Decellularized tissue / polymer multi-component biomaterial

Abstract

The present technology relates to a construct comprising at least one tissue region and at least one polymer region for use as an implant.

Description

Technical Field

[0001] The present invention relates to a novel type of multi-component material and its general use.

Background Art

[0002] There are a wide variety of materials that are foreign to the human body and are used in direct contact with the organs, tissues, and body fluids of the human body. These materials are called biomaterials, and among them, polymers play extremely important roles in all clinical fields such as orthopedics, cardiovascular area, plastic surgery, drug delivery, and wound dressing materials. Decellularized tissue is a kind of unique biomaterial derived from biological tissue. These materials are playing important roles in an increasing number of applications.

[0003] Polymers constitute one of the main types of materials used in implants for human or animal treatment due to the versatility of their chemical, physical, mechanical, and biological properties, and the extremely wide range of values they can achieve. To illustrate this point, it is sufficient to mention the following differences regarding their different properties. [i] Chemically, polymers can be very hydrophilic and water-soluble materials or extremely hydrophobic materials. [ii] Mechanically, the rigidity of polymers, which is reflected in the corresponding Young's modulus value, ranges from several kilopascals to several hundred gigapascals, an eight-digit or more range. On the other hand, the extensibility of polymers can be negligible, or polymers can exhibit fracture strain values in the range of thousands of percent.

[0004] Decellularization (or acellularization) is a technique widely used to produce semi-natural biomaterials, by which cells and genetic materials are separated from the extracellular matrix (ECM) of natural tissues. Decellularization can be achieved by chemical, enzymatic, or physical methods and is performed such that the ECM retains its original chemical and structural properties. The resulting decellularized tissues, typically pericardium, meshes, or small intestinal mucosa, can be used in many fields such as hernia repair, staple line reinforcement, pelvic floor reconstruction, dural closure, membranes in the dental area, heart patches, or heart valve leaflets.

[0005] The pericardium, a collagen-rich membrane that surrounds the heart, is the most widely used type of decellularized tissue and is currently utilized in various clinical applications. Among several applications, its use in dural closure, periosteum, heart valve leaflets, and surgical buttresses are examples of those of greater interest. The most widely used sources of decellularized pericardial tissue are cows and pigs.

[0006] Despite their respective advantageous properties, both decellularized tissues and polymeric materials have limitations in their behavior, particularly in clinical settings, and thus their biological performance is insufficient. SUMMARY OF THE INVENTION

[0007] The inventors of the technology disclosed herein have developed a novel hybrid material comprising a decellularized tissue and a polymeric material, each associated with each other via one or more physical fixation means. The interaction between these two components is not essentially chemical, although random chemical interactions that may form between the tissue and the polymer cannot be excluded.

[0008] Unlike known methods that utilize the polymerization of reactive monomers, the constructs of the present invention are produced by associating a decellularized tissue with a pre-made polymer. Such an assembly process overcomes many of the drawbacks associated with the use of polymerizable monomers. These drawbacks include: [a] the reactivity of the monomers towards the functional groups of the proteins that make up the decellularized tissue affects the properties and biocompatibility of the tissue; [b] the toxicity inherent to the monomers; [c] the effective solubilization of the monomers in organic agents; [d] the high volatility and flammability of the monomers; [e] the inability to properly position the monomers within the decellularized tissue; [f] unwanted polymerization that can occur due to the presence of certain functional groups present on the tissue and the catalyst used to induce the polymerization of the polymer component; [g] the formation of ill-defined polymers with variable average molecular weights, polydispersities, and often their compositions due to unwanted polymerization within the tissue components; [h] the possibility of leaving residual monomers that can remain after polymerization, which can have an adverse effect on the tissue and may be extracted over time after transplantation, causing additional local and even systemic problems.

[0009] By avoiding the use of in situ polymerization of reactive monomers, the inventors were able to achieve a stable and well-defined construct comprising at least one decellularized tissue and at least one polymer component, wherein the polymer component at least partially penetrates at least one surface region of the decellularized tissue.

[0010] Thus, in its broadest scope, the present invention provides a construct comprising at least one decellularized tissue and at least one polymer component, wherein the polymer component at least partially penetrates at least one surface region of the decellularized tissue.

[0011] In other words, the present invention provides a decellularized tissue physically associated with a polymer component, said association comprising or consisting of at least partial penetration of the polymer component into the surface region of the tissue.

[0012] Also provided is a polymer-assembled decellularized tissue in which the association between the polymer and the tissue is physical.

[0013] The present invention further provides a construct of at least one decellularized tissue and at least one polymer, the construct being configured as an implant or drug delivery device in vivo.

[0014] In another aspect, the present invention provides a construct comprising at least one decellularized tissue and at least one polymer component, the polymer component having at least one surface feature that protrudes from one side of the decellularized tissue and traverses the tissue through at least one hole formed in the tissue to the other side.

[0015] As used herein, the term "construct" is used to define a structure, element, device, or arrangement comprising a tissue phase or region in the form of at least one decellularized tissue and a polymer phase or region in the form of at least one polymer component, as defined herein. This term does not imply a particular arrangement or structure of the two phases or regions, and in fact encompasses any arrangement in which the two phases are present and associated with each other by physical interaction. This term excludes structures or elements in which there is no physical association between the two phases.

[0016] In all aspects of the present invention, the polymer component is implemented in the form of a sheet or segment, i.e., a piece of polymer material such as a strip of polymer material, and the polymer material is not formed in situ from monomers or prepolymers of the polymer material. The constructs of the present invention are formed by using prefabricated polymer sheets or segments of a size and shape selected to be suitable for a particular structure or use, as further detailed herein, or can be formed on the surface region of the tissue from a solution or liquid form of the polymer material. In other words, the methods of manufacturing the constructs of the present invention do not include an in situ polymerization step utilizing monomers, oligomers, or prepolymers.

[0017] The decellularized tissue and polymer may be implemented in the manufacture of a construct multi-sheet comprising two or more tissue sheets and several polymer sheets or segments that hold the construct together. Thus, the multi-sheets of the present invention can be provided in various forms each having at least one tissue sheet and at least one polymer sheet or segment. The sheets can be stacked at least partially on top of each other such that each of the polymer sheets or segments associates with another polymer sheet in the stack via at least one hole or pore formed in the decellularized tissue or via welding. Each of the material sheets in the multi-sheet is a solid sheet, but one or more of the sheets disposed internally may be formed from a gel, hydrogel, or may be formed as a liquid or fluid film.

[0018] As used herein, the term "sheet" is given its broadest meaning and can be in the form of a film of material of any size and shape, typically a continuous material or spread of material consisting of a polymeric material or tissue. In the constructs of the present invention where two material sheets are present, each sheet may be of the same material or different materials, and may be of the same or different sizes and shapes. In some embodiments, in the multi-sheet constructs of the present invention, each of the polymer sheets may be the same or different in composition, structure, size, shape, or any other physical, mechanical, or chemical property.

[0019] Typically, the sheet of decellularized tissue is provided as an elongated strip or ribbon of tissue, and its size and shape may vary. Similarly, the polymer component may be provided as an elongated sheet or strip of a size and / or shape similar or identical to the tissue, or as a fabricated (or formed) segment or piece or tag of polymer of a size and shape different (typically smaller) from the tissue sheet.

[0020] The multi-sheet construct of the present invention includes at least one sheet of decellularized tissue and at least one sheet or at least one segment of a polymer component, any sheet of the decellularized tissue being adjacent to or in contact with at least one sheet or segment of the polymer component, and at least two sheets or segments of the polymer component being joined to each other through at least one hole formed in at least one sheet of the decellularized tissue.

[0021] In some embodiments, in the multi-sheet construct, the sheets or segments of the polymer component are joined to each other through at least one hole formed in at least one sheet of the decellularized tissue.

[0022] In some embodiments, in the multi-sheet construct, at least two sheets or segments of the polymer component are joined to each other by welding.

[0023] In some embodiments, either at least one sheet or at least one sheet of the decellularized tissue is confined between any two sheets or segments of the polymer component.

[0024] In some embodiments, the multi-sheet construct comprises a number of sheets of decellularized tissue and the same number of sheets or segments of the polymer component.

[0025] In some embodiments, the construct comprises two or more assemblies of decellularized tissue confined between two sheets or segments of the polymer component, each of the assemblies being joined to each other by welding, if desired.

[0026] In some embodiments, at least two or any two of the assemblies are oriented in opposite directions.

[0027] The interaction or association between the polymer component (sheet or segment) and the tissue is selected and configured such that the polymer is firmly fixed in the tissue and, thus, the mechanical properties of the construct are improved. The improvement of the mechanical properties of the construct can be achieved by a physical non-chemical association that holds the two components together. An association that is not of a chemical nature can be defined as follows. (i) At least partially embedding a polymeric sheet or segment into the surface region of a layer of tissue, the embedding may be made through a single point or two or more points on the surface region of the layer, or (ii) Fixing a polymer sheet or segment into the tissue to a tissue depth that allows for a secure association, the fixing may be made through a single fixing point or two or more fixing points, and the depth of fixation or penetration of the polymer sheet or segment may vary but does not include piercing of the tissue, or (iii) Completely fixing a polymer sheet or segment to the tissue so as to completely penetrate the tissue from one side of the tissue to the other side, the fixing may be made through a single fixing point or two or more fixing points. Typically, such penetration will involve surface features configured to protrude from one side of the decellularized tissue through at least one hole formed in the tissue to the other side. As detailed below, such features may be formed in situ after the holes are formed in the tissue or may be provided on the polymer sheet or segment in a form selected and configured to pierce or perforate the tissue.

[0028] As used herein, the expression "at least partially penetrating" with respect to the surface region of a decellularized tissue implies any of the above interactions, including embedding into the tissue surface, fixation without piercing the tissue surface, and actually penetrating the tissue from one surface to the other. Similarly, the expression "the above association comprises or consists of at least partial penetration of the polymer component" encompasses any of the above associations or interactions, suggesting a single type of association or interaction (consisting of) or a combination of such associations or interactions (including).

[0029] Polymer sheets or segments are said to be associated with each other to ensure association with the decellularized tissue. One type of association or interaction present in the constructs of the present invention is, as detailed herein, through fixing or piercing the decellularized tissue surface, or by forming holes in the tissue through which two polymer sheets or segments can associate. In some embodiments of the present invention where the construct is composed of a plurality of construct assemblies, each assembly comprising, for example, a decellularized tissue restricted between two sheets or segments of a polymer component, the association of the plurality of assemblies can be achieved by welding of polymer to polymer.

[0030] Generally, welding of segments of polymer sheets occurs when the polymer chains on the surface of one sheet or segment are sufficiently mobile to intertwine with the chains of the other sheet or segment. To achieve welding, thermal energy can be applied to raise the temperature of the polymer above the appropriate transition temperature, i.e., the glass transition temperature Tg for amorphous thermoplastic polymers or the melting temperature Tm for semi-crystalline polymers. Bringing two sheets or segments of the polymer component into close contact under these conditions results in the intertwining of the polymer chains and welding. According to an aspect of the present invention, welding does not need to be performed over the entire surface of the sheet or segment. Spot welding in one or more regions of the polymer component may be sufficient to obtain a strong association of a plurality of assemblies or any two polymer sheets or segments.

[0031] In some embodiments, the polymer component is in the form of polymer particles (nanoparticles, microparticles, or larger-sized particles), which are patterned on the surface of the tissue to form a particle sheet or a particle continuum. In such embodiments, the particles may be embedded in the tissue surface (tissue outer layer), in which case the association between the tissue and the particles is strong enough to maintain the association between the two over a long period of time.

[0032] In other embodiments, the association between the polymer component and the tissue may involve at least one feature or functional group that exists on the polymer component, for example as a pendant group (or as a ligand group present on the surface of the polymer particles, or as at least one polymer material in the form of a layer or sheet or coating or polymer sheet or polymer film or polymer fiber or polymer mesh), and penetrates or protrudes through the tissue from one side to the other. The penetration may be through pores present in the tissue, or through at least one hole pre-formed in the tissue, for example by puncture or by stamping, i.e., the polymer material is placed in contact with the tissue and then physical penetration of the polymer material into the tissue surface is induced by applying pressure and / or temperature as desired.

[0033] At least one surface feature or functional group that penetrates or protrudes through the tissue from one side to the other may be in the form of long pins or needles that extend outward from the surface of the polymer material and are oriented perpendicular to or angled with respect to the surface of the polymer material. The size and shape of the holes formed by the long pins or needles (i.e., the size and shape of the pre-formed holes) may vary, and the number or distribution of the holes may cover part or all of the surface of the decellularized tissue.

[0034] The pins or needles, or generally at least one surface feature present on the surface of the polymer material, may have an end that securely holds the feature in place and prevents it from slipping out of the hole formed in the tissue.

[0035] In some embodiments, the construct comprises a polymeric sheet having one or more tissue-penetrating features and a decellularized tissue having one or more holes through which the features protrude, wherein the tissue-penetrating features are composed of the material of the polymeric sheet. The patterning of the tissue-penetrating features and / or the patterning of the holes may follow any patterning profile (hole profile) as defined herein (e.g., size, shape, distribution density, location, etc.).

[0036] In some embodiments, the construct comprises two polymeric sheets or segments, each of which is joined to the other via one or more polymeric features or members extending over the surface regions of each of the sheets, and the decellularized tissue disposed between the two polymeric sheets has one or more holes through which the one or more polymeric members cross.

[0037] In some embodiments, the construct is formed by making one or more holes in the surface region of the tissue and forming a sheet or segment of the polymeric component using the polymeric component in liquid or fluid form, wherein the liquid or fluid form is adapted to penetrate and fill the holes.

[0038] In some embodiments where there are two polymeric sheets or segments with one decellularized tissue disposed therebetween, each of the polymeric sheets is continuous and made of the same polymeric material, but one of the sheets is larger in size than the other.

[0039] In some embodiments, the construct is in the form of a multi-sheet device comprising one or more polymeric sheets and one or more sheets of decellularized tissue.

[0040] The present invention also provides a multi-sheet construct comprising at least one sheet of decellularized tissue and at least one sheet of a polymer component, wherein the decellularized tissue is confined between any two sheets of the polymer component, and any two sheets of the polymer component are joined to each other through at least one hole formed in at least one sheet of the decellularized tissue.

[0041] In some embodiments, one or more of the sheets can be designed as a substance reservoir for releasing active or inactive substances such as bioactive substances and drugs.

[0042] In some embodiments, at least a portion of the polymer component is configured to release a substance such as a bioactive substance or a drug. In some embodiments, all or a particular portion of the polymer component can locally release a bioactive substance or a drug.

[0043] The bioactive substance or drug that will be released from the construct of the present invention after being placed in the body can be selected from any drug or pharmaceutical intended to achieve a medical improvement, prevent the onset of a disease or medical condition (local or systemic), or maintain good health over time. Thus, the bioactive substance or drug can be selected, inter alia, based on the region of the body where the construct is to be implanted or placed, as well as the type of medical complications associated with the implantation site and procedure.

[0044] Generally speaking, an active agent or drug is widely characterized as being non-toxic as regulated by the FDA or EMA, or classified as GRAS (Generally Recognized As Safe). Non-limiting examples of such active agents and drugs include analgesics including non-narcotic and narcotic analgesics; anxiolytics; antiarrhythmics; antibacterial agents; antibiotics including natural, synthetic, broad-spectrum antibiotics; anticoagulants and thrombolytics for arterial or venous thrombosis; anticonvulsants; antidepressants including mood-elevating antidepressants; tricyclics, monoamine oxidase inhibitors, and SSRIs; antidiarrheals including antidiarrheal formulations and drugs that calm the contractions of the intestinal muscles; antiemetics; antifungals including infections affecting hair, skin, nails, and mucous membranes; antihistamines; antihypertensives including diuretics, beta blockers, calcium channel blockers, ACE (angiotensin converting enzyme) inhibitors; anti-inflammatory agents; antineoplastic agents; antipsychotics including major tranquilizers; antipyretics; antivirals including treatment and temporary protection against viral infections; beta blockers; corticosteroids related to immunosuppression, malignancy, or deficiency disorders; antineoplastic agents and further cytotoxic drugs as immunosuppressive agents; hormones including synthetic equivalents and natural hormone extracts; immunosuppressive agents; muscle relaxants and minor tranquilizers including those that relieve muscle cramps; sex hormones (female) including those used for menstrual and menopausal disorders, oral contraceptives, and further those for the treatment of female and male cancers; sex hormones (male) including those used for male hormone deficiency in hypopituitarism or testicular disorders, further those for the treatment of cancer, and anabolic steroids; enzymes such as collagenase or elastase; and vitamins may be mentioned.

[0045] In some embodiments, either the polymer or the tissue component may be porous, i.e., contain surface pores that are limited to a particular region of the component or are distributed along the entire surface of the polymer or tissue. The pores may be pre-existing or may be formed in the decellularized tissue to achieve a particular pore (or porosity) profile. The profile defines at least one surface region of the tissue where the pores are to be formed, the number, shape, and size of the pores, and the density of the pores in a particular surface region. Different patterning profiles may be used to meet the performance or attributes or utility of the desired construct. Once one or more pores are formed, the association of the polymer material can proceed by various methods as disclosed herein. This may include the use of polymer components having protruding features that can be inserted into or penetrate the pores. There may also be methods involving the use of liquid polymers. In such methods, the tissue may be immersed in, sprayed with, or otherwise generally treated with a liquid polymer material or a solution containing a polymer material, causing the polymer material to penetrate the pores and deposit on the tissue surface, thereby forming a polymer sheet or segment on one or both of the tissue surfaces. The polymer sheet or segment formed on one or both of the tissue surfaces can be fused or welded to another polymer component or sheet to form the multi-sheet or construct of the present invention, layer by layer or component by component.

[0046] Alternatively, the decellularized tissue is perforated and a liquid polymer is injected into the holes formed in the tissue. The assembly is constructed by forming a polymer sheet between two tissue sheets, and then the construct is thermally joined. The construct can be attached to another polymer sheet or feature by welding.

[0047] According to yet another approach, the multi-sheet structure is formed by stacking various sheets on top of each other, and then the multi-sheet is perforated and a liquid polymer is injected to obtain a fused construct with the desired pore profile.

[0048] According to yet another approach, a liquid polymer is cast onto the tissue to form a bilayer of tissue and polymer. Subsequently, the bilayer is perforated to obtain a pore profile, and a liquid polymer is injected to obtain a construct. The two-layer construct may be used to form a multi-sheet or may be welded to another polymer sheet.

[0049] In some embodiments, in the constructs of the present invention, the decellularized tissue constitutes 5 wt% to 95 wt% of the construct.

[0050] As used herein, "decellularized tissue" refers to tissue from which inhibitory cells have been removed, leaving the extracellular matrix (ECM) of the tissue. As described above, decellularization can be achieved by chemical, enzymatic, or physical methods as known in the art. Decellularized tissue can be obtained from oral mucosal tissue, small intestinal submucosa, or bladder acellular matrix, which provide natural and optimal integration properties with respect to the extracellular matrix. Other tissues may be used.

[0051] Methods for achieving decellularized tissue are known in the art, such as International Application Publication No. 2005 / 032473 and U.S. patent applications derived therefrom, each incorporated herein by reference, and U.S. Patent No. 5,993,844, incorporated herein by reference.

[0052] In some embodiments, the decellularized tissue is selected from pericardium, mesh, or small intestinal mucosa.

[0053] In some embodiments, the decellularized tissue is pericardium.

[0054] In some embodiments, the decellularized tissue is bovine pericardium or porcine pericardium.

[0055] In some embodiments, the decellularized tissue is small intestinal submucosa.

[0056] In some embodiments, the decellularized tissue is a mesh.

[0057] The polymers used in the constructs of the present invention are polymer materials known in the art. The polymers used can be selected from among thermoplastic polymers and thermosetting polymers. The polymers can be hydrophobic, hydrophilic, or amphiphilic, and can further be selected from among biocompatible polymers or biodegradable polymers.

[0058] In some embodiments, the polymer is a blend of different polymers, an interpenetrating polymer network (IPN), or a semi-interpenetrating polymer network (semi-IPN).

[0059] In some embodiments, the semi-IPN polymer is selected to crosslink via an addition or condensation reaction, a click chemistry reaction, or any other type of reaction, and combinations thereof. In some embodiments, the compound capable of crosslinking is selected from compounds containing two or more carbon double bonds, such as ethylene glycol dimethyl acrylate (EGDMA) and ethylene glycol diacrylate (EGDA), triethylene glycol dimethacrylate (TEGDMA), tetra(ethylene glycol) diacrylate (TEGDA), divinylbenzene (DVB), bisacrylamide, polyethylene glycol dimethacrylate (PEGDMA), polyethylene glycol diacrylate (PEGDA), polypropylene glycol dimethacrylate (PPGDMA), polypropylene glycol diacrylate (PPGDA), polyethylene glycol / polypropylene glycol (PEG / PPG), dimethacrylate (DMA) and diacrylate (DA) of copolymers, polytetramethylene glycol dimethacrylate (PTMGDMA) and polytetramethylene glycol diacrylate (PTMGDA), polydimethylsiloxane DMA and DA, polycaprolactone dimethacrylate (PCLDMA and polycaprolactone diacrylate (PCLDA), dimethacrylate (DMA) and diacrylate (DA) of polycaprolactone / lactide P(CL / LA), dimethacrylate (DMA) and diacrylate (DA) of polycaprolactone / glycolide P(CL / GA), and dimethacrylate (DMA) and diacrylate (DA) of polyglycolide / lactide P(GA / LA), dimethacrylate (DMA) and diacrylate (DA) of polyethylene glycol / caprolactone (PEG / CL), dimethacrylate (DMA) and diacrylate (DA) of polyethylene glycol / lactide PEG / LA, and dimethacrylate (DMA) and diacrylate (DA) of polyethylene glycol / glycolide (PEG / GA), dimethacrylate (DMA) and diacrylate (DA) of polypropylene glycol / caprolactone (PPG / CL), dimethacrylate (DMA) and diacrylate (DA) of polypropylene glycol / lactide (PPG / LA), and dimethacrylate (DMA) and diacrylate (DA) of polypropylene glycol / glycolide (PPG / GA), dimethacrylate (DMA), and DA of polytetramethylene / caprolactone (PTMG / CL), and dimethacrylate (DMA) and diacrylate (DA) of polytetramethylene / lactide (PTMG / LA), and dimethacrylate (DMA) and diacrylate (DA) of polytetramethylene / glycolide (PTMG / GA), etc.

[0060] In some embodiments, the IPN is composed of polymers formed by compounds containing two or more carbon-carbon double bonds selected from the group consisting of EGDMA and EGDA, TEGDMA and TEGDA, DVB, bisacrylamide, PEG DMA and DA, and higher functionality ones with various molecular weights, PPG DMA and DA, and higher functionality ones with various molecular weights, DMA and DA of PEG / PPG copolymers, and higher functionality ones, PTMG DMA and DA, and higher functionality ones with various molecular weights, siloxane DMA and DA, and higher functionality ones with various molecular weights, PCL DMA and DA, and higher functionality ones with various molecular weights, DMA and DA of P(CL / LA), P(CL / GA), and P(GA / LA), and higher functionality ones with various molecular weights, DMA and DA of PEG / CL, PEG / LA, and PEG / GA, higher functionality ones with various molecular weights, DMA and DA of PPG / CL, PPG / LA, and PPG / GA, higher functionality ones with various molecular weights, DMA and DA of PTMG / CL, PTMG / LA, and PTMG / GA, higher functionality ones with various molecular weights, and combinations thereof.

[0061] In some embodiments, the polymer is an acrylic or methacrylic polymer.

[0062] In some embodiments, the polymer is a polyolefin.

[0063] In some embodiments, the polymer is a silicone polymer.

[0064] In some embodiments, the polymer is a polycarbonate, polyurethane, polyurea, or polyamide, and combinations thereof. In some embodiments, the polymer is a polyurethane.

[0065] In some embodiments, the polymer has a glass transition point or melting point below 120°C. In some embodiments, the polymer has a glass transition point or melting point below 85°C.

[0066] In some embodiments, the polymer is selected to flow under a pressure of 50 kPa and at a temperature above 42°C.

[0067] In some embodiments, the polymer is selected from polymethyl methacrylate (PMMA), poly(n-butyl methacrylate) (PBMA), poly(hexyl methacrylate) (PHMA), polystyrene (PST), poly(2-hydroxyethyl methacrylate) (PHEMA), poly(N-(2-hydroxypropyl) methacrylamide) (PHPMA), polycyanoacrylate (PCA), polyethylene / polypropylene copolymer, polyethylene / polybutylene copolymer, polypropylene / polybutylene copolymer, polyisobutylene, polydimethylsiloxane (PDMS), phenyl-containing PDMS, polyester urethane, polyether urethane (e.g., Pellethane, Elastane, Elastolan, Tecoflex, Biomer), polycarbonate urethane (e.g., Chronoflex, Biospan, and Bionate), and silicone-containing polyurethane (e.g., CarboSil, PurSil, Avcothane, and Cardiothane), polyglycolic acid, polylactic acid, polycaprolactone, polylactide-caprolactone copolymer, polyglycolic acid-lactic acid copolymer, polyethylene oxide-polylactic acid copolymer, polyethylene oxide-polycaprolactone copolymer, polytetramethylene oxide-caprolactone copolymer, polyhydroxybutyrate, polyhydroxyvalerate, polyethylene adipate, polybutylene adipate, polyethylene succinate, polybutylene succinate, and polybutylene terephthalate, and polyethylene / butylene terephthalate copolymer, and combinations and copolymers thereof.

[0068] As used herein, the term "ChronoFlex" hereinafter means a family of biodurable aromatic polycarbonate-based thermoplastic polyurethanes. These aromatic polycarbonate urethanes are typically designed for molding, casting, and dip coating applications.

[0069] In some embodiments, the polymer is an aliphatic polyether-based thermoplastic polyurethane and is Tecoflex, if desired.

[0070] In some embodiments, the polymer is an aromatic polycarbonate-based urethane and is Chronoflex, if desired.

[0071] In some embodiments, the polymer is PMMA, PBMA, PHMA, PMA, PHEMA, PHPMA, and combinations thereof.

[0072] In some embodiments, the polymer is a low molecular weight (typically 500 - 10,000 Da) or amorphous or branched chain polyolefin, and combinations thereof.

[0073] In some embodiments, the polymer is PDMS, or phenyl-containing PDMS, or a derivatized PDMS chain containing double bonds and / or hydroxyl groups and / or amine groups and / or thiol groups.

[0074] In some embodiments, the polymer is an aliphatic or aromatic polyurethane containing a soft segment of polyether or polyester.

[0075] The polymer can be selected to have forms and properties that change over time later. These changes can be due to chemical, physical, and / or biological phenomena. In some embodiments, the polymer is selected to undergo secondary chemical changes such as coupling, polymerization or crosslinking, oxidation, or hydrolysis or enzymatic degradation. In some embodiments, apart from or in addition to the chemical changes, the polymer can be selected to undergo physical processes such as crystallization or phase separation or to be affected by physical processes.

[0076] In some embodiments, the polymer is a shape memory polymer that can be actuated by various stimuli such as temperature, pH, ionic strength, hydration, biological triggers, electric or magnetic fields, any kind of radiation, and combinations thereof, and the shape memory polymer can exhibit a one-time response or a cyclic shape memory response.

[0077] In some embodiments, the actuated shape memory polymer is a polymer that can regulate the shape of the tissue / polymer construct.

[0078] In some embodiments, all or part of the polymer components can exhibit shape memory behavior.

[0079] In some embodiments, all or part of the polymer components can be environmentally responsive. In some embodiments, all or part of the polymer components can be reverse thermo-responsive.

[0080] In some embodiments, the polymer is an environmentally responsive polymer that is responsive to temperature, pH, ionic strength, biological triggers, various kinds of radiation, electric or magnetic fields, and combinations thereof. In some embodiments, the environmentally responsive polymer is reverse thermo-responsive.

[0081] The inverse heat responsiveness may be selected from N-alkyl substituted acrylamides (such as poly-N-isopropylacrylamide [PNIPAAm]), or be based on polyethylene oxide / polypropylene oxide segments, or be a cellulose derivative selected from hydroxypropyl methylcellulose and hydroxypropyl cellulose, or an alternating polymer or a random polymer, and various amphiphilic polymers such as poly(ethylene oxide)-polylactic acid block copolymers.

[0082] In some embodiments, the polymer is polybutyl methacrylate.

[0083] In some embodiments, the polymer is a polyolefin such as a polyethylene / polypropylene copolymer, a polyethylene / polybutylene copolymer, a polypropylene / polybutylene copolymer, polyisobutylene, or a combination thereof.

[0084] In some embodiments, the polymer is a silicone polymer which is derivatized PDMS selected to crosslink in situ.

[0085] In some embodiments, the polymer is a polyurethane selected from Pellethane, Elasthane, Elastolan, chronoflex, Tecoflex, Cardiothane, Avcothane, CarboSil, PurSil, Biomer, BioSpan, and Bionate, as defined herein, and combinations thereof.

[0086] In some embodiments, the polyurethane is an aliphatic polyether-based thermoplastic polyurethane, such as Tecoflex.

[0087] In some embodiments, the polyurethane is an aromatic polycarbonate-based urethane, such as Chronoflex.

[0088] In some embodiments, the polymer is a polyester and is optionally selected from polyglycolic acid, polylactic acid, polycaprolactone, polyhydroxybutyrate, polyhydroxyvalerate, polyethylene adipate, polybutylene adipate, polyethylene succinate, polybutylene succinate, polybutylene terephthalate, and polyethylene / butylene terephthalate copolymers, and combinations and copolymers thereof.

[0089] In the constructs of the present invention, the polymers used can be a blend or mixture or combination of two or more polymer materials. In some embodiments, such blends can include two or more polyolefins, or acrylic or methacrylic polymers, or silicone polymers, or polycarbonates, or polyurethanes, or polyureas, or polyamides, and combinations thereof.

[0090] In another aspect, the constructs of the present invention comprise one or more tissue phases (including or consisting of decellularized tissue) and two or more polymer phases (including or consisting of polymers), each of which forms a separate phase of a different geometric shape, and the size of which can range from nanometers to the centimeter scale.

[0091] The polymer phase and / or tissue phase can be constructed or composed of two or more polymers or decellularized tissues. Each of the polymers and / or tissues can form a single construct as further disclosed herein or can form an array of constructs associated with each other. In some embodiments, the polymers and / or tissues can be part of a medical device having at least one polymer region and / or at least one tissue region, and the construct can be associated therewith. Such devices can be composed of any type of material, exemplified by tissues, polymers, metals, ceramics, carbonaceous materials, and combinations thereof.

[0092] In the construct of the present invention, the polymer phase is configured or selected or designed to connect to the tissue phase or two or more of the other phases including materials exemplified by polymers, metals, ceramics, carbonaceous materials, and combinations thereof as disclosed above. In some embodiments, the polymer phase is configured to connect two or more of the tissue phase or other phases, and the connection can maintain a stress exceeding 5% of the cohesive force of the polymer itself. In some embodiments, for these and other purposes, the polymer is selected to exhibit shape memory behavior in this way. In some embodiments, the polymer is environmentally responsive. In some embodiments, the polymer is biodegradable.

[0093] In some embodiments, the polymer is inversely thermoresponsive and forms an aqueous solution that undergoes an LCST transition below 37°C.

[0094] In some embodiments, the construct is a composite material.

[0095] The present invention further provides an implant or medical device that is or comprises the construct of the present invention.

[0096] Regardless of the type of construct used in accordance with the present invention and regardless of the type of device to be manufactured from the construct of the present invention, the construct is configured, designed, or intended to contact human or animal tissue or organs. In some cases, the construct may be a device intended to associate with tissue within the human or animal body or may be constructed as a device to be implanted within the body. For example, the construct may, as appropriate, be a medical device configured to be implanted into the subject's GI tract, into the respiratory system (airway system), along a vascular structure, into the cardiac region, into the urinary system, or into any other organ of the human or animal body, or may be part of a medical device. In particular, depending on the material, shape, and intended use of the construct, and further depending on the organ in which the implant is to be used, the construct may be porous or non-porous, and the pores may range from nanometers to centimeters in scale. Non-limiting examples of devices that utilize or comprise the construct of the present invention include stents, metallic stents, vascular grafts, heart valves (optionally with a metallic frame), membranes, sealing devices, suture or staple lines, hernia meshes or hernia repair devices, pelvic floor reconstruction devices, wound or burn dressings, dural closure materials, heart patches, and the like.

[0097] In some embodiments, the device is implemented in a heart valve that also comprises a metallic frame. Thus, the heart valve may, in some embodiments, comprise a pericardial leaflet, a tissue / polymer phase, and a metallic frame, as defined herein.

[0098] In some embodiments, the construct is a patch disposed in the CV system, along the GI tract, in the bronchial tree, in the urinary and genital regions, in the central or peripheral nervous system, in a vascular graft, an A / V shunt, and the polymer forms films, fibers, particles of any size and shape, porous or solid, hollow or non-hollow, or any other shape, and these are manufactured by any manufacturing technique including weaving methods and procedures for manufacturing non-woven structures, and combinations thereof.

[0099] The device of the present invention containing the construct of the present invention can be used as a transporter for delivering one or more active agents to a site or organ or tissue within the body of a subject. Such active agents can be bioactive substances of cells or molecules. The active substance may be contained throughout the construct, in the polymer component, or in the decellularized tissue. In some embodiments, the decellularized tissue contains an active agent, such as a bioactive substance, as defined herein. In some embodiments, the active substance is released over time.

[0100] The construct of the present invention can be provided in various forms, as exemplified herein. In some embodiments, the polymer associates with two or more tissue regions or segments, thus forming any other structure such as a laminate or multilayer structure. In some embodiments, the multilayer structure may comprise three layers of polymer, two of which are on the outside and one is in the center of the construct, with two layers of tissue sandwiched between the polymer layers.

[0101] In some embodiments, the construct comprises a polymer film restricted between two sheets of decellularized tissue.

[0102] In some embodiments, the construct can be formed with a polymer connection that associates each of the polymer layers through holes formed in the tissue layer. In some embodiments, the polymer connection has the same composition or a different composition from the polymer forming the layer. For synthesizing such a construct, according to the present invention, a polymer or polymer mixture used, for example, pre-formed on a polymer film, is selected to be able to flow under a pressure of 50 kPa to 1 MPa and at a temperature below 120°C to fill the holes formed in the tissue, whereby the tissue and the polymer are connected or associated together. When cooled, the polymer creates a continuous film-like structure distally with respect to the flow direction, contributing to the strength and long-term stability of the association between the tissue and the polymer phase.

[0103] In some embodiments, the polymer is an RTR polymer that flows under a pressure of less than 100 kPa and a temperature of less than 37°C. Upon reaching physiological temperature, the polymer creates a continuous film-like structure distal to the flow direction, contributing to the strength and long-term stability of the association between the tissue and the polymer phase.

[0104] In some embodiments, the polymer in the construct of the present invention is pre-formed into the form or shape of a film as desired. The pre-formed polymer may include two or more materials that are blended together or separated at distances ranging from nanometers to centimeters, and are isotropically or anisotropically organized to produce a layer or any other spatial arrangement, and the additional materials may be selected from polymers, tissues, active ingredients, metals, ceramics, or carbonaceous materials, and combinations thereof.

[0105] In some embodiments, the pre-formed polymer further includes low molecular weight molecules that soften the film or a part thereof. The low molecular weight molecules enable the polymer phase or a part thereof to flow under pressure and temperature conditions. In some embodiments, the low molecular weight molecules are polymerizable or crosslinkable, so that before polymerization or crosslinking, they soften the polymer phase or a part thereof, and upon polymerization or crosslinking, they strengthen or harden the polymer phase. In some embodiments, whether inert or reactive, the low molecular weight molecules may be biocompatible or biodegradable, may exhibit shape memory behavior, or may be environmentally responsive, or combinations thereof.

[0106] In some cases, the construct can be obtained by a polymerization reaction or a crosslinking reaction that occurs via an addition reaction or a condensation reaction, or any other type of reaction, and combinations thereof. The addition polymerization reaction or crosslinking reaction may involve reactive carbon double bonds that follow any type of catalyst, including reactions initiated chemically, thermally, and by radiation.

[0107] In some cases, the construct can be obtained by a polymerization reaction or a crosslinking reaction of a suitable precursor, and the above reactions are carried out partly before contacting with the decellularized tissue and partly during or after the association of the polymer component and the decellularized tissue component. In some embodiments, a partially polymerized or crosslinked polymer film or sheet or construct having any shape can flow under the applied conditions and result in the desired association with any other component, whether it is a decellularized tissue or a polymer. In some embodiments, the partially polymerized or crosslinked polymer is selected to flow under a pressure of 50 kPa and a temperature above 42°C.

[0108] In some embodiments, the partially polymerized or crosslinked polymer has a glass transition point or melting point below 120°C, while in some other embodiments, the partially polymerized or crosslinked polymer has a glass transition point or melting point below 85°C.

[0109] In other embodiments, the partially polymerized or crosslinked polymer has a glass transition point or melting point below 60°C.

[0110] In some embodiments, the partially polymerized or crosslinked polymer that is further polymerized or crosslinked during or after the association with the decellularized tissue or any other component, whether it is a polymer or not, has a higher glass transition point or melting point than the partially polymerized or crosslinked polymer.

[0111] In some embodiments, the polymer member that is further polymerized or crosslinked has a glass transition point or melting point above 50°C, in other embodiments, the polymer member that is further polymerized or crosslinked has a glass transition point or melting point above 60°C, and in still other embodiments, the polymer member that is further polymerized or crosslinked has a glass transition point or melting point above 80°C.

[0112] In some embodiments, the polymer member that has undergone further polymerization or crosslinking has a glass transition point or melting point above 100°C.

[0113] In some embodiments, the polymer in the construct of the present invention comprises a partially polymerized or crosslinked polymer component that is pre-formed in the form or shape of a film, if desired, and has a degree of initial polymerization and / or crosslinking that is different from the degree of final polymerization and / or crosslinking reached in the case of the polymer member that has undergone further polymerization or crosslinking.

[0114] In some embodiments, the pre-formed polymer further comprises low molecular weight molecules (500 - 100,000 DA), which may or may not be polymerizable and / or crosslinkable, and suitable precursors that are partially polymerized or crosslinked, and the polymerization or crosslinking reaction is carried out, in part, before contact with the decellularized tissue and, in part, during or after the association of the polymer component with the decellularized tissue component.

[0115] In other cases, the construct can be obtained by the reaction of compounds selected from methyl methacrylate (MMA), butyl methacrylate (BMA), hexyl MA, styrene (ST), (2-hydroxyethyl methacrylate) (HEMA), acrylamide (AAm), acrylic acid (AAc), N-vinyl pyrrolidone (NVP), cyanoacrylate, N-iso-PAAm, maleic anhydride, EGDMA and EGDA, TEGDMA and TEGDA, DVB, bisacrylamide, PEG DMA and DA, and higher functionality ones of various molecular weights, PPG DMA and DA, and higher functionality ones of various molecular weights, PEG / PPG copolymer DMA and DA, and higher functionality ones, PTMG DMA and DA, and higher functionality ones of various molecular weights, siloxane DMA and DA, and higher functionality ones of various molecular weights, PCL DMA and DA, and higher functionality ones of various molecular weights, P(CL / LA), P(CL / GA), and P(GA / LA) DMA and DA, and higher functionality ones of various molecular weights, PEG / CL, PEG / LA, and PEG / GA DMA and DA, higher functionality ones of various molecular weights, PPG / CL, PPG / LA, and PPG / GA DMA and DA, higher functionality ones of various molecular weights, PTMG / CL, PTMG / LA, and PTMG / GA DMA and DA, higher functionality ones of various molecular weights, and combinations thereof.

[0116] The reaction can be selected from click chemistry reactions, any kind of complexation, host / guest reactions, and non-covalent supramolecular polymerization. In some embodiments, a condensation reaction is involved, whereby urethane groups, urea groups, amide groups, ester groups, carbonate groups, or ether groups, and combinations thereof are formed.

[0117] In some embodiments, an epoxy / amine reaction and hydrolysis and condensation reactions of siloxane-containing molecules may be involved. In some embodiments, a reaction of a thiol group may be involved in the reaction. The epoxy / amine reaction may be carried out between GMA and amines of various functionalities and molecular weights, including amino-terminated PEG, PPG, PTMG, PCL, PDMS, amino-terminated P(CL / LA), P(CL / GA), and P(GA / LA)-containing molecules of various molecular weights, PEG / CL, PEG / LA, and PEG / GA-containing molecules of various molecular weights, PPG / CL, PPG / LA, and PPG / GA-containing molecules of various molecular weights, PTMG / CL, PTMG / LA, and PTMG / GA-containing molecules of various molecular weights containing one or more amine groups, oligopeptides and peptides containing reactive amine moieties, and epoxy-terminated molecules of various functionalities and molecular weights capable of reacting with the above amino-terminated molecules, and combinations thereof.

[0118] In some embodiments, the molecules of the polymer material in the construct of the present invention can unwind, cross the polymer / tissue interphase or any other interphase, and diffuse into the tissue phase or any other phase, thereby mixing with themselves and the molecules of the tissue or any other phase, re-crosslinking, and welding both together. In some embodiments, the molecules crossing the polymer / tissue interface can also react with themselves and / or portions present in the tissue or any other phase, thereby improving the strength and long-term stability of the connection between the polymer phase and any other phase.

[0119] The construct of the present invention can be formed in situ, i.e., in a state where the polymeric material is in direct contact with the tissue phase while in a liquid or semi-liquid state. This in situ generated polymer phase is formed by applying a liquid or semi-liquid polymerization-reactive or crosslinking-reactive precursor of an optimized composition. In some embodiments, the in situ generated polymer phase is formed by applying a solution of the polymer onto and / or into the tissue phase, and combinations thereof. In some embodiments, the polymer solution is an aqueous solution or a solution using another hydrophilic solvent. In some embodiments, the polymer is dissolved in a halogenated solvent such as acetone, THF, dioxane, DMSO, chloroform, and dichloromethane, an alcohol and a polyol, a polyether, acetonitrile, ethyl acetate, dimethylacetamide, DMAC, and an organic solvent such as dimethylformamide (DMF). The polymer solution has a concentration in the range of 1 wt% to 40 wt%, or 1 wt% to 20 wt%, or 2 wt% to 10 wt%.

[0120] In some embodiments, the tissue phase and the polymer phase are connected by applying pressure at a supra-physiological temperature over a time shorter than 1 hour. In some embodiments, the pressure is in the range of 20 kPa to 100 GPa, the temperature is in the range of 40 degrees to 120 degrees, and is applied at time intervals of 1 second to 60 minutes. In some embodiments, the pressure is in the range of 500 kPa to 10 GPa, the temperature is in the range of 40 degrees to 85 degrees, and is applied at time intervals of 2 seconds to 5 minutes.

[0121] Accordingly, in another aspect of the present invention, a method for manufacturing the construct of the present invention is provided.

[0122] According to one aspect, a method for manufacturing a construct according to the present invention is provided, the method comprising - contacting a perforated surface region of at least one decellularized tissue with a liquid polymer, and - Passing the liquid polymer through the perforated part (hole) to form a polymer sheet on the surface area. is included.

[0123] The perforated surface area of the tissue may be an area in which holes are formed that penetrate it, or an area that naturally has such holes or pores. If holes are to be formed, the holes may be formed by using cutting or perforating devices such as needles of various diameters. The position of the holes, the shape of the holes, the size of the holes, and the distribution of the holes across the entire surface area (the hole profile as defined herein) may be predetermined and selected so as to meet, for example, the mechanical properties of the construct or any prerequisite related to its use. Typically, the number of holes is two or more.

[0124] Thus, in some embodiments, the method includes perforating or forming holes in the surface area of at least one decellularized tissue. In some embodiments, the holes have a predetermined hole profile as defined.

[0125] In some embodiments, the liquid polymer is injected into the perforated part (hole).

[0126] When a liquid polymer is used, for example, curing by heat treatment of the polymer may be required. Since different polymers may be used, different curing methods may be used.

[0127] A further method for manufacturing a construct comprising at least one decellularized tissue and at least one polymer component is provided, the method comprising: - Contacting the surface area of at least one decellularized tissue perforated with one or more holes with a liquid polymer, and - Passing the liquid polymer through the one or more holes to form a polymer sheet on the surface area. is included.

[0128] In some embodiments, it includes perforating or forming holes in the surface area of at least one decellularized tissue. In some embodiments, the holes have a predefined hole profile as determined.

[0129] In some embodiments, a liquid polymer is injected into the perforated part (hole).

[0130] In some embodiments, the liquid polymer completely penetrates through one or more holes.

[0131] In some embodiments, the liquid polymer partially penetrates through one or more holes.

[0132] In some embodiments, the liquid polymer completely penetrates through one or more holes and forms a polymer sheet on both sides of the surface area to form a construct assembly.

[0133] In some embodiments, the method includes the step of joining or fusing two or more constructs. The fusion can be by welding.

[0134] There is further provided a method for manufacturing a construct comprising at least one decellularized tissue and at least one polymer component, and this method - stacking a sheet of one or more decellularized tissues and a sheet or segment of one or more polymer materials to obtain a stacked structure, - forming holes in the above stacked structure to form one or more holes in each of one or more sheets of tissue and polymer material, with at least some arranged coaxially as desired, and - treating the above stacked structure with a liquid polymer to penetrate the liquid polymer through one or more holes and fuse the sheets to form a construct, including.

[0135] The present invention provides a construct comprising at least one decellularized tissue, at least one metal element, and at least one polymer component, wherein the polymer component at least partially covers at least one surface region of the decellularized tissue and at least one surface region of the metal element.

[0136] The present invention further provides a decellularized tissue physically associated with a polymer component, said association comprising or consisting of at least partial covering of the polymer component on the surface region of the tissue.

[0137] The present invention further provides a construct comprising at least one decellularized tissue, at least one metal element, and at least one polymer component, wherein the polymer component has at least one feature that at least partially covers at least one surface of the decellularized tissue and at least one surface region of the metal element.

[0138] The present invention further provides the construct as defined above in the form of a multi-sheet construct.

[0139] The present invention further provides the construct as defined above, wherein the coating is performed by at least one method selected from the group consisting of spraying, brush coating, dip coating, and any combination thereof.

[0140] The present invention further provides the construct as defined above, wherein the at least one sheet of the decellularized tissue is dried before being coated by the polymer.

[0141] The present invention further provides the construct as defined above, wherein the drying is freeze-drying (i.e., lyophilization) of the at least one sheet of the decellularized tissue.

[0142] The present invention further provides a construct as defined above, wherein at least one or any of at least one sheet of the decellularized tissue is confined between any two sheets of the polymer component.

[0143] The present invention further provides a construct as defined above, wherein the multi-sheet construct comprises a number of sheets of decellularized tissue and the same number of sheets of the polymer component.

[0144] The present invention further provides a construct as defined above, comprising two or more assemblies of decellularized tissue confined between two sheets or segments of the polymer component, wherein the assemblies are associated with each other.

[0145] The present invention further provides a construct as defined above, wherein at least two assemblies are oriented in opposite directions.

[0146] The present invention further provides a construct as defined above, wherein the metal element is a metal wire.

[0147] The present invention further provides a construct as defined above, wherein the polymer component is in the form of a layer or coating of particles, a polymer sheet, a polymer film, polymer fibers, or a polymer mesh.

[0148] The present invention further provides a construct as defined above, wherein at least one hole is pre-formed or present in the decellularized tissue.

[0149] The present invention further provides a construct as defined above, wherein the decellularized tissue constitutes 5 wt% to 95 wt% of the construct.

[0150] The present invention further provides a construct as defined above, wherein the decellularized tissue is obtained from a tissue selected from oral mucosa, small intestinal submucosa, and bladder decellularized matrix.

[0151] The present invention further provides the construct as defined above, wherein the decellularized tissue is selected from pericardium, mesh, or small intestinal mucosa.

[0152] The present invention further provides the construct as defined above, wherein the decellularized tissue is pericardium.

[0153] The present invention further provides the construct as defined above, wherein the decellularized tissue is bovine pericardium or porcine pericardium.

[0154] The present invention further provides the construct as defined above, wherein the polymer component is a polymer selected from hydrophobic polymers, hydrophilic polymers, and amphiphilic polymers or comprises a polymer selected from them.

[0155] The present invention further provides the construct as defined above, wherein the polymer component is a blend, IPN, or semi-IPN or comprises a blend, IPN, or semi-IPN.

[0156] The present invention further provides the construct as defined above, wherein the polymer component is an acrylic polymer or a methacrylic polymer or comprises an acrylic polymer or a methacrylic polymer.

[0157] The present invention further provides the construct as defined above, wherein the polymer component is a polyolefin or comprises a polyolefin.

[0158] The present invention further provides the construct as defined above, wherein the polymer component is a silicone polymer or comprises a silicone polymer.

[0159] The present invention further provides the construct as defined above, wherein the polymer component is a polycarbonate, polyurethane, polyurea, or polyamide, and combinations thereof, or comprises a polycarbonate, polyurethane, polyurea, or polyamide, and combinations thereof.

[0160] The present invention further provides a construct as defined above, wherein the polymer component is polyurethane or comprises polyurethane.

[0161] The present invention further provides a construct as defined above, wherein the polymer component is a polymer selected from polymethyl methacrylate (PMMA), poly(n-butyl methacrylate) (PBMA), poly(hexyl methacrylate) (PHMA), polystyrene (PST), poly(2-hydroxyethyl methacrylate) (PHEMA), poly(N-(2-hydroxypropyl)methacrylamide) (PHPMA), polycyanoacrylate (PCA), polyethylene / polypropylene copolymer, polyethylene / polybutylene copolymer, polypropylene / polybutylene copolymer, polyisobutylene, polydimethylsiloxane (PDMS), phenyl-containing PDMS, polyester urethane, polyether urethane, polycarbonate, silicone-containing polyurethane, polyglycolic acid, polylactic acid, polycaprolactone, polylactide-caprolactone copolymer, polyglycolic acid-lactic acid copolymer, polyethylene oxide-polylactic acid copolymer, polyethylene oxide-polycaprolactone copolymer, polytetramethylene oxide-caprolactone copolymer, polyhydroxybutyrate, polyhydroxyvalerate, polyethylene adipate, polybutylene adipate, polyethylene succinate, polybutylene succinate, and polybutylene terephthalate, and polyethylene / butylene terephthalate copolymer, and combinations and copolymers thereof, or comprises a polymer selected therefrom.

[0162] The present invention further provides a construct as defined above, wherein the polymer component is a shape memory element or comprises a shape memory element.

[0163] The present invention further provides a construct as defined above, wherein the polymer component is a polyether urethane selected from Pellethane, Elastane, Elastolan, Tecoflex, Biomer or comprises a polyether urethane selected from them.

[0164] The present invention further provides a construct as defined above, wherein the polymer component is a polycarbonate urethane selected from Chronoflex, Biospan, and Bionate or comprises a polycarbonate urethane selected from them.

[0165] The present invention further provides a construct as defined above, wherein the polymer component is a silicone-containing polyurethane selected from CarboSil, PurSil, Avcothane, and Cardiothane or comprises a silicone-containing polyurethane selected from them.

[0166] The present invention further provides a construct as defined above, wherein the polymer component is Chroniflex or Tecoflex or comprises Chroniflex or Tecoflex.

[0167] The present invention further provides a construct as defined above, wherein one or more of the sheets are designed as a substance reservoir for releasing an active substance or an inactive substance.

[0168] The present invention further provides a construct as defined above, wherein the active substance is selected from analgesics; anxiolytics; antiarrhythmics; antibacterial agents; antibiotics; anticoagulants and thrombolytics; antiepileptic drugs; antidepressants; antidiarrheal drugs; antiemetic drugs; antifungal drugs; antihistamines; antihypertensive drugs; anti-inflammatory drugs; antineoplastic drugs; antipsychotic drugs; antipyretics; antiviral drugs; beta blockers; corticosteroids; cytotoxic drugs; hormones and sex hormones; enzymes; and vitamins.

[0169] The present invention further provides a construct as defined above, wherein the metal element is selected from a stent, a metallic stent, a vascular graft, a heart valve, a membrane, a sealing device, a suture or staple line, a hernia mesh or hernia repair device, a pelvic floor reconstruction device, a wound or burn dressing, a dural closure material, and a heart patch.

[0170] The present invention further provides a device comprising a construct according to any of the above.

[0171] The present invention further provides a device as defined above configured as an implant.

[0172] The present invention further provides a device as defined above, wherein the device is selected from a stent, a metallic stent, a vascular graft, a heart valve, a membrane, a sealing device, a suture or staple line, a hernia mesh or hernia repair device, a pelvic floor reconstruction device, a wound or burn dressing, a dural closure material, and a heart patch.

[0173] The present invention further provides a device as defined above, wherein the device is a heart valve.

[0174] The present invention further provides a method for manufacturing a construct according to any one of the above, the method comprising: - contacting at least one surface region of at least one decellularized tissue with a liquid polymer; and - contacting at least one surface region of at least one metal element with at least one liquid polymer; forming the construct by contacting the at least one surface region of the at least one metal element comprising the at least one liquid polymer with the at least one surface region of the at least one decellularized tissue comprising the liquid polymer. comprising.

[0175] The present invention further provides the method as defined above, wherein the liquid polymer coats the at least one surface area of the at least one metal element and / or the at least one surface area of the at least one decellularized tissue.

[0176] The present invention further provides the method as defined above, wherein the liquid polymer is cured.

[0177] The present invention further provides a method for manufacturing a construct comprising at least one decellularized tissue, at least one metal element, and at least one polymer component, the method comprising: - contacting at least one surface area of the at least one decellularized tissue with at least one liquid polymer, and - contacting at least one surface area of the at least one metal element with at least one liquid polymer, forming the construct by contacting the at least one surface area of the at least one metal element with the at least one liquid polymer with the at least one surface area of the at least one decellularized tissue with the at least one liquid polymer. comprising.

[0178] The present invention further provides the method as defined above, wherein the liquid polymer coats the at least one surface area of the at least one metal element and / or the at least one surface area of the at least one decellularized tissue.

[0179] The present invention further provides the method as defined above, wherein the liquid polymer is cured.

[0180] Another object of the present invention is to provide a construct comprising at least one decellularized tissue, at least one metal element, and at least one polymer component, wherein the at least one polymer component physically contacts at least a portion of at least one surface region of the decellularized tissue without penetrating it, and further, the at least one polymer component at least partially coats at least one surface region of the metal element.

[0181] Another object of the present invention is to provide a decellularized tissue physically associated with at least one metal element by at least one polymer component, said association comprising or consisting of at least partial physical contact of the polymer component with at least a portion of the surface region of the tissue without penetrating it, and coating of at least a portion of the at least one metal element by the at least one polymer component.

[0182] Another object of the present invention is to provide a construct comprising at least one decellularized tissue, at least one metal element, and at least one polymer component, wherein the polymer component has at least one surface feature that at least partially and without penetrating at least one surface of the decellularized tissue and at least partially associates with at least one surface region of the metal element.

[0183] Another object of the present invention is to provide the construct as defined above, wherein the at least one decellularized tissue in at least partial contact with the at least one polymer component and the metal element at least partially coated with the at least one polymer component are thermally joined to obtain the construct.

[0184] Another object of the present invention is to provide the construct as defined above, wherein the at least one decellularized tissue at least partially coated with the at least one polymer component and the metal element at least partially coated with the at least one polymer component are thermally joined to obtain the construct.

[0185] Another object of the present invention is to provide the construct as defined above, wherein the thermal joining is achieved by raising the temperature of the polymer component above either (a) its transition temperature in the case of an amorphous thermoplastic polymer; or (b) its melting temperature in the case of a semi-crystalline polymer, resulting in at least partial entanglement of the polymer chains along at least one region of the polymer component.

[0186] Another object of the present invention is to provide the construct as defined above in the form of a multi-sheet construct.

[0187] Another object of the present invention is to provide the construct as defined above, wherein the coating is performed by at least one method selected from the group consisting of spraying, brush painting, dip coating, and any combination thereof.

[0188] Another object of the present invention is to provide the construct as defined above, wherein the at least one decellularized tissue is dried before being physically contacted by the polymer.

[0189] Another object of the present invention is to provide the construct as defined above, wherein at least one or any of the at least one sheet of the decellularized tissue is restricted between any two sheets of the polymer component.

[0190] Another object of the present invention is to provide the construct as defined above, wherein the metal element is a metal wire.

[0191] Another object of the present invention is to provide a construct as defined above, further comprising at least one solvent, wherein (a) the solvent is added to a region where at least one surface region of the decellularized tissue in contact with the polymer component contacts at least one surface region of a metal element coated with the polymer component, (b) the polymer component is dissolved in the at least one solvent, and (c) at least one of any combinations thereof applies.

[0192] Another object of the present invention is to provide a construct as defined above, wherein the solvent is selected from the group consisting of THF, acetone, dioxane, DMSO, halogenated solvents such as chloroform and dichloromethane, alcohols and polyols, polyethers, acetonitrile, ethyl acetate, dimethylformamide (DMF), dimethylacetamide, DMAC, and any combinations thereof.

[0193] Another object of the present invention is to provide a construct as defined above, wherein the polymer component is in the form of a layer or coating of particles, a polymer sheet, a polymer film, polymer fibers or a polymer mesh, a gel, a hydrogel, or a liquid or fluid film, and any combinations thereof.

[0194] Another object of the present invention is to provide a construct as defined above, wherein the decellularized tissue is obtained from a tissue selected from oral mucosa, small intestinal submucosa, and bladder decellularized matrix, pericardium, bovine or porcine pericardium, mesh, or small intestinal mucosa, and any combinations thereof.

[0195] Another object of the present invention is to provide a construct as defined above, wherein the polymer component is a hydrophobic polymer, a hydrophilic polymer, an amphiphilic polymer, a blend, an IPN, or a semi-IPN, an acrylic polymer or a methacrylic polymer, a polyolefin, a silicone polymer, a polycarbonate, a polyurethane, a polyurea, or a polyamide, a polyurethane, a polymethyl methacrylate (PMMA), a poly(n-butyl methacrylate) (PBMA), a poly(hexyl methacrylate) (PHMA), a polystyrene (PST), a poly(2-hydroxyethyl methacrylate) (PHEMA), a poly(N-(2-hydroxypropyl) methacrylamide) (PHPMA), a polycyanoacrylate (PCA), a polyethylene / polypropylene copolymer, a polyethylene / polybutylene copolymer, a polypropylene / polybutylene copolymer, a polyisobutylene, a polydimethylsiloxane (PDMS), a phenyl-containing PDMS, a polyester urethane, a polyether urethane, a polycarbonate, a silicone-containing polyurethane, a polyglycolic acid, a polylactic acid, a polycaprolactone, a polylactide-caprolactone copolymer, a polyglycolic acid-lactic acid copolymer, a polyethylene oxide-polylactic acid copolymer, a polyethylene oxide-polycaprolactone copolymer, a polytetramethylene oxide-caprolactone copolymer, a polyhydroxybutyrate, a polyhydroxyvalerate, a polyethylene adipate, a polybutylene adipate, a polyethylene succinate, a polybutylene succinate, and a polybutylene terephthalate, and a polyethylene / butylene terephthalate copolymer, and combinations and copolymers thereof, a shape memory material, a urethane, Pellethane, Elastane, Elastolan, Tecoflex, Biomer, Chronoflex, Biospan, Bionate, a silicone-containing polyurethane selected from CarboSil, PurSil, Avcothane, and Cardiothane, Chroniflex, Tecoflex, or a polymer selected from the group consisting of any combination thereof, or a polymer containing a polymer selected therefrom.

[0196] Another object of the present invention is to provide the construct as defined above, wherein one or more of the sheets are designed as a substance reservoir for releasing an active or inactive substance.

[0197] Another object of the present invention is to provide the construct as defined above, wherein the active substance is selected from analgesics; anxiolytics; antiarrhythmics; antibacterial agents; antibiotics; anticoagulants and thrombolytics; antiepileptics; antidepressants; antidiarrheals; antiemetics; antifungals; antihistamines; antihypertensives; anti-inflammatories; antineoplastic agents; antipsychotics; antipyretics; antivirals; β-blockers; corticosteroids; cytotoxic agents; hormones and sex hormones; enzymes; and vitamins.

[0198] Another object of the present invention is to provide the construct as defined above, wherein the metal element is selected from stents, metallic stents, vascular grafts, heart valves, membranes, sealing devices, suture or staple lines, hernia meshes or hernia repair devices, pelvic floor reconstruction devices, wound or burn dressings, dural closure materials, and heart patches.

[0199] Another object of the present invention is to provide the construct as defined above, wherein, in different cases, at least one polymer component has a viscosity substantially different from that of at least one second polymer component.

[0200] Another object of the present invention is to provide the construct as defined above, wherein at least one second decellularized tissue is at least partially coated by the at least one polymer component and is in physical contact with the at least one polymer component that is in physical contact with the at least one decellularized tissue, whereby the at least one decellularized tissue and the at least one second decellularized tissue are connected by the application of heat bonding.

[0201] Another object of the present invention is to provide a construct as defined above, wherein the polymer component physically contacting at least a portion of at least one surface region of the decellularized tissue has (a) at least partially penetrated at least one surface region of a second decellularized tissue, and / or (b) at least one surface feature that protrudes from one side of the second decellularized tissue and crosses the tissue through at least one hole formed in the tissue to the other side.

[0202] Another object of the present invention is to provide a construct as defined above in the form of a multi-sheet construct.

[0203] Another object of the present invention is to provide a construct as defined above, wherein the multi-sheet construct comprises at least one sheet of a second decellularized tissue and at least one sheet or segment of a polymer component, and any sheet of the second decellularized tissue is adjacent to or in contact with at least one sheet or segment of the polymer component.

[0204] Another object of the present invention is to provide a construct as defined above, wherein the multi-sheet construct comprises a number of sheets of a second decellularized tissue and the same number of sheets of a polymer component.

[0205] Another object of the present invention is to provide a construct as defined above, wherein the at least one second decellularized tissue is dried before being coated with the polymer.

[0206] Another object of the present invention is to provide a construct as defined above, wherein the at least one hole is pre-formed or present in the decellularized tissue.

[0207] Another object of the present invention is to provide a device comprising a construct according to any one of the above.

[0208] Another object of the present invention is to provide a device configured as an implant, as defined above.

[0209] Another object of the present invention is to provide a device as defined above, the device being selected from a stent, a metallic stent, a vascular graft, a heart valve, a membrane, a sealing device, a suture or staple line, a hernia mesh or hernia repair device, a pelvic floor reconstruction device, a wound or burn dressing, a dural closure material, and a heart patch.

[0210] Another object of the present invention is to provide a method for manufacturing a construct according to any one of the above, the method comprising: - contacting at least one surface region of at least one decellularized tissue with at least one polymer, and - contacting at least one surface region of at least one metallic element with at least one polymer, forming the construct by contacting the at least one surface region of the at least one metallic element with the at least one polymer with the at least one surface region of the at least one decellularized tissue with the polymer. comprises.

[0211] Another object of the present invention is to provide a method as defined above, wherein the polymer coats at least one surface region of at least one metallic element and / or at least one surface region of at least one decellularized tissue.

[0212] Another object of the present invention is to provide a method as defined above, wherein the polymer is cured.

[0213] Another object of the present invention is to provide a method as defined above, further comprising the step of adding at least one solvent, wherein (a) the solvent is added to a region where at least one surface region of the decellularized tissue in contact with the polymer component contacts at least one surface region of a metal element coated with the polymer component, (b) the polymer component is dissolved in the at least one solvent, and (c) at least one of any combinations thereof applies.

[0214] Another object of the present invention is to provide a method as defined above, wherein the solvent is selected from the group consisting of THF, acetone, dioxane, DMSO, halogenated solvents such as chloroform and dichloromethane, alcohols and polyols, polyethers, acetonitrile, ethyl acetate, dimethylformamide (DMF), dimethylacetamide, DMAC, and any combinations thereof.

[0215] Another object of the present invention is to provide a method as defined above, contacting at least one perforated surface region of the decellularized tissue with at least one polymer, and penetrating the at least one polymer through the perforation (hole) to form a polymer sheet on the surface region, further comprising.

[0216] Another object of the present invention is to provide a method as defined above, contacting at least one surface region of the decellularized tissue with at least one polymer, and coating at least partially the at least one polymer with the at least one decellularized tissue to form a polymer sheet thereon, further comprising.

[0217] Another object of the present invention is to provide a method as defined above, further comprising at least one step selected from: (a) a step of perforating or forming holes in a surface area of at least one decellularized tissue; (b) a step of injecting a polymer into the perforated part (holes); (c) a step of curing the polymer; and (d) any combination thereof.

[0218] Another object of the present invention is to provide a method as defined above, further comprising at least one step selected from: (a) a step of thermally bonding at least one decellularized tissue and at least one polymer by raising the temperature of the polymer component above either (i) its transition temperature in the case of an amorphous thermoplastic polymer or (ii) its melting temperature in the case of a semi-crystalline polymer, thereby at least partially causing entanglement of polymer chains along at least one region of the polymer component; (b) a step of selecting the polymer to be in the form of a layer or coating of particles, a polymer sheet, a polymer film, polymer fibers or a polymer mesh, a gel, a hydrogel, or a liquid or flowable film, and any combination thereof; and (c) any combination thereof.

[0219] Another object of the present invention is to provide a method for manufacturing a construct comprising at least one decellularized tissue, at least one metal element, and at least one polymer component, the method comprising: - contacting at least one surface area of at least one decellularized tissue with at least one polymer, and - contacting at least one surface area of at least one metal element with at least one polymer, forming the construct by contacting the at least one surface area of the at least one metal element provided with the at least one polymer with the at least one surface area of the at least one decellularized tissue provided with the polymer. comprising.

[0220] Another object of the present invention is to provide the method defined above, wherein the polymer coats at least one surface region of the at least one metal element and / or at least one surface region of the at least one decellularized tissue.

[0221] Another object of the present invention is to provide the method defined above, wherein the polymer is cured.

[0222] Another object of the present invention is to provide the method defined above, further comprising the step of adding at least one solvent, wherein (a) the solvent is added to a region where at least one surface region of the decellularized tissue in contact with the polymer component contacts at least one surface region of the metal element coated with the polymer component, (b) the polymer component is dissolved in the at least one solvent, and (c) at least one of any combination thereof applies.

[0223] Another object of the present invention is to provide the method defined above, wherein the solvent is selected from the group consisting of THF, acetone, dioxane, DMSO, halogenated solvents such as chloroform and dichloromethane, alcohols and polyols, polyethers, acetonitrile and ethyl acetate, dimethylformamide (DMF), dimethylacetamide, DMAC, and any combination thereof.

[0224] Another object of the present invention is to provide the method defined above, a. contacting at least one surface region of at least one decellularized tissue having one or more holes formed therein with at least one polymer; and, b. passing the at least one polymer through the one or more holes to form a polymer sheet on the surface region. is further included.

[0225] Another object of the present invention is to provide the method defined above, c. contacting a surface region of at least one decellularized tissue with at least one polymer, and d. coating at least partially the at least one decellularized tissue with the at least one polymer to form a polymer sheet thereon, further comprising.

[0226] Another object of the present invention is to provide a method as defined above, comprising at least one step selected from (a) forming holes in a surface region of at least one decellularized tissue; (b) injecting a polymer into the perforated portion (holes); (c) curing the polymer; (d) joining or fusing two or more constructs; (e) any combination thereof.

[0227] Another object of the present invention is to provide a method as defined above, wherein at least one of the following applies: (a) the polymer completely penetrates one or more holes; (b) the polymer partially penetrates one or more holes; (c) the polymer completely penetrates one or more holes to form polymer sheets on both sides of the surface region, thereby forming a construct assembly; and (d) any combination thereof.

[0228] Another object of the present invention is to provide a method as defined above, comprising (a) thermally bonding the at least one decellularized tissue and the at least one polymer by raising the temperature of the polymer component above (i) its transition temperature in the case of an amorphous thermoplastic polymer or (ii) its melting temperature in the case of a semi-crystalline polymer, to at least partially cause entanglement of polymer chains along at least one region of the polymer component; (b) selecting the polymer in the form of a layer or coating of particles, a polymer sheet, a polymer film, polymer fibers or a polymer mesh, a gel, a hydrogel, or a liquid or fluid film, and any combination thereof; (c) further comprising at least one step selected from any combination thereof.

[0229] The present invention further provides a method as defined above, further comprising associating or fusing two or more constructs.

[0230] For a better understanding of the subject matter disclosed herein and to exemplify how it may be actually implemented, embodiments will now be described, by way of non-limiting example only, with reference to the accompanying drawings.

Brief Description of the Drawings

[0231]

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Mode for Carrying Out the Invention

[0232] Accordingly, for the purpose of providing an excellent type of biomedical construct and medical device without the drawbacks associated with currently available equivalents, the inventors of the technology disclosed herein have developed a novel and unique type of construct comprising [i] a decellularized tissue and [ii] a polymer component, each component forming spatially distinct phases, wherein the decellularized tissue and the polymer component are associated with each other via one or more physical fixation means. Unexpectedly, the constructs and medical devices disclosed herein exhibit properties that have not been achievable heretofore.

[0233] The decellularized tissue / polymer constructs taught by the present invention constitute or are part of a medical device, and examples of such constructs include, inter alia, vascular grafts, heart patches, stents, heart valves, wound or burn dressings, membranes, sealing devices, or devices for reinforcing suture or staple lines, devices used for hernia repair, pelvic floor reconstruction, or dural closure.

[0234] The teachings of the present invention are readily applicable to a variety of decellularized tissues and polymers. However, for the sake of clarity, brevity, and simplicity, and without diminishing the generality of the scope of the present invention in any form or manner, the inventors have chosen to illustrate the present invention disclosed herein by focusing on constructs comprising decellularized pericardium, and more precisely decellularized bovine pericardium. For the sake of clarity, brevity, and simplicity, and without diminishing the generality of the scope of the present invention in any form or manner, the inventors have chosen to illustrate the present invention disclosed herein by focusing on the decellularized pericardium / polymer construct as part of a larger medical device, more precisely a heart valve. For the sake of clarity, brevity, and simplicity, and without diminishing the generality of the scope of the present invention in any form or manner, the inventors have chosen to illustrate the present invention disclosed herein by focusing on polyurethane as the polymer phase of the decellularized pericardium / polymer construct, more precisely when the polyurethane is Tecoflex.

[0235] Among these, but not limited to, the decellularized bovine pericardium / polymer construct disclosed by the present invention was developed in the inventors' laboratory. The mechanical properties of the decellularized pericardium vary to some extent depending on the batch and the decellularization technique used. Typically, the breaking stress of the decellularization technique falls within the range of 10 - 80 MPa, its typical Young's modulus value is in the range of 80 - 300 MPa, and thereby exhibits a breaking strain value of 20 - 50%. The values of breaking stress, Young's modulus, and breaking strain measured in the inventors' laboratory for freeze-dried decellularized bovine pericardium were 75 MPa, 280 MPa, and 43%, respectively.

[0236] Tecoflex polyurethane is an aliphatic polyether urethane containing polytetramethylene oxide soft segments and methylene dicyclohexane diisocyanate (MDI) (Richards JM, McClennen WH, Meuzelaar HLC, Shockcor JP, Lattimer RP: Determination of the structure and composition of clinically important polyurethanes by mass spectrometric techniques. Journal of Applied Polymer Science 1987, 34:1967 - 1975). TIFF2025516509000001.tif26170

[0237] The mechanical properties of Tecoflex were measured in the inventors' laboratory and are shown in Table 1. TIFF2025516509000002.tif49170

[0238] As is evident from the mechanical data shown, Tecoflex is a very strong material and will not be a weak component of the construct. Other polyurethanes such as Elastolan, and even other polymers showing similar rheology and mechanical properties may be used. In the case of constructs that require the polymer component to be biodegradable, among many others, polymers such as polylactic acid (PLA), poly(lactic / glycolic acid) (PLGA), polycaprolactone (PCL), and their copolymers, and further, hydrophilic polyethers such as polyethylene oxide (PEO) or their hydrophobic counterparts such as polypropylene oxide (PPO) or polytetramethylene oxide (PTMO), or soft aliphatic polyesters such as amorphous polycaprolactone, or biodegradable polymers that are softer, such as plastic or elastomeric biodegradable block copolymers containing silicone-based segments such as polydimethylsiloxane (PDMS). The molecular weight of the soft segments typically varies from 600 to 20,000 Daltons.

[0239] For purposes of clarity, brevity, and simplicity, and without limiting the generality of the scope of the invention in any form or manner, the inventors have chosen to illustrate the invention disclosed herein by focusing on constructs in which the polymer phase of the construct can connect two or more of the other phases, where the other phases include any type of material selected from the group consisting of tissue phases or polymers, metals, ceramics, carbonaceous materials, and combinations thereof. More specifically, the inventors have chosen to illustrate the invention disclosed herein by focusing on constructs in which the polymer Tecoflex phase of the construct connects two decellularized bovine pericardial tissue samples. Even more specifically, the inventors have chosen to illustrate the invention disclosed herein by focusing on constructs that are part of a heart valve and also include a metal frame, where the leaflets are connected to each other and to the metal frame of the valve. In some embodiments of the invention, the tissue / polymer construct is part of a decellularized pericardial leaflet. In still other embodiments of the invention, the polymer Tecoflex phase of the construct connects two tissue leaflets, thereby forming a multi-layer integrated tissue / polymer construct having three polymer layers, two of which are on the outside of the construct and one of which is in the center, and two tissue layers on the inside of the two outer layers of the polymer layers to form a laminate. In other embodiments, the multi-layer integrated tissue / polymer construct formed consists of polymer layers connected by polymer connectors ranging in size from nanometers to the centimeter scale.

[0240] Materials and Methods Lyophilization of Pericardium In most cases, since the decellularized tissue component of the construct needs to be lyophilized prior to the manufacture of the construct, the decellularized tissue component was examined after lyophilization. The structure of the lyophilized decellularized bovine pericardium (DBP) component is shown in the SEM cross-sectional micrographs presented in FIGS. 1A - 1D below.

[0241] The magnification is increased from a relatively low magnification of ×500, where the SEM micrograph shows the entire thickness of the DBP, to a high magnification of ×120,000, where the known structure of the collagen fibrils can be easily observed.

[0242] Figure 2 shows a high magnification (×200,000) SEM micrograph of the as-received DBP.

[0243] Preparation of the Polymer A selected polymer such as Tecoflex was added to the weld in two different states: a solution of Tecoflex in THF and a pre-prepared film. The Tecoflex solution was Tecoflex in the range of 5 - 25% in THF (hereinafter referred to as the "polymer solution"). For the preparation of the film, 3.78 g of Tecolfex was added to 100 ml of THF and completely dissolved. This solution was poured into a glass Petri dish and the THF was slowly evaporated. The film had an average thickness of several microns.

[0244] Welding of the Pericardium For the welding process, two dry pericardium strips were prepared. For each, a region 2 mm from the end was marked and six holes were drilled. The holes were introduced with the polymer solution using a syringe with a 25G needle at the tip. The needle was inserted from the rough side of the fabric to the smooth side. The solution was poured while pulling the needle back out. The holes were linearly aligned parallel to the edge line in the center of the welding area.

[0245] The perforated fabric was left for several minutes to further evaporate the solvent and then welded with a sealer while pressing the rough sides against each other. Immediately before performing the welding process, a sheet of Tecoflex film was inserted between the pressed fabrics.

[0246] In the welding process, three sealings were performed with a precise strategy combining the temperature, pressure, and time of the press, with relaxation inserted between each cycle of the three cycles. After welding, the welded material was dried at room temperature.

[0247] Welding Property Evaluation Method The strength of the welded strip was analyzed by an Instron device in a tensile test at 10 mm / min, while the leaflet grasped a position 10 mm from the welding area on both sides thereof.

[0248] Also, the structure of the welded strip was examined by SEM to confirm that there were no significant changes in the structure due to pressing and heat.

[0249] The results are shown in Tables 2 to 4 below. TIFF2025516509000003.tif68170TIFF2025516509000004.tif62170TIFF2025516509000005.tif63170

[0250] The data shown in Tables 2 to 4 supports the understanding that the more holes filled with polymer are formed in the tissue, the stronger the association or welding becomes.

[0251] Discussion In some embodiments of the present invention, it may be necessary to apply a mild pressure, particularly when the polymer phase connects two or more DBP and / or polymer phases. The purpose of applying pressure in these embodiments of the present invention is to flow one or more polymer phases in contact with DBP in some embodiments, in contact with other polymer phases in other embodiments, and in combinations thereof.

[0252] In some embodiments of the present invention, the polymer phase can flow through holes or pores formed in the tissue phase, and polymer molecules can also diffuse and entangle with molecules of other phases. In some embodiments, due to the flow of polymer molecules, the interface between the phases disappears and they are welded together.

[0253] Figures 3A - 3C show the structure of the tissue after applying gradually increasing pressures of 100 MPa, 200 MPa, and 500 MPa, respectively, which are far beyond the pressures actually required for the design of DBP / polymer constructs, typically less than 1 MPa, preferably less than 0.5 MPa, and even more preferably less than 0.3 MPa. The fact that the collagen fibrils maintained their structure completely even after these very high pressures demonstrates the robustness of DBP and its remarkable resistance to very high pressures.

[0254] Several polymers capable of forming the polymer phase of the DBP / polymer construct have been demonstrated to have the ability to flow under physiologically acceptable pressure and temperature conditions. Two biodegradable poly(ester - urethane)s consisting of polycaprolactone (PCL) and hexamethylene diisocyanate (HDI) with different molecular weights (2,000 daltons and 14,000 daltons), and the thermograms obtained by differential scanning calorimetry (DSC) of these welded phases showed a new broader endotherm shifted to the low - temperature side. This suggests that the molecules of both polymers diffuse into each other and one hinders the crystallization of the other.

[0255] This phenomenon and the welding connection strength were further investigated. Here, two films of two different polyurethanes, Tecoflex and PEU, were welded together. The results demonstrate the ability of the chains of Tecoflex and PEU to flow across the interface between the two films, providing a strong connection with long - term clinical importance. In addition, although the former is a poly(ester - urethane) and the latter is a poly(ether - urethane), they are similar in that they are both polyurethanes. To illustrate a wide range of this phenomenon, the DSC thermograms of the two polymers were examined.

[0256] It was shown by DSC that two polymers, one being a soft poly(ester-urethane) and the other being a hard polymethacrylate, are very different. The soft polyurethane consists of PCL2000 segments and HDI as its coupling agent, while the hard polymethacrylate is poly(ethyl methacrylate) (PEMA). Also in this case, although the polymers are very different in their composition and mechanical properties, it is clear from their thermograms of the welded phase that this phenomenon involves the mixing and entanglement of the chains of both components at the molecular level. In some embodiments of the present invention, the mobility of the polymer chains and the ability of the polymer chains to unwind, cross the interface with another polymer phase or decellularized tissue phase, diffuse into the second phase, and then entangle again play an important role in manufacturing the constructs or medical devices taught by the present invention.

[0257] The connection strength between DBP and a polymer phase exemplified by Tecoflex was quantitatively determined using an Instron device. As a result, it was found that the construct cohesively failed within the tissue and no peel failure was observed.

[0258] In some embodiments, when aiming for a particularly strong connection between the DBO phase and the polymer phase, holes were formed in the DBP phase to maximize the flow of the polymer phase through them. In some embodiments, needles in the range of 16G to 27G were used to form the holes. FIGS. 4A - 4B show the holes formed with 25G and 27G needles, respectively, filled with a polymer phase which is Tecoflex in this case.

[0259] The DBP phase and the polymer phase may be of any size ranging from nanometers to centimeters and may take any shape including, but not limited to, spherical, fibrous, strip, ribbon, porous or non-porous films, and combinations thereof. One or more polymer phases may also be present in a phase having a size ranging from the nanoscale to the centimeter scale, and they may be present on the surface of and / or within the body of the DBP phase, each of these cases being of a size ranging from nanometers to centimeters and taking any shape.

[0260] The object of the present invention is to connect a DBP / polymer construct or a device of which the construct is a part to a further component of the device. In the case of a heart valve which is a highly preferred embodiment of the present invention, the said further component is the metal frame of the heart valve.

[0261] In some embodiments of the present invention, when welding a DBP / polymer construct or a device of which the construct is a part to a metal stent, the struts of the stent are coated with a weldable polymer which may be the same as or different from the polymer constituting the polymer phase of the DBP / polymer construct. Figure 5 shows the coated struts of a metal stent, with the left side in the compressed state and the right side in the expanded state. In this embodiment, due to the large expansion of the stent, a weldable polymer that is particularly soft and exhibits a high fracture strain value was selected to coat its struts. In the case shown below, the weldable polymer used consisted of PCL and HDI. Starting from a thin coating of 5 micrometers, the thickness was increased as required to prepare coatings of different thicknesses.

[0262] Other high-softness polymers were also used. Among them, various polyurethanes were used, and their soft segments composed of polyethers or polyesters had various molecular weights, hydrophilicities, and in the case of biodegradable polymers, degradation rates as well. One of the polymers used consisted of poly(tetramethylene oxide) (PTMO) (MW = 650) segments chain-extended via HDI. As reported in Table 5, the polymer was welded within 20 seconds at a temperature of 47 - 48 °C. TIFF2025516509000006.tif60170

[0263] The same polymer was also used for coating the struts of the stent, and also for fabricating the polymer phase of the DBP / polymer construct and further polymer phases of medical devices of which the polymer is a part. To illustrate not only the speed of the welding process but also the strength of the formed polymer / polymer connection, a patch of PTMO650 / HDI polymer was welded to the strut of a stent coated with the same polymer in 20 seconds. Further, as shown in Figure 6, the patch was welded only to a very small portion of the coated strut of the metal stent (about 15% of the stent area).

[0264] First, a manual and forceful attempt was made to peel the patch from the coated stent, but as shown in Figure 7, the stent was stretched to three times its original length and the patch did not peel off from the stent.

[0265] Finally, when the patch was peeled from the stent using an Instron device, unexpectedly and surprisingly, it was the metal stent that broke, while the welded connection remained unaffected. This convincingly demonstrates an advantageous feature resulting from the ability of the polymer phase of the DBP / polymer construct, or the polymer phase of a medical device of which the construct is a part, to rapidly and strongly form a long-term stable connection between different phases.

[0266] Figure 8 shows a structure comprising DBP and a two-component construct comprising a DBP and a polymer phase which in this case is Tecoflex. Six holes formed in the DBP to facilitate the flow of Tecoflex, for the purpose of achieving a strong connection between the two DBP phases and the polymer phase, are readily visible. By doing so, in addition to the diffusion of Tecoflex molecules in the DBP phase, a laminate is formed comprising two outer thin Tecoflex films continuously connected to a central Tecoflex film through the holes formed in the two DBP phases.

[0267] In a highly preferred embodiment, as shown in FIGS. 9A-9C, the DCP leaflets of a heart valve which also comprises a metal frame are connected together via a polymer connection, preferably polyurethane, more preferably Tecoflex. In this case, the DBP leaflets are welded together via the polymer phase of the DBP / polymer construct and also to the metal frame of the heart valve via its coated struts.

[0268] As is commonly done and routinely reported in the literature, the long-term dynamic stability of a heart valve is characterized in vitro under accelerated conditions by an accelerated wear tester (AWT).

[0269] Among many studies describing the use of AWT in heart valves, references to three representative papers are shown below: {1} A correlation between long-term in vitro dynamic calcification and abnormal flow patterns past bioprosthetic heart valves, Oleksandr Barannyk, Robert Fraser and Peter Oshkai, J Biol Phys (2017) 43:279-296); {2} Pitfalls and outcomes from accelerated wear testing of mechanical heart valves, A Campbell, T Baldwin, G Peterson, J Bryant and K Ryder, J Heart Valve Dis, 1996 Jun; 5 Suppl 1:S124-32;discussion 144-8; {3} A study in the design of an Accelerated Wear Tester that is compatible with a particle image velocimetry and high-speed camera setup, Edward A. Brown, Master of Science thesis, The Pennsylvania State University, The Graduate School, College of Engineering, 2015.).

[0270] During these tests, a maximum repetitive stress of 100 mmHg was applied, which is equal to 0.0133 MPa (12.3 kPa). The failure values measured using an Instron device typically fell in the range of 5 - 10 MPa, which was more than 300 times higher than the maximum repetitive stress applied during AWT measurements.

[0271] The advantageous features of the DBP / polymer construct taught by the present invention become even more remarkable and surprising when compared to alternative suturing techniques that are commonly used procedures.

[0272] The connection between the phases of the construct and the connection between the construct and the other phases of the medical device of which the construct is a part have the following important advantages compared to suturing: [a] being very fast, in contrast to the extremely cumbersome and time-consuming suturing process through a large number of suture points; [b] generating a strong connection; [c] being reproducible and not dependent on technicians; [d] achieving a much better compliance fit between the phases of the construct and the device, in marked contrast to the use of very much stiffer suturing threads; [e] achieving a much more homogeneous stress distribution, in contrast to the stress concentration effect of suturing threads, which is extremely harmful and sometimes life-threatening; and [f] being low-cost.

[0273] In some embodiments, any phase of the construct and / or the medical device of which the construct is a part, or any element of the present invention, may include at least one additional substance, in particular, to improve any aspect of the clinical performance of any of the embodiments of the present invention and combinations thereof, including, among other things, its biocompatibility, its blood compatibility, the cellular responses it induces, etc. The at least one additional substance may be selected from among active substances and inactive substances. In some embodiments, the active substance is selected from various bioactive agents. Exemplary bioactive agents include, among others, anticoagulants such as heparin and chondroitin sulfate; fibrinolytic agents such as tPA, plasmin, streptokinase, urokinase, and elastase; steroid and non-steroid anti-inflammatory agents such as hydrocortisone, dexamethasone, prednisolone, methylprednisolone, promethazine, aspirin, ibuprofen, indomethacin, ketoralac, meclofenamic acid, tolmetin; calcium channel blockers such as diltiazem, nifedipine, verapamil; antioxidants such as ascorbic acid, carotene and α-tocopherol, allopurinol, trimetazidine; antibiotics such as n-acetylcysteine and other anti-infective antibiotics; gastrointestinal motility improving agents that promote intestinal motility; collagen cross-linking inhibitors such as cis-hydroxyproline and D-penicillamine; and mast cell degranulation inhibitors such as disodium chromolglycate.

[0274] In addition to the above agents that generally exhibit favorable pharmacological activities related to promoting wound healing, reducing infection, maintaining hemostatic properties, or improving blood compatibility, for example, among numerous additional agents, amino acids, peptides, proteins including enzymes, carbohydrates, growth factors, antibiotics (for treating specific microbial infections), anti-cancer agents, neurotransmitters, hormones, immunizing agents including antibodies, nucleic acids including antisense agents, ovulation-inducing agents, psychotropic drugs, and other bioactive agents including local anesthetics can be delivered by the constructs or medical devices of the present invention. The delivery of these and other agents depends on, among numerous factors, the pharmacological activity of the agent, the site of action in the body, the physicochemical properties of the agent being delivered, the therapeutic index of the agent, etc. One skilled in the art would be able to easily adjust the physicochemical properties of the polymers of the present invention and the hydrophobicity / hydrophilicity of the agent being delivered so as to bring about the intended effect. In this aspect of the present invention, the bioactive agent is administered at an effective concentration or amount to bring about the intended result. It should be noted that the chemistry of the polymer phase according to the present invention can be modified to accommodate a wide range of hydrophilic and hydrophobic bioactive agents and their delivery to sites within the patient's body.

[0275] In some embodiments, the inactive substance is selected from among dyes, polymeric materials, thickeners, plasticizers, agents that affect hydrophilicity, agents that affect lubricity, and the like.

[0276] The constructs and medical devices taught by the present invention can be manufactured by any of the existing manufacturing techniques such as extrusion, compression molding, injection molding, dip coating, solvent casting, welding, any of numerous 3D printing techniques, etc., and in each case, the particular manufacturing technique used will be adapted to be compatible with the constructs and medical devices taught by the present invention.

[0277] In some embodiments where at least one of the polymer phases is biodegradable, the biodegradable polymer is selected from the group consisting of lactic acid, lactide, glycolic acid, glycolide, or β-propiolactone, ε-caprolactone, δ-glurolactone, δ-valerolactone, β-butyrolactone, pivalolactone, α,α-diethylpropionolactone, ethylene carbonate, trimethylene carbonate, γ-butyrolactone, p-dioxanone, 1,4-dioxepan-2-one, 3-methyl-1,4-dioxane-2,5-dione, 3,3-dimethyl-1,4-dioxane-2,5-dione, cyclic esters of α-hydroxybutyric acid, α-hydroxyvaleric acid, α-hydroxyisovaleric acid, α-hydroxycaproic acid, α-hydroxy-α-ethylbutyric acid, α-hydroxyisocaproic acid, α-hydroxy-α-methylvaleric acid, α-hydroxyheptanoic acid, α-hydroxystearic acid, α-hydroxylignoceric acid, salicylic acid, and mixtures thereof, and related aliphatic hydroxycarboxylic acids or esters (lactones) selected from the group consisting thereof.

[0278] The polymer phase according to the present invention may optionally contain low molecular weight molecules that improve the fluidity of the polymer and / or enable the polymer phase or a part thereof to flow under milder temperature and pressure conditions. A further object of the present invention is to provide low molecular weight molecules that are polymerizable or crosslinkable, whereby the low molecular weight molecules soften the polymer phase or a part thereof before polymerization or crosslinking and strengthen or harden the polymer phase after polymerization or crosslinking. In this case, the low molecular weight molecules can polymerize or crosslink according to any mechanism including, but not limited to, addition polymerization reactions and condensation polymerization reactions, as well as further reactions including all kinds of click chemistry and combinations thereof. Among others, examples of the polymerizable or crosslinkable low molecular weight molecules include precursors containing one or more double bonds. Some examples are shown in the following figures.

[0279] In Fig. 11, the polymer phase consists of PCL / HDI poly(ester-urethane), and the low molecular weight polymerizable molecule is hydroxyethyl methacrylate (HEMA).

[0280] In Fig. 11, when CLUR2k is not plasticized (100:0), it has a tensile modulus of around 180 MPa. As the addition amount of the smart HEMA component is gradually increased, the tensile modulus decreases significantly, showing a value of about 60 MPa in the presence of 20% monomer HEMA, and it is reported to decrease to 6 MPa at a composition of 50:50. As is clear from the presented data, the polymerized PHEMA results in a significant increase in the elastic modulus of CLUR, reaching a value exceeding 250 MPa at a composition of 50:50 of CLUR2K:PHEMA.

[0281] Fig. 12 shows the behavior of CLUR2K when the low molecular weight component is not only polymerizable but also crosslinkable, as in the case of triethylene glycol dimethacrylate containing two carbon-carbon double bonds.

[0282] Some additional chemistries, such as the epoxy-amine reaction, may be used to polymerize or crosslink the low molecular weight component. This chemistry is exemplified by blending very hard and somewhat brittle polyglycidyl methacrylate (PGMA) (see Fig. 13) with polyethyleneimine molecules containing several amine groups. When first blended with PGMA, the PEI molecules lower the rigidity of PGMA, but when reacting with the epoxide rings (see Fig. 13), they crosslink PGMA and harden the polymer.

[0283] Figure 14 schematically shows the manufacture of a tissue / polymer construct. The first step is to form holes in the tissue component, the size, number, and arrangement of which are optimized. In this case, a liquid polymer phase is added, which may be, for example, a polymer solution in a suitable solvent, or even when the polymer is above its glass transition point or melting point, provided that these transitions occur at suitably low temperatures. The polymer penetrates mainly through the holes formed in the tissue, and the length, size, number, and arrangement of the holes are controlled and vary widely. In some cases, the polymer phase contains holes of different penetration depths. In some cases, the holes only partially penetrate the tissue phase, and in other embodiments, the depth of the holes traverses the tissue phase from face to face. In the case illustrated below, the holes traverse the entire thickness of the tissue, and two polymer layers are formed on both sides of the tissue component. Since they are connected via polymer connectors that fill the holes, the two polymer films form one integrated polymer phase. In this case, the mechanical properties of the construct produced by the physical association of the tissue phase and the polymer phase are particularly high because it is derived from the cohesive force of the polymer itself.

[0284] In other embodiments of the present invention, a pre-formed polymer phase is first manufactured, the composition and form of which are such that it can flow under suitable temperature and pressure conditions as described above.

[0285] In some embodiments, the pre-formed polymer phase has different properties in its surface layer as contrasted with its body, such that the polymer phase in contact with the tissue is enabled to flow into the holes formed in the tissue. In some embodiments, the difference between the surface layer with optimized thickness and the body of the polymer phase is compositional, such that the surface layer exhibits the required fluidity under the applied temperature and pressure conditions. In some embodiments, the difference between the surface layer with optimized thickness and the body of the polymer phase is morphological. In this embodiment, the surface layer is less crystalline and in some embodiments is amorphous, while the polymer in the body of the polymer phase exhibits enhanced crystallinity and thus higher rigidity at an appropriate temperature. The morphological differences encoded in the polymer phase can be realized according to various strategies. In particular, this can be realized by performing a spatially limited heat treatment that renders the surface layer less crystalline or amorphous as contrasted with the body of the polymer phase. In some embodiments, the difference between the surface layer and the body of the polymer phase is realized by the addition of a mobility component that tends to migrate and concentrate in the surface layer of the polymer phase. In still other embodiments, the difference between the surface layer and the body of the polymer phase is realized by judiciously selecting the contact surface on which the polymer phase is manufactured. These and other similar effective techniques may be used separately or in combination.

[0286] In yet further embodiments of the present invention, the polymer phase, which is initially liquid, and the pre-formed polymer phase are combined in different ways. In some embodiments, for example, they are added simultaneously, or they may be arranged sequentially or in any other way that creates an optimal association of tissue / polymer derived from its clinical use.

[0287] A procedure for manufacturing an organization / polymer composed of at least two tissue phases using a polymer solution is exemplified herein and shown in FIG. 14B. In some cases, the term "welding" is used interchangeably with "connection" and similar terms. Using two freeze-dried pericardial strips, the tissue / polymer association resulted in the connection of two tissue phases through the polymer phase. Six holes were formed at intervals across the entire width of the tissue strip within a region 2 mm from the end on each pericardial strip. In this embodiment, hole formation and polymer solution filling were performed simultaneously. In this case, an 8% Tecoflex / THF solution was prepared and the solution was filled into a syringe with a 25G needle at the tip (in other experiments for other purposes, Tecoflex at a concentration of 1% or 4% was used). The needle was inserted from the rough side of the pericardial tissue strip, punctured to form a hole, the polymer solution was first poured from the distal end, then the hole was filled while pulling back, and finally a polymer film was formed on the tissue proximal surface. By doing so, a laminate composed of two outer polymer phases connected through a polymer connection part (through the holes formed in the tissue) was manufactured. The holes were linearly aligned parallel to the edge line at the center of the welding area. After leaving the tissue / polymer construct composed of the laminate described above for 2 minutes to optimally evaporate the solvent (THF in this specific case), the two processed tissue / polymer constructs of each tissue strip were overlapped as needed and welded together using a sealer while pressing the rough surfaces against each other under appropriate temperature, pressure, and time conditions. Three rounds with the sealer were applied at 85°C. Each round with the sealer consisted of a 2 bar, 12 second step, and a 1 second relaxation between cycles. After connection, the welded tissue / polymer construct was dried at room temperature for 5 hours and then immersed in physiological saline for 24 hours. The connection strength between the two connected tissue strips was examined using an Instron device with a crosshead speed of 10 mm / min, grasping the strip 10 mm away from the connection area.

[0288] The strength of the formed connection part was specified even under peeling conditions, and for this purpose, samples were connected accordingly. All other details of the procedure were the same. The mechanical tests of these samples were carried out using the same conditions as those shown previously, and the only difference was that a portion 20 mm from the welding area was grasped.

[0289] Some of the various conditions and connection parameters tested are listed below. 1) For 27G needles - 2 / 4 / 6 / 8 holes For 25G needles - 2 / 4 / 6 holes For 23G needles - 2 / 4 / 6 holes For 21G needles - 2 / 4 holes For 19G needles - 2 / 4 holes For 16G needles - 2 holes 2) Cycles performed with a sealer - 1 to 6 3) Connection pressure - 2 bar to 10 bar 4) Connection time - 8 seconds to 12 seconds

[0290] A procedure for manufacturing a tissue / polymer composed of at least two tissue phases by combining a polymer solution and a pre-formed polymer phase is exemplified herein. The process involving the Tecoflex / THF solution was the same as described above.

[0291] The tissue / polymer construct composed of the laminate formed as described above was left for 2 minutes to optimally evaporate the solvent (THF in this particular case). Before welding the two fabricated tissue / polymer constructs, a 2×6 mm Tecoflex film (thickness approximately 160 μm) was placed between the two tissue / polymer constructs, and then a welding process using a sealer was performed. The Tecoflex film was pre-fabricated by a solvent casting method using 3.78 g of Tecoflex in 100 ml of chloroform, although other solvents were also used. The same process as described previously was performed with the sealer, and the connected samples were tested with an Instron device as described above.

[0292] Some of the various conditions and connection parameters tested are listed below. 1) For 27G needles - 2 / 4 / 6 / 8 holes 2) For 25G needles - 2 / 4 / 6 holes 3) For 23G needles - 2 / 4 / 6 holes 4) For 21G needles - 2 / 4 holes 5) For 19G needles - 2 / 4 holes 6) For 16G needles - 2 holes 2) The film prepared in advance was prepared from THF or chloroform solvent using 1.5 - 4 g of Tecoflex in 100 ml of the described solvent. 3) Cycles performed with a sealer - 1 - 6 4) Welding pressure - 2 bar to 10 bar 5) Welding time - 8 seconds to 12 seconds

[0293] Thus, according to one embodiment of the present invention, a process for connecting tissue to tissue becomes possible.

[0294] According to a preferred embodiment, this process includes the following two main steps: (a) Place two tissue sheets 10 from both sides of the polymer film 20 (see FIGS. 15a and 15b). (b) By injecting the polymer from one side to the other side of the tissue - polymer - tissue "sandwich structure", form a rivet 30 that functions as a polymer tube (see FIGS. 16a - 16c). Such a rivet functions similarly to a conventional "wall flange" or rivet. According to a preferred embodiment, a polymer - tissue - polymer - tissue - polymer sandwich structure is produced (as illustrated in FIG. 14b).

[0295] In other words, the connection / welding of two tissues is a superposition of the following two mechanisms: 1. Surface adhesion (like an adhesive), and 2. Physical connection like a rivet.

[0296] It should be noted that each tissue sheet can be coated with a polymer on both sides thereof. Thus, a sandwich-structured construct is polymer-tissue-polymer-tissue-polymer.

[0297] According to another embodiment of the present invention, laser cutting is used to cut the tissue in the connection ( "welding") region (i.e., the region of connection with the polymer).

[0298] According to another embodiment of the present invention, the tissue is frozen and dried (i.e., the freeze-drying process) at a temperature of -70 to 20 °C, with or without vacuum, for about 5 to 20 hours before being welded to the polymer.

Example

[0299] The following examples are given to evaluate various parameters and their influence on the strength of the weld.

[0300] The tests were mainly divided into three groups. · Welding-related parameters: Heat, pressure, and time were examined to see their influence on the welding strength. It should be noted that the sensitivity of the pericardium to high temperatures is limited to 85 °C. Thus, the main experiments applied were at pressures of 2, 4, and 6 bar, and heating times of 36 seconds (3 pulses), 50 seconds, and 120 seconds (1 pulse). · Hole-related parameters: The number of holes, the distance between the holes, the size of the holes, and the depth. · Polymer-related parameters: Two types of polymers and different thicknesses of the polymer films used.

[0301] It should be noted that all the holes were formed with a laser cutting machine and were arranged in a row. Generally, the protocol used was as follows. The pericardial sheet was divided into 6 pairs of strips, and each pair was designated as one welding sample.

[0302] X holes were formed at a distance of Y from each other in the upper 1 mm of each strip (X and Y are disclosed in each experiment detailed below).

[0303] The laser cutting process was carried out using the following parameters: 100% output, 50% speed, 1000 points per inch (PPI).

[0304] Next, tensile tests were performed. In each test, the tissue was securely fastened to a metal handle covered with plastic to increase surface resistance (Testometric X250-2.5 device, maximum cell load 250 N) and stretched at a rate of 10 mm / min until the breaking point.

[0305] In the following example, the polymer used was Chronoflex.

[0306] Welding-related parameters: 1) Influence of pressure on welding strength: Pressures of 2, 4, and 6 bar and 3 pulses with heating for 12 seconds were applied. In the test, the strength required to "break" the weld was measured. The measurement was carried out using a Testometric X250-2.5 device and stretching at a rate of 10 mm / min until the breaking point.

[0307] Next, refer to Figure 17 showing the influence of pressure on the stress of the weld. It should be noted that stress rather than strength was measured, taking into account the difference in the thickness of the pericardial tissue.

[0308] As can be seen, the higher the applied pressure, the stronger the weld results (more force is required to break the weld). In the figure, "W" represents the breakage of the weld site, "PW" represents the breakage of the pericardium at the weld site, and "P" represents the breakage of the pericardium.

[0309] Using a high pressure forces the polymer chains to move at the weld site, thereby resulting in a higher entanglement. Thus, as shown in the figure, the welding strength increases.

[0310] 2) Influence of heating time and number of pulses on welding strength: Due to the sensitivity of the pericardium to high temperatures, higher temperatures could not be used. However, tests could be conducted to heat at 85 °C for a longer time without relaxation. Therefore, in this example, the heating time was tested as follows: 36 seconds with 3 pulses, 50 seconds with 1 pulse, and 120 seconds with 1 pulse.

[0311] Next, refer to Figure 18 showing the effect of temperature on the welding strength. As disclosed above, due to the difference in the thickness of the pericardial tissue, stress rather than strength was measured.

[0312] As can be seen from the figure, the maximum heating time is 50 seconds, and after that, the welding strength decreases.

[0313] In the figure, "W" represents the failure of the welded part, "PW" represents the failure of the pericardium at the welded part, and "P" represents the failure of the pericardium.

[0314] This result can be explained by the fact that some damage was caused to the pericardium, leading to a decrease in its mechanical properties. In other words, an extended heating time has the same effect as using a higher temperature.

[0315] Hole-related parameters: 1) Effect of hole size on welding strength: Next, refer to Figure 19 showing the effect of hole size on the welding strength. As disclosed above, due to the difference in the thickness of the pericardial tissue, stress rather than strength was measured.

[0316] As can be seen from the figure, up to a hole size of 0.4 mm, the larger the hole diameter, the higher the welding strength. After that, the larger the hole diameter, the lower the welding strength. In the figure, "W" represents the failure of the welded part, "PW" represents the failure of the pericardium at the welded part, and "P" represents the failure of the pericardium.

[0317] 2) Effect of the number of holes on welding strength: Next, refer to FIG. 20 showing the influence of the number of holes on the welding strength. In the figure, "W" represents the failure of the welded part, "PW" represents the failure of the pericardium at the welded part, and "P" represents the failure of the pericardium. As disclosed above, due to the difference in the thickness of the pericardial tissue, stress rather than strength was measured.

[0318] For the purpose of this example, holes of 0.2 were tested. As can be seen from this figure, the maximum strength is obtained for four holes at a distance of 1.5 mm from each other, and then the strength decreases as the number of holes increases.

[0319] On the one hand, the less the number of holes, the less the polymer, and on the other hand, the more the number of holes, the less the tissue. In other words, the welding became weaker in the case of additional holes because the tissue at the welded part decreased.

[0320] Polymer-related parameters: Influence of film thickness on welding strength: Next, refer to FIG. 21 showing the influence of the film thickness on the welding strength. In the figure, "W" represents the failure of the welded part, "PW" represents the failure of the pericardium at the welded part, and "P" represents the failure of the pericardium. As disclosed above, due to the difference in the thickness of the pericardial tissue, stress rather than strength was measured.

[0321] As can be seen, this result shows that the thicker the polymer, the weaker the welding area. A thicker polymer means that a higher energy (temperature in the case of the present inventors) is required to break the entanglement of the polymer chains. Since there is a temperature limit, the use of a thicker polymer is not preferred, and as a result, the welding strength will increase.

[0322] In the following example, the polymer used is polycaprolactone 43K, PCL 43K. It should be noted that PCT is a biodegradable polyester having a low melting point of around 60 °C.

[0323] 1) Influence of hole size on welding strength: Since the melting point of PCL is about 60 °C, the welding process of PCL was carried out at 55 °C, a pressure of 4 bar, and for 50 seconds with 1 pulse.

[0324] As described above, the insertion of holes strengthens the welding site because the holes contain polymer and increase the occurrence of welding.

[0325] The 8% solution of PCL is not as viscous as Chronoflex, so the amount of polymer contained in the holes can be reduced. In addition, the temperature used in this process essentially melts the PCL (because PCL has a low melting temperature), and thus, as shown in Figure 22, the welding becomes weaker (in the figure, "W" represents the failure of the welding site, "PW" represents the failure of the pericardium at the welding site, and "P" represents the failure of the pericardium). As disclosed above, due to the difference in the thickness of the pericardial tissue, stress rather than strength was measured.

[0326] 2) Influence of the number of holes on welding strength: In this section, the number of holes was tested, and the size of the holes used was 0.2 mm.

[0327] Next, refer to Figure 23 showing the influence of film thickness on welding strength. In the figure, "W" represents the failure of the welding site, "PW" represents the failure of the pericardium at the welding site, and "P" represents the failure of the pericardium. As disclosed above, due to the difference in the thickness of the pericardial tissue, stress rather than strength was measured.

[0328] As can be seen from Figure 23, the welding strength increased as the number of holes increased.

[0329] Conclusion: All the experiments conducted demonstrated the importance of holes for strengthening the welding site and a better understanding of the welding process and all parameters for achieving improved welding with mechanical properties.

[0330] In summary, the insertion of both PCL 43K and Chronoflex into the holes improved the welding strength and enhanced their mechanical properties. However, even when using the same set of parameters, the best results were not obtained. For PCL, the best results were obtained by forming six 0.2-mm holes, and the peak stress was almost twice as high. On the other hand, for Chronoflex, the best results were obtained by inserting it into four 0.4-mm holes, and the strength was almost three times that of the result without holes.

[0331] Next, refer to FIGS. 24 to 33 as follows. FIG. 24 shows a dry pericardial tissue without holes. FIG. 25 shows a wet pericardial tissue without holes after a tensile test. FIG. 26 shows a dry pericardial tissue with four 0.6-mm holes. FIG. 27 shows a wet pericardial tissue with four 0.6-mm holes that was wetted after a tensile test. FIG. 28 shows a dry pericardial tissue with three 0.2-mm holes. FIG. 29 shows a wet pericardial tissue with three 0.2-mm holes that was wetted after a tensile test. FIG. 30 shows a dry pericardial tissue with four 0.2-mm holes. FIG. 31 shows a wet pericardial tissue with four 0.2-mm holes that was wetted after a tensile test. FIG. 32 shows a dry pericardial tissue with six 0.2-mm holes. FIG. 33 shows a wet pericardial tissue with six 0.2-mm holes that was wetted after a tensile test.

[0332] The following examples are given to evaluate various steps in the manufacture of a multi-sheet construct and its effect on the strength of the weld.

[0333] 1. Effect of freeze-drying (lyophilization) on tissue strength (before welding): In the following tests, the effect of rapid freezing on the mechanical properties of the tissue was evaluated. The mechanical properties of the dry tissue were compared with and without liquid nitrogen (N2).

[0334] The following test protocol was used and implemented: 1. Laser cut a tissue area with dimensions of 6 mm × 40 mm and a thickness of 250 microns ± 50 microns. 2a. Measure the tensile strength at the central part with a width of 6 mm, both ends along 40 mm, and the central part. 2b. In the liquid nitrogen (N2) test, the tissue was immersed in liquid nitrogen. 3. Next, the tissue sample was placed in a freeze dryer under vacuum at a temperature in the range of -70 to 0 °C for 7 to 24 hours. 4. Next, the dry tissue thickness was measured to compare the differences in treatment with and without liquid nitrogen. 5. Subsequently, the tissue sample was placed in physiological saline for 12 hours for recovery. 6. Next, the dry tissue thickness was measured again. 7. Finally, the tissue was subjected to a tensile test to determine the force required to rupture it, and the stress results were calculated. The measured mechanical properties are as follows. TIFF2025516509000007.tif77170

[0335] As can be seen from the table, freeze-drying the tissue with N2 yielded fairly good results, but the maximum stress was obtained from the treatment of the tissue after freeze-drying without using N 2 It was obtained from the treatment of the tissue after freeze-drying without using it.

[0336] When water is frozen at a very fast cooling rate (an uncontrolled cooling rate), the water molecules in the tissue cells expand and damage the collagen matrix of the tissue. Therefore, when nitrogen is used, the mechanical properties of the tested tissue are inferior to those without using nitrogen. Conversely, by controlling the cooling rate of the water molecules, tissue damage is reduced and better mechanical properties can be obtained.

[0337] 2. Welding strength: a) In this section, the drying time in the freeze dryer was tested and its effect on the welding strength was measured. TIFF2025516509000008.tif56170

[0338] There was no significant difference, but as can be seen from the table, the maximum stress was obtained after 4 days of evaporation.

[0339] b) Test the adhesion strength of the connection between tissues (do not punch holes in the tissue and let the polymer cross the tissue; that is, do not use polymer rivets): Since the pericardial tissue has different characteristics on each side, such as one side being smooth and the other side being rough, the mechanical properties of both sides were tested. TIFF2025516509000009.tif136170

[0340] As can be seen, the best results were obtained when the rough surfaces were facing each other or when indenting into the tissue. All the break points were at the welded connections.

[0341] c) Test of "rivets" In the following tests, the mechanical properties of the rivets were measured according to the following protocol. Measure the tissue thickness area of 250 microns ± 50 microns. 1. Laser cut an appropriate number of holes at a position 1 mm from the end of the tissue with dimensions of 6 mm × 40 mm and a thickness of 250 microns ± 50 microns, and the tensile model. 2. Measure the tensile strength at the central part with a width of 6 mm, both ends and the central part along 40 mm. 3. Next, place the tissue sample in a freeze-dryer under vacuum at a temperature in the range of -70 to 0 °C for 7 to 24 hours. 4. Next, attach the dried tissues together and inject polymer rivets (or inject polymer) into the formed holes to produce a dried tissue polymer film. 5. Thermo-bond the construct (tissue-polymer-tissue) at 60 °C and 2 bar by hot pressing. That is, evaporate the solvent (e.g., THF) for 4 days. The main purpose is to melt the polymer components and connect them together to perform welding. 6. Subsequently, place the tissue sample in physiological saline for 12 hours to recover. 7. Next, the dry tissue thickness was measured again. 8. Finally, the tissue was subjected to a tensile test to determine at what force it would rupture, and the stress results were calculated. TIFF2025516509000010.tif201170

[0342] As can be seen from the above, the results obtained when evaporation was used were not good. Also, the better the results, the greater the number of holes with smaller diameters within the tissue.

[0343] d) Hole size and shape, and holes in different arrays TIFF2025516509000011.tif160170

[0344] e) Different polymers: In the following table, the measured stress results for the different polymers used are summarized. TIFF2025516509000012.tif164170

[0345] As can be seen from the above table, Chronoflex C 75D yields the best results.

[0346] f) Hole alignment, offset test between holes pre-formed in two tissues: In this test, the holes in the tissue are not aligned. The holes overlap to some extent but are not concentric. TIFF2025516509000013.tif45170

[0347] It should be noted that according to another embodiment of the present invention, the tissue-polymer-tissue connection can be achieved by coating / physically contacting the two tissues at least partially with the polymer (without the need to form holes). According to this embodiment, each tissue is at least partially coated (or contacted with the polymer), and then heat bonding is performed to weld the two tissues together.

[0348] In the mechanical property test, two tissue samples of 6 mm × 6 mm were coated with a polymer (the coating thickness was 0.25 - 0.26 mm). TIFF2025516509000014.tif98170

[0349] As can be seen from the above table, such tissue-to-tissue connection (welding) is realized to withstand a force of 4.5 MPa.

[0350] Therefore, according to one embodiment of the present invention, the construct is: Comprising at least two decellularized tissues and at least one polymer component, the polymer component at least partially coating at least one surface region of each of the two decellularized tissues. The welding between the two decellularized tissues is realized by thermally bonding the layers together.

[0351] As described above, heat energy is applied to raise the temperature of the polymer above the appropriate transition temperature, i.e., the glass transition temperature Tg in the case of an amorphous thermoplastic polymer or the melting temperature Tm in the case of a semi-crystalline polymer. When two sheets or segments of the polymer component are brought into intimate contact under these conditions, entanglement of the polymer chains occurs, resulting in welding. According to an aspect of the present invention, the welding does not need to be performed over the entire surface of the sheet or segment. Spot welding in one or more regions of the polymer component may be sufficient to obtain a strong association of multiple assemblies or any two polymer sheets or segments.

[0352] According to another embodiment, a connection / welding between tissue and metal is provided. According to this embodiment, a single attachment mechanism is provided. According to that, the tissue (which is in physical contact with the polymer) is in contact with the polymer-coated metal in order to facilitate welding. In this embodiment, the construct is an assembly of (a) n acellular tissues, at least one of which is at least partially associated with at least one polymer, and m metal elements, at least one of which is at least partially coated with a polymer. n and m are integers greater than 0. According to one embodiment, m is equal to n, and according to another embodiment, m and n are substantially different.

[0353] According to one embodiment, the association between the at least one acellular tissue and the polymer means physical contact / interaction, coating, adhesion, welding, firm fixation, and any combination thereof. As another option or in addition, this association is referred to as follows: (i) At least partially embedding a polymer sheet or segment in the surface area of a layer of tissue, and the embedding may be made through a single point or two or more points on the surface area of the layer, with or without penetrating the tissue, or (ii) Fixing a polymer sheet or segment into the tissue to a tissue depth that allows for secure association, and the fixing may be made through a single fixing point or two or more fixing points, and the depth of fixation or penetration of the polymer sheet or segment may vary but does not include piercing the tissue, or (iii) To completely fix the polymer sheet or segment to the tissue so as to penetrate the tissue completely from one side of the tissue to the other side, and the fixing may be made through a single fixing point or through two or more fixing points. Typically, such penetration will involve surface features configured to protrude from one side of the decellularized tissue through at least one hole formed in the tissue to the other side. As detailed below, such features may be formed in situ after the holes are formed in the tissue, or may be provided on the polymer sheet or segment in a form selected and configured to pierce or perforate the tissue.

[0354] The polymer sheet or segment is said to be associated with the tissue so as to ensure association with the decellularized tissue. One type of association or interaction present in the constructs of the present invention is, as detailed herein, through fixing or perforating the decellularized tissue surface, or by forming in the tissue holes through which two polymer sheets or segments can associate. In some embodiments of the present invention where the construct is composed of a plurality of construct assemblies, each assembly comprising, for example, decellularized tissue confined between two sheets or segments of a polymer component, the association of the plurality of assemblies can be achieved by welding polymer to polymer.

[0355] According to this embodiment, welding between the tissue (which is associated with the polymer) and the polymer-coated metal is made possible by bringing together (e.g., physically contacting) the tissue (which is already associated with the polymer) and the metal element that is at least partially polymer-coated in order to facilitate the welding.

[0356] According to another embodiment, at least one solvent (e.g., THF, DMF) is dropped onto the contact area between the tissue (associated with the polymer) and the metal element that is at least partially coated. According to another embodiment, the solvent partially dissolves the coating polymers (both the polymer associated with the tissue and the polymer-coated metal in the welding area) and fuses them, resulting in the connection / welding of the tissue and the metal.

[0357] According to another embodiment, the use of the solvent at least partially dissolves the top surface of the polymer-coated metal and the top surface of the polymer associated with the tissue, and thus the polymer chains on the top surface are sufficiently mobile to intertwine with the chains of other sheets or segments (and thereby fuse the tissue and the metal together). Such an effect is the reflow effect. It is within the scope of the present invention that the term "reflow" is referred to as a process in which at least partial dissolution of the polymer occurs and welding occurs by rearrangement of the polymer chains.

[0358] According to another embodiment, the polymer is first dissolved in at least one solvent and then applied for the first time to coat the tissue / metal element.

[0359] According to one embodiment, the solvent may be THF, acetone, dioxane, DMSO, halogenated solvents such as chloroform and dichloromethane, alcohols and polyols, polyethers, acetonitrile, ethyl acetate, dimethylformamide (DMF), dimethylacetamide, DMAC, and any combination thereof.

[0360] According to another embodiment, the use of the solvent, when applied to heating, at least partially melts / dissolves the top surface of the metal-coated polymer and the top surface of the polymer associated with the tissue. The terms "thermal reflow" or "thermal bonding" are within the scope of the present invention when referring to the process in which dissolution occurs and then the entire assembly is subjected to controlled heat (as a result, at least a part of the polymer is in a molten state), resulting in its permanent casting (and welding). The coating of the metal element can be carried out by dip coating and / or brushing and / or spraying.

[0361] Next, refer to FIG. 34c showing the polymer-encapsulated frame 40 (implemented by dip coating or spraying) and the tissue 10 associated with the polymer on a specific connection region with the metal frame.

[0362] Thus, according to this embodiment, the metal frame is coated with a polymer, and the tissue is coated with the same polymer in the connection region / welding region.

[0363] The metal frame can be any element selected from a stent, a metal stent, a vascular graft, a heart valve, a membrane, a sealing device, a suture or staple line, a hernia mesh or hernia repair device, a pelvic floor reconstruction device, a wound or burn dressing, a dural closure material, and a heart patch.

[0364] According to another embodiment, the tissue is freeze-dried before the polymer is applied to it.

[0365] It should be pointed out that the above and below disclose the connection between the tissue and the metal, but are equally applicable to any other material (e.g., polymer material) having substantially the same mechanical properties.

[0366] The following measurements provide a test of the mechanical strength of the polymer and its welding to the tissue / metal frame. 1. Strength of the polymer thickness against welding to each other on the top of the metal stent: TIFF2025516509000015.tif242170TIFF2025516509000016.tif104170

[0367] As can be seen from the table, the best mechanical properties were obtained with the combination of Chronoflex 75A and Chronoflex 75A, and the combination of Chronoflex 75A and Chronoflex 75D.

[0368] As an example of a medical device, a stichless heart valve using either the tissue-to-tissue welding and / or tissue-to-metal welding described in any of the above is provided as follows.

[0369] According to this example, the heart valve includes four components.

[0370] A metal stent (or frame) 11 made of nitinol, cobalt chrome, or stainless steel, usually three leaflets 12, an inner skirt 13, and an outer skirt 14 (see Figure 35a). In addition, as disclosed above and below, the heart valve includes a polymer-based component. As disclosed below, the polymer component is used in both its liquid form and as a sheet or thin film.

[0371] According to one embodiment of the present invention, first, the entire tissue portion is laser cut, and holes created by 2D / 3D laser cutting are formed in the leaflets, inner skirt, and outer skirt (see Figure 36a).

[0372] Next, the polymer is either dissolved in at least one solvent (e.g., THF) or used as is, cast as a thin film (10 - 100 microns thick), and subsequently laser cut to create polymer sheets and rivets (hereinafter, to connect different parts of the tissue).

[0373] According to one embodiment, the polymer sheet is cast and laser cut in the shape of a 2D projection view of the assembled heart valve (see Figure 35b).

[0374] According to one embodiment, the polymer sheet is perforated, and according to another embodiment, the polymer sheet is maintained without perforation.

[0375] Next, freeze-drying of the tissue is carried out overnight at a temperature of -54 °C with the vacuum pressure reduced from 110 KPa to 2 - 3 Pa.

[0376] Subsequently, coating of the stent (frame) is initiated. At least one layer of coating is applied onto the stent using Chronoflex in tetrahydrofuran (THF), preferably more than 10 layers of coating are applied onto the frame. Preferably, the metal stent (frame) is coated by means selected from brushing, spraying, or dip coating. According to one embodiment of the present invention, heating for drying the coating layer is applied between at least one layer of coating and another coating.

[0377] Next, welding different tissue components to each other is achieved by welding the polymer sheet to the tissue on both of its sides. Welding is achieved by injecting a solution of Chronoflex dissolved in THF into the holes of the tissue. The solution of Chronoflex dissolved in THF results in adhering the tissues to each other. To further strengthen the adhesion, thermo-bond heating is applied. Thermo-bonding is performed at least 6 bar and a heating temperature of 80 °C to promote and strengthen the welding for all welded parts.

[0378] It should be noted that all holes, taking the holes of the leaflets and skirts as an example, always need to be substantially aligned in order to obtain improved welding.

[0379] Thermo-bond can be used to promote and strengthen welding.

[0380] Next, after welding, the assembly can be placed in physiological saline for at least 14 hours for tissue recovery.

[0381] Next, start the process of tissue and metal to obtain a completed heart valve. It should be noted that the welding of tissue to tissue was performed with the tissue in a dry state, while the welding of tissue to metal is obtained with the tissue in a wet state (after recovery in physiological saline).

[0382] In the welding process of tissue and metal, the tissue-polymer-tissue assembly is placed inside the stent / frame, and the welding of the tissue-polymer-tissue and the polymer-coated stent is also obtained by using a polymer here. That is, the polymer is used to coat the corresponding regions of the tissue-polymer-tissue assembly and the polymer-coated stent. Next, thermal bonding is applied. As described above, the welding of polymer sheets occurs when the polymer chains on the surface of one sheet or segment are sufficiently mobile to intertwine with the chains of the other sheet or segment. To achieve welding, thermal energy can be applied to raise the temperature. When two sheets or segments of the polymer component are brought into close contact under these conditions, the intertwining of the polymer chains occurs, resulting in welding. According to an aspect of the present invention, the welding does not need to be performed over the entire surface of the sheet or segment. Spot welding in one or more regions of the polymer component may be sufficient to obtain a strong association of multiple assemblies or any two polymer sheets or segments.

[0383] It should be pointed out that the welding can be realized by arranging a large number along the circumferential direction of the stent.

[0384] According to another embodiment, the coated metal (i.e., polymer-metal-polymer) is encapsulated within a tissue-polymer-tissue assembly, i.e., a tissue-polymer-tissue-polymer-metal-polymer-tissue-polymer-tissue construct is obtained (see FIGS. 36b-36c). As can be seen from FIG. 36c, a portion of the metal frame is encapsulated within the tissue-polymer-tissue assembly.

[0385] According to one embodiment, at least one polymer component may be used, and as another option, it should be noted that two different polymers each having a different viscosity may be used.

[0386] The all-welded stitchless valve assembly was subjected to hemodynamic and durability tests. Next, refer to FIG. 37 showing the hemodynamic behavior of the valve according to the present invention.

[0387] In the test, two main parameters were presented: EOA - Effective Orifice Area, which provides a numerical value for how well the valve opens and how well blood flows through the valve (the higher the numerical value, the better the function), and R.F - Regurgitation Fraction, which provides the percentage of how much blood leaks due to valve regurgitation per heartbeat (i.e., how much leaks out of the valve). As can be seen from the figure, the valve according to the present invention reached 130M cycles out of the required 200M cycles.

[0388] The terms "comprises," "comprising," "includes," "including," "having," and their conjugations mean "including but not limited to."

[0389] The term "consisting of" means "including and limited to."

[0390] The term "consisting essentially of" means that a composition, method, or structure may include additional ingredients, steps, and / or parts, provided that such additional ingredients, steps, and / or parts do not substantially change the basic and novel characteristics of the claimed composition, method, or structure.

[0391] As used herein, the singular forms "a," "an," and "the" include plural references unless the context clearly dictates otherwise. For example, the terms "a compound" or "at least one compound" can include multiple compounds, including mixtures thereof.

[0392] Throughout this application, various embodiments of the invention may be presented in a range format. It should be understood that the description in range format is merely for convenience and brevity and should not be construed as an immutable limitation on the scope of the invention. Accordingly, a range description should be considered to specifically disclose not only the individual numerical values within that range but also all possible sub-ranges. For example, a range description such as 1-6 should be considered to specifically disclose not only the individual numbers within that range, e.g., 1, 2, 3, 4, 5, and 6, but also sub-ranges such as 1-3, 1-4, 1-5, 2-4, 2-6, 3-6, etc. This applies regardless of the width of the range.

[0393] When numerical ranges are indicated herein, it is intended that each recited numerical value within the indicated ranges (fractional or integral) be included. The terms "ranging between" and "ranging from" a first recited number to a second recited number are used interchangeably herein and are intended to include the first and second recited numbers, and all fractional and integral values therebetween. As used herein, the term "method" means a method, means, technique, and procedure for achieving a given objective, including, but not limited to, those techniques, means, techniques, and procedures known to or readily developed from known techniques, means, techniques, and procedures by those skilled in the fields of chemistry, pharmacy, biology, biochemistry, and medicine. As used herein, the term "treating" includes preventing the progression of a medical condition, substantially inhibiting, delaying or reversing, substantially alleviating the clinical or aesthetic symptoms of a medical condition, or substantially preventing the appearance of the clinical or aesthetic symptoms of a medical condition.

[0394] For clarity, it is understood that certain features of the invention described in the context of separate embodiments may also be provided in combination in a single embodiment. Conversely, for brevity, various features of the invention described in the context of a single embodiment may also be provided separately, or in any suitable subcombination, or as suitable in any other embodiment of the invention described. Certain features described in the context of various embodiments should not be considered essential features of those embodiments, except where the embodiment cannot be practiced without those elements. The various embodiments and aspects of the invention detailed above herein and claimed in the following claims sections are experimentally supported in the following examples.

Claims

**Claim 1** A construct comprising at least one decellularized tissue, at least one element, and at least one polymer component, wherein the at least one polymer component physically contacts at least a portion of at least one surface region of the at least one decellularized tissue, the at least one polymer component at least partially coats at least one surface region of the element, and further, the at least one surface region of the decellularized tissue in physical contact with the polymer component contacts the at least one surface region of the element coated by the polymer component. **Claim 2** A decellularized tissue physically associated with at least one element by at least one polymer component, the association including or consisting of at least partial physical contact of the polymer component with at least a portion of a surface region of the at least one decellularized tissue and coating of at least a portion of the at least one element by the at least one polymer component, and the at least one surface region of the decellularized tissue associated with the polymer component contacts the at least one surface region of the element coated by the polymer component. **Claim 3** A construct comprising at least one decellularized tissue, at least one element, and at least one polymer component, wherein the polymer component has at least one surface feature that at least partially associates with at least one surface of the decellularized tissue and at least partially associates with at least one surface region of the element, and the at least one surface region of the decellularized tissue in physical contact with the polymer component contacts the at least one surface region of the element coated by the polymer component. **Claim 4** The construct according to any one of claims 1 to 3, wherein the at least one element is selected from the group consisting of a metal element, a polymer material, any material having mechanical properties substantially different from the at least one decellularized tissue, and any combination thereof. **Claim 5** The construct according to claim 4, wherein the at least one decellularized tissue in at least partial contact with the at least one polymer component is thermally joined to obtain the association.

6. The construct according to claim 5, wherein the thermal joining is achieved by raising the temperature of the polymer component above either (a) its transition temperature in the case of an amorphous thermoplastic polymer or (b) its melting temperature in the case of a semi-crystalline polymer, resulting in at least partial entanglement of polymer chains along at least one region of the polymer component.

7. The construct according to claim 4, which is in the form of a multi-sheet construct.

8. The construct according to claim 4, wherein the coating is performed by at least one method selected from the group consisting of spraying, brushing, dip coating, and any combination thereof.

9. The construct according to claim 4, wherein the at least one decellularized tissue is dried before being physically contacted with the polymer.

10. The construct according to claim 9, wherein the drying is freeze-drying the at least one decellularized tissue.

11. The construct according to any one of claims 1 to 10, wherein at least one or any of the at least one sheet of the decellularized tissue is confined between any two sheets of the polymer component.

12. The construct according to claim 11, wherein the multi-sheet construct comprises a certain number of sheets of the decellularized tissue and the same number of sheets of the polymer component.

13. The construct according to any one of claims 1 to 3, comprising two or more assemblies of decellularized tissue confined between two sheets or segments of the polymer component, the assemblies being associated with each other.

14. The construct according to claim 13, wherein at least two assemblies are oriented in opposite directions.

15. The construct according to claim 4, wherein the metal element is a metal wire.

16. Furthermore, the construct according to any one of claims 1 to 15, comprising at least one solvent, wherein the at least one surface region of the decellularized tissue in physical contact with the polymer component is added to a region in contact with the at least one surface region of the element coated by the polymer component.

17. The construct according to claim 16, wherein the addition of the solvent results in at least partial dissolution of (a) at least a portion of the polymer component in physical contact with the decellularized tissue and (b) at least a portion of the polymer component covering at least one surface region of the element.

18. The construct according to claim 17, obtained by a reflow effect of entanglement of chains along at least one region of the polymer component of (a) at least a portion of the polymer component in physical contact with the decellularized tissue and (b) at least a portion of the polymer component covering at least one surface region of the element.

19. Furthermore, the construct according to any one of claims 1 to 18, comprising at least one solvent, wherein the polymer component is dissolved in the at least one solvent.

20. The construct according to any one of claims 16 to 19, wherein the solvent is selected from the group consisting of THF, acetone, dioxane, DMSO, halogenated solvents such as chloroform and dichloromethane, alcohols and polyols, polyethers, acetonitrile, ethyl acetate, dimethylformamide (DMF), dimethylacetamide, DMAC, and any combination thereof.

21. The construct according to any one of claims 1 to 20, wherein the polymer component is in the form of a layer or coating of particles, a polymer sheet, a polymer film, polymer fibers or a polymer mesh, a gel, a hydrogel, or a liquid or fluid film, and any combination thereof.

22. The construct according to any one of claims 1 to 21, wherein the decellularized tissue constitutes 5 wt% to 95 wt% of the construct.

23. The construct according to any one of claims 1 to 22, wherein the decellularized tissue is obtained from a tissue selected from oral mucosa, small intestinal submucosa, and bladder decellularized matrix, pericardium, mesh, or small intestine mucosa, bovine pericardium, porcine pericardium, and any combination thereof.

24. The construct according to any one of claims 1 to 23, wherein the polymer component is a polymer selected from hydrophobic polymers, hydrophilic polymers, and amphiphilic polymers or comprises a polymer selected from among them.

25. The construct according to any one of claims 1 to 23, wherein the polymer component is a blend, IPN, or semi-IPN or comprises a blend, IPN, or semi-IPN.

26. The construct according to any one of claims 1 to 23, wherein the polymer component is an acrylic polymer or a methacrylic polymer or comprises an acrylic polymer or a methacrylic polymer.

27. The construct according to any one of claims 1 to 23, wherein the polymer component is a polyolefin or comprises a polyolefin.

28. The construct according to any one of claims 1 to 23, wherein the polymer component is a silicone polymer or comprises a silicone polymer.

29. The construct according to any one of claims 1 to 23, wherein the polymer component is a polycarbonate, polyurethane, polyurea, or polyamide, and combinations thereof, or comprises a polycarbonate, polyurethane, polyurea, or polyamide, and combinations thereof.

30. The construct according to any one of claims 1 to 23, wherein the polymer component is a polyurethane or comprises a polyurethane.

31. The polymer component is a polymer selected from polymethyl methacrylate (PMMA), poly(n-butyl methacrylate) (PBMA), poly(hexyl methacrylate) (PHMA), polystyrene (PST), poly(2-hydroxyethyl methacrylate) (PHEMA), poly(N-(2-hydroxypropyl)methacrylamide) (PHPMA), polycyanoacrylate (PCA), polyethylene / polypropylene copolymer, polyethylene / polybutylene copolymer, polypropylene / polybutylene copolymer, polyisobutylene, polydimethylsiloxane (PDMS), phenyl-containing PDMS, polyester urethane, polyether urethane, polycarbonate, silicone-containing polyurethane, polyglycolic acid, polylactic acid, polycaprolactone, polylactide-caprolactone copolymer, polyglycolic acid-lactic acid copolymer, polyethylene oxide-polylactic acid copolymer, polyethylene oxide-polycaprolactone copolymer, polytetramethylene oxide-caprolactone copolymer, polyhydroxybutyrate, polyhydroxyvalerate, polyethylene adipate, polybutylene adipate, polyethylene succinate, polybutylene succinate, and polybutylene terephthalate, and polyethylene / butylene terephthalate copolymer, and combinations and copolymers thereof, or a construct according to any one of claims 1 to 23 comprising a polymer selected from them.

32. The polymer component is a shape memory material or comprises a shape memory material, a construct according to any one of claims 1 to 23.

33. The polymer component is a polyether urethane selected from Pellethane, Estane, Elastolan, Tecoflex, Biomere or comprises a polyether urethane selected from them, a construct according to any one of claims 1 to 23.

34. The polymer component is a polycarbonate urethane selected from Chronoflex, Biospan, and Bionate or comprises a polycarbonate urethane selected from them, a construct according to any one of claims 1 to 23.

35. The construct according to any one of claims 1 to 23, wherein the polymer component is a silicone-containing polyurethane selected from Carbosi, PurSil, Avcothane, and Cardiothane or comprises a silicone-containing polyurethane selected from them.

36. The construct according to any one of claims 1 to 23, wherein the polymer component is Chronoflex, Tecoflex, or a combination of any of these or comprises Chronoflex, Tecoflex, or a combination of any of these.

37. The construct according to any one of claims 1 to 36, wherein one or more of the sheets are designed as a substance reservoir for releasing an active substance or an inactive substance.

38. The construct according to claim 37, wherein the active substance is selected from analgesics; anxiolytics; antiarrhythmics; antibacterial agents; antibiotics; anticoagulants and thrombolytics; anticonvulsants; antidepressants; antidiarrheals; antiemetics; antifungal agents; antihistamines; antihypertensives; anti-inflammatory agents; antineoplastic agents; antipsychotics; antipyretics; antivirals; beta-blockers; corticosteroids; cytotoxic agents; hormones and sex hormones; enzymes; and vitamins.

39. The construct according to claim 4, wherein the metal element is selected from stents, metallic stents, vascular grafts, heart valves, membranes, sealing devices, suture or staple lines, hernia meshes or hernia repair devices, pelvic floor reconstruction devices, wound or burn dressings, dural closure materials, and heart patches.

40. The construct according to any one of claims 1 to 39, wherein at least two polymer components are used, and further, at least one polymer component has a viscosity substantially different from that of at least one second polymer component.

41. At least a portion of at least one surface region of at least one second decellularized tissue is (a) at least partially physically in contact with said at least one polymer component, and (b) physically in contact with said at least one polymer component that is physically in contact with said at least one decellularized tissue, whereby, by application of heat bonding, said at least one decellularized tissue and said at least one second decellularized tissue are connected, the construct according to any one of claims 1 to 40.

42. The polymer component that is physically in contact with at least a portion of at least one surface region of the decellularized tissue (a) at least partially penetrates at least one surface region of the second decellularized tissue, and / or (b) protrudes from one side of the second decellularized tissue and has at least one surface feature that crosses the tissue through at least one hole formed in the tissue to the other side, the construct according to any one of claims 1 to 41.

43. The construct according to claims 41 to 42, which is in the form of a multi-sheet construct.

44. The multi-sheet construct comprises at least one sheet of a second decellularized tissue and at least one sheet or segment of a polymer component, and any sheet of the second decellularized tissue is adjacent to or in contact with at least one sheet or segment of the polymer component, the construct according to claim 43.

45. The multi-sheet construct according to claim 43, comprising a certain number of sheets of the second decellularized tissue and the same number of sheets of the polymer component.

46. The at least one second decellularized tissue is dried before being coated with the polymer, the construct according to claims 41 to 42.

47. The drying is freeze-drying the at least one decellularized tissue, the construct according to claim 46.

48. The polymer component is in the form of a layer or coating of particles, a polymer sheet, a polymer film, polymer fibers or a polymer mesh, a gel, a hydrogel, or a liquid or a fluid film, and any combination thereof, the construct according to claim 49.

49. The construct according to any one of claims 40 to 48, wherein the at least one hole is pre-formed or present in the second decellularized tissue.

50. The construct according to any one of claims 1 to 49, wherein the metal element is selected from a stent, a metallic stent, a vascular graft, a heart valve, a sealing device, a suture or staple line, a pelvic floor reconstruction device, and any combination thereof.

51. A device comprising the construct according to any one of claims 1 to 50.

52. The device according to claim 51, configured as an implant.

53. The device according to claim 51 or 52, selected from a stent, a metallic stent, a vascular graft, a heart valve, a membrane, a sealing device, a suture or staple line, a hernia mesh or hernia repair device, a pelvic floor reconstruction device, a wound or burn dressing, a dural closure material, and a heart patch.

54. A method for manufacturing a construct according to any one of claims 1 to 4, comprising: - contacting at least one surface area of at least one decellularized tissue with at least one polymer, and - contacting at least one surface area of at least one metal element with at least one polymer, - forming the construct by contacting the at least one surface area of the at least one metal element with the at least one polymer with the at least one surface area of the at least one decellularized tissue with the at least one polymer. The method comprising.

55. The method according to claim 54, wherein the polymer coats at least one surface area of at least one metal element and / or at least one surface area of at least one decellularized tissue.

56. The method according to claim 55, wherein the polymer is cured.

57. The method according to claim 55, further comprising the step of adding at least one solvent, wherein the solvent is added to an area where at least one surface area of the decellularized tissue in contact with the polymer component contacts at least one surface area of the metal element coated with the polymer component.

58. The method according to claim 57, wherein the step of adding the solvent results in at least partial dissolution of (a) at least a portion of the polymer component in physical contact with the decellularized tissue and (b) at least a portion of the polymer component covering at least one surface region of the element.

59. The method according to claim 58, wherein the construct is obtained by a reflow effect of entanglement of chains along at least one region of the polymer component of (a) at least a portion of the polymer component in physical contact with the decellularized tissue and (b) at least a portion of the polymer component covering at least one surface region of the element.

60. The method according to any one of claims 54 to 59, further comprising at least one solvent, wherein the polymer component is dissolved in the at least one solvent.

61. The method according to any one of claims 54 to 60, wherein the solvent is selected from the group consisting of THF, acetone, dioxane, DMSO, halogenated solvents such as chloroform and dichloromethane, alcohols and polyols, polyethers, acetonitrile, ethyl acetate, dimethylformamide (DMF), dimethylacetamide, DMAC, and any combination thereof.

62. a. contacting at least one perforated surface region of the decellularized tissue with at least one polymer; and b. passing the at least one polymer through the perforation(s), The method according to claim 54, further comprising.

63. a. contacting at least one surface region of the decellularized tissue with at least one polymer; and b. at least partially covering the at least one polymer with the at least one decellularized tissue, The method according to claim 54, further comprising.

64. The method according to claim 60, further comprising at least one step selected from (a) a step of perforating or forming a hole in at least one surface region of the decellularized tissue; (b) a step of inserting the polymer into the perforation(s); (c) a step of curing the polymer; (d) a step of passing the at least one polymer through the perforation(s) to form a polymer sheet on the surface region; (e) any combination thereof.

65. Step of manufacturing the construct by thermally bonding (a) the at least one decellularized tissue and the at least one polymer by raising the temperature of the polymer component beyond either (i) its transition temperature in the case of an amorphous thermoplastic polymer; or (ii) its melting temperature in the case of a semi-crystalline polymer, to at least partially cause entanglement of polymer chains along at least one region of the polymer component; (b) selecting the polymer to be in the form of a layer or coating of particles, a polymer sheet, a polymer film, polymer fibers or a polymer mesh, a gel, a hydrogel, or a liquid or a flowable film, and any combination thereof; (c) further comprising at least one step selected from any combination thereof, the method according to claim 55.

66. The method according to claim 55, wherein at least one of the following applies: (a) the polymer completely penetrates the one or more holes; (b) the polymer partially penetrates the one or more holes; (c) the polymer completely penetrates the one or more holes to form polymer sheets on both sides of the surface region, thereby forming a construct assembly; and (d) any combination thereof.

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