Three-dimensional biocompatible matrices and their use in wound management - Patents.com

Abstract

The present invention relates to a three-dimensional biocompatible matrix comprising a three-dimensional scaffold made of a polymeric agent and further comprising at least one component. The present invention also relates to a composition comprising such a three-dimensional biocompatible matrix. Furthermore, the present invention also relates to the use of such a three-dimensional biocompatible matrix and / or composition. In particular, the present invention also relates to such a three-dimensional biocompatible matrix and / or composition for use in a method of wound treatment or management. Furthermore, the present invention also relates to a method for producing the three-dimensional biocompatible matrix according to the present invention. Furthermore, the present invention also relates to a three-dimensional (3D) printing ink composition and its use in producing the three-dimensional biocompatible matrix according to the present invention.

Description

[Technical field]

[0001] The present invention relates to a three-dimensional biocompatible matrix comprising a three-dimensional scaffold made of a polymeric agent and comprising at least one further component. The present invention also relates to a composition comprising such a three-dimensional biocompatible matrix. Furthermore, the present invention also relates to the use of such a three-dimensional biocompatible matrix and / or composition. In particular, the present invention also relates to such a three-dimensional biocompatible matrix and / or composition for use in a method of wound treatment or management. Furthermore, the present invention also relates to a method for producing the three-dimensional biocompatible matrix according to the present invention. Furthermore, the present invention relates to a three-dimensional (3D) printing ink composition and its use in producing the three-dimensional biocompatible matrix according to the present invention. [Background technology]

[0002] Due to the complexity of the processes involved in the healing process, wounds may take a significant period of time to heal and close. Often, wounds are painful and their healing may be complicated by infection that may cause further complications. Medical treatment of wounds, also called "wound management", should ideally support wound healing, prevent wound infection and relieve wound pain. Additionally, scar formation should be limited or prevented altogether. An important aspect of wound management is to facilitate the natural healing process with medical intervention while not adversely affecting it. Burn wounds are of particular interest due to their frequency. Causes of burns include fire, radiation, electrical current, chemicals and friction. Depending on the severity of the burn, burns / wounds are classified into different categories:

[0003] First-degree burns involve only the epidermis and cause only minor damage to the skin. These burns usually heal spontaneously and do not form scars. A typical example of such a first-degree burn is a minor sunburn. Conservative treatment with skin care creams is recommended but not strictly necessary. Antibiotics and wound dressings are not usually required for these burns.

[0004] Second-degree burns, also known as partial thickness burns, affect the epidermis and part of the dermis. The burn area appears red, blistered, swollen, and painful. The wound may ooze or bleed. These burns usually heal in one to three weeks, but the skin may become discolored after healing. These burns generally do not leave a raised scar, but treatment depends on the depth of the burn. Ointments and special dressings may be used. Very deep second-degree burns may require surgery. In some cases, treatment includes removal of dead tissue and antibiotic treatment.

[0005] Third-degree burns involve destruction of the epidermis and dermis, and may extend to the innermost layer of skin, the subcutaneous tissue. The burned area may look whitish or charred. Sensation in the skin may be impaired because nerve endings have been destroyed. Treatment involves removal of dead tissue and antibiotic treatment, as there is a high risk of bacterial infection. Treatment may also involve skin grafts or temporary coverage with a skin substitute.

[0006] Fourth-degree burns involve both layers of the skin, the subcutaneous tissue, deeper tissues, and sometimes even muscle and bone. Treatment requires surgery, skin grafts, and often amputation of the affected limb.

[0007] In general, there are various options for treating burns, with different combinations depending on the severity of the burn. These include wound cleansing, removal of necrotic tissue, moisturizing, wound care with antibiotics, either alone or in combination with appropriate dressings or bandages, surgery, skin grafting, and coverage with skin substitutes. Skin grafts can be from the patient's own skin (autografts), from healthy individuals (allografts), or from animals (xenografts). The use of skin grafts is limited, especially in patients with large areas of the body affected, due to the limited availability of suitable healthy skin. One approach to solving this problem has been to artificially create skin tissue patches by in vitro culture or use artificial skin substitutes that mimic the properties of natural skin and the extracellular matrix, and these methodologies have been used with varying degrees of success.

[0008] Therefore, there is a need in the art to provide an alternative in the treatment and management of wounds, especially burns.There is also a need in the art for a skin substitute for wound management that can alleviate the pain and / or reduce inflammation of such wounds.There is also a need in the art to provide a skin substitute for wound management that can promote wound healing and / or remodel the skin in and / or around the wound.There is also a need in the art to provide a skin substitute that can safely and effectively deliver painkillers to such wounds. [Summary of the Invention] [Brief description of the drawings]

[0009] [Figure 1] FIG. 1 shows schematic diagrams of exemplary structures A and B of compositions in embodiments of the present invention, in which the biocompatible matrix comprises two different types of active ingredients (= ingredients selected from non-opioid analgesics, extracellular vesicles, and artificially constructed lipid vesicles). [Diagram 2]FIG. 2 shows schematic diagrams of exemplary structures A and B of compositions according to embodiments of the present invention, in which the biocompatible matrix contains one active ingredient (= an ingredient selected from non-opioid analgesics, extracellular vesicles and artificially constructed lipid vesicles). DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0010] In a first aspect, the present invention relates to a three-dimensional biocompatible matrix comprising: - a three-dimensional scaffold made of a polymeric agent, and at least one component selected from a non-opioid analgesic, extracellular vesicles derived from animal cells, preferably derived from mammalian cells, more preferably derived from human cells, and artificially constructed lipid vesicles having a size in the range of 20 nm to 150 nm, wherein said at least one component is incorporated into said scaffold.

[0011] In one embodiment, the scaffold is dimensionally stable and is not a spreadable semi-solid or liquid such as a cream, lotion, ointment, salve, paste, ointment, or balm.

[0012] In one embodiment, the scaffold is manufactured by three-dimensional (3D) printing, preferably by three-dimensional (3D) screen printing.

[0013] In one embodiment, the scaffold is not knitted, woven or non-woven and / or does not have a thread-like structure, and preferably the scaffold is resorbable.

[0014] In one embodiment, the three-dimensional scaffold and the three-dimensional biocompatible matrix do not contain cells, particularly amniotic cells, and are not derived from amniotic membrane or solubilized amniotic membrane (SAM).In one embodiment, the three-dimensional scaffold and the three-dimensional biocompatible matrix are acellular.The term "acellular" as used in this context means that such scaffolds and matrices are acellular and do not contain any biological cells before being used (in wound management or wound treatment).

[0015] In another embodiment, the scaffold also does not have a thread-like structure, is not knitted, woven or nonwoven, but preferably in such another embodiment, the scaffold is not resorbable.In such an embodiment, the scaffold is typically made of a polymer that is chemically inert under physiological conditions (as would be encountered in a wound) and is preferably not susceptible to degradation or other processes that lead to its disappearance, such as enzymatic processes.Such polymers may be synthetic polymers, such as poly(methyl)methacrylate, polyurethane, polycaprolactone, polyamide, polydioxanone, and mixtures or combinations of such synthetic polymers, or silicone-based polymers, such as polysiloxanes, such as polydimethylsiloxanes, or mixtures or combinations thereof.

[0016] In one embodiment, the scaffold has a thickness in the range of 100 μm to 20 mm, preferably in the range of 100 μm to 15 mm, more preferably in the range of 100 μm to 12 mm, even more preferably in the range of 100 μm to 10 mm, and even more preferably, the scaffold has a minimum thickness in the range of 100 μm to 200 μm, allowing it to accommodate a monolayer of cells, in particular fibroblasts and / or keratinocytes. Typically, when the scaffold contains cells, for example in the form of a continuous monolayer or a patch of such a monolayer, this only occurs when the three-dimensional biocompatible matrix is ​​applied to the wound and the cells contained within the scaffold originate from the wound tissue or tissue surrounding the wound. That is, in such an embodiment before the application of the three-dimensional biocompatible matrix to the wound, such a matrix does not contain cells. It should also be noted that in a preferred embodiment, the three-dimensional biocompatible matrix before application to the wound is sterile and does not contain microorganisms capable of replicating and / or growing.

[0017] In one embodiment, said three-dimensional biocompatible matrix is ​​in dry form, i.e. does not contain a liquid phase. If such a three-dimensional biocompatible matrix is ​​in dry form, it is preferably rendered in such dry form after production by a suitable drying process, such as evaporation or freeze-drying. Such "dry" embodiments are particularly useful when the three-dimensional biocompatible matrix is ​​to be stored for long periods of time and / or shipped and / or is not (yet) subjected to any (medical) use. In one embodiment, said three-dimensional biocompatible matrix further comprises a liquid phase, such that the three-dimensional scaffold is in a gel state, preferably a hydrogel state. Such an embodiment is particularly useful and is in fact a preferred embodiment when the three-dimensional biocompatible matrix is ​​to be used for medical purposes, for example in methods of treating and / or managing wounds. When the three-dimensional scaffold is in a hydrogel state, the liquid phase is water or an aqueous solution. Also, when the three-dimensional scaffold is in a hydrogel state and the three-dimensional biocompatible matrix is ​​nevertheless stored for some time, it is preferred that the three-dimensional biocompatible matrix is ​​enclosed in a sealed compartment, for example a sealed bag or pouch, so that the liquid phase does not evaporate.

[0018] In one embodiment, the three-dimensional biocompatible matrix comprises: a) - a scaffold made of a polymeric agent, and - a non-opioid analgesic incorporated into said scaffold; or b) - a scaffold made of a polymeric agent, and - extracellular vesicles derived from animal cells, preferably derived from mammalian cells, more preferably derived from human cells, incorporated into said scaffold, or c) - a scaffold made of a polymeric agent, and - artificially constructed lipid vesicles having a size in the range of 20 nm to 150 nm, incorporated into said scaffold; or d) - a scaffold made of a polymeric agent, and - a non-opioid analgesic drug and extracellular vesicles derived from animal cells, preferably derived from mammalian cells, more preferably derived from human cells, incorporated into said scaffold; or e) - a scaffold made of a polymeric agent, and - an artificially constructed lipid vesicle having a size ranging from 20 nm to 150 nm and a non-opioid analgesic drug incorporated into said scaffold; or f) - a scaffold made of a polymeric agent, and - a non-opioid analgesic drug incorporated into said scaffold, and extracellular vesicles derived from animal cells, preferably derived from mammalian cells, more preferably derived from human cells, and artificially constructed lipid vesicles having a size in the range of 20 nm to 150 nm.

[0019] In one embodiment, the polymeric agent is selected from the group comprising: a) naturally occurring polymers selected from polysaccharides, such as agarose, chitin, chitosan, dextran, alginates, carrageenan, cellulose, starch, fucoidan, laminaran, glycosaminoglycans, copolymers of glycosaminoglycans with collagen; gums selected from xanthan gum, gum arabic, gum ghatti, guar gum, locust bean gum, gum tragacanth, gum karaya; and inulin; polypeptides, such as collagen, and hydrolyzed forms of such polypeptides, such as gelatin; polyamino acids, such as polylysine; polynucleotides; b) synthetic polymers, such as poly (α-hydroxy acids) such as polylactic acid (PLA), polyglycolic acid (PGA), polylactic-co-glycolic acid (PLGA), poly(alkyl)methacrylates such as poly(methyl)methacrylate, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), polyurethanes, polycaprolactones, polyamides, polydioxanones, polyglycerins, and mixtures or combinations of such synthetic polymers b) with such naturally occurring polymers a); c) silicon-based polymers such as polysiloxanes, e.g. polydimethylsiloxane; and combinations or mixtures of any of the above a)-c).

[0020] Preferred embodiments of glycosaminoglycans include hyaluronic acid, heparin, heparan sulfate, chondroitin sulfate, and keratan sulfate.

[0021] As used herein, the term "cellulose" is meant to include cellulose derivatives such as cellulose ether derivatives (e.g., methyl cellulose, ethyl cellulose, propyl cellulose, hydroxypropyl cellulose, hydroxypropyl methyl cellulose, hydroxyethyl cellulose, carboxymethyl cellulose, etc.) and cellulose ester derivatives (e.g., cellulose acetate (CA), cellulose acetate phthalate (CAP), cellulose acetate butyrate (CAB), cellulose acetate trimellitate (CAT), and hydroxypropyl methyl cellulose phthalate (HPMCP), etc.).

[0022] In one embodiment, the polymeric agent is selected from naturally occurring polymers, for example selected from polysaccharides such as agarose, chitin, chitosan, dextran, alginates, carrageenan, cellulose, starch, fucoidan, laminaran, glycosaminoglycans, copolymers of glycosaminoglycans and collagen, gums selected from xanthan gum, gum arabic, gum ghatti, gum guar, locust bean gum, gum tragacanth, gum karaya, and inulin, polypeptides such as collagen, and hydrolyzed forms of such polypeptides, for example gelatin, polyamino acids such as polylysine, polynucleotides, wherein preferably the polymeric agent is selected from alginates, gelatin, cellulose, collagen, chitosan, and mixtures of any of the foregoing.

[0023] In one embodiment, the non-opioid analgesic is selected from aminoester analgesics, aminoamide analgesics, fomocaine, and carbonate adducts of the aminoester analgesics or the aminoamide analgesics or the fomocaine, where preferably the aminoester analgesics include procaine, chloroprocaine, oxybuprocaine, benzocaine, tetracaine, and proxymethacaine; where preferably the aminoamide analgesics include lidocaine, mepivacaine, prilocaine, articaine, bupivacaine, ropivacaine, scinticaine, and etidocaine; and where preferably the fomocaines include fomocaine and its C-alkylmorpholine derivatives.

[0024] In one embodiment, the non-opioid analgesic is an aminoester analgesic or its carbonate adduct, preferably selected from procaine, chloroprocaine, oxybuprocaine, benzocaine, tetracaine, and proxymethacaine, and their respective carbonate adducts.

[0025] In one embodiment, the non-opioid analgesic is an aminoamide analgesic or its carbonate adduct, preferably selected from lidocaine, mepivacaine, prilocaine, articaine, bupivacaine, ropivacaine, scinticaine, and etidocaine, and their respective carbonate adducts.

[0026] Carbonate adducts of the aforementioned analgesics are known and are described, for example, in WO2019 / 048590A1. Particularly preferred analgesics are procaine, chloroprocaine, and lidocaine, and their respective carbonate adducts.

[0027] In one embodiment, the non-opioid analgesic is selected from acetylsalicylic acid, ibuprofen, diclofenac, naproxen, indomethacin, paracetamol, metamizole, phenazone, propyphenazone, parecoxib, celecoxib, etoricoxib, ketamine, capsaicin, ziconotide, cannabinoids, and flupirtine.

[0028] In one embodiment, the extracellular vesicles are derived from animal platelets or stem cells, preferably mammalian platelets or stem cells, more preferably human platelets or stem cells, and contain at least one component selected from cytokines, growth factors, transcription factors, RNA, particularly microRNA, and mRNA; wherein preferably, the stem cells are selected from mesenchymal stem cells and induced stem cells, and wherein more preferably, the mesenchymal stem cells are derived from bone marrow, umbilical cord blood, adipose tissue, or amniotic fluid; wherein preferably, the artificially constructed lipid vesicles contain at least one component selected from cytokines, growth factors, transcription factors, RNA, particularly microRNA, mRNA, and non-opioid analgesics as defined herein. It should be noted that the extracellular vesicles are not derived from the extracellular matrix and do not contain extracellular matrix or extracellular matrix components.

[0029] In one embodiment, a) said extracellular vesicles are derived from animal platelets, preferably mammalian platelets, more preferably human platelets, and are positive for at least one cell marker selected from CD9, CD41a, CD41b, CD42b, CD61, CD62P, CD63 and syntenin, and / or said extracellular vesicles are positive for at least one cell marker selected from CD81, CD3, CD4, CD19, CD20, CD2, CD8, CD11a and CD25. or b) the extracellular vesicles are derived from stem cells and are positive for CD81, CD9, CD63, Tsg101, and HSP70, and are positive for one, some, or all of CD105, CD90, CD73, and CD44, and / or the extracellular vesicles are negative for one, some, or all of CD45, CD34, CD31, CD19, CD79α, CD14, CD11b, and HLA-DR.

[0030] In one embodiment, said three-dimensional biocompatible matrix further comprises cells, in particular fibroblasts and / or keratinocytes, obtained from living or cadaveric tissue. If such cells are comprised within the three-dimensional biocompatible matrix, they are typically already incorporated during the manufacture of the three-dimensional biocompatible matrix, i.e., they may already be comprised by such three-dimensional biocompatible matrix prior to its application / use.

[0031] In another embodiment different from the embodiment of the previous paragraph, said three-dimensional biocompatible matrix does not further comprise cells such as fibroblasts and / or keratinocytes. In particular, in this embodiment, said three-dimensional biocompatible matrix before application / use does not comprise cells. This means that no cells are included or incorporated into the manufacture of such a three-dimensional biocompatible matrix, and such a biocompatible matrix does not comprise any biological cells before application / use. Only after its application, for example after being placed on a wound, can the three-dimensional biocompatible matrix be colonized by biological cells from the wound or its surroundings.

[0032] The present invention also relates to a kit configured for producing a three-dimensional biocompatible matrix as defined herein, said kit comprising a three-dimensional scaffold made of a polymeric agent as defined herein and a container comprising said at least one component as defined herein, preferably said container comprising a solution or dispersion or suspension of said at least one component in a suitable solvent.

[0033] The present invention also relates to a composition comprising a three-dimensional biocompatible matrix according to the invention as defined herein.

[0034] In one embodiment, a composition according to the invention is prepared as a patch, dressing, pad, plaster, bandage, band-aid, patch, or pouch.

[0035] In one embodiment of the composition according to the invention, the composition, in particular when the composition is used, has a layered arrangement, for example a laminate or a multilayer arrangement, preferably one of the layers of the layered arrangement is formed by the three-dimensional biocompatible matrix. In such an embodiment, the composition preferably comprises at least one additional layer attached to the three-dimensional biocompatible matrix. Such an additional layer attached to the three-dimensional biocompatible matrix may be a carrier layer providing mechanical support to the biocompatible matrix, or a semi-permeable backing layer preventing diffusion of the at least one component from one side of the biocompatible matrix, or a solvent-impermeable backing layer, in particular a water-impermeable backing layer, sealing the biocompatible matrix on one side and preventing diffusion of the at least one component from the biocompatible matrix on such side. In such an embodiment, it is particularly preferred that such a layered arrangement has a thickness in the range of 5 mm to 20 mm, preferably 5 mm to 15 mm, more preferably 8 mm to 12 mm, more preferably 9 mm to 11 mm, even more preferably about 10 mm. In one embodiment, the three-dimensional biocompatible matrix has a thickness in the range of 100 μm to 20 mm, preferably 100 μm to 15 mm, more preferably 100 μm to 12 mm, and even more preferably 100 μm to 10 mm; and the additional layer has a thickness in the range of 50 μm to 2 mm, preferably 100 μm to 2 mm, more preferably 100 μm to 1 mm, and even more preferably 100 μm to 500 μm. The embodiment(s) of the composition may also include additional components, for example, when the composition is not in use but is stored or transported; for example, the composition may be attached to a removable carrier sheet for transport and / or storage. Such a removable carrier sheet may serve as a means for safe transport and / or storage of the composition, but is removed from the composition prior to the intended use of the composition.

[0036] In another embodiment of the composition, the composition has a layered arrangement, but comprises only a monolayer formed by the three-dimensional biocompatible matrix, especially when the composition is used. In such an embodiment, when used, the composition has a monolayer arrangement, such monolayer being made of a polymeric agent as defined herein, or a combination or mixture of polymeric agents as defined herein; such monolayer constitutes a three-dimensional biocompatible matrix. In such an embodiment, the composition preferably does not comprise an additional layer or sheet attached to the three-dimensional biocompatible matrix. In such an embodiment, it is particularly preferred that such a monolayer arrangement has a thickness in the range of 0.5 mm to 3 mm, preferably in the range of 0.5 mm to 2 mm, more preferably in the range of 1 mm to 2 mm. However, it should be noted that in the above (monolayer) embodiment of the composition, when the composition is not in use but is stored or transported, it may also comprise additional components; for example, it may be attached to a re-peelable carrier sheet for transport and / or storage of the composition. Such a re-peelable carrier sheet may serve as a means for safe transport and / or storage of the composition, but is removed from the composition before the intended use of the composition. It should also be noted that when such a monolayer constituting a three-dimensional biocompatible matrix is ​​produced by three-dimensional (3D) screen printing, due to the inherent modality of the screen printing process, such a screen-printed monolayer may consist of several sublayers, however all of these sublayers are made of the same polymeric agent as defined herein, or the same combination or mixture of such polymeric agents as defined herein, and such sublayers collectively constitute a monolayer that is a three-dimensional biocompatible matrix.

[0037] In one embodiment of the composition, the composition has one of the following two structures A or B:

[0038] Structure A: the three-dimensional biocompatible matrix is ​​disposed on a first layer, and the composition comprises an additional layer attached to the first layer, which additional layer is a carrier layer providing mechanical support to the biocompatible matrix, or a semipermeable backing layer preventing diffusion of the at least one component from one side of the biocompatible matrix, or a solvent-impermeable backing layer, in particular a water-impermeable backing layer, sealing the biocompatible matrix on one side and preventing diffusion of the at least one component out of the biocompatible matrix on that side, or the additional layer is both a carrier layer and a semipermeable backing layer or a solvent-impermeable backing layer; the first layer has a thickness in the range of 100 μm to 20 mm, preferably 100 μm to 15 mm, more preferably 100 μm to 12 mm, even more preferably 100 μm to 10 mm; and the additional layer has a thickness in the range of 50 μm to 2 mm, preferably 100 μm to 2 mm, more preferably 100 μm to 1 mm, even more preferably 100 μm to 500 μm; or Structure B: the three-dimensional biocompatible matrix is ​​disposed as a sole layer within the composition, with no additional layers attached to the layer of the three-dimensional biocompatible matrix; and wherein the sole layer has a thickness in the range of 500 μm to 3 mm, preferably in the range of 500 μm to 2 mm, more preferably in the range of 1 mm to 2 mm; wherein, optionally, said composition of Structure A and said composition of Structure B may be attached to a removable carrier sheet for transport and / or storage of said compositions, and when said compositions are in use, e.g. during wound treatment and / or management, they are not attached to or do not comprise such a removable carrier sheet.

[0039] In one embodiment of the composition having Structure A or B, during transport and / or storage, the composition of Structures A and B is attached to a removable carrier sheet for transport and / or storage of the composition, and when the composition is in use, e.g., during wound treatment and / or management, it is not attached to or does not contain such a removable carrier sheet.

[0040] Exemplary structures A and B of compositions in accordance with embodiments of the present invention are shown in FIGS.

[0041] In a further aspect, the present invention also relates to a three-dimensional biocompatible matrix as defined herein or a composition according to the invention for use in a method of treating and / or managing a wound.

[0042] It should be noted that the three-dimensional biocompatible matrices and compositions according to the present invention are prefabricated matrices and prefabricated compositions. The term "prefabricated" used in this context preferably means that such matrices and compositions are ready-made and complete, separated from the wound before being applied to the wound. Preferably, such prefabricated matrices and such prefabricated compositions do not require in situ printing on the wound, in situ crosslinking on the wound, or in situ polymerization on the wound; instead, the prefabricated matrices and prefabricated compositions according to the present invention are printed and manufactured at a site and time separate from the wound to be treated and from the time when the wound treatment is performed. More specifically, the prefabricated matrices and compositions according to the present invention are not injectable solutions or injectable scaffolding materials, or free-flowing solutions or scaffolding materials. Furthermore, the prefabricated matrices and compositions according to the present invention are not polymerized, cured, crosslinked or hardened in situ, i.e. in the wound to be treated or managed, but are prepared at a site and time separate from such wound and application to such wound.

[0043] In one embodiment of the three-dimensional biocompatible matrix or composition for use according to the present invention, the wound is an acute wound or a chronic wound, preferably the acute wound is a burn, a skin lesion, a skin injury, a surgical wound, a cut or a puncture wound, preferably the chronic wound is a pressure sore or a diabetic ulcer; wherein, more preferably, the burn is selected from thermal burns, radiation burns, chemical burns, burns due to electrical current exposure, and friction burns, wherein, even more preferably, the burn is a first, second or third degree burn, even more preferably, a second or third degree burn.

[0044] In one embodiment of the three-dimensional biocompatible matrix or composition for use according to the present invention, the method of treating and / or managing a wound comprises applying the three-dimensional biocompatible matrix or composition to the wound and leaving it in contact with the wound for a defined period of time.

[0045] In one embodiment of the three-dimensional biocompatible matrix or composition for use according to the invention, said method is for relieving pain and / or reducing inflammation in said wound.

[0046] In one embodiment of the three-dimensional biocompatible matrix or composition for use according to the invention, the method is for increasing the rate at which wound healing is achieved compared to a wound not treated by the method, and / or the method is for the delivery of a pain relieving agent to the wound, and / or the method is for the reconstruction of skin in and / or around the wound.

[0047] In a further aspect, the present invention also relates to the use of the three-dimensional biocompatible matrix or composition according to the present invention for the manufacture of a medicament for treating and / or managing a wound. In such aspect, the three-dimensional biocompatible matrix, the composition and the wound are as defined herein. In such aspect, said medicament is used in a method for treating and / or managing a wound. More particularly, in such aspect, said method for treating and / or managing a wound comprises applying said three-dimensional biocompatible matrix or said composition to said wound and leaving it in contact with such wound for a defined period of time.

[0048] In one embodiment of this aspect, the method in which the medicament is used is for relieving pain and / or reducing inflammation of the wound.

[0049] Further, in addition to or in the alternative to the above embodiments, the method is for increasing the rate at which wound healing is achieved compared to a wound not treated by the method, and / or the method is for delivery of a pain relieving agent to the wound, and / or the method is for remodeling the skin in and / or around the wound.

[0050] In a further aspect, the present invention also relates to a method of treating and / or managing a wound, such method comprising applying to said wound a three-dimensional biocompatible matrix as defined herein or a composition according to the present invention and leaving it in contact with said wound for a defined period of time, and in this aspect, in one embodiment, said method is for relieving pain and / or reducing inflammation of said wound.

[0051] Further, in addition to or in the alternative to the previous embodiments of this aspect, the method is for increasing the rate at which wound healing is achieved compared to a wound not treated by the method, and / or the method is for delivering a pain relieving agent to the wound, and / or the method is for remodeling the skin in and / or around the wound.

[0052] In a further aspect, the present invention also relates to a method for producing a three-dimensional biocompatible matrix according to the present invention as defined herein, said method comprising: One of the following: a) providing a polymeric agent and 3D printing, preferably 3D screen printing, such agent onto a three-dimensional scaffold; b) applying at least one component selected from a non-opioid analgesic, extracellular vesicles derived from animal cells, preferably derived from mammalian cells, more preferably derived from human cells, and artificially constructed lipid vesicles having a size ranging from 20 nm to 150 nm to said three-dimensional scaffold, and allowing said at least one component to be incorporated into said scaffold, thereby producing said three-dimensional biocompatible matrix; said polymeric agent and said at least one component are as defined herein; or a*) providing at least one component selected from a polymeric agent, a non-opioid analgesic agent, extracellular vesicles derived from animal cells, preferably derived from mammalian cells, more preferably derived from human cells, and artificially constructed lipid vesicles having a size in the range of 20 nm to 150 nm, and mixing the polymeric agent and the at least one component; and then b*) 3D printing, preferably 3D screen printing, the mixture obtained from step a*) onto a three-dimensional scaffold such that said at least one component is incorporated into said scaffold, thereby producing said three-dimensional biocompatible matrix; The polymeric agent and the at least one component are as defined herein.

[0053] In one embodiment, said step b) may be carried out by any suitable technique including spraying, dipping, coating, soaking, impregnation, application followed by freeze-drying, nanosol techniques, application followed by exposure of the scaffold / matrix to negative pressure or vacuum, etc.

[0054] In a further aspect, the present invention also relates to an ink composition for three-dimensional (3D) printing, in particular for three-dimensional (3D) screen printing, said ink composition comprising: i. A polymeric agent selected from the group including: a) naturally occurring polymers selected from polysaccharides, such as agarose, chitin, chitosan, dextran, alginates, carrageenan, cellulose, starch, fucoidan, laminaran, glycosaminoglycans, copolymers of glycosaminoglycans and collagen; gums selected from xanthan gum, gum arabic, gum ghatti, guar gum, locust bean gum, gum tragacanth, gum karaya; and inulin; polypeptides, such as collagen, and hydrolyzed forms of such polypeptides, such as gelatin; polyamino acids, such as polylysine; polynucleotides; b) synthetic poly synthetic polymers such as poly(α-hydroxy acids), e.g. polylactic acid (PLA), polyglycolic acid (PGA), polylactic-co-glycolic acid (PLGA), poly(alkyl)methacrylates, e.g. poly(methyl)methacrylate, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), polyurethanes, polycaprolactones, polyamides, and mixtures or combinations of such synthetic polymers b) with such naturally occurring polymers a); c) silicone-based polymers, e.g. polysiloxanes, e.g. polydimethylsiloxane; and combinations or mixtures of any of the foregoing a)-c); ii. a solvent for i, preferably said solvent is selected from water, an aqueous solution, an alcohol, in particular methanol, ethanol, n-propanol, or isopropanol, an alcohol solution, tetrahydrophane, acetone, and ethyl acetate; iii. at least one component selected from a non-opioid analgesic, an extracellular vesicle derived from an animal cell, preferably derived from a mammalian cell, more preferably derived from a human cell, and an artificially constructed lipid vesicle having a size in the range of 20 nm to 150 nm; wherein said non-opioid analgesic, the extracellular vesicle derived from an animal cell, preferably derived from a mammalian cell, more preferably derived from a human cell, and the artificially constructed lipid vesicle are as defined herein; iv. Optionally, one or more further ingredients selected from fillers, crosslinkers, binders, surfactants, additives, diluents, thickeners, colorants, dyes, stabilizers, buffers, humectants, emulsifiers, dispersants, and preservatives.

[0055] In a preferred embodiment, the ink composition for three-dimensional (3D) printing, in particular for three-dimensional (3D) screen printing, has a viscosity in the range of 100 to 300000 mPas, preferably 1000 to 30000 mPas. Preferably, such viscosity range values ​​are measured and determined according to the methodologies of DIN EN ISO 3219-1 and / or ISO 3219.

[0056] In one embodiment of the ink composition, the at least one component is present in the ink composition at a concentration ranging from 0.1 to 20 wt.%, based on the weight of the ink composition. It should be noted that in an embodiment in which the at least one component is applied to a three-dimensional (3D) biocompatible matrix, after such matrix is ​​formed, the at least one component is applied as part of a solution or dispersion or suspension in which the at least one component is present at a concentration ranging from 0.1 to 20 wt.%, based on the weight of such solution or dispersion or suspension.

[0057] [Detailed Description] As used herein, terms such as "of the [present] invention," "in accordance with the invention," "according to the invention," and the like, are intended to refer to all aspects and embodiments of the invention as described and / or claimed herein. As used herein, the term "comprising" is to be construed as embracing both "including" and "consisting of," both meanings being specifically and explicitly intended and therefore separately disclosed embodiments in accordance with the present invention. As used herein, "and / or" is to be considered as specifically disclosing each of the two specified features or components, with or without the other. For example, "A" and / or "B" is to be considered as a specific disclosure of each of (i) A, (ii) B, and (iii) A and B, as if each were individually defined herein. When an indefinite or definite article is used to refer to a singular noun, such as "a," "an," or "the," the plural of that noun is also included unless something else is specifically stated. Similarly, such disclosure is meant to be considered a specific disclosure of a single individual entity preceded by "a," "an," or "the."

[0058] The inventors have surprisingly found that the inclusion of at least one component selected from a non-opioid analgesic, an extracellular vesicle derived from an animal cell, preferably an extracellular vesicle derived from a mammalian cell, more preferably an extracellular vesicle derived from a human cell, and an artificially constructed lipid vesicle in a three-dimensional scaffold made of a polymeric agent results in a three-dimensional biocompatible matrix that can be advantageously used for medical purposes, in particular for methods of treating and / or managing wounds. In a preferred embodiment, said three-dimensional scaffold is manufactured by three-dimensional (3D) printing, preferably three-dimensional (3D) screen printing. These 3D printing techniques, in particular 3D screen printing techniques, are very versatile and very gentle, since harsh conditions such as high pressure or high temperature can be avoided. As a result, the use of 3D printing techniques, in particular 3D screen printing techniques, in the manufacture of such three-dimensional biocompatible matrices allows the inclusion and / or incorporation directly during the manufacturing process of delicate or sensitive components such as active pharmaceutical ingredients (APIs) or lipid vesicles, as well as biological cells. The direct inclusion and / or incorporation of such delicate or sensitive components during the manufacturing process allows these components to become an integral part of the three-dimensional scaffold, rather than being loosely contained in the pore spaces surrounding the three-dimensional scaffold. Furthermore, 3D printing techniques, and in particular 3D screen printing techniques, allow a high degree of precision, reproducibility, and extremely high resolution, such that structures in the micrometer range can be reliably and reproducibly generated, i.e., printed. Such 3D printing techniques, and in particular 3D screen printing techniques, avoid production at different locations, the transportation of intermediate products between such different locations, and possibly the use of complex manufacturing schemes involving different production stages or multiple processes.

[0059] The three-dimensional biocompatible matrix thus produced is also structurally different from other matrices such as bandages, dressings, patches, plasters, pouches, pads or gels commonly used in wound management. In particular, the three-dimensional scaffold of the three-dimensional biocompatible matrix according to the present invention (and therefore the matrix itself) does not have a classical woven structure in that it does not have a thread-like structure and / or is not knitted, woven or nonwoven. Furthermore, the three-dimensional scaffold of the three-dimensional biocompatible matrix according to the present invention (and therefore the matrix itself) is also structurally different from commercially available wound gels in that the three-dimensional scaffold is dimensionally stable and is not a spreadable semi-solid or liquid such as a cream, lotion, ointment, salve, paste, angulent or balm. It is also not a spreadable or free-flowing gel, nor is it soaked into a gauze pad, nonwoven, woven or other textile. Furthermore, it is not a gel that is structurally supported or anchored by a textile or fibrous mesh or the like. The term "dimensionally stable" as used herein preferably refers to a scenario in which said three-dimensional scaffold does not substantially change any of its dimensions, i.e. length, width or height, respectively, under conditions of storage or when placed in its intended use. In a preferred embodiment, "dimensionally stable" refers to a scenario in which said three-dimensional scaffold substantially maintains its dimensions when exposed to the conditions of its intended use, in particular the temperature conditions of its intended use, in particular temperature conditions ranging from -20°C to +50°C, in particular 0°C to +45°C, and / or humidity conditions of relative humidity ranging from 0% to 100%. This term particularly refers to a three-dimensional scaffold that is not free-flowing and free-diffusing. It is also preferably meant to refer to a scaffold that does not assume a completely different shape, extension, width and / or height when subjected to changes in the external conditions to which said scaffold is exposed. The term "dimensionally stable" is preferably meant to include swelling of the gel upon hydration or rehydration upon exposure of the gel to water and / or a moist environment, so long as the gel / hydrogel is not free-flowing and resists changes in overall shape even after swelling.More particularly, said three-dimensional scaffold is preferably "dimensionally stable" in that its dimensions do not change substantially when included in a wound management or wound treatment composition. More specifically, it is preferably "dimensionally stable" in that its dimensions do not change substantially when stored as part of a wound management composition, such as a patch, dressing, pad, bandage, dressing, band-aid, tape or pouch, or when applied to a wound. The term "dimensionally stable" is however preferably meant to include swelling when the three-dimensional scaffold is transferred from a dry storage state to a wet state ready for application to a wound or already applied, again with the proviso that in such wet state, such a three-dimensional scaffold is still not free-flowing, but retains its shape and provides resistance to changes in its overall shape, even after being in such wet state.

[0060] Furthermore, by appropriate selection of the at least one component contained by the three-dimensional biocompatible matrix, the quality of such a three-dimensional biocompatible matrix can be tailored to specific needs and / or requirements. For example, if pain caused by a wound needs to be relieved, the at least one component may be a non-opioid analgesic. However, if the treatment and management of the inflammatory aspect of the wound is of primary importance, the three-dimensional biocompatible matrix may contain extracellular vesicles that contain appropriate cytokines, growth factors and / or mRNA that can address inflammation. One advantage of such extracellular vesicles is that they are easily taken up by cells that form part of the affected tissue. In cases where extracellular vesicles are not available due to the lack of appropriate cells from which such extracellular vesicles originate, one may nevertheless consider including artificially constructed lipid vesicles in the three-dimensional biocompatible matrix loaded with the desired components to produce the desired effect. For example, such artificially constructed lipid vesicles may be loaded with non-opioid analgesic drugs as defined herein.

[0061] In a preferred embodiment, the "polymeric agent" as used herein is a gel-forming agent. As used herein, the term "gel-forming agent" refers to an agent capable of forming a gel when exposed to a suitable liquid phase. As used herein, the term "gel" refers to a solid three-dimensional network structure, such three-dimensional network structure having pores or interstitial spaces filled by a liquid phase. When such liquid phase is water or an aqueous solution, the resulting gel is also called a "hydrogel". When the liquid is an organic solvent, the gel is also called an "organogel". Examples of suitable organic solvents are ethanol, acetone, isopropanol, glycerol, PEG-400, 3-methoxy-3-methyl-1-butanol (MBB), propylene carbonate, and vegetable oils. In a preferred embodiment herein, the gel-forming agent is a hydrogel-forming agent, and the liquid phase used and included by the three-dimensional biocompatible matrix in such preferred embodiment is water or an aqueous solution. In such an aqueous solution, additional components that may be included are buffer components, preservatives, chelating agents, humectants, dexpanthenol, and the like. When water or such an aqueous solution is contained by the three-dimensional biocompatible matrix, the three-dimensional scaffold is in a hydrogel state or is a hydrogel.

[0062] The term "extracellular vesicles" as used herein refers to particles excreted or released by biological cells. In accordance with the present invention, said extracellular vesicles are not derived from the extracellular matrix and do not contain extracellular matrix or extracellular matrix components. Typically, the extracellular vesicles in the embodiments of the present invention have a lipid bilayer membrane and have a size in the range of 20 nm to 500 nm, preferably 20 nm to 300 nm, more preferably 20 nm to 200 nm, even more preferably 20 nm to 150 nm, and even more preferably 20 nm to 100 nm. In a particularly preferred embodiment, such extracellular vesicles have a lipid bilayer membrane and have a size in the range of 30 nm to 100 nm. In the expression "extracellular vesicles derived from animal cells, preferably from mammalian cells, more preferably from human cells", the term "derived from animal cells, preferably from mammalian cells, more preferably from human cells" refers to a scenario in which such extracellular vesicles are released from such animal cells, preferably from mammalian cells, more preferably from human cells. In a preferred embodiment, such extracellular vesicles are released from human cells. For example, such extracellular vesicles may be released from human platelets or stem cells. Depending on the respective cellular origin, the respective extracellular vesicles may differ in the composition and / or content of the respective membrane (and associated cell surface markers). Typically, extracellular vesicles have a content and surface that reflect the cellular origin of the vesicles. They may contain proteins and / or fragments thereof, as well as nucleic acids, all of which are derived from the vesicles. In an embodiment according to the invention, the extracellular vesicles may contain one or several components selected from cytokines, growth factors, transcription factors, proteolytic enzymes, RNA, in particular microRNA, and mRNA. Such components may have anti-inflammatory or pro-inflammatory effects, for example anti-inflammatory or pro-inflammatory cytokines. Proteolytic enzymes may contribute to wound excision. Methods for the production of extracellular vesicles are known to those skilled in the art and are described, for example, in Doyle et al., Cells, 2019, vol. 8, 727, doi:10.3390 / cells8070727.

[0063] The term "artificially constructed lipid vesicles" as used herein refers to vesicles that have a lipid membrane, usually also a lipid bilayer, and are artificially synthesized rather than released from living cells. Such artificially constructed lipid vesicles are sometimes called "liposomes". Such artificially constructed lipid vesicles have a size ranging from 20 nm to 150 nm, preferably from 20 nm to 100 nm. The outer shell is composed of a lipid membrane, typically a lipid bilayer, surrounding a core formed in a liquid phase. In one embodiment of the present invention, such artificially constructed lipid vesicles may contain proteins and / or fragments thereof as well as nucleic acids. In one embodiment, such artificially constructed lipid vesicles may contain one or several components selected from cytokines, growth factors, transcription factors, proteolytic enzymes, RNA, particularly microRNA, and mRNA. In one embodiment, such artificially constructed lipid vesicles may also contain a non-opioid analgesic.

[0064] The term "biocompatibility" as used herein in conjunction with said three-dimensional matrices, in the context of wound management, refers to the quality of such a matrix not eliciting undesirable local or systemic effects in a recipient of such matrix, and generating a beneficial cellular or tissue response in a recipient of such matrix, preferably an appropriate beneficial cellular or tissue response, more preferably the most appropriate beneficial cellular or tissue response.

[0065] The term "resorbable" as used herein in the context of a three-dimensional scaffold according to the invention refers to the property of the scaffold (and thus the matrix comprising the scaffold) to be resorbed by the biological tissue to which it is applied. Resorption can occur, for example, by degradation and subsequent metabolism of the scaffold.

[0066] Further aspects of the present invention are illustrated by the following schemes, examples, tables, figures and procedural descriptions, which are provided for illustrative purposes only and not for the purpose of limiting the present invention. The scope of protection of the present invention is limited only by the appended claims.

[0067] Additionally, please refer to the following figure.

[0068] In the following, reference is made to examples, which are given to illustrate but not to limit the invention. EXAMPLES

[0069] Example 1: Preparation of various inks (1.1-1.8) for three-dimensional (3D) screen printing of biocompatible matrices and preparation of ink 1.9 for three-dimensional (3D) screen printing of backing layers

[0070] 1.1 Dissolve 12 g of gelatin in 200 ml of PBS at 40 °C under stirring at 200 rpm. After complete dissolution, add 10 g of cellulose and stir for 10 min. Increase the stirring to 400 rpm and add 2 g of alginate to the ink and stir for 2 h. The ink is stored at 4° C. until further use.

[0071] 1.2 Add 2 g of collagen to 200 ml of H2O and stir at 200 rpm for 1 hour. After complete dispersion or dissolution, add 5 g of cellulose and 0.5 g of xanthan gum and stir for 1 hour. The ink is stored at 4° C. until further use.

[0072] 1.3 Add 100ml of 1.2% collagen dispersion to 100ml H2O. Control the pH and adjust to pH<7. After complete dispersion, add 2g alginate and stir for 2 hours. The ink is stored at 4° C. until further use.

[0073] 1.4 Add 2 g cellulose to 200 ml H2O and incubate overnight at 4 °C. The next day, stir the soaked cellulose for 1 h at room temperature. Add 10 ml glycerol and 1 g PEG and stir at 400 rpm for 1 h. Add 1.5 g collagen and stir at 400 rpm for 2 h. Mix the bioink further in a speed mixer for 10 min, increasing the speed from 800 rpm to 1500 rpm. The ink is stored at 4° C. until further use.

[0074] 1.5 Solubilize 10 g of nanocellulose in 100 ml of PBS, ddH2O or CO2-enriched water under constant stirring at 300 rpm. Further, add 1 g of alginate and stir at 450 rpm for 2 h. Incubate the bioink overnight at 4 °C. The next day, mix the bioink further for 10 min in a speed mixer and increase the speed from 800 rpm to 1300 rpm under vacuum. The ink is stored at 4° C. until further use.

[0075] 1.6 2 g of alginate is dissolved in 50 ml of PBS or ddH2O and mixed with 20 ml of 0.9% collagen suspension under constant stirring at 200 rpm. To the obtained bioink, additionally a non-opioid analgesic and / or EVs are added. For this purpose, 2.5 g of a non-opioid analgesic and 2.5 g of PVP are solubilized in 20 ml of CO2-enriched water. This non-opioid analgesic / PVP suspension is added to the bioink and mixed immediately. Furthermore, an EV concentration of 2.5x10 9 Add 10 ml of 1 / ml EV / PBS suspension (extracellular vesicles in phosphate buffered saline). The ink is stored at 4° C. until further use.

[0076] 1.7 Dissolve 2.5 g of alginate and 5.5 g of gelatin in 100 ml of PBS, ddH2O or CO2-enriched water under stirring at 400 rpm at 40 °C. Additionally, solubilize 5 g of non-opioid analgesic and 5 g of PVP in the resulting bioink. The ink is stored at 4°C until further use.

[0077] 1.8 Solubilize 3.5 g of non-opioid analgesic and 3.5 g of PVP in 100 ml of 7.5% nanocellulose gel under constant stirring at 400 rpm. The ink is stored at 4° C. until further use.

[0078] 1.9 Add 180g of silicone component A and mix with 20g of silicone component B. Add 0.1-0.5w% of thixotropic thickener and 0.5-4w% of setting time inhibitor and mix. Finally add 1-4w% of color paste. Mix this silicone paste in a speed mixer for 10 minutes at a speed of 800-1500 rpm.

[0079] Example 2: Fabrication of three-dimensional biocompatible matrices

[0080] A) The 3D printing ink compositions 1.1. to 1.8 are used to produce a three-dimensional biocompatible matrix in a 3D screen printing device. The inks 1.1. to 1.8 are preheated or precooled to a temperature of 4 to 50 ° C, preferably 20 ° C. Screen printing squeegees are used with an angle of 50 ° to 80 °, preferably 65 °, and a hardness of 65 to 85 Shore, preferably 75 Shore for submersion and 40 to 90 Shore, preferably 55 Shore for printing. The screen mesh is made of polyamide, polyester, or steel, with 8 to 400 threads / cm, preferably 48 threads / cm, and a thread thickness of 27 to 300 μm, preferably 55 μm. The EOM ("Emulsion over Mesh") of the screen varies between 8 to 150 μm, preferably 20 to 25 μm. The printing process to build the shape of the z-axis is a continuous repetition of submersion, printing, and curing. The ink is flood-printed through the screen mesh using a squeegee at a temperature of 4-65 °C, preferably 20-40 °C. The humidity in the printing chamber during the printing process is adjusted to 20-100%, preferably 60%. Curing of the printed ink is carried out at 0-180 °C using IR, convection drying, UV, or a cooling system. Preferably, the bioinks 1.1. to 1. are cured at a temperature of 4-40 °C. The humidity in the curing chamber can be adjusted to 10-60%, preferably 40%. The printing process is repeated until the final size of the three-dimensional biocompatible matrix is ​​achieved. The printing process can include the combination and / or rotation of the inks 1.1 to 1.8. Furthermore, the printing process can include multiple screens with different mesh compositions and geometries. When ink 1.6 or 1.8 is used, the active ingredient, i.e. "at least one ingredient selected from a non-opioid analgesic, an animal cell derived, preferably a mammalian cell derived, more preferably a human cell derived, and an artificially constructed lipid vesicle", becomes incorporated into the three-dimensional biocompatible matrix by and during the printing process itself.However, when inks 1.1-1.5 are used, such active ingredients, i.e. "at least one ingredient selected from...", have to be added separately to the already printed scaffold / matrix, as described below:.

[0081] B) The 3D printing ink compositions 1.1. to 1.5 are used to produce a three-dimensional biocompatible matrix in a 3D screen printing device. The inks 1.1. to 1.5 are preheated or precooled to a temperature of 4 to 50 ° C, preferably 20 ° C. The screen printing squeegee is used with an angle of 50 ° to 80 °, preferably 65 °, and a hardness of 65 to 85 Shore, preferably 75 Shore for submersion and 40 to 90 Shore, preferably 55 Shore for printing. The screen mesh is made of polyamide, polyester, or steel, with 8 to 400 threads / cm, preferably 48 threads / cm, and thread thickness of 27 to 300 μm, preferably 55 μm. The EOM ("Emulsion over Mesh") of the screen varies between 8 to 150 μm, preferably 20 to 25 μm. The printing process that builds the shape of the z-axis is a continuous repetition of submersion, printing, and curing. The ink is flood-printed through the screen mesh using a squeegee at a temperature of 4-65 °C, preferably 20-40 °C. The humidity in the printing chamber during the printing process is adjusted to 20-100%, preferably 60%. Curing of the printed ink is carried out at 0-180 °C using IR, convection drying, UV, or a cooling system. Preferably, the bioinks 1.1. to 1.5. are cured at a temperature of 4-40 °C. The humidity in the curing chamber can be adjusted to 10-60%, preferably 40%. The printing process is repeated until the final size of the three-dimensional biocompatible matrix is ​​achieved. The printing process can include the combination and / or rotation of the inks 1.1 to 1.5. Furthermore, the printing process can include multiple screens with different mesh compositions and geometries.

[0082] Thereafter, the desired active ingredient(s), i.e. "at least one ingredient selected from...", e.g., a non-opioid analgesic / EV, is introduced into the three-dimensional biocompatible scaffold / matrix by applying such "at least one ingredient selected from..." to or onto such scaffold / matrix. The step of applying such ingredient can be by any suitable technique, including spraying, dipping, coating, soaking, impregnation, application and subsequent freeze-drying, nanosol techniques, application and subsequent exposure of the scaffold / matrix to negative pressure or vacuum, etc.

[0083] Example 3: Fabrication of backing layers and their application to three-dimensional (3D) biocompatible matrices

[0084] To maximize the stability of the 3D screen printed three-dimensional biocompatible matrix, a crosslinker can be used during or after the printing process.

[0085] A three-dimensional backing layer is made using the 3D printing silicon-based composition 1.9 of Example 1 above. The paste of 1.9 is processed in a screen printing device under room temperature conditions. A screen printing squeegee is used with an angle of 50° to 80°, preferably 65°, and a hardness of 65 to 85 Shore for submersion, preferably 75 Shore, and 40 to 90 Shore for printing, preferably 55 Shore. The screen mesh is made of polyamide, polyester or steel, with 8 to 400 threads / cm, preferably 48 threads / cm, and thread thickness of 27 to 300 μm, preferably 55 μm. The screen EOM varies between 8 to 150 μm, preferably 20 to 25 μm. The printing process to build the shape of the z-axis is a continuous repetition of submersion, printing, and curing. The silicon paste is submerged and printed through the screen mesh with a squeegee at room temperature. Curing of the printed silicone paste is performed using infrared and / or convection drying at 120-180°C for 10-50 seconds, preferably 20-30 seconds. The printing process is repeated until the final size of the three-dimensional backing layer is obtained. The printing process can include multiple screens with different mesh compositions and shapes to vary the backing layer. Such a backing layer can then be applied to any of the three-dimensional (3D) biocompatible matrices of Example 2 and attached thereto by gluing, pasting, laminating, etc., or with one or several additional adhesive layers.

[0086] During the 3D screen printed steps 1.1-1.8 and 1.9, there is the possibility of applying one to several adhesive layers for the final finish of the product. For this, a pressure-sensitive adhesive paste is processed under room temperature conditions in a screen printing device as described above. Thus, 5-20 layers each of pressure-sensitive adhesive with a thickness of 5-25 μm, preferably 10-20 μm, are printed as separate carrier matrices.

[0087] The features of the invention disclosed in the specification, the claims and / or the accompanying drawings may, both separately and in any combination thereof, be material for realizing the invention in diverse forms thereof.

Claims

1. A three-dimensional biocompatible matrix comprising: - a three-dimensional scaffold made of polymeric agents, and - at least one component selected from a non-opioid analgesic, extracellular vesicles derived from animal cells, preferably from mammalian cells, more preferably from human cells, and artificially constructed lipid vesicles having a size ranging from 20 nm to 150 nm, wherein said at least one component is incorporated into said scaffold.

2. 10. The three-dimensional biocompatible matrix of claim 1, wherein the scaffold is dimensionally stable and is not a spreadable semi-solid or liquid such as a cream, lotion, ointment, salve, paste, ointment, or balm.

3. The three-dimensional biocompatible matrix according to any of claims 1 to 2, wherein the scaffold is manufactured by three-dimensional (3D) printing, preferably by three-dimensional (3D) screen printing.

4. 3. The three-dimensional biocompatible matrix according to any of claims 1 to 2, wherein the scaffold is not knitted, woven or nonwoven and / or does not have a thread-like structure, and preferably the scaffold is resorbable.

5. 3. The three-dimensional matrix according to any of claims 1 to 2, wherein the scaffold has a thickness in the range of 100 μm to 20 mm, preferably in the range of 100 μm to 15 mm, more preferably in the range of 100 μm to 12 mm, even more preferably in the range of 100 μm to 10 mm, and even more preferably the scaffold has a minimum thickness in the range of 100 μm to 200 μm, making it possible to accommodate a monolayer of cells, in particular fibroblasts and / or keratinocytes.

6. The three-dimensional biocompatible matrix according to claims 1 to 2, comprising: a) - a scaffold made of polymeric agents, and - a non-opioid analgesic incorporated into said scaffold; or b) - a scaffold made of a polymeric agent, and - extracellular vesicles derived from animal cells, preferably derived from mammalian cells, more preferably derived from human cells, incorporated into said scaffold, or c) - a scaffold made of a polymeric agent, and - artificially constructed lipid vesicles having a size ranging from 20 nm to 150 nm incorporated into said scaffold; or d) - a scaffold made of a polymeric agent, and - a non-opioid analgesic and extracellular vesicles derived from animal cells, preferably from mammalian cells, more preferably from human cells, incorporated into said scaffold; or e) - a scaffold made of a polymeric agent, and - a non-opioid analgesic and artificially constructed lipid vesicles having a size ranging from 20 nm to 150 nm, incorporated into said scaffold; or f) - a scaffold made of a polymeric agent, and - incorporated into said scaffold: a non-opioid analgesic, extracellular vesicles derived from animal cells, preferably derived from mammalian cells, more preferably derived from human cells, and artificially constructed lipid vesicles having a size ranging from 20 nm to 150 nm.

7. The three-dimensional biocompatible matrix according to any one of claims 1 to 2, wherein the polymeric agent is selected from the group comprising: a) naturally occurring polymers selected from the following polysaccharides, such as agarose, chitin, chitosan, dextran, alginate, carrageenan, cellulose, starch, fucoidan, laminaran, glycosaminoglycans, copolymers of glycosaminoglycans with collagen; gums selected from xanthan gum, gum arabic, gum ghatti, guar gum, locust bean gum, tragacanth gum, karaya gum; and inulin; polypeptides, such as collagen, and hydrolyzed forms of such polypeptides, such as gelatin; polyamino acids, such as polylysine; polynucleotides; b) synthetic polymers, such as poly poly(α-hydroxy acids), such as polylactic acid (PLA), polyglycolic acid (PGA), polylactic-co-glycolic acid (PLGA), poly(alkyl)methacrylates, such as poly(methyl)methacrylate, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), polyurethanes, polycaprolactone, polyamides, polydioxanones, polyglycerols, and mixtures or combinations of such synthetic polymers b) with such naturally occurring polymers a); c) silicone-based polymers, such as polysiloxanes, e.g., polydimethylsiloxane; and combinations or mixtures of any of the foregoing a)-c).

8. 8. The three-dimensional biocompatible matrix of claim 7, wherein the polymeric agent is selected from naturally occurring polymers selected from polysaccharides, such as agarose, chitin, chitosan, dextran, alginate, carrageenan, cellulose, starch, fucoidan, laminaran, glycosaminoglycans, copolymers of glycosaminoglycans and collagen; gums selected from xanthan gum, gum arabic, gum ghatti, guar gum, locust bean gum, tragacanth gum, and karaya gum; and inulin; polypeptides, such as collagen, and hydrolyzed forms of such polypeptides, such as gelatin; polyamino acids, such as polylysine; and polynucleotides, wherein preferably the polymeric agent is selected from alginate, gelatin, cellulose, collagen, chitosan, and mixtures of any of the foregoing.

9. 2. The three-dimensional biocompatible matrix of claim 1, wherein the non-opioid analgesic is selected from aminoester analgesics, aminoamide analgesics, fomocaine, and carbonate adducts of the aminoester analgesics or the aminoamide analgesics or the fomocaine, preferably, the aminoester analgesics include procaine, chloroprocaine, oxybuprocaine, benzocaine, tetracaine, and proxymetacaine; preferably, the aminoamide analgesics include lidocaine, mepivacaine, prilocaine, articaine, bupivacaine, ropivacaine, sinticaine, and etidocaine; and preferably, the fomocaines include fomocaine and its C-alkylmorpholine derivatives.

10. 2. The three-dimensional biocompatible matrix of claim 1, wherein the non-opioid analgesic is an aminoester analgesic or a carbonate adduct thereof, preferably selected from procaine, chloroprocaine, oxybuprocaine, benzocaine, tetracaine, and proxymetacaine, and their respective carbonate adducts.

11. 2. The three-dimensional biocompatible matrix of claim 1, wherein the non-opioid analgesic is an aminoamide analgesic or a carbonate adduct thereof, preferably selected from lidocaine, mepivacaine, prilocaine, articaine, bupivacaine, ropivacaine, sinticaine, and etidocaine, and their respective carbonate adducts.

12. 2. The three-dimensional biocompatible matrix of claim 1, wherein the non-opioid analgesic is selected from acetylsalicylic acid, ibuprofen, diclofenac, naproxen, indomethacin, paracetamol, metamizole, phenazone, propyphenazone, parecoxib, celecoxib, etoricoxib, ketamine, capsaicin, ziconotide, cannabinoids, and flupirtine.

13. The extracellular vesicles are derived from animal platelets or stem cells, preferably mammalian platelets or stem cells, more preferably human platelets or stem cells, and contain at least one component selected from cytokines, growth factors, transcription factors, RNA, particularly microRNA, and mRNA; wherein preferably, the stem cells are selected from mesenchymal stem cells and induced stem cells, more preferably, the mesenchymal stem cells are derived from bone marrow, umbilical cord blood, adipose tissue, or amniotic fluid; wherein preferably, the artificially constructed lipid vesicles contain at least one component selected from cytokines, growth factors, transcription factors, RNA, particularly microRNA, mRNA, and non-opioid analgesics; the non-opioid analgesic is selected from aminoester analgesics, aminoamide analgesics, fomocaine, and carbonate adducts of the aminoester analgesics or the aminoamide analgesics or the fomocaine, preferably, the aminoester analgesics include procaine, chloroprocaine, oxybuprocaine, benzocaine, tetracaine, and proxymetacaine; preferably, the aminoamide analgesics include lidocaine, mepivacaine, prilocaine, articaine, bupivacaine, ropivacaine, sinticaine, and etidocaine; preferably, the fomocaines include fomocaine and its C-alkylmorpholine derivatives; or the non-opioid analgesic is an aminoester analgesic or a carbonate adduct thereof, preferably selected from procaine, chloroprocaine, oxybuprocaine, benzocaine, tetracaine, and proxymetacaine, and their respective carbonate adducts; or the non-opioid analgesic is an aminoamide analgesic or a carbonate adduct thereof, preferably selected from lidocaine, mepivacaine, prilocaine, articaine, bupivacaine, ropivacaine, scinticaine, and etidocaine, and their respective carbonate adducts; or the non-opioid analgesic is selected from acetylsalicylic acid, ibuprofen, diclofenac, naproxen, indomethacin, paracetamol, metamizole, phenazone, propyphenazone, parecoxib, celecoxib, etoricoxib, ketamine, capsaicin, ziconotide, cannabinoids, and flupirtine; The three-dimensional biocompatible matrix of claim 1.

14. a) the extracellular vesicles are derived from animal platelets, preferably mammalian platelets, more preferably human platelets, and are positive for at least one cell marker selected from CD9, CD41a, CD41b, CD42b, CD61, CD62P, CD63, and syntenin, and / or the extracellular vesicles are negative for at least one cell marker selected from CD81, CD3, CD4, CD19, CD20, CD2, CD8, CD11a, and CD25; or b) the extracellular vesicles are derived from stem cells and are positive for CD81, CD9, CD63, Tsg101, and HSP70, and positive for one, some, or all of CD105, CD90, CD73, and CD44; and / or the extracellular vesicles are negative for one, some, or all of CD45, CD34, CD31, CD19, CD79α, CD14, CD11b, and HLA-DR; The three-dimensional biocompatible matrix of claim 1.

15. A composition comprising the three-dimensional biocompatible matrix of claim 1.

16. 16. The composition of claim 15, manufactured as a patch, dressing, pad, plaster, bandage, band-aid, tape, or pouch.

17. The composition of any one of claims 15 to 16, having one of the following two structures A or B: Structure A: the three-dimensional biocompatible matrix is ​​disposed on a first layer, and the composition comprises an additional layer attached to the first layer, the additional layer being a carrier layer that provides mechanical support to the biocompatible matrix, or a semipermeable backing layer that prevents diffusion of the at least one component from one side of the biocompatible matrix, or a solvent-impermeable backing layer, in particular a water-impermeable backing layer, that seals the biocompatible matrix on one side and prevents diffusion of the at least one component out of the biocompatible matrix on that side, or wherein the additional layer is both a carrier layer and a semipermeable or solvent-impermeable backing layer; the first layer has a thickness in the range of 100 μm to 20 mm, preferably 100 μm to 15 mm, more preferably 100 μm to 12 mm, even more preferably 100 μm to 10 mm; and wherein the additional layer has a thickness in the range of 50 μm to 2 mm, preferably 100 μm to 2 mm, more preferably 100 μm to 1 mm, even more preferably 100 μm to 500 μm; or Structure B: the three-dimensional biocompatible matrix is ​​disposed as the only layer within the composition, with no additional layers attached to the layer of three-dimensional biocompatible matrix; and wherein the only layer has a thickness in the range of 500 μm to 3 mm, preferably in the range of 500 μm to 2 mm, and more preferably in the range of 1 mm to 2 mm; wherein, optionally, the composition of Structure A and the composition of Structure B may be attached to a re-peelable carrier sheet for transport and / or storage of the composition, and when the composition is in use, e.g., during wound treatment and / or management, it is not attached to or does not include such a re-peelable carrier sheet.

18. 18. The composition of claim 17, wherein the composition of structures A and B is attached to a re-peelable carrier sheet for transport and / or storage of the composition, and wherein the composition is not attached to or does not contain a re-peelable carrier sheet when in use, e.g., during wound treatment and / or management.

19. A three-dimensional biocompatible matrix according to any one of claims 1 to 2 or a composition according to any one of claims 15 to 16 for use in a method for treating and / or managing a wound.

20. The three-dimensional biocompatible matrix or composition of claim 19, wherein the wound is an acute wound or a chronic wound, preferably wherein the acute wound is a burn, skin lesion, skin injury, surgical wound, incision wound, or puncture wound, and preferably wherein the chronic wound is a pressure ulcer or diabetic ulcer; and more preferably wherein the burn is selected from thermal burns, radiation burns, chemical burns, burns caused by exposure to electric current, and friction burns, and even more preferably wherein the burn is a first-, second-, or third-degree burn, and even more preferably a second- or third-degree burn.

21. 20. The three-dimensional biocompatible matrix or composition of claim 19, wherein the method of treating and / or managing the wound comprises applying the three-dimensional biocompatible matrix or composition to the wound and leaving it in contact with the wound for a defined period of time.

22. 20. The three-dimensional biocompatible matrix or composition of claim 19, wherein the method is for relieving pain and / or reducing inflammation in the wound.

23. 20. The three-dimensional biocompatible matrix or composition of claim 19, wherein the method is for increasing the rate at which wound healing is achieved compared to a wound not treated with the method, and / or the method is for delivering an analgesic to the wound, and / or the method is for reconstructing skin in and / or around the wound.

24. A method for producing the three-dimensional biocompatible matrix according to any one of claims 1 to 2, comprising: One of the following: a) providing a polymeric agent and 3D printing, preferably 3D screen printing, such agent onto a three-dimensional scaffold; b) applying at least one component selected from a non-opioid analgesic, extracellular vesicles derived from animal cells, preferably derived from mammalian cells, more preferably derived from human cells, and artificially constructed lipid vesicles having a size ranging from 20 nm to 150 nm to the three-dimensional scaffold, and incorporating the at least one component into the scaffold, thereby producing the three-dimensional biocompatible matrix; The polymeric agent and the at least one component are defined in any of claims 1-2; or a*) providing a polymeric agent and at least one component selected from a non-opioid analgesic, extracellular vesicles derived from animal cells, preferably mammalian cells, more preferably human cells, and artificially constructed lipid vesicles having a size in the range of 20 nm to 150 nm, and mixing the polymeric agent and the at least one component; and then b*) 3D printing, preferably 3D screen printing, the mixture obtained from step a*) onto a three-dimensional scaffold so that said at least one component is incorporated into said scaffold, thereby producing said three-dimensional biocompatible matrix; The polymeric agent and the at least one component are as defined in any of claims 1-2.

25. 1. An ink composition for three-dimensional (3D) printing, in particular three-dimensional (3D) screen printing, the ink composition comprising: i. a polymeric agent selected from the group including: a) naturally occurring polymers selected from polysaccharides, such as agarose, chitin, chitosan, dextran, alginate, carrageenan, cellulose, starch, fucoidan, laminaran, glycosaminoglycans, copolymers of glycosaminoglycans with collagen; gums selected from xanthan gum, gum arabic, gum ghatti, guar gum, locust bean gum, tragacanth gum, karaya gum; and inulin; polypeptides, such as collagen, and hydrolyzed forms of such polypeptides, such as gelatin; polyamino acids, such as polylysine; polynucleotides; b) synthetic polysaccharides polymers, such as poly(α-hydroxy acids), e.g., polylactic acid (PLA), polyglycolic acid (PGA), polylactic-co-glycolic acid (PLGA), poly(alkyl)methacrylates, e.g., poly(methyl)methacrylate, polyvinylpyrrolidone (PVP), polyethylene glycol (PEG), polyurethanes, polycaprolactone, polyamides, and mixtures or combinations of such synthetic polymers b) with such naturally occurring polymers a); c) silicone-based polymers, e.g., polysiloxanes, e.g., polydimethylsiloxane; and combinations or mixtures of any of the foregoing a)-c); ii. a solvent for i, preferably said solvent is selected from water, an aqueous solution, an alcohol, in particular methanol, ethanol, n-propanol or isopropanol, an alcohol solution, tetrahydrophane, acetone and ethyl acetate; iii. at least one component selected from a non-opioid analgesic, an extracellular vesicle derived from animal cells, preferably derived from mammalian cells, more preferably derived from human cells, and an artificially constructed lipid vesicle having a size ranging from 20 nm to 150 nm; wherein said non-opioid analgesic, an extracellular vesicle derived from animal cells, preferably derived from mammalian cells, more preferably derived from human cells, and an artificially constructed lipid vesicle are as defined in any of claims 9 to 14; iv. Optionally, one or more additional ingredients selected from fillers, crosslinkers, binders, surfactants, additives, diluents, thickeners, colorants, dyes, stabilizers, buffers, humectants, emulsifiers, dispersants, and preservatives.

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