect coupling (at least one additional element is located between the two elements). Thus, the terms "coupled to" and "coupled with" are used interchangeably.

[0051]

[0052] It will be apparent to those skilled in the art that many more modifications besides those already described are possible without departing from the inventive concepts herein. Accordingly, the inventive subject matter is not limited except as by the scope of the appended claims. Moreover, in interpreting both the specification and the claims, all terms should be interpreted in the broadest possible manner consistent with the content. In particular, the terms "comprises" and "comprising" should be interpreted as referring to elements, components, or steps in a non-exclusive manner, indicating that a referenced element, component, or step may be present or may be utilized in conjunction with or combined with other elements, components, or steps not specifically mentioned. When the specification or claims refer to at least one of something selected from the group consisting of A, B, C, and N, the sentence should be interpreted as requiring only one element from the group, and not A+N, or B+N, etc.

Claims

1. a first container configured to receive and sealingly enclose an object; a second container configured to sealingly surround the first container; A kit comprising: the first container comprises a nitric oxide-permeable microbial barrier; the nitric oxide-permeable microbial barrier has sufficient nitric oxide permeability to allow a sterile nitric oxide concentration in the first container when the second container contains nitric oxide at or above a sterile nitric oxide concentration; the nitric oxide permeable microbial barrier is made from a synthetic polymer; kit.

2. 10. The kit of claim 1, wherein the first container is sealable via an adhesive seal, a heat seal, a snap-fit ​​seal, a clamp seal, or a screw seal.

3. The kit of claim 1 , wherein the first container is flexible and / or at least a portion of the first container is transparent.

4. The first and / or second container is 10 cm ３ ~1,000 cm ３ The kit of claim 1 having an internal volume of

5. 10. The kit of claim 1, wherein the second container is configured to sealingly enclose another first container and / or the second container is configured to enclose the first container via an adhesive seal, a heat seal, a snap-fit ​​seal, a clamp seal, or a threaded seal.

6. The kit of claim 1 , wherein the object is a medical device, a surgical or dental tool, an implant, a catheter, an IV set, or a biological product.

7. The kit of any one of claims 1 to 6, wherein the nitric oxide permeable microbial barrier comprises a porous, breathable film.

8. 8. The kit of claim 7, wherein the porous breathable film comprises a perforated film, a composite film of a microporous polymer and an inorganic filler, or a nonwoven polymeric fibrous web.

9. 10. The kit of claim 1, wherein the first container is configured as a pouch comprising a nonwoven polymeric fibrous web on at least one side as the nitric oxide permeable microbial barrier and a transparent PET / LDPE polyester film on another side.

10. The kit of any one of claims 1 to 6, wherein the nitric oxide-permeable microbial barrier comprises a non-porous, breathable film.

11. 11. The kit of claim 10, wherein the non-porous, breathable film comprises a polymer composite comprising a polyurethane polymer, a poly(N-isopropylacrylamide) polymer, a side-chain crystalline polymer, or a paraffin wax.

12. The nitric oxide permeable microbial barrier is at least 0.2 x 10 －５ cm ２ 10. The kit of claim 1, having an apparent diffusion coefficient of nitric oxide of 1000 ppm / sec.

13. The nitric oxide permeable microbial barrier is at least 1.0 x 10 －５ cm ２ 10. The kit of claim 1, having an apparent diffusion coefficient of nitric oxide of 1000 ppm / sec.

14. 2. The kit of claim 1, wherein the sterilizing nitric oxide concentration is a steady state concentration of 1 to 50 ppb nitric oxide.

15. 10. The kit of claim 1, wherein the second container contains a nitric oxide-releasing source.

16. 16. The kit of claim 15, wherein the nitric oxide emitting source is in communication with the second container and / or the nitric oxide emitting source is replaceable upon depletion.

17. 16. The kit of claim 15, wherein the nitric oxide releasing source comprises S-nitroso-N-acetyl-D-penicillamine (SNAP), nitrite, S-nitrosothiol, S-nitrosocysteine, S-nitrosoglutathione, a diazeniumdiolate compound, arginine, an organic nitrite, or a biological source adapted to produce nitric oxide, and optionally, the nitric oxide releasing source is linked to a polymer.

18. 16. The kit of claim 15, wherein the nitric oxide emitting source releases nitric oxide upon irradiation with visible or UV light.

19. 10. The kit of claim 1, wherein the first and / or second container further comprises a carrier containing a chromophore that undergoes a color change in the presence of nitric oxide.

20. 20. The kit of claim 19, wherein the chromophore is 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid), methyl orange, or thymol blue.

21. 1. A method of sterilizing an object, comprising: placing the object in a first container and then sealing the first container, the first container comprising a nitric oxide-permeable microbial barrier; placing the sealed first container within a second container and sealing the second container, thereby enclosing the sealed first container within the second container; providing or generating nitric oxide into said second container to at least a sterile nitric oxide concentration; the nitric oxide-permeable microbial barrier has sufficient nitric oxide permeability to allow a sterilizing nitric oxide concentration in the first container when the second container contains nitric oxide at or above the sterilizing nitric oxide concentration, and the nitric oxide-permeable microbial barrier is made from a synthetic polymer; exposing the object in the first container to the sterilizing nitric oxide concentration for a time sufficient to sterilize the object; A method comprising:

22. 22. The method of claim 21, wherein the first container is sealed using an adhesive seal, a heat seal, a snap fit seal, a clamp seal, or a screw seal.

23. 22. The method of claim 21, wherein the first container is flexible and / or at least a portion of the first container is transparent.

24. The first and / or second container is 10 cm ３ ~1,000 cm ３ 22. The method of claim 21 , having an internal volume of

25. 22. The method of claim 21, wherein the second container is configured to enclose another first container and / or the second container is sealed using an adhesive seal, a heat seal, a snap fit seal, a clamp seal, or a screw seal.

26. 22. The method of claim 21, wherein the object is a medical device, a surgical or dental tool, an implant, a catheter, an IV set, or a biological product.

27. 22. The method of claim 21, wherein the nitric oxide permeable microbial barrier comprises a porous, breathable film.

28. 28. The method of claim 27, wherein the porous breathable film comprises a perforated film, a composite film of a microporous polymer and an inorganic filler, or a nonwoven polymeric fibrous web.

29. 22. The method of claim 21, wherein the first container is configured as a pouch comprising a nonwoven polymeric fibrous web on one side as the nitric oxide permeable microbial barrier and a transparent PET / LDPE polyester film on another side.

30. 22. The method of claim 21, wherein the nitric oxide permeable microbial barrier comprises a non-porous breathable film.

31. 31. The method of claim 30, wherein the non-porous breathable film comprises a polymer composite comprising a polyurethane polymer, a poly(N-isopropylacrylamide) polymer, a side chain crystalline polymer, or a paraffin wax.

32. The nitric oxide permeable microbial barrier is at least 0.2 x 10 －５ cm ２ 22. The method of claim 21, having an apparent diffusion coefficient of nitric oxide of 1 / sec.

33. The nitric oxide permeable microbial barrier is at least 1.0 x 10 －５ cm ２ 22. The method of claim 21, having an apparent diffusion coefficient of nitric oxide of 1 / sec.

34. 22. The method of claim 21, wherein the sterilizing nitric oxide concentration is a steady state concentration of 1 to 50 ppb of nitric oxide, and / or the time sufficient to sterilize the object is from 20 to 240 minutes.

35. 22. The method of claim 21, wherein the second container contains a nitric oxide emitting source.

36. 36. The method of claim 35, wherein the nitric oxide emitting source is in communication with the second container or the nitric oxide emitting source is replaceable upon depletion.

37. 36. The method of claim 35, wherein the nitric oxide releasing source comprises S-nitroso-N-acetyl-D-penicillamine (SNAP), nitrite, S-nitrosothiol, S-nitrosocysteine, S-nitrosoglutathione, a diazeniumdiolate compound, arginine, an organic nitrite, or a biological source adapted to produce nitric oxide, and optionally, the nitric oxide releasing source is linked to a polymer.

38. 36. The method of claim 35, wherein the nitric oxide emitting source releases nitric oxide when irradiated with visible or UV light.

39. 39. The method of any one of claims 21 to 38, wherein the first and / or second container further comprises a carrier containing a chromophore that undergoes a color change in the presence of nitric oxide.

40. 40. The method of claim 39, wherein the chromophore is 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid), methyl orange, or thymol blue.

41. 1. A method for non-contact sterilization of an object, comprising: exposing the object to a sterilizing nitric oxide concentration for a time sufficient to sterilize the object; During the exposing step, (a) the object is sealed in a first container containing a nitric oxide-permeable microbial barrier, the nitric oxide-permeable microbial barrier being made from a synthetic polymer, and (b) nitric oxide is delivered to the object across the nitric oxide-permeable microbial barrier. A method comprising:

42. 42. The method of claim 41, wherein the sterilizing nitric oxide concentration is a steady state concentration of 1 to 50 ppb nitric oxide.

43. 42. The method of claim 41, wherein the time period is from 20 to 240 minutes.

44. 42. The method of claim 41, wherein the object is exposed to the sterilizing nitric oxide concentration at a temperature of 20-50°C.

45. 42. The method of claim 41, wherein the nitric oxide permeable microbial barrier comprises a porous, breathable film.

46. 46. ​​The method of claim 45, wherein the porous breathable film comprises a perforated film, a composite film of a microporous polymer and an inorganic filler, or a nonwoven polymeric fibrous web.

47. 42. The method of claim 41, wherein the first container is configured as a pouch comprising a nonwoven polymeric fibrous web on one side as the nitric oxide permeable microbial barrier and a transparent PET / LDPE polyester film on another side.

48. 42. The method of claim 41, wherein the nitric oxide permeable microbial barrier comprises a non-porous breathable film.

49. 49. The method of claim 48, wherein the non-porous breathable film comprises a polymer composite comprising a polyurethane polymer, a poly(N-isopropylacrylamide) polymer, a side chain crystalline polymer, or a paraffin wax.

50. 42. The method of claim 41, wherein during the exposing step, the first container is contained within a second sealed container.

51. 51. The method of claim 50, further comprising providing or generating nitric oxide in the second container.

52. 52. The method of claim 51, wherein the nitric oxide is generated in the second container from a nitric oxide emitting source, and / or the nitric oxide emitting source is replaceable upon depletion.

53. 42. The method of claim 41, further comprising detecting a color change of a chromophore associated with a carrier and located in the first container, wherein the color change indicates the presence of nitric oxide.

54. 54. The method of claim 53, wherein the chromophore is 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid), methyl orange, or thymol blue.

55. 53. The method of any one of claims 41 to 52, wherein the object is sterile according to ASTM E1766-15.

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