Container and method for nitric oxide sterilization

Abstract

Devices and methods for sterilizing an object are presented in which the object is sealed within a container that includes a nitrogen monoxide permeable microbial barrier that allows nitrogen monoxide to penetrate into the container to achieve sterilization conditions within the container. In a typical embodiment, a sealed container is placed in a sealable second container to which nitric oxide is introduced or in which nitric oxide is generated. From the sealed second container, nitric oxide enters the container containing the object through the nitric oxide permeable microbial barrier to provide the nitric oxide concentration required for sterilization. Advantageously, the contemplated devices and methods allow for rapid sterilization at low temperatures and do not harm operators or the environment.

Description

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims priority to co-pending U.S. Provisional Patent Application No. 63 / 438,073, filed January 10, 2023, and is incorporated herein by reference. Technical Field

[0003] The present invention relates to an apparatus and method for sterilizing objects in a closed environment using nitric oxide as a sterilizing agent, and more particularly to a container having a nitric oxide permeable microbial barrier that allows the nitric oxide to penetrate into the container to achieve sterilization conditions within the container. Background Art

[0004] The Background Description contains information that is helpful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, nor is it an admission that any publication specifically or implicitly referenced is prior art.

[0005] All publications and patent applications cited 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. If a definition or use of a term in a cited document is inconsistent or contrary to the definition of that term provided herein, the definition of that term provided herein controls and the definition of that term in the cited document does not apply.

[0006] Sterilization of medical devices and equipment is crucial for their safe use. The most common sterilization methods include steam autoclaving and irradiation. However, not all medical devices, equipment, and biologics can withstand such harsh conditions, necessitating the use of alternative sterilization methods. Among other options, certain gases can be used for sterilization. For example, ethylene oxide is the most commonly used sterilization gas for items such as surgical kits, catheters, cardiac implants, stents, and IV devices. Unfortunately, ethylene oxide presents several physical and health hazards that require special attention. According to the U.S. Department of Labor's Occupational Safety and Health Administration, acute exposure to ethylene oxide gas can cause respiratory irritation and lung damage, headaches, nausea, vomiting, diarrhea, shortness of breath, and cyanosis. Chronic exposure has been linked to cancer, reproductive effects, mutagenic changes, neurotoxicity, and sensitization. In fact, the U.S. Environmental Protection Agency has classified ethylene oxide as a carcinogen.

[0007] To avoid some of the disadvantages of ethylene oxide, other sterilizing gases can be used. For example, formaldehyde can be produced from formalin and has been used as a sterilizing gas at relatively high concentrations (e.g., 8 mg / L to 16 mg / L). Although its flammability is significantly lower than that of ethylene oxide, formaldehyde gas is produced / used at operating temperatures of approximately 70°C to 75°C, which precludes its suitability for use with heat-sensitive materials or equipment. Alternatively, hydrogen peroxide vapor can be used as a sterilizing gas. Hydrogen peroxide vapor is typically produced by vacuum vaporization. Among other advantages, hydrogen peroxide vapor typically has a fast cycle time (e.g., 30 to 45 minutes), can be used effectively at low temperatures (e.g., 20°C), and produces environmentally safe byproducts (water, oxygen). In addition, hydrogen peroxide vapor generally has good material compatibility and is easy to operate. However, hydrogen peroxide vapor may react with certain polymers and has not yet been approved by the FDA for sterilization of medical devices. On the other hand, ozone can be used as a sterilizing gas and is relatively effective even at low temperatures. Unfortunately, ozone is chemically very unstable and can react with biological products. Furthermore, producing sufficient quantities for sterilization often requires specialized equipment.

[0008] Furthermore, currently known sterilization systems and methods are not readily adaptable for safe field use or point-of-care use, as these systems typically require specialized equipment and / or pose health hazards to operators or the environment. Furthermore, in most cases, currently known sterilization equipment typically requires electricity to operate.

[0009] Thus, although various devices and methods for sterilizing objects are known in the prior art, all or almost all of them have several disadvantages, particularly when the sterilizing agent is toxic and / or delivered at high temperatures. Therefore, there is still a need for improved devices and methods to achieve safe and effective sterilization. Summary of the Invention

[0010] The present subject matter relates to an apparatus and method for sterilizing an object in a safe and conceptually simple manner, wherein the object is sealed within a container comprising a nitric oxide permeable microbial barrier that allows nitric oxide to permeate into the container to achieve sterile conditions within the container.

[0011] In one aspect of the present subject matter, the inventors contemplate a kit comprising a first container configured to receive and sealingly enclose an object, and a second container configured to sealingly enclose the first container. It is also generally contemplated that the first container comprises a nitric oxide permeable microbial barrier, wherein the barrier is sufficiently permeable to nitric oxide to achieve a sterilizing nitric oxide concentration within the first container when nitric oxide in the second container reaches or exceeds a sterilizing nitric oxide concentration.

[0012] In some embodiments, the first container can be sealed by adhesive sealing, heat sealing, snap sealing, clamping sealing or thread sealing, and / or preferably (but not necessarily), the first container is flexible. As required, at least a portion of the first container can be transparent or include a transparent portion. In addition, it should be appreciated that the second container can be configured to seal at least one other first container. Although not limiting the subject matter of the present invention, the object of special envision will relate to the medical field, and therefore may include medical equipment, surgical tools or dental tools, various implants, catheters and / or trocars, IV devices or biological products.

[0013] In a further embodiment, the nitric oxide permeable microbial barrier comprises a porous breathable membrane, and particularly contemplated breathable membranes include perforated membranes, composite membranes of microporous polymers and inorganic fillers, and non-woven polymer fiber webs. For example, the first container can be configured as a bag, one side of which includes a non-woven polymer fiber web as a nitric oxide permeable microbial barrier and the other side includes a transparent PET / LDPE polyester film. Alternatively, or additionally, the nitric oxide permeable microbial barrier may also include a non-porous breathable membrane. Other suitable options envisioned non-porous breathable membranes include polyurethane polymers, poly (N-isopropylacrylamide) polymers, side chain crystalline polymers, and polymer composites containing paraffin wax.

[0014] Regardless of the type of nitric oxide permeable microbial barrier, it is generally preferred that the barrier have a nitric oxide permeability of at least 0.2 × 10 - 5 cm 2 / s, or at least 1.0×10 -5 cm 2 Furthermore, a sterilizing nitric oxide concentration is envisioned to be a steady-state concentration of nitric oxide between 1 ppb and 50 ppb.

[0015] When needed, the second container may contain a nitric oxide release source, which may or may not be connected to the second container or form a part of the second container. For example, a suitable nitric oxide release source can include S-nitroso-N-acetyl-D-penicillamine (SNAP), nitrites, S-nitrosothiols, S-nitrosocysteine, S-nitrosoglutathione, diazepine diol compounds, arginine, organic nitrites (organitrite) or a biological source suitable for generating nitric oxide. It will be understood that such a nitric oxide release source can be contained in a separate container, or can be connected to a carrier, or can be covalently bound to a polymer. Depending on the type of nitric oxide release source, it will be understood that the source can release nitric oxide under light.

[0016] In still further embodiments, the first container and / or the second container may further comprise a carrier comprising a chromophore that changes color in the presence of nitric oxide, and exemplary chromophores include 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid), methyl orange, or thymol blue.

[0017] Therefore, in another aspect of the subject matter of the present invention, the inventors have also envisioned a method for sterilizing an object, the method comprising placing the object in a first container and then sealing the first container, wherein the first container contains a nitric oxide permeable microbial barrier. This method will also include placing the sealed first container in a second container and sealing the second container so as to enclose the sealed first container in the second container. In yet another step, nitric oxide is then introduced into or generated in the second container to at least a sterilizing nitric oxide concentration. Most typically, when the nitric oxide in the second container reaches or exceeds the sterilizing nitric oxide concentration, the permeability of the nitric oxide permeable microbial barrier to nitric oxide is sufficient to achieve a sterilizing nitric oxide concentration in the first container. Therefore, in another step of the envisioned method, the object is exposed to a sterilizing nitric oxide concentration in the first container for a sufficient time to achieve sterilization.

[0018] In some embodiments, the first container and / or the second container are sealed using adhesive seals, heat seals, snap seals, clamp seals or thread seals. In addition, it is envisioned that the first container and / or the second container are flexible. As mentioned above, at least a portion of the first container can be transparent, and / or the second container can be configured to seal at least one other first container. Although not limiting the subject matter of the present invention, the object of special envision will relate to the medical field, and can therefore include medical equipment, surgical tools or dental tools, various implants, catheters and / or trocars, IV devices or biological products.

[0019] With respect to nitric oxide permeability of microbial barriers, sterilizing nitric oxide concentrations, sources of nitric oxide release, and carriers comprising chromophores, the same considerations provided above apply and will not be repeated here.

[0020] Therefore, the inventors have also contemplated a method for contactless sterilization of an object comprising the step of exposing the object to a sterilizing concentration of nitric oxide for a sufficient time to achieve sterilization, wherein during the exposing step, (a) the object is sealed in a first container comprising a nitric oxide permeable microbial barrier, and (b) the nitric oxide is delivered to the object through the nitric oxide permeable microbial barrier.

[0021] Most typically, but not necessarily, the sterilizing nitric oxide concentration is a steady-state concentration of 1 ppb to 50 ppb of nitric oxide for a period of 20 minutes to 240 minutes, and / or the object is exposed to the sterilizing nitric oxide concentration at a temperature of 20° C. to 50° C. The same considerations provided above regarding nitric oxide permeability of microbial barriers and sterilizing nitric oxide concentrations apply and are not repeated here.

[0022] In addition, in the most typical embodiment, the first container is contained in the second sealed container during the exposure step.It will be appreciated that nitric oxide can be passed into or generated in the second container from a nitric oxide release source as described above to the second container subsequently.As required, the color change of the chromophore (usually connected to a carrier and positioned in the first container) can be detected to confirm the existence of nitric oxide.Use the above-mentioned device to realize the sterile state of object by the above method, and can verify or test according to ASTME1766-15.

[0023] Various objects, features, aspects and advantages of the present subject matter will become more apparent from the following detailed description of preferred embodiments and the accompanying drawings in which like numerals represent like components. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] Figure 1 1 and 2 are photographs of the nitric oxide indicator sealed within a polymer film / Tyvek sealed bag as seen through the transparent film of the bag, and the nitric oxide indicator outside the sealed bag before exposure to nitric oxide.

[0025] Figure 2 Sealed in a polymer film / Tyvek bag Figure 1 Nitric oxide indicator and the outside of the sealed bag after exposure to nitric oxide Figure 1 Photo of the indicator. DETAILED DESCRIPTION

[0026] The inventors have discovered that objects within the container can be sterilized with nitric oxide when the container comprises a microbially permeable barrier that allows sufficient nitric oxide to permeate into the container to achieve a steady-state concentration of nitric oxide that is effective in sterilizing the objects.

[0027] Advantageously, the systems and methods contemplated herein allow for a simple and effective sterilization method in which objects can be sterilized in a non-contact manner and in which the objects can be maintained in a closed container to prevent contamination of the objects after sterilization. Furthermore, sterilization of objects with nitric oxide using such systems and methods can be performed under desired low temperature conditions (e.g., 20° C. to 50° C.) and ambient pressure (e.g., approximately 1 bar), and can be completed in a relatively short time (e.g., 30 to 180 minutes).

[0028] For this reason, it is generally envisaged that the object is sterilized contactlessly in a sealed container containing a nitric oxide permeable microbial barrier. Most typically, the permeability of the nitric oxide permeable microbial barrier to nitric oxide is sufficient to allow a sterilizing nitric oxide concentration to be achieved within the container when the container is in an environment containing nitric oxide at a concentration that reaches or exceeds the sterilizing concentration. With respect to the sterilizing nitric oxide concentration, it should be understood that the specific value depends on a variety of 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 believed that the sterilizing nitric oxide concentration is a steady-state concentration of nitric oxide in the range of 1 ppb to 50 ppb or above.

[0029] Therefore, in order to achieve a sterilizing concentration of nitric oxide in a closed container in a relatively short time, the nitric oxide permeable microbial barrier should generally have a concentration of at least 0.01 × 10 -5 cm 2 / s, and more typically at least 0.05×10 -5 cm 2 / s, or at least 0.75×10 -5 cm 2 / s, or at least 0.1×10 -5 cm 2 / s, or at least 0.25×10 -5 cm 2 / s, or at least 0.40×10 -5 cm 2 / s, or at least 0.50×10 -5 cm 2 / s, or at least 0.60×10 -5 cm 2 / s, or at least 0.70×10 -5 cm 2 / s, or at least 0.8×10 -5 cm 2 / s, or at least 0.9×10 -5 cm 2 / s, or at least 1.0×10 -5 cm 2 / s, or at least 1.2×10 -5 cm 2 / s, or at least 1.4×10 - 5 cm 2 / s, or at least 1.6×10 -5 cm 2 / s, or at least 1.8×10 -5 cm 2 / s, or at least 2.0×10 -5 cm 2 / s, or at least 2.25×10-5 cm 2 / s, or at least 2.5×10 -5 cm 2 / s, or at least 2.75×10 -5 cm 2 / s, or at least 3.0×10 -5 cm 2 / s for nitric oxide. Therefore, a suitable nitric oxide permeable microbial barrier typically has an apparent diffusion coefficient of 0.05×10 -5 cm 2 / s to 0.5×10 -5 cm 2 / s, or 0.5×10 -5 cm 2 / s to 1.5×10 -5 cm 2 / s, or 1.0×10 -5 cm 2 / s to 3.0×10 -5 cm 2 / s for the apparent diffusion coefficient of nitric oxide.

[0030] It is easy to understand that as long as the material has the above-mentioned nitric oxide permeability and can form a microbial barrier, the type of suitable material can vary greatly. Regarding nitric oxide permeability testing, there are various known methods in the art, and exemplary methods suitable for this article are shown in ACS Biomater.Sci.Eng.2016, 2, 1483-1492, which are incorporated herein by reference. Similarly, regarding microbial barrier performance testing, there are various known methods in the art, and exemplary methods suitable for this article are ASTM F2638 (Standard Test Method for Measuring Porous Packaging Materials as Alternative Microbial Barrier Performance Using Aerosol Filtration).

[0031] Therefore, suitable nitric oxide permeable microbial barrier for this article include various porous breathable membranes and non-porous breathable membranes. For example, porous breathable membranes generally include (laser or microneedle) perforated membranes, composite membranes made of one or more fillers in microporous polymers, and various non-woven polymer fiber webs. Similarly, when the nitric oxide permeable microbial barrier is a non-porous breathable membrane, suitable membranes include various polyurethane polymers, poly (N-isopropylacrylamide) polymers, side chain crystalline polymers and paraffin-containing polymer composites. Additional polymers and suitable parameters are described in Trends in Food Science & Technology, Volume 76, 2018, Pages 15-27, which are incorporated herein by reference.

[0032] From a different perspective, suitable nitric oxide permeable microbial barriers can be made from a 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. These polymers can be microporous, form porous networks, or be permeable to nitric oxide at elevated temperatures or mechanical stress, etc. In addition, all microbial barriers suitable for use herein are contemplated that are suitable for packaging materials for contents sterilized with ethylene oxide.

[0033] It will be readily appreciated that the particular nature of the objects to be sterilized may vary widely. However, it is particularly envisaged that the objects are used in the medical field and will typically include, for example, various medical instruments, surgical or dental instruments, implants, catheters, IV devices, wound dressings, sutures, staples, and biological products (which may or may not be connected to implantable or injectable materials). Other non-medical uses include the sterilization of cosmetic formulations, containers, food items, and / or items used to prepare food, as well as common household items (such as toothbrushes and toothbrush heads) and baby care items (such as feeding bottles, nipples, pacifiers, etc.).

[0034] Therefore, as long as container comprises above-mentioned nitric oxide permeable microbial barrier and as long as this container can be sealed to keep sterile object, and there is not the risk that object is accidentally contaminated, then the configuration of the container comprising sterilized article can change significantly.In some embodiments, for example, for household products, container can be the relatively rigid container comprising nitric oxide permeable microbial barrier with screw cap.On the other hand, when object is for medical treatment, container can be configured as disposable bag, and it will include on one side as non-woven polymer fiber web as microbial barrier, and on the transparent PET / LDPE polyester film (it can be heat-sealed together and can be sealed closed after object is put into bag) of opposite side.In view of the above, therefore it should be understood that nitric oxide permeable microbial barrier can form a part (such as the wall or one side of bag) of container structure, or nitric oxide permeable microbial barrier can be connected to frame or carrier or otherwise fixed (such as gluing, sewing, welding etc.) to at least the part of container.In addition, it should be noted that nitric oxide permeable microbial barrier can be permanently connected (such as in most disposable embodiments) or detachably connected (such as in most reusable embodiments) with container. For example, where the nitric oxide permeable microbial barrier is detachably connected to the container, the nitric oxide permeable microbial barrier may be reused or replaced with a new nitric oxide permeable microbial barrier.

[0035] The volume of the appropriate container (after placing the item) will be 1cm 3 Up to 10cm 3, or 10cm 3 Up to 50cm 3 , or 50cm 3 Up to 500cm 3 , or 500cm 3 Up to 5000cm 3 , and even greater than 5000cm 3 Likewise, contemplated containers may be flexible (deformable using manual force, which will not retain its shape after the force is applied) or rigid (not deformable using manual force, which will retain its shape after the force is applied), or include flexible portions. Furthermore, it is contemplated that in some embodiments, the container will include a visually transparent portion through which the contents of the container are at least partially visible and / or through which a color change of the chromogen can be observed. Thus, the containers contemplated herein will be configured to receive a single object or multiple objects for sterilization.

[0036] It will be readily appreciated that suitable means of sealingly enclosing a container will vary widely, and for example, after the object is received in the container, the container may be sealed using an adhesive seal, heat seal, snap seal, clamp seal, or thread seal. The seal may be permanent or reusable, may form an integral part of the container, or may be external to the container.

[0037] It will be appreciated that nitric oxide can be provided to the object within the sealed container in a variety of ways, depending on the specific application. However, most typically, the sealed container containing the object is placed in a second external container, and nitric oxide can then be delivered to the second external container from an external nitric oxide source, or the nitric oxide source can be placed inside the second container and, after the second container is sealed, generated or released from the second external container. Therefore, it should be noted that the container containing the object need not include any nitric oxide source or valve structure for receiving nitric oxide; instead, nitric oxide is delivered to the object from outside the container through the nitric oxide permeable microbial barrier.

[0038] Among other suitable nitric oxide sources, it is generally contemplated that the nitric oxide is provided in the form of a (generally pure, (e.g., at least 90 mole % or at least 95 mole % or at least 98 mole %)) gas from a gas storage device such as a compressed gas cylinder, or is generated in situ by the decomposition of a precursor chemical substance, which can be decomposed by heat, by irradiation with light (photolytic cleavage), by a change in pH, by an electrochemical reaction, or by an enzymatic process to produce nitric oxide as a reaction product. Furthermore, it is generally preferred that the nitric oxide source produces only nitric oxide as the active species to avoid other undesirable chemical reactions with the object to be sterilized.

[0039] Therefore, suitable nitric oxide sources include nitric oxide donor polymers using different nitric oxide moieties and different polymer base materials. In some embodiments, the nitric oxide donor can be covalently attached to a polymer or mixed into a polymer. In addition, the nitric oxide donor can also be used in solid, liquid or gel form. In other selections, particularly envisioned nitric oxide sources include S-nitroso-N-acetyl-D-penicillamine (SNAP), nitrites, S-nitrosocysteine, S-nitrosoglutathione, diazepine diol compounds, arginine (by enzyme action) and various organic nitrites. Non-limiting examples of S-nitroso-N-acetyl-D-penicillamine and other photosensitive S-nitrosothiol covalently attached polymers suitable for the present invention are described in U.S. Patent No. 9884943B2 and International Publication No. WO 2020 / 018488 A1, both of which are incorporated herein by reference.

[0040] Other examples of nitric oxide sources include gas-phase delivery from polymers, acidified nitrites or nitrates, nitric oxide donor molecules such as diazenediolates, nitrosothiols, nitrosyl compounds, or other NO generation methods such as enzymatic production of nitric oxide, ascorbic acid or metal-catalyzed chemical production of nitric oxide, electrochemical production of nitric oxide, photolytic cleavage to release nitric oxide, direct delivery of nitric oxide gas, etc. Other applicable considerations and compositions are described in WO 2022 / 164894, which is incorporated herein by reference.

[0041] No matter the specific source of nitric oxide, it is generally envisioned that the configuration of this source and / or container should be able to produce at least sterilizing nitric oxide concentration in the inner container of encapsulating object and the outer container of encapsulating this inner container.In most embodiments, sterilizing nitric oxide concentration will be 1ppb to 500ppb or 1ppb to 10ppb or 10ppb to 50ppb or 50ppb to 250ppb or 250ppb to 500ppb, or even higher than the steady-state concentration of nitric oxide of 500ppb.Therefore, suitable sterilizing nitric oxide concentration will be 1ppb or at least 5ppb or at least 10ppb or at least 50ppb or at least 100ppb or at least 200ppb and even higher than 200ppb.

[0042] In view of the above, it should be noted that the second (outer) container can be configured, for example, as an inner container that encloses only a single enclosed object, or the second (outer) container can be configured, for example, as an inner container that encloses only a plurality of inner containers that each enclose one or more objects to be sterilized. Thus, the second container can be a room that can be closed to maintain the sterilizing nitric oxide concentration, or a container that is an enclosure, box, or other configuration with a sealed door. In further contemplated aspects, the second container can also be a bag or bag that receives the inner container containing the object, wherein the outer container can be sealed or closed with a clamp. Thus, the internal volume ratio of the outer container to the single inner 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 even greater than 100:1.

[0043] From a different perspective, it should be understood that the nature and type of sterilization will determine, at least to some extent, the configuration of the inner and outer containers. For example, when sterilizing in a hospital sterilization facility, the outer container may be a room or a larger container (e.g., having at least 5 m 3 On the other hand, when sterilizing in a dental or medical office, the outer container can be a medium-sized shell (e.g., having an inner volume of less than 2m). 3 In addition, when sterilizing at the point of care or in the field, the outer container can be a bag or package (e.g., an inner volume of 10 cm) that encloses a moderate number (e.g., 10 to 50) of inner containers. 3 Up to 500cm 3 Similarly, when sterilizing household items in a home environment, the outer container may be a box (e.g., with a 100 cm 3 Up to 5000cm 3 internal volume).

[0044] In further aspects, the outer container may or may not include one or more elements to control one or more environmental parameters that will at least partially affect the duration required to ensure sterility of the object. For example, the outer container may include insulation, heating and / or cooling circuits, an energy source to promote the release / cleavage of nitric oxide from the precursor material, and / or be connected to a system for increasing the pressure in the outer container.

[0045] It is readily understood that the sterilization of objects in the inner container typically depends, at least in part, on the nitric oxide concentration, temperature, pressure, and exposure time in the inner and outer containers. However, it is generally contemplated that sterilization using the systems and methods proposed herein can be carried out at temperatures ranging from 10°C to 20°C, or from 15°C to 25°C, or from 15°C to 30°C, or from 20°C to 40°C, or from 15°C to 45°C, or from 15°C to 50°C, or from 20°C to 55°C. Thus, suitable sterilization temperatures will typically be 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 sterilization pressure will be ambient pressure, but elevated pressures, such as at least 0.1 bar, or at least 0.2 bar, or at least 0.5 bar, or at least 1.0 bar, or at least 2 bar, are also expressly contemplated herein. Furthermore, it should be noted that the time required to sterilize the objects in the inner container may 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.

[0046] Although a specific sterilization protocol can be used to confirm sterility in certain embodiments, it is also conceivable to add an indicator indicating sterility in an internal (and sometimes external) container. For example, a particularly contemplated sterility indicator includes a dye reacting with nitric oxide, especially a chromophore that is exposed to nitric oxide and changes color, such as a suitable chromophore including 2,2'-azinobis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), methyl orange (MeORG), thymol blue (ThBlu) and any reasonable combination thereof. This chromophore can be connected to a carrier, or the carrier can be impregnated with the chromophore. It is readily understood that the carrier can also include a layer covering the chromophore to control the diffusion of nitric oxide to the chromophore, and / or allow the carrier to be attached to the base layer (which may or may not include a low-viscosity adhesive) inside the object or inner container. In other selections, particularly suitable sterility indicator devices and compositions are described in WO 2022 / 164905, which is incorporated herein by reference.

[0047] Of course, it should be recognized that the object sterility achieved using the above-mentioned device and method can be verified or tested according to ASTM E1766-15. Therefore, from a different perspective, compared with before sterilization, the number of viable microorganisms of the object sterilized using the system, device and method proposed herein will be reduced by at least one order of magnitude and 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. Advantageously, and as mentioned above, sterilization can be carried out in ambient pressure, low temperature (such as 20 ℃ to 24 ℃) and a relatively short time (such as 20 minutes to 60 minutes). In addition, once sterilized, articles can remain in the container and will be protected from external microbial contamination.

[0048] Example

[0049] To validate the concept of contactless sterilization of objects with nitric oxide, the inventors tested a commercially available packaging container (Oliver Healthcare Packaging, Grand Rapids, MI 49504). The container was configured as a bag, with one side made of a nonwoven polymer mesh (Tyvek 1073B) serving as a porous, breathable membrane and acting as a barrier to nitric oxide-permeable microorganisms. The other side was made of a flexible, transparent PET / LDPE polyester film (TPF-0501A48PET / 200LDPE) heat-sealed to the nonwoven polymer mesh, allowing visualization of the container contents. To test the permeability of nitric oxide through the nonwoven polymer mesh, the inventors placed a test strip treated with ABTS (2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid)) as a chromogen, which turns green upon contact with nitric oxide. The test container was then heat-sealed to enclose the test strip, and a second test strip was provided as an external control. The sealed container and the outer test paper were then enclosed in a second plastic package containing a PDMS polymer tape containing S-nitroso-N-acetyl-D-penicillamine (SNAP-PDMS) as a nitric oxide release source.

[0050] Figure 1 This is a photo of the indicator test paper sealed inside the bag and the second test paper outside the bag before being placed into the second package. Figure 1 As can be seen, the two test strips have the same color, indicating no exposure to nitric oxide. SNAP-PDMS was then irradiated with light to generate nitric oxide within the secondary plastic bag, and the device was kept at room temperature (25°C for 16 hours). Figure 2As can be clearly seen in the image above, both test strips display a dark green color, indicating exposure to nitric oxide. In this case, it should be noted that the first test strip remains sealed within the packaging bag, and the color intensity of both test strips is identical. This experiment clearly demonstrates that objects contained within the bag described herein can be sterilized using a non-contact method.

[0051] In certain embodiments, the numerical values ​​used to describe and claim characteristic parameters such as the quantity and concentration of ingredients and reaction conditions of specific embodiments of the present invention should be understood to be modified by the word "about" in some cases. As used herein, the terms "about" and "approximately" when referring to specific measurable values ​​(such as parameters, quantities, time lengths, etc.) are intended to cover the specified value and its deviation range of ±10% or less, or ±5% or less, or ±1% or less, or ±0.1% or less, as long as the deviation is applicable to the disclosed embodiment. Therefore, the numerical values ​​modified by "about" or "approximately" themselves also constitute explicit disclosure. The numerical ranges listed herein are merely a shorthand way of referring individually to each discrete value within the range. Unless otherwise indicated, each discrete value should be considered to be included in the specification as if it were listed separately herein.

[0052] All methods described herein can be performed in any suitable order unless otherwise indicated or clearly contradicted by context. The use of any and all examples or exemplary language (e.g., "for example") provided with respect to specific embodiments is intended solely to better illuminate the invention and does not limit the scope of the invention as defined by the appended claims. No language in this specification should be construed as indicating any non-claimed element is essential to the practice of the invention.

[0053] As used in this specification and the claims that follow, the meaning of a noun without a quantifier includes plural reference unless the context clearly dictates otherwise. Similarly, as used in this specification, the meaning of "in" includes "in" and "on" unless the context clearly dictates otherwise. Furthermore, unless the context clearly dictates otherwise, the term "connected to" is intended to encompass both direct connections (where two interconnected elements are in contact with each other) and indirect connections (where at least one additional element is located between the two elements). Thus, "connected to" is synonymous with "connected with."

[0054] It will be apparent to those skilled in the art that, in addition to what has been described, further modifications may be made without departing from the present invention. Therefore, the subject matter of the present invention shall not be limited except as defined by the appended claims. In addition, when interpreting the specification and claims, all terms should be interpreted in the broadest manner consistent with the context. In particular, "comprising" and "including" should be interpreted as referring to elements, components or steps in a non-exclusive manner, indicating that the referenced elements, components or steps may exist, be used, or be combined with other elements, components or steps that are not explicitly referenced. When the specification or claims mention at least one selected from the group consisting of A, B, C... and N, the expression should be interpreted as requiring only one element in the group, rather than A plus N or B plus N, etc.

Claims

1. A kit comprising: a first container configured to receive and sealingly enclose an object; a second container configured to sealingly enclose the first container; wherein the first container comprises a nitric oxide permeable microbial barrier; and The nitric oxide permeable microbial barrier has a permeability to nitric oxide sufficient to allow a sterile nitric oxide concentration to be achieved in the first container when the second container contains nitric oxide at or above a sterile nitric oxide concentration.

2. The kit of claim 1, wherein the first container is sealable by adhesive sealing, heat sealing, snap sealing, clamping sealing, or thread sealing.

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

4. The kit according to claim 1, wherein the first container and / or the second container has a 10 cm 3 Up to 1000cm 3 internal volume.

5. The kit of claim 1 , wherein the second container is configured to sealingly enclose another first container, and / or wherein the second container is configured to enclose the first container by adhesive sealing, heat sealing, snap sealing, clamping sealing, or thread sealing.

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 biologic.

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

8. The kit according to claim 7, wherein the porous breathable membrane comprises a perforated membrane, a composite membrane of a microporous polymer and an inorganic filler, or a nonwoven polymer fiber web.

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

10. The kit of any one of claims 1 to 6, wherein the nitric oxide permeable microbial barrier comprises a non-porous, gas-permeable membrane. 11 . The kit according to claim 10 , wherein the non-porous, breathable membrane comprises a polyurethane polymer, a poly(N-isopropylacrylamide) polymer, a side-chain crystalline polymer, or a polymer composite containing paraffin wax.

12. The kit of claim 1, wherein the nitric oxide permeable microbial barrier has a microbial barrier size of at least 0.2 x 10 -5 cm 2 / s for the apparent diffusion coefficient of nitric oxide.

13. The kit of claim 1, wherein the nitric oxide permeable microbial barrier has a microbial barrier size of at least 1.0 x 10 -5 cm 2 / s for the apparent diffusion coefficient of nitric oxide.

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

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

16. The kit of claim 15, wherein the nitric oxide releasing source is connected to the second container, and / or wherein the nitric oxide releasing source is replaceable when exhausted.

17. 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 diazenediol compound, arginine, an organic nitrite, or a biological source suitable for generating nitric oxide, and optionally wherein the nitric oxide-releasing source is linked to a polymer.

18. The kit of claim 15, wherein the nitric oxide-releasing source releases nitric oxide upon exposure to visible light or UV light.

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

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

21. A method for sterilizing an object, comprising: placing the object into a first container and then sealing the first container, wherein the first container comprises a nitric oxide permeable microbial barrier; placing the sealed first container into a second container and sealing the second container, thereby enclosing the sealed first container within the second container; introducing nitric oxide into the second container or generating nitric oxide in the second container to achieve at least a sterilizing nitric oxide concentration; wherein the nitric oxide permeable microbial barrier has a permeability to nitric oxide sufficient to achieve a sterilizing concentration of nitric oxide in the first container when the second container contains nitric oxide at or above a sterilizing concentration of nitric oxide; and The objects in the first container are exposed to the sterilizing nitric oxide concentration for a time sufficient to sterilize the objects.

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

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

24. The method according to claim 21, wherein the first container and / or the second container has a diameter of 10 cm 3 Up to 1000cm 3 internal volume.

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

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

27. The method of claim 21, wherein the nitric oxide permeable microbial barrier comprises a porous gas permeable membrane.

28. The method according to claim 27, wherein the porous breathable membrane comprises a perforated membrane, a composite membrane of a microporous polymer and an inorganic filler, or a nonwoven polymer fiber web.

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

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

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

32. The method of claim 21, wherein the nitric oxide permeable microbial barrier has a microbial barrier size of at least 0.2 x 10 -5 cm 2 / s for the apparent diffusion coefficient of nitric oxide.

33. The method of claim 21, wherein the nitric oxide permeable microbial barrier has a -5 cm 2 / s for the apparent diffusion coefficient of nitric oxide.

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

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

36. The method of claim 35, wherein the nitric oxide releasing source is connected to the second container, or wherein the nitric oxide releasing source is replaceable when exhausted.

37. 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 diazenediol compound, arginine, an organic nitrite, or a biological source suitable for generating nitric oxide, and optionally wherein the nitric oxide-releasing source is linked to a polymer.

38. The method of claim 35, wherein the nitric oxide-releasing source releases nitric oxide upon exposure to visible light or UV light.

39. The method of any one of the preceding claims, wherein the first container and / or the second container further comprises a carrier comprising a chromophore that changes color in the presence of nitric oxide.

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

41. A method for contactless sterilization of an object, comprising: exposing the object to a sterile concentration of nitric oxide 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 through the nitric oxide permeable microbial barrier.

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

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

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

45. The method of claim 41, wherein the nitric oxide permeable microbial barrier comprises a porous gas permeable membrane.

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

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

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

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

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

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

52. The method of claim 51, wherein the nitric oxide is generated by a nitric oxide-releasing source within the second container, and / or wherein the nitric oxide-releasing source is replaceable upon depletion.

53. The method of claim 41, further comprising detecting a color change of a chromophore attached to a carrier and located within the first container, wherein the color change indicates the presence of nitric oxide.

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

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

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