In vivo and in vitro use of graphene

Graphene in vivo and in vitro uses

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This document requests the rights and interests of U.S. Provisional Petition 61/951,926 filed on March 12, 2014, which is incorporated herein by reference in its entirety.

[0003] Background of the Invention

[0004] The present disclosure generally relates to transporting and delivering substances in a biological environment, and more specifically, to methods and devices for transporting and delivering substances using carbon nanomaterials.

[0005] The delivery of drugs and cells in immunocompetent organisms and immune incompetent organisms is a practical current problem in medical research and practice today. This study used polymer devices and hydrogels as delivery vehicles. Some examples include polytetrafluoroethylene with a nonwoven polyester mesh backing, silica gel, hydrogel, alginate, cellulose sulfate, collagen, gelatin, agarose, chitosan, and the like. Current delivery vehicles and devices are challenged by biofouling, biocompatibility issues, and delayed response. The thickness of prior art devices may limit efficacy because the limited diffusion of nutrients can kill the cells contained within or delay the two-way transport of drugs or molecules that are being sensed. Low permeability can also be problematic, due at least in part to thickness and mechanical stability in view of physical stress and osmotic stress.

[0006] In view of the foregoing, improved technologies for transporting and delivering substances under various conditions, especially in biological environments, will have considerable benefits in the art. The present disclosure satisfies this need and also provides related advantages.

[0007] Summary of the invention

[0008] The present disclosure describes encapsulants formed from porous graphene or other porous two-dimensional materials. The encapsulation can contain a variety of substances, which allows selected substances to move back and forth in the encapsulation, keep other selected substances in it, and prevent other selected substances from entering the encapsulation. The encapsulation of the present invention can be used to release one or more selected substances into the environment outside the encapsulation, allowing one or more selected substances to enter the encapsulation from the environment outside the encapsulation, and inhibit Preferably, one or more selected substances are prevented from entering the encapsulation from the external environment, one or more selected substances are retained (inhibited or preferably prevented from exiting) in the encapsulation, or a combination of these applications. Based on the specific application of the encapsulant, the size or size range of the hole or opening is selected. The term "encapsulation" refers to a space used to contain one or more substances, which is at least partially formed of a porous two-dimensional material such as graphene-based materials, wherein one or more substances in the encapsulation can pass through The channel of the porous two-dimensional material leaves the encapsulation. Likewise, in certain embodiments, one or more substances from the external environment can enter the encapsulation through the passage of the porous two-dimensional material. In a specific embodiment, the external environment is a biological environment, which may be a biological environment in the body or a biological environment in vitro.

[0009] In an embodiment, the encapsulant comprises one or more than one sub-compartment, each sub-compartment comprising a porous two-dimensional material such that at least a portion of the wall or side surface forming the sub-compartment is a porous two-dimensional material . The fluid communication is achieved by selectively penetrating one or more substances into and/or out of the enclosure or its sub-compartments. The fluid may be liquid or gas, and includes fluids with entrained gas. The substance can be dissolved or suspended, or otherwise carried in a fluid. The fluid can be aqueous. The sub-compartment can be in direct fluid communication with the adjacent sub-compartment or the external environment (where the adjacent sub-compartments share at least one wall or side). In an embodiment, one or more sub-compartments may be in direct fluid communication with an adjacent sub-compartment, but not in direct fluid communication with the external environment. At least one sub-compartment in the enclosure is in direct fluid communication with the external environment. The enclosure may have sub-compartments of different configurations. The sub-compartment can have any shape. The sub-compartments may be spherical, cylindrical or linear, for example. In an embodiment, the subcompartments may be nested. In an embodiment, the encapsulant may have a central sub-compartment, which is compatible with

Multiple surrounding sub-compartments share one wall or side. In an embodiment, the sub-compartments may be arranged linearly within the enclosure. In an embodiment, the encapsulant contains two sub-compartments. In embodiments, the encapsulant contains three, four, five or six sub-compartments. In an embodiment, a sub-compartment may be completely contained within another sub-compartment, wherein the inner sub-compartment is in direct fluid communication with the outer sub-compartment, and the outer sub-compartment is in direct fluid communication with the external environment. In this embodiment, the internal sub-compartment is in indirect rather than direct fluid communication with the external environment. In embodiments where the enclosure contains multiple sub-compartments, at least one sub-compartment is in direct fluid communication with the external environment, and the remaining sub-compartments are in direct fluid communication with adjacent sub-compartments, but not all sub-compartments are in direct fluid communication with the adjacent sub-compartments. The chamber is in direct fluid communication with the external environment. In embodiments where the enclosure contains multiple sub-compartments, all sub-compartments are in direct fluid communication with the external environment.

[0010] The encapsulant encapsulates at least one substance. In embodiments, the encapsulant may contain more than one different substance. Different substances can be in the same or different sub-compartments. In an embodiment, the different substances in the encapsulation are not all released to the environment outside the encapsulation. In an embodiment, all the different substances in the encapsulation are released to the external environment. In embodiments, the rate of release of different substances from the encapsulation into the external environment is the same. In embodiments, the rate of release of different substances from the encapsulation into the external environment is different. In an embodiment, the relative amounts of the different substances released from the encapsulation are the same or different. The rate of release of the substance from the encapsulation can be controlled by the selected pore size, the functionalization of the pore, or both.

[0011] Also described herein are methods for transporting and delivering substances in a biological environment. In some embodiments, the method may include: introducing an encapsulation formed of graphene or other two-dimensional materials into a biological environment, and releasing at least a part of the substances in the encapsulation to the biological environment. In some or other embodiments, the method may include introducing an encapsulant formed of graphene into a biological environment, and migrating a substance from the biological environment into the encapsulant.

[0012] In an embodiment, the invention provides a method comprising the following steps:

[0013] introducing into the environment an encapsulation containing a porous two-dimensional material, the encapsulation containing at least one substance; and

[0014] At least a part of the at least one substance is released to the environment outside the encapsulation through the pores of the two-dimensional material. Any of the encapsulants herein can be used in this method.

[0015] In an embodiment, the present invention provides a method comprising the following steps:

[0016] Introducing an encapsulation containing a porous two-dimensional material into the environment, the encapsulation containing at least one first substance; and migrating the second substance from the environment into the encapsulation. In an embodiment, the first substance is cells, the second substance is nutrients, and the other second substance is oxygen.

[0017] The foregoing provides a fairly broad overview of the features of the present disclosure so that the following detailed description may be better understood. Additional features and advantages of the present disclosure will be described below. According to the following description, these and other advantages and features will become more apparent.

[0018] Brief Description of the Drawings

[0019] In order to have a more complete understanding of the present disclosure and its advantages, reference is now made to the following description in conjunction with the accompanying drawings describing specific embodiments of the present disclosure, in which:

[0020] Figure 1 shows a schematic diagram showing the thickness of graphene-based materials compared to common drug delivery vehicles and devices. The figure also illustrates an embodiment of the present invention in contact with biological tissues in a biological environment, where an encapsulant is provided together with one or more supporting materials, which are external to the porous two-dimensional material. ), and indicates possible capillary vessel formation into this type of support material.

[0021] Figures 2A-D show the structure of an encapsulant made from useful two-dimensional materials according to various embodiments of the present disclosure.

Schematic diagram of the same structure.

[0022] Figures 3A and 3B are schematic diagrams of implementing the encapsulant of the present invention for immune isolation of living cells.

[0023] Figures 4A-C illustrate exemplary preparations of encapsulates of the present invention.

[0024] Detailed description of the invention

[0025] The present disclosure relates in part to methods of transporting and delivering substances using graphene-based materials and other two-dimensional materials in a biological environment. The present disclosure also partially relates to encapsulants formed by graphene-based materials and other two-dimensional materials on a suitable substrate or substrates, or suspended on a suitable substrate or substrates, and the substrate may be Porous or non-porous, the graphene-based materials and other two-dimensional materials can be used as delivery vehicles in the environment outside the encapsulation, especially in the biological environment. The present disclosure also partially relates to encapsulants containing cells, drugs, and other drugs formed from graphene-based materials or other two-dimensional materials.

[0026] Graphene has received extensive attention for its use in many applications due to its good mechanical and electronic properties. Graphene represents a thin layer of atomic carbon, in which carbon atoms exist as closely spaced atoms in regular lattice positions. The regular crystal lattice position may have multiple defects present therein, and the defects may exist naturally or be intentionally introduced to the basal plane of graphene. Such defects are also equivalently referred to herein as "openings", "perforations" or "holes." The term "porous graphene" is used herein to mean that the graphene thin layer has defects in its base surface, regardless of whether the defects are naturally occurring or intentionally generated. In addition to such openings, graphene and other two-dimensional materials can represent impermeable layers for many substances. Therefore, when the size is appropriate, the openings in the impermeable layer of such materials can be used to enter and exit the enclosure formed by the impermeable layer.

[0027] The present disclosure considers a variety of graphene-based encapsulants capable of delivering targets to in vivo or in vitro locations in an organism or similar biological environment while maintaining a barrier (eg, an immune isolation barrier). Encapsulation of molecules or cells that are transported through semipermeable membranes (such as porous graphene or other two-dimensional materials) in both directions, while isolating cells in the biological environment (such as living organisms), can enable treatment to overcome transplant rejection and drug resistance. Repeated dose requirements and excessive surgical intervention. The above can be achieved by providing technologies that allow xenogeneic and allogeneic tissue grafts, long-term low-dose treatment levels of drugs, and even perceptual response paradigms to process nutrients after surgical intervention, thereby reducing the amount of food from the same location. Complications of multiple surgical operations. It should be appreciated that the foregoing only represents specific advantages of the present disclosure, and should not be considered as limiting the scope of the embodiments described herein.

[0028] The present inventors have recognized that porous graphene and other two-dimensional materials can easily promote the foregoing while being superior to current delivery vehicles and devices, especially immune isolation devices. Graphene can achieve the foregoing due to its unique thinness, strength, conductivity (for potential electrical stimulation), and permeability in the form of internal perforations. Compared to the long-term diffusion seen with thicker polymer films with comparable size properties, the subsequent sieve-like transport characteristics that are thin and across the surface of the graphene film can allow for a disruptive time response.

[0029] Two-dimensional materials are most often atomically thin materials, ranging in thickness from a single layer of sub-nanometer thickness to a few nanometers, and they generally have a high surface area. Two-dimensional materials include metal chalcogenides (e.g. transition metal dichalcogenides), transition metal oxides, hexagonal boron nitride, graphene, silicone, and zirconene (see: Xu et al. (2013) Graphene-like TwoDimensional Materials''Chemical Reviews 113:3766-3798). Graphene represents a form of carbon in which carbon atoms are present in a single atom-level thin layer or several thin layers (for example, about 20 layers or less) that form a fused six-membered ring that extends the sp2-hybrid carbon plane lattice Inside. Among its many forms, graphene has gained wide attention for its use in many applications, mainly due to its high electrical conductivity and thermal conductivity value, good in-plane mechanical strength, and unique optical and electrical properties. Advantageous combination of features. Other two-dimensional materials with a thickness of a few nanometers or less and an extended planar lattice,

It has also gained attention for a variety of applications. In an embodiment, the two-dimensional material has a thickness of 0.3 nm to 1.2 nm. In other embodiments, the two-dimensional material has a thickness of 0.3 nm to 3 nm.

[0030] In various embodiments, the two-dimensional material comprises a thin layer of graphene-based material. In an embodiment, the thin layer of graphene-based material is a thin layer of single or multilayer graphene or a thin layer including multiple interconnected single or multilayer graphene domains. In an embodiment, the multilayer graphene domain has 2 to 5 layers, or 2 to 10 layers. In an embodiment, the layer including a thin layer of graphene-based material further includes a non-graphene carbon-based material on the surface of the thin layer of graphene-based material. In an embodiment, the amount of the non-graphene carbon-based material is less than the amount of graphene. In an embodiment, the amount of graphene in the graphene-based material is 60% to 95%, or 75% to 100%.

[0031] In an embodiment, the characteristic size of the perforations is 0.3nm to 10nm, 1nm to 10nm, 5nm to 10nm, 5nm to 20nm, 10nm to 50nm, 50nm to 100nm, 50nm to 150nm, 100nm to 200nm, or 100nm to 500nm. In an embodiment, the average pore size is within the specified range. In an embodiment, 70% to 99%, 80% to 99%, 85% to 99%, or 90% to 99% of the perforations in the thin layer or layer fall within the specified range, but other holes fall within the specified range Outside.

[0032] The technology used to form graphene or graphene-based materials in the embodiments described herein is not considered to be specifically limited. For example, in some embodiments, CVD graphene or graphene-based materials may be used. In various embodiments, the CVD graphene or graphene-based material can be released from the substrate (eg, Cu) on which it is grown and transferred to the polymer backing. Likewise, in addition to selecting this technique to produce perforations within a desired size range, the technique used to introduce perforations into graphene or graphene-based materials should not be considered to be specifically limited. Determining the appropriate size of the perforations as described herein provides the desired selective permeability of substances (atoms, molecules, proteins, viruses, cells, etc.) for a given application. Selective permeability refers to the tendency of porous materials or porous two-dimensional materials to allow one or more substances to pass (or transport) more easily or faster than others. Selective permeability allows the separation of substances exhibiting different passage or transport rates. In two-dimensional materials, selective permeability is related to the size or size (for example, diameter) of the opening and the relative effective size of the substance. The selective permeability of perforations in two-dimensional materials, such as graphene-based materials, can also depend on the functionalization (if any) of the perforations and the specific substance to be separated. The separation of two or more substances in the mixture includes the change in the ratio (weight ratio or molar ratio) of the two or more substances in the mixture after the mixture passes through the porous two-dimensional material.

[0033] Graphene-based materials include, but are not limited to: single-layer graphene, multilayer graphene, or interconnected single-layer or multilayer graphene domains, and combinations thereof. In an embodiment, the graphene-based material further includes a material formed by stacking a single layer or a multilayer graphene thin layer. In an embodiment, the multilayer graphene includes 2 to 20 layers, 2 to 10 layers, or 2 to 5 layers. In an embodiment, graphene is the main material among graphene-based materials. For example, the graphene-based material contains at least 30% graphene, or at least 40% graphene, or at least 50% graphene, or at least 60% graphene, or at least 70% graphene, or at least 80% graphene, or At least 90% graphene, or at least 95% graphene. In an embodiment, the graphene-based material includes graphene selected from the range of 30% to 95%, 40% to 80%, 50% to 70%, 60% to 95%, or 75% to 100%.

[0034] As used herein, "crystal domain" refers to a region of material in which atoms are uniformly arranged in a crystal lattice. The crystal domains are uniform within their boundaries, but are different from adjacent regions. For example, the ordered atoms of a single crystalline material have a single crystal domain. In an embodiment, at least some of the graphene domains are nanocrystals, which have a domain size of 1 to 100 nm, or 10-100 nm. In an embodiment, at least some of the graphene crystal domains have a crystal domain size greater than 100 nm to lum, or 200 nm to 800 nm, or 300 nm to 500 nm. The "grain boundaries" formed by crystal defects at the edges of each crystal domain distinguish adjacent crystal lattices. In some implementation

In the solution, by rotating around an axis perpendicular to the plane of the thin layer, the first crystal lattice can be rotated relative to the second crystal lattice, so that the two crystal lattices are different in "lattice orientation".

[0035] In an embodiment, the thin layer of graphene-based material includes a single layer or a multilayer graphene thin layer or a combination thereof. In an embodiment, the thin layer of graphene-based material is a single-layer or multi-layer graphene thin layer or a combination thereof. In another embodiment, the thin layer of graphene-based material is a thin layer comprising a plurality of interconnected single-layer or multi-layer graphene domains. In an embodiment, interconnected crystal domains are covalently bonded together to form a thin layer. When the crystal domains in the thin layer differ in the crystal lattice orientation, the thin layer is polycrystalline.

[0036] In an embodiment, the thickness of the thin layer of the graphene-based material is 0.34 to 10 nm, 0.34 to 5 nm, or 0.34 to 3 nm. In an embodiment, the thin layer of graphene-based material contains inherent defects. In contrast to the selective introduction of a thin layer of graphene-based material or perforation of a thin layer of graphene, inherent defects are produced by the preparation of graphene-based materials. Such inherent defects include, but are not limited to: lattice abnormalities, small holes, tears, cracks or wrinkles. Lattice anomalies can include, but are not limited to: having carbon rings other than 6 members (for example, 5-membered, 7-membered, or 9-membered rings), vacancies, interstitial defects (which include the incorporation of non-carbon atoms in the crystal lattice), and grain boundaries .

[0037] In an embodiment, the layer comprising a thin layer of graphene-based material further comprises a non-graphene-based carbon-based material on the surface of the thin layer of graphene-based material. In embodiments, non-graphene carbon-based materials do not have long-range order and can be classified as amorphous. In an embodiment, the non-graphene carbon-based material further contains elements other than carbon and/or billet. Non-carbon elements that can be incorporated into non-graphene carbon include, but are not limited to: hydrogen, oxygen, silicon, copper, and iron. In an embodiment, the non-graphene carbon-based material comprises a billet. In an embodiment, carbon is the main material in non-graphene carbon-based materials. For example, a non-graphene carbon-based material contains at least 30% carbon, or at least 40% carbon, or at least 50% carbon, or at least 60% carbon, or at least 70% carbon, or at least 80% carbon, or at least 90% carbon , Or at least 95% carbon. In an embodiment, the non-graphene carbon-based material includes a carbon range selected from: 30% to 95%, or 40% to 80%, or 50% to 70%.

[0038] Such nanomaterials in which pores are intentionally generated are referred to herein as "porous graphene", "porous graphene-based materials" or "porous two-dimensional materials". The present disclosure also partially relates to porous graphene, porous graphene-based materials, and other porous two-dimensional materials containing multiple pores of a size (or size range) suitable for a given encapsulant application. The size distribution of the holes may be narrow, for example, limited to a size deviation of 1T0%, or a size deviation of 1-20%. In an embodiment, the reference size of the hole is selected for the application. For round holes, the reference size is the diameter of the hole. In embodiments related to non-circular holes, the reference size can be considered as the maximum distance across the hole, the minimum distance across the hole, the average of the maximum and minimum distance across the hole, or based on the in-plane of the hole The equivalent diameter of the area. As used herein, porous graphene-based materials include materials in which non-carbon atoms are incorporated at the edges of the holes.

[0039] In various embodiments, the two-dimensional material includes graphene, sulfide buttons, or boron nitride. In a more specific embodiment, the two-dimensional material may be graphene. The graphene described in the embodiment of the present disclosure may include single-layer graphene, multi-layer graphene, or any combination thereof. Other nanomaterials with extended two-dimensional molecular structures can also form the two-dimensional materials in various embodiments of the present disclosure. For example, the sulfide button is a representative chalcogenide having a two-dimensional molecular structure, and various other chalcogenides can constitute the two-dimensional material in the embodiment of the present disclosure. A variety of factors can determine the selection of a suitable two-dimensional material for a specific application, including the chemical or physical environment in which graphene or other two-dimensional materials will be finally deployed. For the application in the present invention, the materials used in preparing the encapsulation are preferably biocompatible or can be made biocompatible.

[0040] The process of forming holes in graphene and other two-dimensional materials is referred to herein as "perforated", and such nanomaterials are referred to herein as "porous". In the graphene thin layer, the structure of each six-carbon atom ring in the thin layer forms a gap

The gap opening, and the gap opening has a span less than one nanometer. Specifically, the gap opening is considered to be about 0.3 nanometers across its longest dimension (the center-to-center distance between carbon atoms is about 0.28 nm, and the opening is slightly smaller than this distance). The perforation of a thin layer containing a two-dimensional network structure generally refers to the formation of holes larger than the gap opening in the network structure.

[0041] Due to the atomic level thinness of graphene and other two-dimensional materials, it is possible to achieve high liquid flux flows during separation or filtration processes, even with pores in the 1-20 nm range.

[0042] Chemical techniques can be used to create pores in graphene and other two-dimensional materials. Exposure of graphene or another two-dimensional material to ozone or atmospheric pressure plasma (eg, oxygen/nitrogen, or nitrogen/nitrogen plasma) can affect the perforation. Physical techniques such as ion bombardment can also be used to remove substances from the planar structure of the two-dimensional material in order to create pores. All such physical or chemical methods can be applied to prepare the porous two-dimensional material used herein, depending on the pore size or range of pore sizes desired for a given application.

[0043] In various embodiments of the present disclosure, the size of the pores generated in graphene or other two-dimensional materials may range from about 0.3 nm to about 50 nm. In a more specific embodiment, the size of the pores may range from 1 nm to 50 nm. In a more specific embodiment, the size of the pores may range from 1 nm to 10 nm. In a more specific embodiment, the size of the pores may range from 5 nm to 10 nm. In a more specific embodiment, the size of the pores may range from 1 nm to 5 nm. In a more specific embodiment, the pore size ranges from about 0.5 nm to about 2.5 nm. In another embodiment, the size of the pores is 0.3nm to 0.5nm_oIn other embodiments, the size of the pores is 0.5 nm to 10 nm. In other embodiments, the size of the pores is 5 nm to 20 nm. In other embodiments, the size of the pores is 0.7nm to 1.2nm_oIn other embodiments, the size of the pores is 10 nm to 50 nm. In embodiments where a larger pore size is preferred, the pore size is 50 nm to 100 nm, 50 nm to 150 nm, or 100 nm to 200 nm_o

[0044] The term "substance" is generally used herein to refer to atoms, molecules, viruses, cells, particles, and aggregates thereof. The substances of specific concern are molecules of different sizes, including biomolecules such as proteins and nucleic acids. Substances can include drugs, drugs, medicaments and therapeutic agents, including biological agents and small molecule drugs.

[0045] FIG. 1 shows a schematic diagram showing the thickness of graphene compared to common drug delivery vehicles and devices. The biocompatibility of graphene can also be specifically promoted by functionalizing graphene to be compatible with a specific biological environment (for example, via available edge bonding, overall surface functionalization of W bonds, etc.) to promote this application. Functionalization can provide membranes with increased complexity for the treatment of local and systemic diseases. Figure 1 illustrates the wall of an encapsulant formed with a porous two-dimensional material having a pore size in the range of 400-700 nm, which will trap living cells. Use an optional porous support structure (polymer or ceramic) adjacent to and outside the porous two-dimensional material and an optional textile support material outside the porous two-dimensional material. The enclosure is not shown) adjacent to the external biological environment. As shown, the implantation of such encapsulants allows for vascularization into any such external support material. In embodiments intended to provide immune isolation, smaller pore sizes are generally preferred to prevent antibodies from entering the encapsulant.

[0046] In various embodiments, the present disclosure describes sealed enclosures formed primarily of two-dimensional materials, such as graphene, that retain the ability to transport materials in a bidirectional manner. In various embodiments, at least one cross-section or surface of the encapsulation contains perforations of appropriate size in the two-dimensional material to allow materials of a desired size to go in and out from the inside of the encapsulation, respectively.

[0047] In some embodiments, a two-dimensional material such as graphene can be adhered to a suitable porous substrate. Suitable porous substrates can include, for example, thin film polymers and ceramics.

[0048] In an embodiment, the enclosure may have multiple sub-compartments within the main enclosure, each sub-compartment comprising a porous two-dimensional material to allow one or more substances to pass into or out of the sub-compartment . In such embodiments, the sub-compartment can have any

Meaning useful shape or size. In specific embodiments, there are 2 or 3 sub-compartments. Several examples of encapsulation subcompartments are illustrated in Figures 2A-2D. In Figure 2A, the nested structure is illustrated, the main enclosure B completely contains the smaller enclosure A, The substance in the most central enclosure A can enter the main enclosure B, and may react with the main compartment or react in the main compartment during entry and exit from it. In this embodiment, one or more substances in A can enter B, and one or more substances in A can be retained in A without entering B. The two sub-compartments through which one or more substances can pass directly between the sub-compartments are in direct fluid communication. The channels between the sub-compartments and between the encapsulation and the external environment are channels through the pores of the porous two-dimensional material. The barrier (membrane, porous two-dimensional material) between the compartments A and B can penetrate all substances in A or selectively permeate certain substances in A. The barrier (membrane) between B and the external environment can permeate all substances in B or selectively permeate certain substances in B. In FIG. 2A, sub-compartment A is in direct fluid communication with sub-compartment B, and sub-compartment B is in direct fluid communication with the external environment. The compartment A in this nested configuration can only be in indirect fluid communication with the external environment via the intermediate passage into the sub-compartment B. The two-dimensional materials used in different sub-compartments of a given encapsulant can be the same or different materials, and the size of the perforations and holes in the two-dimensional materials of different sub-compartments can be the same or different. It depends on the substance and application involved.

[0049] In Figure 2B, the encapsulant is divided into two by an impermeable wall (for example, formed by a non-porous or impermeable sealant) to form sub-compartments A and B, allowing the two parts to enter independently Exit position, but there is no direct or indirect passage for the substance from A to B (however, it should be understood that the substance leaving A or B can indirectly enter another sub-compartment via the external environment).

[0050] In FIG. 2C, the main enclosure is also divided into two sub-compartments A and B, but a porous material is used to form a barrier between the sub-compartments. In an embodiment, the two sub-compartments not only enter the exit position independently, but can also interact with each other, that is, the sub-compartments are in direct fluid communication. In an embodiment, the barrier (membrane) between compartments A and B is selectively permeable, for example it allows at least one substance in A to enter B, but does not allow substances originating in B to pass to A .

[0051] Figure 2D illustrates an enclosure with three compartments. The enclosure is exemplified by sub-compartment A with an outlet into sub-compartment B, which in turn has an outlet to sub-compartment C, which in turn has an outlet into the external environment. The compartments A and B do not have an outlet to the external environment, i.e. they are not in direct fluid communication with the external environment. The adjacent sub-compartments A and B and the adjacent sub-compartments B and C are each separated by a porous two-dimensional material, and thus are in direct fluid communication with each other. Sub-compartment A can only be in indirect fluid communication with compartment C and the external environment via sub-compartment B, or B and C, respectively. Various other combinations of semi-permeable barriers (membranes) or impermeable barriers can be used to separate the compartments in the enclosure herein. Different perforation size limits can be changed, depending on how the encapsulation is finally shaped (for example, whether one encapsulation is inside the other relative to the side-by-side format). Regardless of the selected configuration, the boundary of the encapsulant or at least a part thereof can be constructed from a two-dimensional material in order to realize its benefits, in particular to make the thickness of the active membrane smaller than the diameter of the target to be selectively passed through the membrane. In some embodiments, the pore size of the two-dimensional material may range from about 0.3 nm to about 1 Onm in size. Larger hole sizes are also possible. [0052] It should also be noted that in some embodiments, the enclosure may be supported by one or more support structures. In implementation In the solution, the supporting structure itself may have a porous structure, wherein the pores are larger than the pores of the two-dimensional material. In an embodiment, the support structure is completely porous. In an embodiment, the support structure is at least partially non-porous.

[0053] The multiple physical embodiments of the encapsulant and its use described herein may allow for different levels of interaction and proportional complexity of the problem to be solved. For example, a single encapsulant can provide drug elution for a given period of time, or there can be multiple sized perforations to limit or allow the movement of different targets, each of a specific size.

[0054] The increased complexity of the embodiments described herein with multiple subcompartments may allow for interaction between target compounds to catalyze or activate secondary responses (ie, the "perceived response" paradigm). For example, if there are two parts of the encapsulant that enter the outlet independently, the exemplary compound A can undergo constant diffusion into the body, or after a period of time or only

It undergoes constant diffusion into the body in the presence of irritants from the body. In such embodiments, the exemplary compound A can activate the exemplary compound B, or inactivate its function to block the exemplary compound B to prevent spillage. The combination that produces the above effects can be reversible or irreversible. In addition, in other embodiments, the exemplary compound A can interact with a chemical cascade generated outside the encapsulant, and the metabolite after the interaction can release the exemplary compound B (by inactivating the functionalization). Using other examples that produce effects in a similar manner, including the use of source cells (non-host, allogeneic) contained in the encapsulation in which secretions from the cells can produce a "sensory response" paradigm .

[0055] In other embodiments, growth factors may be loaded in the encapsulant to promote blood vessel formation (see Figure 1). In the foregoing embodiments, cell survival can be far superior to the results of two-way transport of nutrients and waste.

[0056] In other embodiments, the relative thinness of graphene enables bidirectional transport through membrane encapsulants that are very close to blood vessels (especially capillaries) and other target cells. The present embodiment using the graphene-based encapsulant can provide a difference from other solutions that achieve the same effect because the graphene film does not significantly restrict permeability. On the contrary, the diffusion of molecules through the media or gap junctions will limit the movement of the target.

[0057] Regarding the foregoing, any "perceptual response" paradigm is possible with graphene through an excellent time response. The biocompatibility of graphene can also enhance this application. In the treatment of local and systemic diseases, functionalized graphene membranes with increased complexity are expected to have a lower degree of biofouling (due to functionalization or charging). In addition, the mechanical stability of graphene can make it suitable for physical stress and osmotic stress within the receptor.

[0058] Figures 3A and 3B provide schematic diagrams of the immunoisolation of the encapsulant of the present invention. The enclosure is exemplified as having a single compartment. It should be understood that the enclosure may have multiple sub-compartments, for example, two or three sub-compartments. The encapsulant (30) of FIG. 3A is formed of an inner thin layer or inner layer (31) containing a porous two-dimensional material (such as a graphene-based material) and an outer thin layer or outer layer (32) of a supporting material The cross section is shown. The support material may be porous, selectively permeable, or non-porous and impermeable. However, at least a part of the support material is porous or selectively permeable, which is suitable for the application of the encapsulant. The supporting thin layer or supporting layer can be, for example, a polymer or ceramic. The enclosure contains selected living cells (33) for a given use. Figure 3B provides an alternative cross-section of the encapsulant of Figure 3A, which shows the space or cavity formed between the first and second composite layers (32/31), where the sealant 34 is exemplified as a sealing composite layer the edge of. It should be understood that a physical method of clamping or crimping can form a seal at the edge of the composite layer. The methods and materials used to form the seal at the edge are not specifically limited, but a non-porous and impermeable seal or closure must be provided.

[0059] If the cells are contained within the closure, at least a portion of the encapsulant is permeable to oxygen and nutrients sufficient for cell growth and maintenance, and permeable to waste products. The encapsulant is impermeable to cells, especially immune cells. Cells from the external environment cannot enter the encapsulation, and the cells in the encapsulation are retained. The encapsulation cannot penetrate viruses or bacteria. The encapsulant is impermeable to antibodies. In contrast, depending on the application, the encapsulant can penetrate desired products, such as growth factors produced by cells. The cells in the encapsulation are immunoisolated. In a specific embodiment, the pore size in the porous two-dimensional material used for immunoisolation is in the range of 1-1 Onm, more preferably 3-1 Onm, and still more preferably 3-5 nm.

[0060] Figures 4A-4C illustrate an exemplary method for forming an encapsulant of the present invention and introducing a selected substance (e.g., cell) therein. The method is illustrated together with the use of a sealant to form an encapsulant. The exemplary enclosure has no sub-compartments. The encapsulation with sub-compartments (such as nested or adjacent sub-compartments) can be easily prepared using the method of the example. As illustrated in Figure 4A, the second layer is formed by laying a thin layer or layer of two-dimensional material in contact with the support layer (42), especially a thin layer of graphene-based material or a thin layer of graphene (41). A composite layer or thin layer. At least a portion of the support layer (42) of the first composite material is porous or permeable. The pore size of the supporting layer is usually larger than the pores or openings in the two-dimensional material used, and can be adjusted to the environment (for example, the body cavity). Apply a layer of sealant (eg silicone) (44) on the wheel that draws the encapsulation compartment

On a thin layer or layer of porous two-dimensional material, where the sealant will form an impermeable seal near the perimeter of the encapsulant. The formation of a single compartment is exemplified in FIGS. 4A-4C, however, it should be understood that multiple independent compartments within the enclosure can be formed by a similar method. Then use a thin layer or layer of porous two-dimensional material in contact with the sealant, prepare the second composite layer in the same manner as the first composite layer, and determine its position (optionally, the sealant can be applied to the composite layer And the layer can be folded upward in contact with the sealant to form an encapsulant). Then a seal is formed between the two composite layers. Appropriate pressure can be applied to promote sealing without damaging the two-dimensional material or its supporting layer. It should be understood that an alternative encapsulant can be formed by applying a thin layer or layer of non-porous and impermeable support material by contact with the sealant. In this case, only part of the encapsulant is porous and permeable. The sealed composite layer is exemplified in Figure 4B, which shows that the sealing layer can be trimmed to the size around the sealant to form an encapsulant. The formed encapsulant shows an outer porous support layer 42, a thin layer or layer of porous two-dimensional material (41) placed as an inner layer, and a sealant 44 around the periphery of the encapsulant. As illustrated in FIG. 4C, after the encapsulant is formed, injection through the sealant layer can introduce cells or other substances excluded by the porous two-dimensional thin layer or layer of channels into the encapsulant. If necessary, any perforations formed by such injection can be sealed. It should be understood that The substance and cells are introduced into the enclosure before the seal is formed. Those skilled in the art will understand that during or after the preparation of the encapsulant, the conceived sterilization method suitable for the application can be used.

[0061] In an embodiment, the present invention provides an encapsulation of a porous two-dimensional material encapsulating a substance such that the substance is released to the environment outside the encapsulation through the passage of the pores in the porous two-dimensional material. In an embodiment, the encapsulant encapsulates more than one different substance. In an embodiment, the different substances are not all released to the environment outside the encapsulation. In an embodiment, all the different substances are released into the environment outside the enclosure. In embodiments, different substances are released into the environment outside the enclosure at different rates. In embodiments, different substances are released into the environment outside the enclosure at the same rate.

[0062] In an embodiment, the enclosure comprises two or more sub-compartments, wherein at least one sub-compartment is in direct fluid communication with the environment outside the enclosure through a hole in the two-dimensional material of the sub-compartment. In an embodiment, each sub-compartment contains a porous two-dimensional material, and each sub-compartment is in direct fluid communication with the environment outside the encapsulation through the pores in the two-dimensional material of each sub-compartment.

[0063] In an embodiment, the encapsulant is subdivided into two sub-compartments, which are at least partially separated from each other by a porous two-dimensional material, so that the two sub-compartments are in direct fluid communication with each other through pores in the two-dimensional material . In an embodiment, the encapsulation is subdivided into two sub-compartments each containing a two-dimensional material, and the sub-compartments are in direct fluid communication with each other through holes in the two-dimensional material, and there is only one sub-compartment and the encapsulation The external environment is in direct fluid communication. In an embodiment, the encapsulant is subdivided into two sub-compartments each containing a two-dimensional material, and the sub-compartments are in direct fluid communication with each other through pores in the two-dimensional material, and the two sub-compartments are also connected to the encapsulation. The environment outside the object is in direct fluid communication.

[0064] In an embodiment, the encapsulant has an inner sub-compartment and an outer sub-compartment each containing a porous two-dimensional material, wherein the inner sub-compartment is completely enclosed in the outer sub-compartment, and the inner and outer compartments The chambers are in direct fluid communication with each other through the pores in the two-dimensional material, and the inner sub-compartments are not in direct fluid communication with the environment outside the encapsulation.

[0065] In an embodiment, wherein the encapsulant has a plurality of sub-compartments each containing a two-dimensional material, the sub-compartments are nested sub-compartments within another, and the sub-compartments each pass through the two-dimensional material The pores are directly in fluid communication with the adjacent sub-compartment, the outermost sub-compartment is in direct fluid communication with the environment outside the enclosure, and the remaining multiple sub-compartments are not in direct fluid communication with the environment outside the enclosure.

[0066] In an embodiment, wherein the encapsulant is subdivided into a plurality of sub-compartments each containing a two-dimensional material, each sub-compartment is in direct fluid communication with one or more adjacent sub-compartments, and there is only one sub-compartment. The environment outside the compartment and the enclosure is directly

Fluid communication.

[0067] In any embodiment of the encapsulation configuration herein, at least one substance within the encapsulation that is released through the pores in the two-dimensional material into the environment outside the encapsulation is a drug, a therapeutic agent, or a drug. In an embodiment, where the released substance is a drug, a therapeutic agent, or a drug, the two-dimensional material of the encapsulation used to release the substance contains pores with a size range of 1-50 nm. In an embodiment, where the released substance is a drug, a therapeutic agent, or a drug, the two-dimensional material of the encapsulation used to release the substance contains pores in the size range of 1-1 Onm.

[0068] In any of the encapsulation embodiments herein, the material in the encapsulation is cells, and the size of the pores in the two-dimensional material is selected to keep the cells in the encapsulation and deny immune cells and antibodies from the encapsulation. The environment outside the enclosure enters the enclosure. In a specific embodiment, for the use of cells, the encapsulant is divided into a plurality of sub-compartments, and one or more sub-compartments contain cells. The encapsulant can contain different cells in one sub-compartment, or different cells in different sub-compartments of the same encapsulant. In a specific embodiment, for the use of cells, the encapsulant is a nested encapsulant, where the cells are in an inner subcompartment.

[0069] In an embodiment, the encapsulant has an inner sub-compartment and an outer sub-compartment each containing a porous two-dimensional material, wherein the inner sub-compartment is completely enclosed in the outer sub-compartment, and the inner and outer compartments The chamber is in direct fluid communication through the holes in the two-dimensional material of the inner sub-compartment, the inner sub-compartment is not in direct fluid communication with the environment outside the enclosure, and the outer compartment is in direct fluid communication with the environment outside the enclosure.

[0070] In an embodiment, for the use of cells, the encapsulant has a plurality of sub-compartments each containing a porous two-dimensional material, and each of the sub-compartments is connected to one or more adjacent The sub-compartments are in direct fluid communication, the cells are in one or more cell-containing sub-compartments, and each of the one or more cell-containing sub-compartments is not in direct fluid communication with the environment outside the encapsulation.

[0071] In embodiments of the cell-containing enclosure, the cell is a yeast cell or a bacterial cell. In the embodiment of the cell-containing encapsulation, the cell is a mammalian cell. In the embodiment of the cell-containing encapsulant, the size of the pores in the two-dimensional material of the encapsulant or subcompartment ranges from 1-1 Onm>3-1 Onm> or 3-5nm.

[0072] In any of the encapsulation embodiments herein, the two-dimensional material in the encapsulation is supported on a porous substrate. In an embodiment, the porous substrate may be a polymer or ceramic.

[0073] In any of the encapsulant embodiments herein, the two-dimensional material is a graphene-based material. In any embodiment of the encapsulant herein, the two-dimensional material is graphene.

[0074] In any embodiment of the encapsulant herein, at least a portion of the pores in the two-dimensional material of the encapsulant are functionalized.

[0075] In any of the encapsulant embodiments herein, at least a portion of the two-dimensional material is conductive, and a voltage can be applied to at least a portion of the conductive two-dimensional material. The voltage can be AC or DC voltage. The voltage can be applied from an external power source of the encapsulation. In an embodiment, the encapsulant device of the present invention further includes a connector, and guides the application of a voltage from an external power source to the two-dimensional material.

[0076] The present invention provides a method for delivering one or more substances to a selected environment using any of the encapsulants herein. In a specific embodiment, the environment is a biological environment. In an embodiment, the encapsulant is implanted in a biological tissue. In an embodiment, an encapsulant is used for the delivery of drugs, drugs, or therapeutic agents.

[0077] In an embodiment, the present invention provides a method comprising: introducing an encapsulation containing a porous two-dimensional material into the environment, the encapsulation containing at least one substance; and The pores of the material release at least a part of the at least one substance to the environment outside the enclosure. In an embodiment, the encapsulation contains fine particles that are not released from the encapsulation.

The cells, and part of the at least one substance released are substances produced by the cells in the encapsulation.

[0078] In an embodiment, the present invention provides a method comprising: introducing into the environment an encapsulation comprising a porous two-dimensional material, the encapsulation containing at least one first substance; and The second substance from the environment migrates into the enclosure. In an embodiment, the first substance is a cell, the second substance is a nutrient, and the other second substance is oxygen.

[0079] In embodiments, the support layer may be a polymer or ceramic material. Useful exemplary ceramics include nanoporous silica or SiN. Useful porous polymer supports include track-etched polymers, foamed polymers, or non-woven polymers. The support material can be porous or permeable. A part of the enclosure or sub-compartment, for example, the wall, side or part thereof, may be a non-porous polymer or ceramic. Biocompatible polymers and ceramics are preferred. A part of the encapsulant can be formed by a sealant, such as silicone, epoxy, polyurethane, or similar materials. A biocompatible sealant is preferred.

[0080] In addition, the conductivity of graphene-based films or films of other two-dimensional materials may allow charging to occur from an external power source. In an exemplary embodiment, an AC or DC voltage may be applied to the conductive two-dimensional material of the encapsulant. The conductive properties of graphene can provide additional gating to charged molecules. The charging can occur permanently or only for a while to affect the gating. Directed gating of charged molecules can not only be guided through pores (or restricted through pores), but also can be guided to the surface of graphene to adsorb or bind and promote growth, promote the formation of protective layers, or other biochemical effects on the body Provide the basis or mechanism.

[0081] In such embodiments, both permanent and temporary bonding with graphene are possible. In addition to the aforementioned advantages, the embodiments described herein may also have the advantage that they not only represent disruptive technologies for existing technology vehicles and other devices, but they also allow for new ways to use These vehicles and devices. For example, cell line development, therapeutic release agents, sensing paradigms (e.g., MRSw, NMR-based magnetic relaxation switch technology, see; Koh et al. (2008) Ang. Chem. Int'l Ed. Engl, 47 ( 22) 4119-4121) is used in the encapsulation described herein to reduce biofouling and oxidative damage (bioreactivity), transmit excellent permeability and less response delay, and provide mechanical stability. That is, the encapsulant described herein can allow existing technologies to be implemented in new ways that are currently impossible.

[0082] In addition to the in vivo and in vitro uses described above, the embodiments described herein can also be utilized in other fields. The encapsulates described herein can also be used for non-therapeutic applications, for example, the dosage of probiotics in dairy products (as opposed to currently used microencapsulation techniques that increase the vitality of delivery to the gastrointestinal tract during processing). In this and other aspects, it should be noted that the resulting enclosures and devices described herein can span several orders of magnitude in size, depending on manufacturing technology and different target usage requirements. However, it is believed that the encapsulant can be made small enough to circulate through the bloodstream. On the other hand, the encapsulant can be made large enough to be implanted (on the order of a few inches or more). These properties can result from the two-dimensional nature of graphene and its growth on a large surface area.

[0083] Although the present disclosure has been described with reference to the disclosed embodiments, those skilled in the art will readily understand that these are only for exemplifying the present disclosure. It should be understood that various modifications can be made without departing from the spirit of the present disclosure. The present disclosure can be modified to incorporate any number of variations, changes, substitutions, or equivalent arrangements not described so far, but these correspond to the spirit and scope of the present disclosure. In addition, although various embodiments of the present disclosure have been described, it should be understood that aspects of the present disclosure may include only some of the described embodiments. Therefore, the present disclosure should not be construed as being limited to the foregoing description.

[0084] Each concept or combination of components described or illustrated can be used to practice the present invention, unless otherwise specified. The specific names of the compounds are intended to be exemplary, as it is known that those skilled in the art can name the same compound differently. When a compound is described herein by, for example, a formula or chemical name without specifying the specific isomer or enantiomer of the compound

In the case of isomers, the description is intended to include each isomer and isomer of the compound described separately or in any combination. Those skilled in the art will understand that the methods, device elements, raw materials, and synthesis methods can be used in the practice of the present invention without undue experimentation, except for those explicitly exemplified. All functional equivalents of any such methods, device elements, raw materials, and synthesis methods known in the art are intended to be included in the present invention. Whenever a range is given in, for example, a temperature range, a time range, or a composition range, all intermediate ranges and subranges, and all individual values included in the given range are intended to be included in the present disclosure. When a Markush group or other grouping is used herein, all individual members of the group and all combinations and possible subcombinations of the group are intended to be individually included in this disclosure.

[0085] As used herein, "comprising" and "including", "containing" or "characterized by" are synonymous, and are inclusive or open-ended, and do not exclude other unlisted elements Or method steps. As used herein, consisting of excludes any element, step or ingredient not specified in the claim element. As used herein, consisting essentially of does not exclude materials or steps that do not substantially affect the basic and new features of the claims. Especially in the description of the components of the composition or in the description of the elements of the device, the term comprising (comprising) any enumeration herein should be understood as encompassing these combinations consisting essentially of the listed components or elements And methods, and these compositions and methods consisting of the listed components or elements. The invention exemplarily described herein can be implemented without any element or any limitation that is not specifically disclosed herein.

[0086] The terms and expressions that have been adopted are used as descriptive terms rather than as limiting terms, and the use of such terms and expressions does not exclude any equivalents or parts of the features shown or described. It is intended, but it should be recognized that various modifications are possible within the scope of protection required by the present invention. Therefore, it should be understood that although the present invention has been specifically disclosed through preferred embodiments and optional features, the concepts disclosed herein can be modified and changed by those skilled in the art, and such modifications and changes are considered It is within the scope of the invention as defined by the appended claims.

[0087] Generally, the terms and phrases used herein have their field-recognized meanings, which can be found by referring to standard texts, journal references, and backgrounds known to those skilled in the art. The foregoing definition is provided to clarify its specific use in the context of the present invention.

[0088] All references in this application, such as patent documents (including published or authorized patents or equivalents; patent application publications); and non-patent literature documents or other original materials, are hereby incorporated by reference in their entirety. , As if separately incorporated by reference, to the extent that each reference is not at least partially inconsistent with the disclosure of this application (for example, the reference will be partially inconsistent with the disclosure of this application, except for the partially inconsistent part of the reference Incorporated by reference).

All patents and publications mentioned in [0089] indicate the level of technical personnel in the field to which the invention belongs. The references cited herein are incorporated herein by reference in their entirety to indicate the prior art in the field, in some cases from their filing date, and the intention is that the information can be used herein (if necessary) to exclude (e.g., , Abandon) the specific implementation of the prior art. For example, when protecting compounds, it should be understood that compounds known in the art, including certain compounds disclosed in references disclosed herein (especially in cited patent documents), are not intended to be included in the claims.