Decellularized tissue / polymeric multi-component biomaterials

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 generally relates to a new class of multi-component materials and their uses. Background of the Invention

[0002] There are many materials that are foreign to the human body and are used in direct contact with the organs, tissues and fluids of the human body. These materials are called biomaterials, and among them, polymers play a key role in all clinical fields, such as orthopedic, cardiovascular fields, plastic surgery, drug delivery, wound dressings and many other fields. Decellularized tissue is a unique class of biomaterials derived from living tissue. These materials play an important role in an increasing number of applications.

[0003] Polymers constitute one of the main classes of materials used in implants for human or animal treatment, due to the diversity of their chemical, physical, mechanical and biological properties and the extremely wide range of values ​​they can obtain. To illustrate this, it is sufficient to mention the following differences related to their different properties: [i] Chemically - polymers can be highly hydrophilic and water-soluble materials as well as extremely hydrophobic materials, [ii] Mechanically - their hardness, as reflected by their corresponding Young's modulus values, ranges from a few kilopascals to hundreds of gigapascals, spanning eight or more orders of magnitude, while their ductility can be negligible or they can show fracture strain values ​​in the range of several thousand percent.

[0004] Decellularization (or acellularization) is a technique widely used to produce semi-natural biomaterials, whereby cells and genetic material are separated from the extracellular matrix (ECM) of native tissues. Decellularization can be achieved by chemical, enzymatic or physical methods, and is performed in a manner that allows the ECM to retain its original chemical and structural properties. The resulting decellularized tissue, typically pericardium, omentum or small intestinal mucosa, can be used in many fields, such as for hernia repair, for staple reinforcement, for pelvic floor reconstruction, for dural closure, as a membrane in the dental field, as a cardiac patch or as a leaflet of a heart valve.

[0005] The pericardium, the collagen-rich membrane that surrounds the heart, is the most widely used type of decellularized tissue and is currently used for a variety of clinical applications. Among other indications, the use of the pericardium in dural closure, as periosteum, as heart valve leaflets, and for surgical support are some of the more interesting applications. The most widely used sources of decellularized pericardial tissue are bovine and porcine.

[0006] Although both decellularized tissues and polymeric materials each have advantageous properties, both decellularized tissues and polymeric materials are limited in their behavior, particularly in clinical scenarios, rendering their biological performance insufficient. SUMMARY OF THE INVENTION

[0007] The inventors of the technology disclosed herein have developed a new hybrid material comprising decellularized tissue and a polymer material, each of which is associated with the other via one or more physical anchoring means. The interaction between the two components is not chemical in nature, but it is not possible to exclude random chemical interactions that may form between the tissue and the polymer.

[0008] Unlike known methods that utilize polymerizable reactive monomers, the constructs of the present invention are prepared by associating decellularized tissue with preformed polymers. Such an assembly process overcomes many of the disadvantages associated with the use of polymerizable monomers. These disadvantages may be related to: [a] the reactivity of the monomers toward the functional groups of the proteins that make up the decellularized tissue, affecting tissue properties and biocompatibility, [b] the inherent toxicity of the monomers, [c] the effective dissolution of the monomers in organic agents, [d] the high volatility and flammability of the monomers, [e] the inability to properly locate the monomers within the decellularized tissue, [f] the undesired polymerization that may occur due to the presence of certain functional groups present on the tissue and due to the one or more catalysts used to trigger the polymerization of the polymer components, [g] the undesired polymerization within the tissue components, which may result in undefined polymers that differ in their average molecular weight and polydispersity and generally in their composition, [h] the remnants of residual monomers that may remain after polymerization and may have adverse effects on the tissue and may also be separated over time after implantation, causing additional local problems as well as systemic problems.

[0009] By avoiding the use of in situ polymerization of reactive monomers, the present inventors have been able to obtain stable and well-defined constructs 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 having a physical association with a polymer component, said association comprising or consisting of at least partial penetration of the polymer component into a surface region of the tissue.

[0012] Also provided are polymer-associated decellularized tissues, wherein the association between the polymer and the tissue is physical.

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

[0014] In another aspect, the present invention provides a construct comprising at least one decellularized tissue and at least one polymer component, wherein the polymer component has at least one surface feature extending from one side of the decellularized tissue, and the surface feature passes through one side of the decellularized tissue to another side through at least one hole formed in the tissue.

[0015] As used herein, the term "construct" is used to define a structure or element or apparatus or arrangement that includes 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, each as defined herein. The term does not imply a specific arrangement or configuration of the two phases or regions, and in fact encompasses any arrangement whereby the two phases exist and are associated with each other by way of physical interaction. The term does not include such a structure or element whereby there is no physical association between the two phases.

[0016] In all aspects of the 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, wherein the polymer material is not formed in situ from monomers or prepolymers of the polymer material. As will be further described in detail herein, the constructs of the invention are formed by using preformed polymer sheets or segments whose size and shape are selected to meet a specific structure or purpose, or can be formed from a solution or liquid form of the polymer material on the surface area of ​​the tissue. In other words, the method for preparing the constructs of the invention does not require an in situ polymerization step utilizing monomers, oligomers or prepolymers.

[0017] Decellularized tissue and polymer can also be used to produce construct multisheets (multisheets), and this construct multisheet includes two or more sheets of tissue and multiple polymer sheets or sections that construct is kept together.Therefore, multisheets of the present invention can be provided in various forms, and each form has at least one tissue sheet and at least one polymer sheet or section.Sheets can be stacked on top of each other at least in part, so that each in polymer sheet or section is associated with another polymer sheet in stacking via at least one hole (hole) or hole (pore) formed in decellularized tissue or via engagement (welding).Although in multisheets, each in material sheet is solid sheet, one or more of the sheets positioned inside can be formed by gel, hydrogel or as liquid film or fluid film.

[0018] As used herein, the term "sheet", in its broadest sense, is a continuous material or a spread of a material, which may be in the form of a film of material, of any size and shape, and generally composed of a polymeric material or tissue. In the construct of the present invention, where there are two sheets of material, each sheet may be the same or different material, and may have the same or different sizes and shapes. In some embodiments, in the multi-sheet construct of the present invention, each of the polymeric sheets may be the same or different in composition, structure, size, shape, or any other physical, mechanical, or chemical properties.

[0019] Typically, sheets of decellularized tissue are provided as elongated strips or ribbons of tissue, the size and shape of which may vary. Similarly, polymer components may be provided as elongated sheets or strips of a size and / or shape similar or identical to that of the tissue, or may be provided as preformed (or formed) segments or fragments or tags of polymer that are different in size and shape (typically smaller) than the tissue sheet.

[0020] In the multi-sheet construct of the present invention, there is at least one sheet of decellularized tissue and at least one sheet or at least one segment of a polymer component, wherein any sheet of the decellularized tissue is adjacent to or in contact with at least one sheet or segment of the polymer component, and wherein at least two sheets or segments of the polymer component are associated with each other via at least one hole formed in at least one sheet of the decellularized tissue.

[0021] In some embodiments, in a multi-sheet construct, sheets or segments of polymeric components are associated with each other via at least one hole formed in at least one sheet of decellularized tissue.

[0022] In some embodiments, in a multi-sheet construct, at least two sheets or segments of a polymer component are associated with each other by bonding.

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

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

[0025] In some embodiments, a construct comprises two or more components having decellularized tissue confined between two sheets or sections of a polymer component, wherein each of the components are associated with one another, optionally by bonding.

[0026] In some embodiments, at least two or any two components are oppositely oriented.

[0027] The interaction or association between the polymer component (sheet or segment) and the tissue is selected and configured to provide a robust anchoring of the polymer in the tissue, thereby improving the mechanical properties of the construct. The improvement in the mechanical properties of the construct is achieved by a physical, non-chemical association that holds the two components together. The association (which is not chemical in nature) can be defined as

[0028] (i) embedding a polymer sheet or segment at least partially into a surface region of a layer of tissue, whereby the embedding may be through a single point on the surface region of the layer, or through two or more points; or (ii) anchoring the polymer sheet or segment into tissue to a depth of tissue that allows for secure association, whereby the anchoring may be through a single anchoring site, or through two or more anchoring sites, whereby the depth of anchoring or penetration of the polymer sheet or segment may vary, which does not include puncture of the tissue; or (iii) completely anchoring the polymer sheet or segment into the tissue to completely penetrate the tissue from one face of the tissue to the other face, whereby the anchoring may be through a single anchoring site, or through two or more anchoring sites. Typically, such penetration will involve surface features that are configured to extend from one face of the decellularized tissue to another face through at least one hole formed in the tissue. As will be described in detail below, such features may be formed in situ after the hole is formed in the tissue, or may be provided on the polymer sheet or segment in a form selected and configured to puncture or penetrate the tissue.

[0029] As used herein, the expression "at least partially penetrates" the surface area of ​​the decellularized tissue to indicate any of the above interactions, ranging from embedding into the surface of the tissue, anchoring without piercing the surface of the tissue, to actual face-to-face penetration of the tissue. Similarly, the expression "the association includes or consists of at least partial penetration of the polymer component" encompasses any of the above associations or interactions, indicating a single type of association or interaction (consisting of) or a combination of such associations or interactions (comprising).

[0030] The polymer sheets or segments are said to be associated with each other in a manner that ensures association with the decellularized tissue. One type of association or interaction present in the constructs of the invention is via anchoring or piercing the decellularized tissue surface as detailed herein, or by forming a hole in the tissue through which two polymer sheets or segments can be associated. In some embodiments of the invention in which the construct is made of more than one construct component, each component includes, for example, a decellularized tissue confined between two sheets or segments of a polymer component, and the association of more than one component can be achieved by polymer-to-polymer bonding.

[0031] In general, when the polymer chains at the surface of one sheet or section are movable enough to entangle with the chains in another sheet or section, the joining of polymer sheets or sections will occur. In order to achieve joining, thermal energy can be applied to raise the temperature of the polymer to 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. When two sheets or sections of polymer components are brought into close contact under these conditions, polymer chain entanglement will occur, resulting in a weld. According to aspects of the present invention, joining does not need to be achieved on the entire surface of a sheet or section. The site joining at one or more regions of the polymer component can be sufficient to provide a robust association of more than one component or any two polymer sheets or sections.

[0032] In some embodiments, the polymer component is in the form of polymer particles (nanoparticles, microparticles or larger sized particles) that are patterned on the surface of the tissue to form a particle sheet or particle continuity. In such embodiments, the particles can be embedded in the tissue surface (tissue outer layer), wherein the association between the tissue and the particles is strong enough to maintain the association between the two over a long period of time.

[0033] In other embodiments, the association between the polymer component and the tissue may involve at least one feature or functional group present on the polymer component, for example as a pendent group (or as a ligand group present on the surface of a polymer particle, or at least one polymer material in the form of a particle layer or particle sheet or particle coating or polymer sheet or polymer film or polymer fiber or polymer mesh), which penetrates or extends from one face of the tissue to another face. The penetration may be via a hole present in the tissue or via at least one hole preformed in the tissue, for example by puncturing or by punching, i.e., by placing the polymer material in contact with the tissue, and then optionally applying pressure and / or temperature to cause the polymer material to physically penetrate into the tissue surface.

[0034] At least one surface feature or functional group that penetrates or extends from one face of the tissue to another face of the tissue can be in the form of an elongated spin or needle that extends outward from the surface of the polymeric material and is oriented perpendicular to or at an angle to the surface of the polymeric material. The size and shape of the hole formed by the elongated spin or needle (i.e., the size and shape of the pre-formed hole) can vary, and the number or distribution of holes can cover part or all of the surface of the decellularized tissue.

[0035] The pins or needles, or at least one surface feature typically present on the surface of a polymeric material, may have a tip that secures the features in their location and prevents them from sliding out of the hole or holes formed in the tissue.

[0036] In some embodiments, the construct comprises a polymer sheet having one or more tissue penetrating features and a decellularized tissue decorated with one or more holes, through which the features extend, the tissue penetrating features comprising the material of the polymer sheet. The patterning of the tissue penetrating features and / or the patterning of the holes can follow any patterning profile (hole profile) as defined herein (e.g., size, shape, distribution density, location, etc.).

[0037] In some embodiments, the construct comprises two polymer sheets or segments, each of which is associated with the other polymer sheet or segment via one or more polymer features or members extending the surface area of ​​each sheet, wherein the decellularized tissue disposed between the two polymer sheets has one or more holes through which the one or more polymer members pass transversely.

[0038] In some embodiments, the construct is formed by piercing one or more holes in a surface area of ​​the tissue and forming a sheet or segment of the polymeric component by utilizing a liquid or fluid form of the polymeric component, wherein the liquid or fluid form is allowed or caused to penetrate and fill the hole.

[0039] In some embodiments, where there are two polymer sheets or sections and a decellularized tissue is disposed between them, each of the polymer sheets is continuous and made of the same polymer material, however one of the sheets is larger in size compared to the other.

[0040] In some embodiments, the construct is in the form of a multi-sheet kit that includes one or more polymer sheets and one or more decellularized tissue sheets.

[0041] 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 wherein any two sheets of the polymer component are associated with each other via at least one hole formed in at least one sheet of the decellularized tissue.

[0042] In some embodiments, one or more of the sheets may be designed as a material reservoir for releasing active or inactive materials such as bioactive substances and drugs.

[0043] In some embodiments, at least a portion of the polymer component is configured to release a material such as a bioactive substance or a drug. In some embodiments, all or a specific portion of the polymer component can locally release a bioactive substance or a drug.

[0044] The bioactive substance or drug to be released from the construct of the invention after positioning in vivo can be selected from any drug or pharmaceutical intended to achieve medical improvement, prevent the development of a disease or condition (local or systemic), or maintain a state of good health over time. Thus, the bioactive substance or drug can be selected based on, among other things, the area of ​​the body in which the construct is to be implanted or positioned and the type of medical complications that may be associated with the implantation site and the procedure.

[0045] Generally, active substances or drugs are broadly characterized as non-toxic and regulated by the FDA or EMA, or are classified as GRAS (Generally Recognized as Safe). Non-limiting examples of such active substances and drugs may include analgesics, including non-narcotic and narcotic analgesics; antianxiety drugs; antiarrhythmic drugs; antibacterial agents; antibiotics, including naturally occurring antibiotics, synthetic antibiotics, broad-spectrum antibiotics; anticoagulants and thrombolytics for arterial or venous thrombosis; anticonvulsants; antidepressants, including mood-enhancing antidepressants: tricyclics, monoamine oxidase inhibitors and SSRIs; antidiarrheal drugs, including antidiarrheal preparations and drugs that slow intestinal muscle contractions; antiemetics; antifungals, including antifungals for infections affecting the hair, skin, nails, mucous membranes; antihistamines; antihypertensive drugs, including diuretics, beta-blockers, calcium channel blockers, ACE (angiotensin converting enzyme) inhibitors; anti-inflammatory drugs; antineoplastic drugs; antipsychotics, including strong tranquilizers; antipyretics; antivirals, including treatment and temporary protection against viral infections; beta-blockers; in immunosuppression, malignancy or deficiency disorders (deficiency disorders); cytotoxins, as anti-tumor agents and also as immunosuppressants; hormones, including synthetic equivalents and natural hormone extracts; immunosuppressants; muscle relaxants, including those that relieve muscle spasms, and weak tranquilizers; sex hormones (female), including those used for menstrual and menopausal disorders, oral contraceptives, and also for the treatment of female and male cancers; sex hormones (male), including those used for androgen deficiency in hypopituitarism or testicular disorders, and also for the treatment of cancers, and anabolic steroids; enzymes, such as collagenase or elastase; and vitamins.

[0046] In some embodiments, any of the polymer components or tissue components can be porous, i.e., include surface pores limited to specific areas of the component or surface pores that can be distributed along the entire surface of the polymer or tissue. The pores can be inherently present in the decellularized tissue or formed in the decellularized tissue to achieve a specific hole (or porosity) profile. The profile defines at least one surface area of ​​the tissue where holes are to be formed; the number, shape and size of the holes; the density of holes at certain surface areas, etc. Different patterning profiles can be used to meet the desired construct capabilities or properties or utilities. Once one or more holes are formed, the association of the polymer material can be performed by a variety of methods, as disclosed herein. Some methods may involve the use of a polymer component with an extension feature that can be inserted into or penetrate the hole. Other methods may involve the use of a liquid polymer. In such methods, tissue can be immersed in a liquid polymer material or a solution containing a polymer material, sprayed with a liquid polymer material or a solution containing a polymer material, or substantially treated in a liquid polymer material or a solution containing a polymer material, or immersed, sprayed or substantially treated with a liquid polymer material or a solution containing a polymer material, allowing the polymer material to penetrate one or more holes and further deposit on the tissue face, thereby forming a polymer sheet or segment on either or both tissue faces. The polymer sheet or segment formed on either or both faces of the tissue can be fused or joined with another polymer component or sheet, thereby building up the multiple sheets or constructs of the present invention layer by layer or component by component.

[0047] Alternatively, the decellularized tissue is punctured and a liquid polymer is injected into the hole created in the tissue. The assembly is constructed by forming a polymer sheet between two tissue sheets and then thermally bonding the construct. The construct can be attached to another polymer sheet or feature by bonding.

[0048] According to yet another approach, a multi-sheet structure is formed by stacking multiple sheets on top of each other, which are then pierced to provide the desired hole profile and injected with a liquid polymer to provide a fused construct.

[0049] According to yet another method, a liquid polymer is cast onto the tissue to form a bilayer of tissue and polymer. Subsequently, the bilayer is pierced to provide a hole profile and injected with a liquid polymer to provide a construct. The binary construct can be used to form multiple sheets or can be joined to another polymer sheet.

[0050] In some embodiments, in the constructs of the invention, the decellularized tissue constitutes between 5% wt and 95% wt of the construct.

[0051] As used herein, "decellularized tissue" is a tissue from which inhibitory cells have been removed, leaving the extracellular matrix (ECM) of the tissue. As mentioned above, decellularization can be achieved by chemical methods, enzymatic methods, or physical methods as known in the art. Decellularized tissue can be obtained from the tissue of the oral mucosa, from the submucosa of the small intestine, or from a decellularized matrix of the bladder, which provides natural and optimal integration properties to the extracellular matrix. Other tissues can also be used.

[0052] Methods for achieving decellularized tissue are known in the art, for example, in International Application No. WO 2005 / 032473 and US applications derived therefrom, each of which is incorporated herein by reference; US Patent No. 5,993,844, incorporated herein by reference; and others.

[0053] In some embodiments, the decellularized tissue is selected from the group consisting of pericardium, omentum, or small intestinal mucosa.

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

[0055] In some embodiments, the decellularized tissue is bovine pericardium or porcine pericardium.

[0056] In some embodiments, the decellularized tissue is small intestinal submucosa.

[0057] In some embodiments, the decellularized tissue is omentum.

[0058] The polymers used in the constructs of the present invention are polymeric materials as known in the art. The polymers used may be selected from thermoplastic polymers and thermosetting polymers. The polymers may be hydrophobic, hydrophilic or amphiphilic, and may also be selected from bioinert polymers or biodegradable polymers.

[0059] In some embodiments, the polymer is a blend of different polymers, an interpenetrating polymer network (IPN), or a semi-interpenetrating polymer network (semi-IPN).

[0060] In some embodiments, the semi-IPN polymer is selected to be cross-linked via addition or condensation reactions, click chemistry reactions, or any other type of reaction and combinations thereof. In some embodiments, the cross-linkable compound is selected from compounds containing two or more carbon double bonds, such as ethylene glycol dimethacrylate (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) copolymer dimethacrylate (DMA) and diacrylate (DA), polytetramethylene glycol dimethacrylate (PTMGDMA) and polytetramethylene glycol diacrylate (PTMGDA), polydimethylsiloxane DMA and DA, polycaprolactone dimethacrylate (PCLDMA) and polycaprolactone diacrylate (PCLDA), polycaprolactone / lactide P (CL / LA) DMA and DA, polycaprolactone / glycolide P(CL / GA) DMA and DA and polyglycolide / lactide P(GA / LA) DMA and DA, polyethylene glycol / caprolactone (PEG / CL) DMA and DA, polyethylene glycol / lactide PEG / LA DMA and DA and polyethylene glycol / glycolide (PEG / GA) DMA and DA, polypropylene glycol / caprolactone (PPG / CL) DMA and DA, polypropylene glycol / lactide (PPG / LA) DMA and DA and polypropylene glycol / glycolide (PPG / GA) DMA and DA, polytetramethylene / caprolactone (PTMG / CL) DMA and DA, polytetramethylene / lactide (PTMG / LA) DMA and DA, and polytetramethylene / glycolide (PTMG / GA) DMA and DA.

[0061] In some embodiments, the IPN is composed of a polymer formed by a compound containing two or more carbon double bonds, the compound being selected from the group consisting of EGDMA and EGDA, TEGDMA and TEGDA, DVB, bisacrylamide, PEG DMA and DA and higher functionality species of various molecular weights, PPG DMA and DA and higher functionality species of various molecular weights, PEG / PPG copolymer DMA and DA and higher functionality species, PTMG DMA and DA and higher functionality species of various molecular weights, siloxane DMA and DA and higher functionality species of various molecular weights, PCL DMA and DA and higher functionality species of various molecular weights, P(CL / LA) DMA and DA, P(CL / GA) DMA and DA and P(GA / LA) DMA and DA and higher functionality species of various molecular weights, PEG / CL DMA and DA, PEG / LA DMA and DA and PEG / GA DMA and DA and higher functionality species of various molecular weights, PPG / CL DMA and DA, PPG / LA DMA and DA and PPG / GA DMA and DA and higher functionality species of various molecular weights, PTMG / CL DMA and DA, PTMG / LA DMA and DA and PTMG / GA DMA and DA and higher functionality species of various molecular weights, and combinations thereof.

[0062] In some embodiments, the polymer is an acrylic polymer or a methacrylic polymer.

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

[0064] In some embodiments, the polymer is a silicone polymer.

[0065] In some embodiments, the polymer is a polycarbonate, polyurethane, polyurea, or polyamide, and combinations thereof. In some embodiments, the polymer is a polyurethane.

[0066] In some embodiments, the polymer has a glass transition temperature or melting point below 120°C. In some embodiments, the polymer has a glass transition temperature or melting point below 85°C.

[0067] In some embodiments, the polymer is selected to flow at a pressure of 50 kPa and a temperature above 42°C.

[0068] 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, Polyurethanes such as Chronoflex, Biospan and Bionate and polyurethanes containing silicones such as CarboSil, PurSil, Avcothane and Cardiothane, polyglycolic acid, polylactic acid, polycaprolactone, polylactide-caprolactone copolymers, polyglycolic acid-lactic acid copolymers, polyethylene oxide-polylactic acid copolymers, polyethylene oxide-polycaprolactone copolymers, polytetramethylene oxide-caprolactone copolymers, polyhydroxybutyrate, polyhydroxyvalerate, polyethylene adipate, polybutylene adipate, polyethylene succinate, polybutylene succinate and polybutylene terephthalate and polyethylene terephthalate / butylene terephthalate copolymers, and combinations and copolymers thereof.

[0069] As used herein, the term "ChronoFlex" hereinafter refers to a family of thermoplastic urethanes based on biodurable aromatic polycarbonates. These aromatic polycarbonate urethanes are generally designed for molding, casting and dip coating applications.

[0070] In some embodiments, the polymer is a thermoplastic polyurethane based on an aliphatic polyether, optionally Tecoflex.

[0071] In some embodiments, the polymer is an aromatic polycarbonate based urethane, optionally Chronoflex.

[0072] In some embodiments, the polymer is PMMA, PBMA, PHMA, PMA, PHEMA, PHPMA, and combinations thereof.

[0073] In some embodiments, the polymer is a low molecular weight (typically between 500 Da and 10,000 Da) or amorphous or branched polyolefin, and combinations thereof.

[0074] In some embodiments, the polymer is PDMS or PDMS containing phenyl groups or derivatized PDMS chains containing double bonds and / or hydroxyl and / or amine and / or thiol groups.

[0075] In some embodiments, the polymer is an aliphatic or aromatic polyurethane containing polyether or polyester soft segments.

[0076] Polymer can be selected to have the morphology and property that change with time.These changes may be due to chemical phenomena, physical phenomena and / or biological phenomena.In some embodiments, polymer is selected to experience secondary chemical changes, such as coupling, polymerization or cross-linking, oxidation or hydrolysis or enzymatic degradation.In some embodiments, individually or in addition to chemical changes, polymer can be selected to experience physical process or be affected by physical process, physical process such as crystallization or phase separation.

[0077] In some embodiments, the polymer is a polymer that exhibits shape memory, which can be actuated by a variety of stimuli such as temperature, pH, ionic strength, hydration, biological cues, electric or magnetic fields, any type of radiation, and combinations thereof, wherein the polymer that exhibits shape memory can exhibit a one-time response or a cyclic shape memory response.

[0078] In some embodiments, an actuated shape memory polymer is a polymer that is capable of adjusting the shape of a tissue / polymer construct.

[0079] In some embodiments, all or a portion of the polymer component may exhibit shape memory behavior.

[0080] In some embodiments, all or a portion of the polymer component may be environmentally responsive. In some embodiments, all or a portion of the polymer component may be reverse thermally responsive.

[0081] In some embodiments, the polymer is an environmentally responsive polymer that responds to temperature, pH, ionic strength, biological signals, various types of radiation, electric or magnetic fields, and combinations thereof. In some embodiments, the environmentally responsive polymer is reverse thermoresponsive.

[0082] The reverse thermoresponsive material can be selected from N-alkyl substituted acrylamides (such as poly-N-isopropylacrylamide [PNIPAAm]), or based on polyethylene oxide / polypropylene oxide segments, or cellulose derivatives selected from hydroxypropyl methylcellulose and hydroxypropyl cellulose, or alternating polymers or random polymers, and various amphiphilic polymers such as poly(ethylene oxide)-polylactic acid block copolymers.

[0083] In some embodiments, the polymer is polybutyl methacrylate.

[0084] 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.

[0085] In some embodiments, the polymer is a silicone polymer that is a derivatized PDMS selected to crosslink in situ.

[0086] In some embodiments, the polymer is a polyurethane selected from the group consisting of Pellethane, Elasthane, Elastolan, chronoflex, Tecoflex, Cardiothane, Avcothane, CarboSil, PurSil, Biomer, BioSpan, and Bionate (each as defined herein)) and combinations thereof.

[0087] In some embodiments, the polyurethane is a thermoplastic polyurethane based on an aliphatic polyether, such as Tecoflex.

[0088] In some embodiments, the polyurethane is an aromatic polycarbonate based urethane, such as Chronoflex.

[0089] 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 and polybutylene terephthalate and polyethylene terephthalate / butylene terephthalate copolymers and combinations and copolymers thereof.

[0090] In the construct of the present invention, the polymer used can be a blend or mixture or combination of two or more polymeric materials. In some embodiments, such a blend can include two or more polyolefins, or acrylic polymers or methacrylic polymers, or silicone polymers, or polycarbonates, or polyurethanes, or polyureas, or polyamides and combinations thereof.

[0091] In another aspect, the constructs of the invention comprise one or more tissue phases (including or consisting of decellularized tissue) and more than one polymer phase (including or consisting of polymers), each of which forms separate phases of different geometries and whose sizes can span from the nanoscale to the centimeter scale.

[0092] The polymer phase and / or tissue phase can be constructed or constituted by more than one type of polymer or decellularized tissue. As further disclosed herein, each of the polymer and / or tissue can form a single construct or form an array of constructs associated with each other. In some embodiments, the polymer and / or tissue can be a part of a medical equipment having at least one polymer region and / or at least one tissue region, and the construct can be associated with it. Such equipment can include all types of materials, such as tissues, polymers, metals, ceramics, carbonaceous materials and combinations thereof.

[0093] In construct of the present invention, polymer phase is configured or selected or designed to be connected to two or more in tissue phase or other phases comprising material as disclosed above, such as polymer, metal, ceramic, carbonaceous material and combination thereof. In some embodiments, polymer phase is configured to connect two or more in tissue phase or other phases, and the connection can maintain 5% stress higher than the cohesive strength of polymer itself. In some embodiments, for these and other purposes, polymer is therefore selected to display shape memory behavior. In some embodiments, polymer is environmentally responsive. In some embodiments, polymer is biodegradable.

[0094] In some embodiments, the polymer is reverse thermoresponsive and forms an aqueous solution that undergoes a LCST transition below 37°C.

[0095] In some embodiments, the construct is a composite material.

[0096] The present invention also provides an implant or a medical device which is or comprises a construct of the present invention.

[0097] According to the present invention, no matter the type of the construct used, and no matter the kind of the equipment to be manufactured by the construct of the present invention, the construct is configured or designed or intended to contact with human or animal tissue or organ. In some cases, the construct is intended to be used for the equipment associated with the tissue in the human or animal body, or can be constructed as equipment to be implanted in the body. For example, the construct can constitute a part of medical equipment or medical equipment, configured to be used for implanting in the gastrointestinal tract of the experimenter, implanting in the respiratory system (airway system), implanting along the vascular system, implanting in the heart region, implanting in the urinary system or implanting in any other organ in the human or animal body, and this can be determined as the case may be. Especially depending on the construct material, shape and intended use, and further depending on the organ to be used therein for implant, the construct can be porous or non-porous, wherein the hole can span from nanometer scale to centimeter scale. Non-limiting examples of devices utilizing or including the constructs of the invention include stents, metal stents, vascular grafts, heart valves (optionally including a metal frame), membranes, sealing devices, sutures or staples, hernia meshes or hernia repair devices, pelvic floor reconstruction devices, wound or burn dressings, dural closures, cardiac patches, and others.

[0098] In some embodiments, the device is implemented in a heart valve further comprising a metal frame.Thus, in some embodiments, the heart valve may comprise pericardial leaflets, a tissue / polymer phase and a metal frame, as defined herein.

[0099] In some embodiments, the construct is a patch deployed in the CV system, along the gastrointestinal tract, in the airway tree, in the urinary and reproductive areas, in the central or peripheral nervous system, a vascular graft, an A / V shunt, wherein one or more of said polymers are formed into films, fibers, particles of any size and geometry, which are porous or solid, hollow or non-hollow or any other geometry, produced by any manufacturing technique including textile methods and procedures to produce non-woven structures and combinations thereof.

[0100] The equipment of the present invention including the construct of the present invention can be used as a vehicle for delivering one or more active agents to a site or organ or tissue in the body of a subject. Such an active agent can be a cellular bioactive material or a molecular bioactive material. The active material can be contained in the construct as a whole, in a polymer component, or in a decellularized tissue. In some embodiments, the decellularized tissue contains an active substance as defined herein, such as a bioactive material. In some embodiments, the active material is released over time.

[0101] The constructs of the present invention can be provided in various forms as exemplified herein. In some embodiments, a polymer is associated with two or more tissue regions or sections to form a laminate or any other structure such as a multilayer structure. In some embodiments, a multilayer structure can include three polymer layers, two of which are outside the construct and one polymer layer in the middle of the construct, with two tissue layers sandwiched between the polymer layers.

[0102] In some embodiments, a construct is provided with a polymer membrane confined between two sheets of decellularized tissue.

[0103] In some embodiments, the construct may be formed with polymeric connections that associate each of the polymer layers via holes made in the tissue layers. In some embodiments, the polymeric connections have the same or different composition as the polymer forming the layers. According to the present invention, in order to synthesize such a construct, the polymer or polymer mixture used and preformed, for example, preformed as a polymer film is selected to be able to flow at a pressure between 50 kPa and 1 MPa and a temperature below 120°C, filling the holes made in the tissue, thereby connecting or associating the tissue and the polymer together. Once cooled, the polymer produces a continuous membranous structure at the far end of the flow direction, which contributes to the strength and long-term stability of the association between the tissue phase and the polymer phase.

[0104] In some embodiments, the polymer is an RTR polymer that flows at pressures below 100 kPa and temperatures below 37° C. Once at physiological temperature, the polymer creates a continuous membranous structure distal to the direction of flow, contributing to the strength and long-term stability of the association between the tissue phase and the polymer phase.

[0105] In some embodiments, the polymer in the construct of the present invention is preformed, optionally in the form or shape of a film. The preformed polymer may include more than one material, blended together or spatially separated by distances spanning from nanometer scale to centimeter scale, isotropically or anisotropically organized, produced in layers or in any other spatial arrangement, wherein the additional material may be selected from polymers, tissues, active components, metals, ceramics or carbonaceous materials and combinations thereof.

[0106] In some embodiments, the preformed polymer also includes low molecular weight molecules that soften the film or its part. The low molecular weight molecules enable the polymer phase or its part to flow under pressure and temperature conditions. In some embodiments, the low molecular weight molecules are polymerizable or cross-linkable, so the low molecular weight molecules soften the polymer phase or its part before it is polymerized or cross-linked, and once polymerized or cross-linked, strengthen or harden the polymer phase. In some embodiments, the inert or reactive low molecular weight molecules can be biologically inert or biodegradable, can show shape memory behavior or can be environmentally responsive, and combinations thereof.

[0107] In some cases, the construct is obtainable by a polymerization or cross-linking reaction that occurs via an addition reaction or a condensation reaction or any other type of reaction and combinations thereof. The addition polymerization or cross-linking reaction may involve reactive carbon double bonds following any type of catalysis, including chemically initiated reactions, thermally initiated reactions, and radiation initiated reactions.

[0108] In some cases, the construct is obtainable by a polymerization reaction or a cross-linking reaction of a suitable precursor, wherein the reaction is performed in part before contact with the decellularized tissue and in part during or after the polymer component is associated with the decellularized tissue component. In some embodiments, a partially polymerized or cross-linked polymer film or sheet or construct having any geometry can flow under the applied conditions to produce the desired association with the decellularized tissue or any other component (polymeric component or non-polymeric component). In some embodiments, the partially polymerized or cross-linked polymer is selected to flow at a pressure of 50 kPa and a temperature above 42°C.

[0109] In some embodiments, the partially polymerized or cross-linked polymer has a glass transition temperature or melting point below 120°C, while in some other embodiments, the partially polymerized or cross-linked polymer has a glass transition temperature or melting point below 85°C.

[0110] In other embodiments, the partially polymerized or cross-linked polymer has a glass transition temperature or melting point below 60°C.

[0111] In some embodiments, the partially polymerized or cross-linked polymer that is further polymerized or cross-linked during or after association with the decellularized tissue or any other component (polymer or non-polymer, tissue or non-tissue) has a higher glass transition temperature or melting point than the partially polymerized or cross-linked polymer.

[0112] In some embodiments, the further polymerized or cross-linked polymer building blocks have a glass transition temperature or melting point above 50°C, and in other embodiments, the further polymerized or cross-linked polymer building blocks have a glass transition temperature or melting point above 60°C, and in yet other embodiments, the further polymerized or cross-linked polymer building blocks have a glass transition temperature or melting point above 80°C.

[0113] In some embodiments, the further polymerized or cross-linked polymer building block has a glass transition temperature or melting point above 100°C.

[0114] In some embodiments, the polymer in the construct of the invention is preformed, optionally in the form or shape of a film, and comprises partially polymerized or cross-linked polymer components having different initial degrees of polymerization and / or cross-linking and different final degrees of polymerization and / or cross-linking obtained upon further polymerization or cross-linking of the polymer components.

[0115] In some embodiments, the preformed polymer also includes low molecular weight molecules (between 500DA and 100,000DA) that are polymerizable or non-polymerizable and / or cross-linkable or non-cross-linkable, and suitable precursors that are partially polymerized or cross-linked, wherein the polymerization reaction or cross-linking reaction is partially carried out before contact with the decellularized tissue, and partially carried out during or after the polymer component is associated with the decellularized tissue component.

[0116] In other cases, the construct may be obtainable by reaction of a compound selected from the group consisting of methyl methacrylate (MMA), butyl methacrylate (BMA), hexyl methacrylate (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 functional species of various molecular weights, PPG DMA and DA and higher functional species of various molecular weights, PEG / PPG copolymer DMA and DA and higher functional species of various molecular weights, PTMG DMA and DA and higher functional species of various molecular weights, siloxane DMA and DA and higher functional species of various molecular weights, PCL DMA and DA and higher functional species of various molecular weights, P(CL / LA) DMA and DA, P(CL / GA) DMA and DA and P(GA / LA) DMA and DA and higher functionality species of various molecular weights, PEG / CL DMA and DA, PEG / LA DMA and DA and PEG / GA DMA and DA and higher functionality species of various molecular weights, PPG / CL DMA and DA, PPG / LA DMA and DA and PPG / GA DMA and DA and higher functionality species of various molecular weights, PTMG / CL DMA and DA, PTMG / LA DMA and DA and PTMG / GA DMA and DA and higher functionality species of various molecular weights, and combinations thereof.

[0117] The reaction may be selected from click chemistry, any type of complexation, host / guest reaction and non-covalent supramolecular polymerization. In some embodiments, a condensation reaction is involved, thereby forming a carbamate group, a urea group, an amide group, an ester group, a carbonate group or an ether group and combinations thereof.

[0118] In some embodiments, epoxy / amine reactions and hydrolysis and condensation reactions of silicone-containing molecules may be involved. In some embodiments, the reaction may involve the reaction of thiol groups. The epoxy / amine reaction may occur between GMA and amines having a variety of functionalities and molecular weights, including amino-terminated PEGs, PPGs, PTMGs, PCLs, PDMSs containing one or more amino groups per molecule, molecules containing amino-terminated P(CL / LA), P(CL / GA), and P(GA / LA) with a variety of molecular weights, molecules containing PEG / CL, PEG / LA, and PEG / GA with a variety of molecular weights, molecules containing PPG / CL, PPG / LA, and PPG / GA with a variety of molecular weights, molecules containing PTMG / CL, PTMG / LA, and PTMG / GA with a variety of molecular weights containing one or more amine groups, oligopeptides and peptides containing reactive amine moieties, and epoxy-terminated molecules with a variety of functionalities and molecular weights capable of reacting with the amino-terminated molecules, and combinations thereof.

[0119] In some embodiments, in the constructs of the invention, molecules of the polymeric material are able to disentangle, cross the polymer / tissue interface or any other interface, diffuse into the tissue phase or any other phase, mix and re-entangle with themselves and with the molecules of the tissue or any other phase, joining the two together. In some embodiments, the molecules that cross the polymer / tissue interface are also able to react with themselves and / or with moieties present in the tissue or any other phase, enhancing the strength and long-term stability of the connection between the polymer phase and any other phase.

[0120] Construct of the present invention can be formed, wherein polymeric material directly contacts (in situ) with tissue phase when being in liquid or semi-liquid state.The polymer phase produced in situ is formed by applying liquid or semi-liquid, polymerizable or cross-linkable reactive precursors of optimization composition.In some embodiments, the polymer phase produced in situ is formed by applying the solution of the polymer on the surface of tissue phase and / or in tissue phase and its combination.In some embodiments, polymer solution is aqueous solution or uses the solution of other hydrophilic solvents.In some embodiments, polymer is dissolved in organic solvent, and the organic solvent is such as acetone, THF, dioxane, DMSO, halogenated solvent such as chloroform and dichloromethane, alcohol and polyol, polyether, acetonitrile, ethyl acetate, dimethylacetamide, DMAC and dimethylformamide (DMF).Polymer solution has the concentration in the range of from 1 wt% to 40 wt%, or from 1 wt% to 20 wt% or from 2 wt% to 10 wt%.

[0121] In some embodiments, the tissue phase and the polymer phase are connected by applying pressure at a supraphysiological temperature for a period of time shorter than one hour. In some embodiments, the pressure is in the range of from 20 kPa to 100 GPa, and the temperature is from 40 degrees across up to 120 degrees, and the application is continued for a time interval of from 1 second to 60 minutes. In some embodiments, the pressure is in the range of from 500 kPa to 10 GPa, and the temperature is from 40 degrees across up to 85 degrees, and the application is continued for a time interval of from 2 seconds to 5 minutes.

[0122] Thus, in a further aspect of the invention, methods of making the constructs of the invention are provided.

[0123] According to one aspect, there is provided a process for making a construct according to the invention, the process comprising contacting the pierced surface area of ​​at least one decellularized tissue with a liquid polymer, and The liquid polymer is allowed to penetrate into the perforations (holes) and form a polymer sheet over the surface area.

[0124] The pierced surface area of ​​the tissue can be an area through which holes have been made, or an area that naturally includes such holes or holes. In the case of holes to be made, they can be made by using cutting equipment or piercing equipment such as needles with various diameters. The position of the hole, the shape of the hole, their size and their distribution on the surface area (hole profile as defined herein) can be predefined and selected to meet any prerequisites related to the mechanical properties of the construct or its use, for example. Typically, the number of holes is greater than 1.

[0125] Thus, in some embodiments, the process comprises puncturing or forming holes in a surface region of at least one decellularized tissue.In some embodiments, the holes have a predetermined hole profile as defined.

[0126] In some embodiments, liquid polymer is injected into the perforations (holes).

[0127] When a liquid polymer is used, curing may be required, for example by heat treating the polymer. As different polymers may be used, different curing methods may be used.

[0128] Additional processes are provided for making a construct comprising at least one decellularized tissue and at least one polymer component, wherein the process comprises contacting the surface area of ​​at least one decellularized tissue that has been punctured to form one or more holes with a liquid polymer, and The liquid polymer is allowed to penetrate into the one or more holes and form a polymer sheet over the surface area.

[0129] In some embodiments, the process comprises piercing or forming holes in a surface region of at least one decellularized tissue. In some embodiments, the holes have a predetermined hole profile as defined.

[0130] In some embodiments, liquid polymer is injected into the perforations (holes).

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

[0132] In some embodiments, the liquid polymer partially penetrates the one or more holes.

[0133] In some embodiments, the liquid polymer completely penetrates the one or more holes to form a polymer sheet on both sides of the surface area, thereby forming a construct component.

[0134] In some embodiments, the process includes the step of associating or fusing two or more constructs. Fusion can be performed by conjugation.

[0135] Also provided is a process for making a construct comprising at least one decellularized tissue and at least one polymer component, the process comprising stacking one or more sheets of decellularized tissue and one or more sheets or sections of polymer material to obtain a stacked structure; forming holes in the stacked structure to form one or more holes in each of the one or more sheets of tissue and the one or more sheets of polymeric material, wherein optionally at least a number of the one or more holes are coaxially arranged; and The stacked structure is treated with a liquid polymer such that the liquid polymer penetrates into the one or more holes and fuses the sheets to form a construct.

[0136] 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 partially covers at least one surface region of the metal element.

[0137] The present invention also provides a decellularized tissue physically associated with a polymer component, said association comprising or consisting of at least partial coating of a surface region of the tissue with the polymer component.

[0138] The present invention also 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 surface feature that at least partially covers at least one surface of the decellularized tissue and at least partially covers at least one surface area of ​​the metal element.

[0139] The invention also provides a construct as defined above, which is in the form of a multi-sheet construct.

[0140] The present invention also provides a construct as defined above, wherein the coating is performed by at least one method selected from the group consisting of spray coating, brush coating, dip coating and any combination thereof.

[0141] The present invention also provides a construct as defined above, wherein said at least one sheet of decellularized tissue is dried prior to being coated with said polymer.

[0142] The present invention also provides a construct as defined above, wherein said drying is freeze-drying (ie lyophilizing) said at least one sheet of decellularized tissue.

[0143] The invention also provides a construct as defined above wherein at least one or any one of the at least one sheets of decellularized tissue is confined between any two sheets of the polymer component.

[0144] The present invention also provides a construct as defined above, wherein the multi-sheet construct comprises a certain number of sheets of decellularized tissue and an equal number of sheets of a polymer component.

[0145] The present invention also provides a construct as defined above, comprising two or more components having a decellularized tissue confined between two sheets or sections of a polymer component, wherein the components are associated with each other.

[0146] The invention also provides a construct as defined above, wherein at least two components are oppositely oriented.

[0147] The invention also provides a construct as defined above, wherein the metal element is a metal wire.

[0148] The invention also provides a construct as defined above, wherein the polymer component is in the form of a particle layer or coating, a polymer sheet, a polymer film, a polymer fiber or a polymer mesh.

[0149] The invention also provides a construct as defined above, wherein at least one hole is pre-formed or present in the decellularized tissue.

[0150] The invention also provides a construct as defined above, wherein the decellularized tissue constitutes between 5% wt and 95% wt of the construct.

[0151] The present invention also provides a construct as defined above, wherein the decellularized tissue is obtained from a tissue selected from the group consisting of oral mucosa, small intestinal submucosa and urinary bladder decellularized matrix.

[0152] The present invention also provides a construct as defined above, wherein the decellularized tissue is selected from the group consisting of pericardium, omentum or small intestinal mucosa.

[0153] The present invention also provides a construct as defined above, wherein the decellularized tissue is pericardium.

[0154] The present invention also provides a construct as defined above, wherein the decellularized tissue is bovine pericardium or porcine pericardium.

[0155] The invention also provides a construct as defined above, wherein the polymer component is or comprises a polymer selected from the group consisting of a hydrophobic polymer, a hydrophilic polymer and an amphiphilic polymer.

[0156] The invention also provides a construct as defined above, wherein the polymer component is or comprises a blend, an IPN or a semi-IPN.

[0157] The invention also provides a construct as defined above, wherein the polymer component is or comprises an acrylic polymer or a methacrylic polymer.

[0158] The invention also provides a construct as defined above wherein the polymer component is or comprises a polyolefin.

[0159] The invention also provides a construct as defined above, wherein the polymer component is or comprises an organosilicon polymer.

[0160] The invention also provides a construct as defined above, wherein the polymer component is or comprises polycarbonate, polyurethane, polyurea or polyamide and combinations thereof.

[0161] The invention also provides a construct as defined above wherein the polymer component is or comprises a polyurethane.

[0162] The present invention also provides a construct as defined above, wherein the polymer component is or comprises a polymer selected from the group consisting of 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 ...isobutylene, polydimethylsiloxane (PDMS), poly(2-hydroxyethyl methacrylate) (PHEMA), poly(N-(2-hydroxypropyl) methacrylamide) (PHPMA), polycyanoacrylate (PCA), polyethylene / polypropylene copolymer, polyethylene / polybutylene copolymer, polyisobutylene, polydimethylsiloxane (PDMS), poly(2-hydroxyethyl methacrylate) (PHEMA), poly(N-(2-hydroxypropyl) methacrylamide) (PHPMA), polycyanoacrylate (PCA), polyethylene / polypropylene copolymer, polyethylene / polybutylene copolymer, polyisobutylene, polydimethylsiloxane ( MS), 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 terephthalate / butylene terephthalate copolymers and combinations and copolymers thereof.

[0163] The invention also provides a construct as defined above, wherein the polymer component is or comprises a shape memory element.

[0164] The present invention also provides a construct as defined above, wherein the polymer component is or comprises a polyether urethane selected from Pellethane, Elastane, Elastolan, Tecoflex, Biomer.

[0165] The invention also provides a construct as defined above, wherein the polymer component is or comprises a polycarbonate urethane selected from the group consisting of Chronoflex, Biospan and Bionate.

[0166] The invention also provides a construct as defined above wherein the polymer component is or comprises a silicone-containing polyurethane selected from CarboSil, PurSil, Avcothane and Cardiothane.

[0167] The invention also provides a construct as defined above, wherein the polymer component is or comprises Chroniflex or Tecoflex.

[0168] The invention also provides a construct as defined above, wherein one or more of the sheets is designed as a material reservoir for releasing an active material or an inactive material.

[0169] The invention also provides a construct as defined above, wherein the active material is selected from analgesics; antianxiety drugs; antiarrhythmics; antibacterial agents; antibiotics; anticoagulants and thrombolytics; anticonvulsants; antidepressants; antidiarrheals; antiemetics; antifungals; antihistamines; antihypertensives; anti-inflammatory drugs; antineoplastics; antipsychotics; antipyretics; antivirals; beta-blockers; corticosteroids; cytotoxins; hormones and sex hormones; enzymes; and vitamins.

[0170] The present invention also provides a construct as defined above, wherein the metal element is selected from the group consisting of a stent, a metal stent, a vascular graft, a heart valve, a membrane, a sealing device, a suture or staple, a hernia mesh or hernia repair device, a pelvic floor reconstruction device, a wound or burn dressing, a dura mater closure and a cardiac patch.

[0171] The invention also provides a kit comprising a construct according to any of the above items.

[0172] The invention also provides a device as defined above, configured as an implant.

[0173] The present invention also provides a device as defined above, selected from a stent, a metal stent, a vascular graft, a heart valve, a membrane, a sealing device, a suture or staple, a hernia mesh or hernia repair device, a pelvic floor reconstruction device, a wound or burn dressing, a dura mater closure and a cardiac patch.

[0174] The present invention also provides a device as defined above, wherein the device is a heart valve.

[0175] The invention also provides a process for producing a construct according to any one of the above definitions, the process comprising contacting at least one surface region of at least one decellularized tissue with a liquid polymer, and contacting at least one surface area of ​​at least one metal element with at least one liquid polymer; Contacting at least one surface region of the at least one metal element having the at least one liquid polymer with at least one surface region of the at least one decellularized tissue having the liquid polymer; thereby forming the construct.

[0176] The present invention also provides a process as defined above, wherein the liquid polymer coats at least one surface area of ​​the at least one metal element; and / or coats at least one surface area of ​​the at least one decellularized tissue.

[0177] The invention also provides a process as defined above, wherein the liquid polymer is solidified.

[0178] The present invention also provides a process for manufacturing a construct comprising at least one decellularized tissue, at least one metal element and at least one polymer component, the process comprising contacting at least one surface region of at least one decellularized tissue with at least one liquid polymer, and contacting at least one surface area of ​​at least one metal element with at least one liquid polymer; Contacting at least one surface area of ​​the at least one metal element having the at least one liquid polymer with at least one surface area of ​​the at least one decellularized tissue having the liquid polymer; thereby forming the construct.

[0179] The present invention also provides a process as defined above, wherein the liquid polymer coats at least one surface area of ​​the at least one metal element; and / or coats at least one surface area of ​​the at least one decellularized tissue.

[0180] The invention also provides a process as defined above, wherein the liquid polymer is solidified.

[0181] 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 is in physical contact with at least a portion of at least one surface region of the decellularized tissue without penetrating it; and further, wherein the at least one polymer component at least partially covers at least one surface region of the metal element.

[0182] Another object of the present invention is to provide a decellularized tissue physically associated with at least one metal element via at least one polymer component, the association comprising or consisting of: the polymer component being in at least partial physical contact with at least a portion of the surface area of ​​the tissue without penetrating it; and at least a portion of the at least one metal element being coated by the at least one polymer component.

[0183] 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 is at least partially associated with at least one face of the decellularized tissue without penetrating it; and at least partially associated with at least one surface area of ​​the metal element.

[0184] It is another object of the present invention to provide a construct as defined above, wherein said at least one decellularized tissue at least partially in contact with said at least one polymer component and said metal element at least partially coated with said at least one polymer component are thermally bonded to provide said construct.

[0185] It is another object of the present invention to provide the construct as defined above, wherein said at least one decellularized tissue at least partially coated with said at least one polymer component and said metal element at least partially coated with said at least one polymer component are thermally bonded to provide said construct.

[0186] Another object of the present invention is to provide a construct as defined above, wherein said thermal bonding is achieved by raising the temperature of the polymer components to above (a) the transition temperature of the polymer components for amorphous thermoplastic polymers; or (b) the melting temperature of the polymer components for semi-crystalline polymers, so as to at least partially cause entanglement of polymer chains along at least one region of the polymer components.

[0187] It is another object of the present invention to provide a construct as defined above in the form of a multi-sheet construct.

[0188] Another object of the present invention is to provide the construct as defined above, wherein said coating is performed by at least one method selected from the group consisting of spraying, brushing, dipping and any combination thereof.

[0189] It is another object of the present invention to provide the construct as defined above, wherein said at least one decellularized tissue is dried prior to being brought into physical contact with said polymer.

[0190] It is another object of the present invention to provide the construct as defined above, wherein at least one or any one of the at least one sheets of decellularized tissue is confined between any two sheets of polymeric component.

[0191] It is another object of the present invention to provide the construct as defined above, wherein the metal element is a metal wire.

[0192] Another object of the present invention is to provide a construct as defined above, which further comprises at least one solvent; wherein at least one of the following is applicable: (a) adding the solvent to an area of ​​the decellularized tissue where at least one surface area in contact with the polymer component contacts at least one surface area of ​​the metal element coated with the polymer component; (b) dissolving the polymer component in the at least one solvent; (c) any combination thereof.

[0193] Another object of the present invention is to provide the 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 combination thereof.

[0194] Another object of the present invention is to provide a construct as defined above, wherein the polymer component is in the form of a particle layer or particle coating, a polymer sheet, a polymer film, a polymer fiber or a polymer mesh, a gel, a hydrogel or a liquid film or a fluid film and any combination thereof.

[0195] Another object of the present invention is to provide the construct as defined above, wherein the decellularized tissue is obtained from a tissue selected from the group consisting of oral mucosa, small intestinal submucosa and bladder decellularized matrix, pericardium, bovine pericardium or porcine pericardium, omentum or small intestinal mucosa and any combination thereof.

[0196] Another object of the present invention is to provide a construct as defined above, wherein the polymer component is a polymer selected from the group consisting of or comprises a polymer selected from the group consisting of: 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, 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 Esters, silicone-containing polyurethanes, polyglycolic acid, polylactic acid, polycaprolactone, polylactide-caprolactone copolymers, polyglycolic acid-lactic acid copolymers, polyethylene oxide-polylactic acid copolymers, polyethylene oxide-polycaprolactone copolymers, polytetramethylene oxide-caprolactone copolymers, polyhydroxybutyrates, polyhydroxyvalerates, polyethylene adipate, polybutylene adipate, polyethylene succinate, polybutylene succinate and polybutylene terephthalate and polyethylene terephthalate / butylene terephthalate copolymers and combinations and copolymers thereof, shape memory materials, urethanes, Pellethane, Elastane, Elastolan, Tecoflex, Biomer, Chronoflex, Biospan, Bionate, silicone-containing polyurethanes selected from CarboSil, PurSil, Avcothane and Cardiothane, Chroniflex, Tecoflex and any combination thereof.

[0197] It is another object of the present invention to provide the construct as defined above, wherein one or more of said sheets is designed as a material reservoir for releasing active material or inactive material.

[0198] Another object of the present invention is to provide a construct as defined above, wherein the active material is selected from analgesics; anxiolytics; antiarrhythmics; antibacterials; antibiotics; anticoagulants and thrombolytics; anticonvulsants; antidepressants; antidiarrheals; antiemetics; antifungals; antihistamines; antihypertensives; anti-inflammatory drugs; antineoplastics; antipsychotics; antipyretics; antivirals; beta-blockers; corticosteroids; cytotoxins; hormones and sex hormones; enzymes; and vitamins.

[0199] Another object of the present invention is to provide a construct as defined above, wherein said metal element is selected from the group consisting of a stent, a metal stent, a vascular graft, a heart valve, a membrane, a sealing device, a suture or staple, a hernia mesh or hernia repair device, a pelvic floor reconstruction device, a wound or burn dressing, a dura mater closure and a cardiac patch.

[0200] It is another object of the present invention to provide the construct as defined above, wherein the different at least one polymer component has a substantially different viscosity than the at least one second polymer component.

[0201] Another object of the present invention is to provide a 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, such that the application of thermal bonding connects the at least one decellularized tissue and the at least one second decellularized tissue.

[0202] Another object of the present invention is to provide a construct as defined above, wherein the polymer component in physical contact with at least a portion of at least one surface area of ​​a decellularized tissue (a) at least partially penetrates at least one surface area of ​​a second decellularized tissue; and / or (b) has at least one surface feature extending from one surface of the second decellularized tissue, and the at least one surface feature passes through one surface of the second decellularized tissue to another surface through at least one hole formed in the tissue.

[0203] It is another object of the present invention to provide a construct as defined above in the form of a multi-sheet construct.

[0204] It is another object of the present invention 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, wherein any sheet of the second decellularized tissue is adjacent to or in contact with at least one sheet or segment of the polymer component.

[0205] It is another object of the present invention to provide the construct as defined above, wherein the multi-sheet construct comprises a number of sheets of the second decellularized tissue and an equal number of sheets of the polymer component.

[0206] It is another object of the present invention to provide the construct as defined above, wherein said at least one second decellularized tissue is dried before being coated by said polymer.

[0207] It is another object of the present invention to provide the construct as defined above, wherein at least one hole is pre-formed or present in the decellularized tissue.

[0208] Another object of the present invention is to provide a kit comprising a construct according to any one of the items defined above.

[0209] It is another object of the present invention to provide a device as defined above, configured as an implant.

[0210] Another object of the present invention is to provide a device as defined above, selected from the group consisting of a stent, a metal stent, a vascular graft, a heart valve, a membrane, a sealing device, a suture or staple, a hernia mesh or hernia repair device, a pelvic floor reconstruction device, a wound or burn dressing, a dura mater closure and a cardiac patch.

[0211] Another object of the present invention is to provide a process for producing a construct according to any one of the above definitions, the process comprising contacting at least one surface region 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; Contacting at least one surface region of the at least one metal element having the at least one polymer with at least one surface region of the at least one decellularized tissue having the polymer; thereby forming the construct.

[0212] It is another object of the present invention to provide the process as defined above, wherein the polymer coats at least one surface region of said at least one metal element; and / or coats at least one surface region of said at least one decellularized tissue.

[0213] It is another object of the present invention to provide a process as defined above, wherein the polymer is cured.

[0214] Another object of the present invention is to provide a process as defined above, which further comprises the step of adding at least one solvent; wherein at least one of the following is applicable: (a) adding the solvent to the area of ​​the decellularized tissue in contact with the at least one surface area of ​​the polymer component in contact with the at least one surface area of ​​the metal element coated with the polymer component; (b) dissolving the polymer component in the at least one solvent; (c) any combination thereof.

[0215] It is another object of the present invention to provide a process 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 combination thereof.

[0216] Another object of the present invention is to provide a process as defined above, which process further comprises: contacting the pierced surface region of at least one decellularized tissue with at least one polymer, and The at least one polymer is allowed to penetrate into the perforations (holes) and form a polymer sheet over the surface area.

[0217] Another object of the present invention is to provide a process as defined above, which process further comprises: contacting a surface region of at least one decellularized tissue with at least one polymer, and The at least one polymer is allowed to at least partially encapsulate the at least one decellularized tissue to form a polymer sheet thereon.

[0218] Another object of the present invention is to provide a process as defined above, which further comprises at least one step selected from the following items: (a) puncturing or forming holes in the surface area of ​​at least one decellularized tissue; (b) injecting a polymer into the perforation (hole); (c) solidifying the polymer; (d) any combination thereof.

[0219] Another object of the present invention is to provide a process as defined above, which further comprises at least one step selected from the following: (a) thermally bonding the at least one decellularized tissue and the at least one polymer by raising the temperature of the polymer component to above (i) the transition temperature of the polymer component for an amorphous thermoplastic polymer, or (ii) the melting temperature of the polymer component for a semi-crystalline polymer, so as 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 particle layer or particle coating, a polymer sheet, a polymer film, a polymer fiber or a polymer mesh, a gel, a hydrogel or a liquid film or a fluid film and any combination thereof; (c) any combination thereof.

[0220] Another object of the present invention is to provide a process for manufacturing a construct comprising at least one decellularized tissue, at least one metal element and at least one polymer component, the process comprising contacting at least one surface region 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; Contacting at least one surface region of the at least one metal element having the at least one polymer with at least one surface region of the at least one decellularized tissue having the polymer; thereby forming the construct.

[0221] It is another object of the present invention to provide the process as defined above, wherein the polymer coats at least one surface region of said at least one metal element; and / or coats at least one surface region of said at least one decellularized tissue.

[0222] It is another object of the present invention to provide a process as defined above, wherein the polymer is cured.

[0223] Another object of the present invention is to provide a process as defined above, which further comprises the step of adding at least one solvent; wherein at least one of the following is applicable: (a) adding the solvent to the area of ​​the decellularized tissue in contact with the at least one surface area of ​​the polymer component in contact with the at least one surface area of ​​the metal element coated with the polymer component; (b) dissolving the polymer component in the at least one solvent; (c) any combination thereof.

[0224] It is another object of the present invention to provide a process 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 and ethyl acetate, dimethylformamide (DMF), dimethylacetamide, DMAC and any combination thereof.

[0225] Another object of the present invention is to provide a process as defined above, which additionally comprises the following steps: a. contacting at least one surface area of ​​the at least one decellularized tissue that has been punctured to form one or more holes with at least one polymer, and b. allowing the at least one polymer to penetrate into the one or more holes and form a polymer sheet on the surface area.

[0226] Another object of the present invention is to provide a process as defined above, which process further comprises c. contacting at least one surface region of the decellularized tissue with at least one polymer, and d. allowing the at least one polymer to at least partially encapsulate the at least one decellularized tissue to form a polymer sheet thereon.

[0227] Another object of the present invention is to provide a process as defined above, which further comprises at least one step selected from the following items: (a) forming holes in the surface area of ​​at least one decellularized tissue; (b) injecting a polymer into the perforation (hole); (c) solidifying the polymer; (d) associating or fusing two or more constructs; (e) any combination thereof.

[0228] It is another object of the present invention to provide a process as defined above, wherein at least one of the following is true: (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 a polymer sheet on both sides of the surface area to form a construct component; and (d) any combination thereof.

[0229] Another object of the present invention is to provide a process as defined above, which further comprises at least one step selected from the following: (a) thermally bonding the at least one decellularized tissue and the at least one polymer by raising the temperature of the polymer component to above (i) the transition temperature of the polymer component for an amorphous thermoplastic polymer, or (ii) the melting temperature of the polymer component for a semi-crystalline polymer, so as 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 particle layer or particle coating, a polymer sheet, a polymer film, a polymer fiber or a polymer mesh, a gel, a hydrogel or a liquid film or a fluid film and any combination thereof; (c) any combination thereof.

[0230] The invention also provides a process as defined above, further comprising associating or fusing two or more constructs. BRIEF DESCRIPTION OF THE DRAWINGS

[0231] For a better understanding of the subject matter disclosed herein and for the purpose of illustrating how the same may be implemented in practice, embodiments will now be described, by way of non-limiting examples only, with reference to the accompanying drawings, in which: Figure 1A-Figure 1D is a SEM cross-sectional micrograph of a freeze-dried decellularized bovine pericardium (DBP) fraction.

[0232] Figure 2 High magnification (×200,000) SEM micrograph showing DBP Figure 3A-3C The structures of the tissues after application of increasingly high pressures are presented: 100 MPa, 200 MPa, and 500 MPa, far exceeding the pressures typically below 1 MPa or below 0.3 MPa required for designing DBP / polymer constructs. The fact that the collagen fibrils completely retained their structure after extremely high pressures demonstrates the robustness of DBP and its remarkable resistance to extremely high pressures.

[0233] Figure 4A-4B Shown are holes made with a 25G needle and a 27G needle, respectively, which were filled with the polymer phase Tecoflex.

[0234] Figure 5The coated struts of a metal stent when compressed (on the left hand side) and when expanded (on the right hand side) are presented. The bondable polymer used consisted of PCL and HDI.

[0235] Figure 6 The patch is shown to be partially engaged with only a small portion of the coated struts of the metal stent (approximately 15% of the stent area).

[0236] Figure 7 A stent stretched three times its original length is shown with no detachment of the patch from the stent.

[0237] Figure 8 A structure comprising DBP and a two-component construct comprising DBP and a polymer phase (Tecoflex in this case) is shown. Aiming to achieve a strong connection between the two DBP phases and the polymer phase, the six holes made in the DBP to enhance the flow of the Tecoflex are easily observed. By doing so, in addition to the diffusion of the Tecoflex molecules in the DBP phase, a laminate is formed comprising two outer Tecoflex films that are continuously connected to the central Tecoflex membrane through the holes made in the two DBP phases.

[0238] Figure 9A-9C Shown is a DBP leaflet of a heart valve, which also includes a metal frame connected together via a polymer connection, such as polyurethane Tecoflex. In this case, the DBP leaflet is joined together via the polymer phase of the DBP / polymer construct and is also joined to the metal frame of the heart valve via the coated struts of the metal frame.

[0239] Fig.10 A stent graft for treating abdominal aortic aneurysm (AAA) and other indications is shown, the stent graft comprising a metal stent and a fabric, wherein the latter is sutured to the former via a plurality of suture sites.

[0240] Fig.11A polymer phase consisting of PCL / HDI poly(ester-urethane) and a low molecular weight polymerizable molecule, hydroxyethyl methacrylate (HEMA), is presented. It is also reported that CLUR2k has a tensile modulus of about 180 MPa when unplasticized (100:0), and that the tensile modulus decreases significantly with the addition of increasing amounts of the smart HEMA component, showing a value of about 60 MPa in the presence of 20% monomeric HEMA, falling to 6 MPa in a 50:50 composition. As is evident from the presented data, for the CLUR2K:PHEMA 50:50 composition, polymerized PHEMA results in a significant increase in the CLUR modulus, reaching values ​​above 250 MPa.

[0241] Fig.12 The behavior of CLUR2K is presented when the low molecular weight component is not only polymerizable but also cross-linkable, as in the case of triethylene glycol dimethacrylate containing two carbon double bonds.

[0242] Fig.13 Additional cross-linking methods are demonstrated.

[0243] Figure 14A-14B Schematically illustrated is A) the creation of a tissue / polymer construct, wherein the first step is to form holes in the tissue component, the size, number and array of the holes being optimized. In B) is illustrated an embodiment of a "sandwich" of two tissues joined together by a dielectric polymer sheet.

[0244] 15-36 are illustrations of tissue-polymer-tissue multi-sheet constructs.

[0245] Figure 17-Figure 33 An example is given to evaluate different parameters and their effect on the joint strength.

[0246] 34a-34b are examples of different tissue piercing shapes (slot and zigzag).

[0247] Figure 34c shows the connection between tissue and metal.

[0248] 35-36c illustrate a heart valve according to one aspect of the present invention.

[0249] Fig.37 The figure illustrates the hemodynamic behavior of a heart valve according to an aspect of the present invention. DETAILED DESCRIPTION

[0250] Therefore, with the goal of providing a superior class of biomedical constructs and medical devices that will not have the drawbacks associated with currently available equivalents, the inventors of the technology disclosed herein have developed a new and unique class of constructs comprising [i] a decellularized tissue and [ii] a polymer component, wherein each of the components of the construct forms a distinct phase in space, wherein the decellularized tissue and polymer component are associated with each other via one or more physical anchoring means. Unexpectedly, the constructs and medical devices disclosed herein exhibit properties that were previously unattainable.

[0251] The decellularized tissue / polymer constructs taught by the present invention constitute the entire medical device or are a part of it, and include, among others, vascular grafts, cardiac patches, stents, heart valves, wound or burn dressings, membranes, sealing devices or devices to reinforce sutures or staples for hernia repair, pelvic floor reconstruction or dura mater closure.

[0252] The teachings of the present invention are readily applicable to a variety of decellularized tissues and polymers. However, for the sake of clarity, simplicity and simplicity, and without departing in any form or manner from the generality of the scope of the present invention, the inventors have chosen to illustrate the invention disclosed herein by focusing on constructs comprising decellularized pericardium and more precisely decellularized bovine pericardium. For the sake of clarity, simplicity and simplicity, and without departing in any form or manner from the generality of the scope of the present invention, the inventors have chosen to illustrate the invention disclosed herein by focusing on medical equipment in which the decellularized pericardium / polymer construct is part of a larger medical equipment and more precisely part of a heart valve. However, for the sake of clarity, simplicity and simplicity, and without departing in any form or manner from the generality of the scope of the present invention, the inventors have chosen to illustrate the invention disclosed herein by focusing on a polyurethane as the polymer phase of the decellularized pericardium / polymer construct and more precisely wherein the polyurethane is Tecoflex.

[0253] In the above scheme and without any limitation, the decellularized bovine pericardium / polymer construct disclosed by the present invention was developed in the inventor's laboratory. The mechanical properties of the decellularized pericardium vary slightly from batch to batch and depending on the decellularization technique used. Typically, the fracture stress of the decellularization technique falls within the range of 10 MPa-80 MPa, with typical Young's modulus values ​​spanning between 80 MPa and 300 MPa, exhibiting fracture strain values ​​of 20%-50%. The fracture stress values, Young's modulus values, and fracture strain values ​​measured for freeze-dried decellularized bovine pericardium in the inventor's laboratory were 75 MPa, 280 MPa, and 43%, respectively.

[0254] 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).

[0255]

[0256] The mechanical properties of Tecoflex were measured in the inventors' laboratory and are shown in Table 1.

[0257]

[0258] Table 1: Mechanical properties of Tecoflex It is obvious from the presented mechanical data that Tecoflex is a very strong material and will not represent the weak component of the construct. Other polyurethanes such as Elastolan can be used, as well as other polymers showing similar rheological properties and mechanical properties. In the case where the construct requires the polymer component to be biodegradable, polymers such as polylactic acid (PLA), poly(lactic / glycolic acid) (PLGA), polycaprolactone (PCL) and copolymers thereof, and more flexible biodegradable polymers such as biodegradable block copolymers, plastics or elastomers, containing hydrophilic polyethers such as polyethylene oxide (PEO) or its hydrophobic counterparts such as polypropylene oxide (PPO) or polytetramethylene oxide (PTMO), or flexible aliphatic polyesters such as amorphous polycaprolactone, or silicone-based segments including polydimethylsiloxane (PDMS) and many others. The molecular weight of the soft segment is usually changed from 600 dalton to 20,000 dalton.

[0259] However, for the sake of clarity, conciseness and simplicity, and without departing from the generality of the scope of the present invention in any form or manner, the inventors have chosen to illustrate the invention disclosed herein by focusing on a construct wherein a polymer phase of the construct is capable of connecting two or more of a tissue phase or other phase, the other phase comprising all types of materials selected from the group including polymers, metals, ceramics, carbonaceous materials and combinations thereof. More precisely, the inventors have chosen to illustrate the invention disclosed herein by focusing on a construct wherein a 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 a construct that is part of a heart valve that also includes a metal frame and connects the leaflets between them and also connects the leaflets to the metal frame of the valve. In some embodiments of the present invention, the tissue / polymer construct is part of a decellularized pericardial leaflet. In yet other embodiments of the invention, the polymer Tecoflex of the construct connects two tissue leaflets, thereby forming a multilayer integrated tissue / polymer construct comprising three polymer layers, two polymer layers on the outside of the construct and one polymer layer in the middle of the construct, and two tissue layers inside the two outer layers of polymer layers, forming a laminate. In other embodiments, the multilayer integrated tissue / polymer construct formed is composed of polymer layers connected by polymer connectors spanning from nanometer scale to centimeter scale in size. Materials and methods

[0260] Lyophilization of pericardium Since in most cases the decellularized tissue components of the constructs must be lyophilized prior to construct generation, the decellularized tissue components were investigated after lyophilization. The structure of the lyophilized decellularized bovine pericardium (DBP) component is described below. Figure 1A-Figure 1D The SEM cross-sectional micrograph shown in FIG.

[0261] The magnification increases from relatively low values ​​of x500, where the SEM micrographs show the entire thickness of the DBP, up to high values ​​of x120,000, where the well-known structure of collagen fibrils is easily observed.

[0262] Figure 2 Shown is a high magnification (x200,000) SEM micrograph of as-received DBP.

[0263] Preparation of polymers Selected polymers such as Tecoflex are added to the bonding area in two different states: a solution in THF and a pre-prepared Tecoflex film. The Tecoflex solution is a Tecoflex in the range of 5%-25% in THF (hereinafter referred to as a "polymer solution"). For film preparation, 3.78 grams of Tecolfex are added to 100 ml of THF until it is completely dissolved. The solution is poured into a glass petri dish and the THF is slowly evaporated. The film has an average thickness of several microns.

[0264] Coaptation of the pericardium Two dry pericardial strips were prepared for the bonding process. On each of them, an area 2 mm from the edge was marked and 6 holes were created. The holes were introduced into the polymer solution by a syringe with a 25G needle at its tip. The needle was inserted into the fabric from the rough side to the smooth side. Once it was pulled back outside, the solution was poured out at the same time. The holes were aligned in a straight line parallel to the edge line in the middle of the bonding area.

[0265] The pierced constructions were left for several minutes to further evaporate the solvent and then joined at the sealer with the rough sides pressed towards each other. Just before the joining process took place, a sheet of Tecoflex film was inserted between the pressed constructions.

[0266] For the bonding process, three rounds of sealing were applied in an elaborate regimen which was a combination of temperature, pressure and pressing time with relaxation between each of the three cycles.After bonding was performed, the bonded materials were dried at room temperature.

[0267] Junction Characterization Methods The strength of the joined strips was analyzed by 10 mm / min tensile test with an Instron instrument, while the leaflets were gripped at both sides thereof 10 mm from the joined area.

[0268] Furthermore, the structure of the joined strips was tested by SEM to ensure that the structure had not undergone significant changes due to pressing and heating.

[0269] The results are provided in Tables 2-4 below:

[0270] Table 2: Tensile test results of tissue-to-tissue bonding models without holes

[0271] Table 3: Tensile test results of tissue-to-tissue bonding model with 4 holes

[0272] Table 4: Tensile test results of tissue-to-tissue bonding model with 6 holes The data presented in Tables 2-4 support the understanding that the more holes formed in the tissue that are filled with polymer, the stronger the association or bond.

[0273] discuss Applying gentle pressure may be necessary in some embodiments of the present invention, particularly when the polymer phase is connected to more than one DBP and / or polymer phase. The purpose of applying pressure in these embodiments of the present invention is to make one or more polymer phases flow, in some embodiments contact with the DBP, and in other embodiments contact with other polymer phases, and combinations thereof.

[0274] In some embodiments of the invention, the polymer phase can flow through the pores or holes made in the tissue phase, and the polymer molecules can also diffuse and mix with molecules of other phases. In some embodiments, the flow of polymer molecules causes the interface between the phases to disappear, joining them together.

[0275] Figure 3A-3C The structure of the tissue after application of increasingly higher pressures is presented: pressures of 100 MPa, 200 MPa, and 500 MPa, respectively, far exceeding the pressures that are typically below 1 MPa, preferably below 0.5 MPa, and even more preferably below 0.3 MPa that are actually required for designing DBP / polymer constructs. The fact that the collagen fibrils completely retain their structure after these extremely high pressures demonstrates the robustness of DBP and its remarkable resistance to extremely high pressures.

[0276] Several polymers capable of forming the polymer phase of DBP / polymer constructs have demonstrated their ability to flow under physiologically acceptable pressure and temperature conditions. Thermograms of two biodegradable poly(ester-urethane)s and their joined phases, composed of polycaprolactone (PCL) of different molecular weights (2,000 Daltons and 14,000 Daltons) and hexamethylene diisocyanate (HDI), obtained by differential scanning calorimetry (DSC), demonstrated a new, broader endotherm, shifted to lower temperatures. This provided an indication that the molecules of the two polymers had interdiffused, one hindering the crystallinity of the other.

[0277] This phenomenon and the strength of the bonded connection were further explored in the case of joining two films of two different polyurethanes, Tecoflex and PEU. The results demonstrated the ability of Tecoflex chains and PEU chains to flow across the interface between the two films, providing a strong connection with long-term clinical importance. In addition, the two polymers are similar because they are polyurethanes, even though in the first case they are poly(ester-urethane), and in the latter case they are poly(ether-urethane). In order to illustrate the wide range of this phenomenon, the DSC thermograms of the two polymers were explored.

[0278] DSC demonstrates that the two polymers are very different, one is a flexible poly(ester-urethane) and the other is a rigid polymethacrylate. The flexible polyurethane consists of PCL 2000 segments and HDI as its coupling agent, while the rigid polymethacrylate is poly(ethyl methacrylate) (PEMA). Also in this case, even though the polymers are substantially different in their composition and mechanical properties, it is evident from the thermal map of their joined phases that this phenomenon requires mixing and entanglement of the chains of the two components at the molecular level. In several embodiments of the present invention, the mobility of the polymer chains and their ability to disentangle, cross an interface with another polymer phase or a decellularized tissue phase, diffuse into a second phase and then re-entangle plays a key role in producing the constructs or medical devices taught by the present invention.

[0279] The strength of the connection between the DBP and the polymer phase, such as Tecoflex, was quantitatively determined using an Instron machine.The construct was found to fail cohesively within the tissue, but no de-welding failures were observed.

[0280] In some embodiments, where a particularly strong connection between the DBO phase and the polymer phase is intended, a hole is made in the DBP phase to maximize the flow of the polymer phase through the hole. In some embodiments, the hole is made using a needle in the range of 16G to 27G. Figure 4A-4B Shown are holes made with a 25G needle and a 27G needle respectively, which were in these cases filled with the polymer phase Tecoflex.

[0281] The DBP phase and the polymer phase can be any size spanning from nanometers to centimeters and can take any shape, including but not limited to spheres, fibers, strips, ribbons, films, porous or non-porous and combinations thereof. One or more polymer phases can also be present in phases ranging in size from nanometer scale up to centimeter scale, and they can be on the surface of the DBP phase and / or in its bulk, and each of these cases has a size ranging from nanometers to centimeters and takes any geometric shape.

[0282] The object of the present invention is to connect a DBP / polymer construct or a device of which the construct is a part to an additional component of the device. In the case of a heart valve (a very preferred embodiment of the present invention), the additional component is the metal frame of the heart valve.

[0283] In some embodiments of the invention, when a DBP / polymer construct or a device of which the construct is a part is bonded to a metallic stent, the struts of the stent are coated with a bondable polymer that may be the same or different than the polymer that makes up the polymer phase of the DBP / polymer construct. Figure 5 The coated pillar (on the left hand side) of the metal support when compressed and the coated pillar (on the right hand side) of the metal support when expanded are presented. In this embodiment, due to the large expansion of support, the engageable polymer coating its pillar is selected to be particularly flexible and show a high fracture strain value. In the case shown below, the engageable polymer used is composed of PCL and HDI. Prepare the coating of different thicknesses, starting from the coating as thin as 5 microns and increasing as required.

[0284] Other highly flexible polymers have also been used. Among others, a variety of polyurethanes have been used, whose soft segments are composed of polyethers or polyesters, which are different in their molecular weight, their hydrophilicity and in the case of biodegradable polymers, and also in their degradation rates. One of the polymers used consists of poly(tetramethylene oxide) (PTMO) (MW=650) segment chains extended via HDI. As reported in Table 5, the polymers were joined in 20 seconds at 47°C-48°C temperatures.

[0285]

[0286] Table 5: Polymer properties The same polymer was used to coat the struts of the stent and was also used to create the polymer phase of the DBP / polymer construct and the additional polymer phase of the medical device of which the polymer is a part. To illustrate not only the speed of the bonding process but also the strength of the polymer / polymer connection formed, a patch of PTMO650 / HDI polymer was bonded to the struts of a stent coated with the same polymer within 20 seconds. Furthermore, the patch was allowed to bond to only a small portion of the coated struts of the metal stent, such as Figure 6 (approximately 15% of the stent area).

[0287] Initially, a forced manual operation was performed to disengage the patch from the covered stent, but Figure 7 As shown in , the stent was stretched three times its initial length without detachment of the patch from the stent.

[0288] When the patch was finally debonded from the stent using the Instron instrument, unexpectedly and surprisingly, the metal stent failed to remain unaffected, while the bonded connection remained unaffected. This convincingly demonstrates the advantageous features stemming from the ability of the polymer phase of the DBP / polymer construct or the polymer phase of the medical device of which the construct is a part to quickly and securely form a long-term, stable connection between different phases.

[0289] Figure 8 Structures comprising DBP and a two-component construct comprising DBP and a polymer phase (Tecoflex in this case) are shown. Aiming to achieve a strong connection between the two DBP phases and the polymer phase, six holes made in the DBP to enhance the flow of Tecoflex are easily observed. By doing so, in addition to the diffusion of Tecoflex molecules in the DBP phase, a laminate is formed comprising two outer Tecoflex films that are continuously connected to the central Tecoflex membrane through the holes made in the two DBP phases.

[0290] In a more preferred embodiment, the DCP leaflets of the heart valve further comprising the metal frame are connected together via a polymer connection, preferably polyurethane, more preferably Tecoflex, such as Figure 9A-9C In this case, the DBP leaflets are joined together via the polymer phase of the DBP / polymer construct and are also joined to the metal frame of the heart valve via the coated struts of the metal frame.

[0291] As commonly done and routinely reported in the literature, the long-term dynamic stability of heart valves is determined in vitro under accelerated conditions by an accelerated wear tester (AWT).

[0292] Three articles representing many of the studies describing the use of AWT in heart valves are referenced below: {1} A correlation between long-term in vitro dynamic calcification and abnormal flow patterns past bioprosthetic heart valves, OleksandrBarannyk, Robert Fraser and Peter Oshkai, J Biol Phys (2017) 43:279–296); {2} Pitfalls and outcomes from accelerated wear testing of mechanical heartvalves, A Campbell, T Baldwin, G Peterson, J Bryant and K Ryder, J Heart ValveDis, June 1996; 5 Suppl 1:S124-32; discussion144-8; {3}. A study in the design of an Accelerated Wear Tester that is compatible with a particle imagevelocimetry and high-speed camera setup, Edward A. Brown, Master of Sciencethesis, The Pennsylvania State University, The Graduate School, College of Engineering, 2015.

[0293] During these tests, a maximum cyclic stress of 100 mmHg, equal to 0.0133 MPa (12.3 kPa), was applied. Failure values ​​measured using the Instron instrument typically fell in the range of 5 MPa to 10 MPa, which is more than 300 times higher than the maximum cyclic stress applied during the AWT determination.

[0294] The advantageous features of the DBP / polymer constructs taught by the present invention become even more dramatic and surprising when compared to conventionally used alternative suturing techniques.

[0295] Connecting phases of a construct and the construct to other phases of the medical device of which the construct is a part has important advantages over suturing: [a] it is very rapid, relative to the extremely lengthy and time-consuming suturing process through many suture sites, [b] produces strong connections, [c] is reproducible and independent of technicians, [d] achieves a much better compliance match between the phases of the construct and the phases of the device, in stark contrast to the much stiffer sutures used, [e] achieves a much more uniform stress distribution, relative to the extremely harmful and sometimes life-threatening stress concentration effects of sutures, and [f] is inexpensive.

[0296] In some embodiments, any phase of the construct and / or any phase of the medical device of which the construct is a part or any element of the present invention may contain at least one additional material to improve any aspect of the clinical performance of any one and combination of the embodiments of the present invention, including its biocompatibility, its blood compatibility, the cellular response they trigger and other aspects. At least one additional material may be selected from active materials and inactive materials. In some embodiments, the active material is selected from a variety of bioactive agents. Exemplary bioactive agents include, for example, anticoagulants such as heparin and chondroitin sulfate; fibrinolytics such as tPA, plasmin, streptokinase, urokinase, and elastase; steroidal and nonsteroidal anti-inflammatory agents such as hydrocortisone, dexamethasone, prednisolone, methylprednisolone, promethazine, aspirin, ibuprofen, indomethacin, ketorolac, meclofenamic acid, tolmetin; calcium channel blockers such as diltiazem, nifedipine, verapamil; antioxidants such as ascorbic acid, carotenes, and alpha-tocopherol, allopurinol, trimetazidine; antibiotics such as noxythiolin and other antibiotics that prevent infection; prokinetic agents that promote intestinal motility; agents that prevent collagen cross-linking such as cis-hydroxyproline and D-penicillamine; and agents that prevent mast cell degranulation such as disodium cromoglycate, as well as many other bioactive agents.

[0297] In addition to the above agents that usually show favorable pharmacological activity related to promoting wound healing or reducing infection or having hemostatic properties or enhancing blood compatibility, other bioactive agents can also be delivered by constructs or medical equipment of the present invention, and the other bioactive agents include, for example, amino acids, peptides, proteins including enzymes, carbohydrates, growth factors, antibiotics (treating specific microbial infections), anticancer agents, neurotransmitters, hormones, immunological agents including antibodies, nucleic acids including antisense agents, fertility drugs, psychotropic drugs and local anesthetics, and many other agents. The delivery of these agents and other agents will depend on the pharmacological activity of the agent, the active site in the body, and the physicochemical properties of the agent delivered, the therapeutic index of the agent, and other factors. Those of ordinary skill in the art will be able to easily adjust the physicochemical properties of the polymer of the present invention and the hydrophobicity / hydrophilicity of the agent delivered, so as to produce the desired effect. In this aspect of the present invention, bioactive agents are applied with a concentration or amount that effectively produces the desired 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 a patient's body.

[0298] In some embodiments, the inactive material is selected from dyes, polymeric materials, thickeners, plasticizers, agents affecting hydrophilicity, agents affecting lubricity, and other inactive materials.

[0299] 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, bonding, any of a number of 3D printing techniques, and in each case, the specific manufacturing technology being used will be customized to make it compatible with the constructs and medical devices taught by the present invention.

[0300] In some embodiments in which at least one of the polymer phases is biodegradable, the biodegradable polymer is selected from the group comprising lactic acid, lactide, glycolic acid, glycolide, or related aliphatic hydroxycarboxylic acids or aliphatic hydroxycarboxylic acid esters (lactones) selected from the group consisting of β-propiolactone, ε-caprolactone, δ-glutarolactone, δ-valerolactone, β-butyrolactone, pivalolactone, α,α-diethylpropiolactone, ethylene carbonate, trimethylene carbonate, γ-butyrolactone, β-propiolactone, α-diethyl ... lactone, p-dioxanone, 1,4-dioxepan-2-one, 3-methyl-1,4-dioxane-2,5-dione, 3,3-dimethyl-1,4-dioxane-2,5-dione, α-hydroxybutyric acid, α-hydroxyvaleric acid, α-hydroxyisovaleric acid, α-hydroxyhexanoic acid, α-hydroxy-α-ethylbutyric acid, α-hydroxyisocaproic acid, α-hydroxy-α-methylvaleric acid, α-hydroxyheptanoic acid, α-hydroxystearic acid, α-hydroxytetracosanoic acid, cyclic esters of salicylic acid, and mixtures thereof.

[0301] The polymer phase according to the present invention comprises optionally low molecular weight molecules, which can enhance the fluidity of the polymer phase and / or allow to cause the polymer phase or a part thereof to flow under milder temperature and pressure conditions. Another object of the present invention is to provide polymerizable or crosslinkable low molecular weight molecules, so they soften the polymer phase or a part thereof before it is polymerized or crosslinked, and once polymerized or crosslinked, strengthen or harden the polymer phase, wherein the low molecular weight molecules can follow any mechanism polymerization or crosslinking, the mechanism includes but is not limited to addition reaction and polycondensation reaction and all types of other reactions including click chemistry, and combinations thereof. Among others, the polymerizable or crosslinkable low molecular weight molecules include precursors comprising one or more double bonds. Several examples are given in the figure below.

[0302] exist Fig.11 In the present invention, the polymer phase consists of PCL / HDI poly(ester-urethane) and the low molecular weight polymerizable molecule is hydroxyethyl methacrylate (HEMA).

[0303] Fig.11 CLUR2k is reported to have a tensile modulus of about 180 MPa when unplasticized (100:0), and with the addition of increasing amounts of the smart HEMA component, the tensile modulus decreases significantly, showing a value of about 60 MPa in the presence of 20% monomeric HEMA, dropping to 6 MPa at a 50:50 composition. As is evident from the data presented, for the CLUR2K:PHEMA 50:50 composition, polymerized PHEMA results in a significant increase in the CLUR modulus, reaching values ​​above 250 MPa.

[0304] Fig.12The behavior of CLUR2K is presented when the low molecular weight component is not only polymerizable but also cross-linkable, as in the case of triethylene glycol dimethacrylate containing two carbon double bonds.

[0305] Several additional chemical methods can be used to polymerize or crosslink the low molecular weight components, such as epoxy-amine reactions. This chemistry is accomplished by using polyglycidyl methacrylate (PGMA) (see Fig.13 ) (polyglycidyl methacrylate (PGMA) is very hard and slightly brittle) blended with polyethyleneimine molecules containing several amine groups. When initially blended with PGMA, the PEI molecule reduces the rigidity of the PGMA, but once it reacts with the epoxide ring (see Fig.13 ), which cross-links the PGMA and hardens the polymer.

[0306] Figure 14 schematically shows the generation of tissue / polymer construct, wherein the first step is to form a hole in the tissue component, and the size, quantity and array of the hole are optimized. In this case, or when polymer is in a state higher than its glass transition temperature or melting point, when these transitions occur at suitable low temperatures, a liquid polymer phase is added, which can be a polymer solution in a suitable solvent for example. Polymer penetrates tissue mainly through the hole made in tissue, and the length, size, quantity and array of these holes are controlled and change in a significant range. In some cases, the polymer phase comprises a hole of different penetration depths. In some cases, the hole only partially penetrates the tissue phase, and in other embodiments, the depth of the hole passes through the tissue phase face to face. In the case of the following description, the hole passes through the entire tissue thickness, and two polymer layers are produced on both sides of the tissue component. Because they are connected via the polymer connector filling the hole, two polymer films become an 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 they are derived from the cohesive strength of the polymer itself.

[0307] In other embodiments of the invention, a preformed polymer phase is first produced, the composition and morphology of which allows it to flow under appropriate temperature and pressure conditions, as previously described.

[0308] In some embodiments, the preformed polymer phase has different properties in its surface layer relative to its main body (bulk), which allows the polymer phase in contact with the tissue to flow into the hole made in the tissue. In some embodiments, the difference between the surface layer (its thickness is optimized) of the polymer phase and the main body of the polymer phase is compositional, so that the surface layer shows required fluidity under the applied temperature and pressure conditions. In some embodiments, the difference between the surface layer (its thickness is optimized) of the polymer phase and the main body of the polymer phase is morphological. In this embodiment, the surface layer is less crystalline, and in some embodiments is amorphous, and the polymer in the main body of the polymer phase shows enhanced crystallinity and therefore shows higher rigidity at the relevant temperature. The morphological difference encoded in the polymer phase can follow a variety of strategies to achieve. Among others, this can be achieved by carrying out a spatially confined heat treatment, which makes the surface layer of the polymer phase less crystalline or amorphous relative to the main body. In some embodiments, the difference between the surface layer of the polymer phase and the main body is achieved by adding a movable component that tends to migrate and concentrate on the surface layer of the polymer phase. In yet other embodiments, differentiation between the surface layer and the bulk of the polymer phase is achieved by judicious selection of the surface with which the polymer phase is produced in contact.These and other similarly effective techniques may be used alone or may be combined.

[0309] In yet other embodiments of the present invention, the initial liquid polymer phase and the preformed polymer phase are combined in different ways. In some embodiments, for example, they are added simultaneously, or they can also be deployed sequentially or in any other way that will produce the best association of tissue / polymer, as deduced from its clinical use.

[0310] A procedure in which a polymer solution is used to produce a tissue / polymer composed of at least two tissue phases is exemplified herein and is described in Fig. 14BDepicted in. In some cases, the term "joining" and "connection" and similar terms are used interchangeably. Two freeze-dried pericardial strips are used, and the tissue / polymer association causes the two tissue phases to be connected via the polymer phase. On each of the pericardial strips, 6 holes are produced in a region 2 mm from the edge, and these holes are spaced apart over the entire width of the tissue strip. In this embodiment, holes are formed simultaneously and the holes are filled with polymer solution. In this case, 8% Tecoflex / THF solution is prepared, and a syringe with a 25G needle at its tip is filled with the solution (for other purposes, in other experiments, 1% or 4% concentration of Tecoflex is used). The needle is inserted into the pericardial tissue strip from the rough side of the pericardial tissue strip, the pericardial tissue strip is pierced and a hole is produced, and the polymer solution is initially poured toward the distal end, and then the hole is filled when pulled back, and finally a polymer film is produced on the proximal face of the tissue. By doing this, a laminate consisting of two external polymer phases connected via a polymer connector (through the hole made in the tissue) is produced. Hole is aligned with the straight line parallel to the edge line in the middle of the joint area.The tissue / polymer construct composed of the laminate described above is placed for 2 minutes to allow the best solvent (THF in this particular case) to evaporate, and then the two processed tissue / polymer constructs of each tissue strip are overlapped as required, and the sealer is used to join together under appropriate temperature, pressure and time conditions, wherein the rough side is pressed toward each other. Three rounds are applied at 85 ℃ at the sealer. Each round at the sealer consists of the following: 2 bar, 12 seconds step, followed by 1 second relaxation between the cycles. After connecting, the tissue / polymer construct of the joint is dried at room temperature for 5 hours, then soaked in saline solution for 24 hours. The cross-head speed (cross-head speed) of 10 mm / min is used on the Instron instrument to study the connection strength between the tissue strips of two connections, and the strip is clamped from the area 10 mm away from the connection.

[0311] The strength of the formed joint was also determined under peeling conditions and, for this purpose, the samples were connected accordingly. All other details of the procedure remained unchanged. The mechanical investigations of these samples were carried out using the same conditions as previously indicated, with the only difference that the gripping took place 20 mm from the joined area.

[0312] Some of the different conditions and connection parameters investigated are listed below: 1) For 27G needle - 2 / 4 / 6 / 8 holes.

[0313] For 25G needles - 2 / 4 / 6 holes.

[0314] For 23G needles - 2 / 4 / 6 holes.

[0315] For 21G needles - 2 / 4 hole.

[0316] For 19G needles - 2 / 4 hole.

[0317] For 16G needle - 2 holes.

[0318] 2) Cycles performed at the sealer - from 1 to 6 times.

[0319] 3) Connection pressure - from 2 bar to 10 bar.

[0320] 4) Connection time - from 8 seconds to 12 seconds.

[0321] Here is exemplified a procedure in which a tissue / polymer consisting of at least two tissue phases is produced by combining a polymer solution and a preformed polymer phase. The steps involving the Tecoflex / THF solution are the same as described above.

[0322] As described above, the tissue / polymer construct composed of formed laminate is placed for 2 minutes to allow the best solvent (THF in this particular case) to evaporate.Before engaging two prepared tissue / polymer constructs, before the bonding process using sealer occurs, 2*6 mmTecoflex films (thickness~160 μm) are deployed between two tissue / polymer constructs.3.78 g Tecoflex in 100 ml chloroform is used to prepare Tecoflex films in advance, even if other solvents are also used.As described above, the same steps described are carried out at the sealer place, and the sample connected is studied at the Instron machine place.

[0323] Some of the different conditions and connection parameters investigated are listed below: 1) For 27G needle - 2 / 4 / 6 / 8 holes.

[0324] For 25G needles - 2 / 4 / 6 holes.

[0325] For 23G needles - 2 / 4 / 6 holes.

[0326] For 21G needles - 2 / 4 hole.

[0327] For 19G needles - 2 / 4 hole.

[0328] For 16G needle - 2 holes.

[0329] 2) A preformed membrane was prepared from THF or chloroform solvent using 1.5-4 g Tecoflex in 100 ml of the mentioned solvent.

[0330] 3) Cycles performed at the sealer - from 1 to 6 times.

[0331] 4) Engagement pressure - from 2 bar to 10 bar.

[0332] 5) Engagement time - from 8 seconds to 12 seconds.

[0333] Thus, according to one embodiment of the present invention, a tissue-to-tissue joining process is achieved.

[0334] According to a preferred embodiment, the process comprises the following two main steps: (a) Two tissue sheets 10 are positioned from both sides of the polymer film 20 (see FIGS. 15a and 15b ).

[0335] (b) A rivet 30 is created that acts as a polymer tube by injecting polymer from one side of a tissue-polymer-tissue "sandwich" to the other side (see FIGS. 16 a-c ). Such a rivet functions similarly to a classic "wall flange" or rivet. According to a preferred embodiment, a sandwich of polymer-tissue-polymer-tissue-polymer is created (as illustrated in FIG. 14 b ).

[0336] In other words, the connection / joining of two tissues is the superposition of two mechanisms: 1. Surface attachment (e.g. gluing); and 2. Physical connection - similar to rivets.

[0337] It should be noted that each of the tissue sheets can be coated with polymer on both sides thereof. Thus, the construct sandwich structure will be polymer-tissue-polymer-tissue-polymer.

[0338] According to another embodiment of the invention, laser cutting is used to cut tissue at the connecting ("joining") area (ie, at the area of ​​connection with the polymer).

[0339] According to another embodiment of the present invention, the tissue is frozen and dried (ie, lyophilization process) at a temperature of -70 degrees Celsius to 20 degrees Celsius with or without vacuum for a period of ~5-20 hours prior to being joined to the polymer.

[0340] Example The following examples are given to evaluate different parameters and their effect on the joint strength.

[0341] The tests are divided into three main groups: Anastomosis-related parameters: Heat, pressure and time were examined to understand their effect on anastomosis strength. It should be noted that the sensitivity of the pericardium to high temperatures is limited to 85°C. Therefore, the main experiments applied were pressures of 2, 4 and 6 bars, and heating times of 36 seconds (3 pulses), 50 seconds and 120 seconds (1 pulse).

[0342] Hole related parameters: number of holes, distance between holes, size and depth of holes.

[0343] Polymer-related parameters: Two polymers and different thicknesses of the polymer films used.

[0344] It should be noted that all holes are prepared by laser cutting machine and are aligned. Generally, the scheme used is as follows:

[0345] The pericardial sheet was divided into 6 pairs of strips, each pair being referred to as a conjoined sample.

[0346] At 1 mm on the upper side of each strip, X holes were implemented, where the distance between the X holes was Y (X, Y are disclosed in each experiment detailed below).

[0347] The laser cutting process was completed using the following parameters: 100% power, 50% speed, and 1000 parts per inch, PPI.

[0348] Next, tensile tests were performed. In each test, the tissue was fixed to a metal handle covered with plastic to increase surface resistance (in a Testometric X250-2.5 apparatus with a cell load of up to 250 N) and stretched at a rate of 10 mm / min until failure point.

[0349] In the following examples, the polymer used was Chronoflex.

[0350] Joint related parameters: 1) Effect of pressure on bonding strength: Pressures of 2, 4 and 6 bars were applied, as well as three pulses of heating for 12 seconds. In the test, the strength required to 'break' the bond was measured. The measurement was carried out with a Testometric X250-2.5 device and tension was applied at a rate of 10 mm / min until the breaking point.

[0351] Reference now Fig.17 , Fig.17 The graph shows the effect of pressure on coaptation stress. It should be noted that given the differences in thickness of the pericardial tissue, stress rather than strength was measured.

[0352] As can be seen, applying higher pressure results in stronger anastomosis (requiring higher force to break the anastomosis). Wherein, in the figure, 'W' represents anastomosis rupture; 'PW' represents pericardial rupture at the anastomosis; and 'P' represents pericardial rupture.

[0353] The use of high pressure forces the polymer chains to move at the junction, resulting in a higher degree of entanglement. Therefore, as shown, the strength of the junction is greater.

[0354] 2) Effect of heating time and pulse number on bonding strength: Due to the sensitivity of the pericardium to high temperatures, higher temperatures could not be used. However, heating at 85°C for longer periods of time and without relaxation could be tested. Therefore, in this example, the heating times were tested as follows: three pulses of 36 seconds, one pulse of 50 seconds, and one pulse of 120 seconds.

[0355] Reference now Fig.18 , Fig.18 The graph shows the effect of temperature on joint strength. As disclosed above, due to differences in the thickness of the pericardial tissue, stress was measured instead of strength.

[0356] As can be seen in the figure, the higher heating time has a maximum at 50 seconds and then the joint strength becomes weaker.

[0357] Wherein, in the figure, 'W' represents a rupture of the joint; 'PW' represents a rupture of the pericardium at the joint; and 'P' represents a rupture of the pericardium.

[0358] These results could be explained by some damage induced in the pericardium leading to a weakening of its mechanical properties. In other words, higher heating times had the same effect as using higher temperatures.

[0359] Hole related parameters: 1) Effect of hole size on bonding strength Reference now Fig.19 , Fig.19 The figure shows the effect of the size of the hole on the strength of the joint. As disclosed above, due to the difference in thickness of the pericardial tissue, stress was measured instead of strength.

[0360] As seen in the figure, larger hole diameters lead to increased joint strength up to a 0.4 mm hole; thereafter, larger hole diameters lead to decreased joint strength. Wherein, in the figure, 'W' represents joint fracture; 'PW' represents pericardial fracture at the joint; and 'P' represents pericardial fracture.

[0361] 2) Effect of the number of holes on bonding strength Reference now Fig. 20 , Fig. 20The figure shows the effect of the number of holes on the strength of the joint. Wherein, in the figure, 'W' represents a joint fracture; 'PW' represents a pericardial fracture at the joint; and 'P' represents a pericardial fracture. As disclosed above, due to the difference in thickness of the pericardial tissue, stress is measured instead of strength.

[0362] For the purpose of this example, 0.2 holes were tested. As can be seen in the figure, the maximum strength is achieved at four holes with a distance of 1.5 mm between each other; then, the strength decreases as the number of holes increases.

[0363] On the one hand, fewer holes have less polymer; on the other hand, more holes have less tissue. In other words, due to less tissue in the joint, the joint becomes weaker for additional holes.

[0364] Polymer related parameters: Effect of film thickness on bonding strength: Reference now Fig.21 , Fig.21 The figure shows the effect of membrane thickness on the strength of the joint. Wherein, in the figure, 'W' represents the joint site fracture; 'PW' represents the pericardial fracture at the joint site; and 'P' represents the pericardial fracture. As disclosed above, due to the difference in the thickness of the pericardial tissue, stress is measured instead of strength.

[0365] As seen, the results indicate that thicker polymers lead to weaker bond areas. Thicker polymers mean that it requires higher energy (temperature in our case) to disentangle the chains of the polymers, and due to temperature limitations, it is not preferred to use thicker polymers, resulting in increased bond strength.

[0366] In the following examples, the polymer used is polycaprolactone 43K, PCL 43K. It should be noted that PCL is a biodegradable polyester with a low melting point of about 60°C.

[0367] 1) Effect of hole size on bonding strength: The melting point of PCL is about 60° C., so the bonding process of PCL was performed at 55° C., a pressure of 4 bar, and one pulse of 50 seconds.

[0368] As described above, the insertion of the holes strengthens the bonding site because the holes contain a polymer, increasing the incidence of bonding.

[0369] Since PCL (8% solution) is not as viscous as Chronoflex, the hole can accommodate a reduced amount of polymer. In addition, the temperature used in the process essentially melts the PCL (because PCL has a low melting temperature); therefore, the bond becomes weaker, as Fig. 22As shown in the figure (wherein, 'W' represents commissure rupture; 'PW' represents pericardial rupture at commissure; and 'P' represents pericardial rupture). As disclosed above, stress is measured rather than strength due to differences in the thickness of the pericardial tissue.

[0370] 2) The influence of the number of holes on the bonding strength: In this section, the number of holes was tested and the hole size used was 0.2 mm.

[0371] Reference now Fig.23 , Fig.23 The figure shows the effect of membrane thickness on the strength of the joint. Wherein, in the figure, 'W' represents the joint site fracture; 'PW' represents the pericardial fracture at the joint site; and 'P' represents the pericardial fracture. As disclosed above, due to the difference in the thickness of the pericardial tissue, stress is measured instead of strength.

[0372] like Fig.23 As can be seen in Figure 3, increasing the number of holes increases the joint strength.

[0373] in conclusion: All the work performed indicates the importance of holes for strengthening the joint, as well as a better understanding of the joining process and all parameters to achieve a joint with enhanced mechanical properties.

[0374] In conclusion, the insertion of holes in both PCL 43K and Chronoflex improved the strength of the joint and increased its mechanical properties. However, the best results were not obtained using the same set of parameters. The best results were obtained with PCL by creating six holes of 0.2 mm, and the peak stress was almost doubled. Whereas in the case of Chronoflex, the best results were obtained by inserting four holes of 0.4 mm, where the strength was almost three times that of the result without holes.

[0375] Reference now Figure 24-Figure 33 , Figure 24-Figure 33 The diagram shows the following: Fig.24 The image shows dry pericardial tissue without holes.

[0376] Fig.25 The figure shows moist pericardial tissue without holes after tensile testing.

[0377] Fig.26 The image shows dried pericardial tissue with 4 holes of 0.6 mm.

[0378] Fig. 27 The image shows 0.6 mm, 4 holes of wet pericardial tissue after stretching.

[0379] Fig.28The image shows 0.2 mm, 3-hole dried pericardial tissue.

[0380] Fig.29 The image shows 0.2 mm, 3 holes of wet pericardial tissue after stretching.

[0381] Fig.30 The image shows 0.2 mm, 4 holes of dried pericardial tissue.

[0382] Fig.31 The image shows 0.2 mm, 4 holes of wet pericardial tissue after stretching.

[0383] Fig.32 The image shows 0.2 mm, 6-hole dried pericardial tissue.

[0384] Fig.33 The image shows 0.2 mm, 6-hole wet pericardial tissue after stretching.

[0385] The following examples are presented to evaluate different steps in the production of multi-sheet constructs and their effect on joint strength.

[0386] 1. Effect of freeze drying (lyophilization) on tissue strength (before joining): In the following tests, the effect of shock freeze on the mechanical properties of tissues was evaluated. The mechanical properties of dried tissues were compared with / without liquid nitrogen N2.

[0387] The following test protocol was adopted and conducted: 1. Laser cutting of tissue areas with dimensions of 6 mm × 40 mm and a thickness of 250 microns + / - 50 microns; 2a. The tensile strength is measured in the middle of the 6 mm width and at the ends and along the middle along 40 mm; 2b. For liquid nitrogen N2, immerse the test tissue in liquid nitrogen; 3. Next, the tissue sample is placed in a freeze dryer in a vacuum at a temperature range of -70 degrees Celsius to 0 degrees Celsius for a period of 7-24 hours; 4. Next, measure the thickness of the dried tissue to compare the difference between the treatment with liquid nitrogen and the treatment without liquid nitrogen; 5. Then, the tissue samples were placed in saline for 12 hours for recovery; 6. Next, measure the thickness of the dried tissue again; 7. Finally, the tissue is tensile tested to determine the force at which it breaks (tears), and the resulting stress is calculated.

[0388] The mechanical properties measured are as follows:

[0389] As can be seen from the table, while freeze drying the tissue with N2 gave fairly good results, the highest stresses were obtained by treating the tissue after it had been freeze dried without the use of N2.

[0390] When water is frozen at a very fast cooling rate (uncontrolled cooling rate), the water molecules within the tissue cells expand and destroy the collagen matrix of the tissue. Therefore, when nitrogen is used, the mechanical properties of the tested tissue are not as good as those without nitrogen. In contrast, a controlled cooling rate of the water molecules will cause less damage to the tissue and will therefore result in excellent mechanical properties.

[0391] 2. Bonding strength: a) In this section, the drying time in the freeze dryer was tested and the effect of drying time on the joint strength was measured:

[0392] Although there are no substantial differences, as can be seen from the table, the maximum stress is obtained after 4 days of evaporation.

[0393] b) Testing the adhesion strength of tissue-to-tissue connections (without piercing the tissue and allowing the polymer to pass through the tissue; i.e., using a polymer-based rivet): Since pericardial tissue has different properties on each surface, being smooth on one side and rough on the other, the mechanical properties of both sides were tested.

[0394]

[0395] As can be seen, the best results were achieved with rough side against rough side or tissue engraving. All break points were at the joint connections.

[0396] c) Testing the "rivets" In the following tests, the mechanical properties of the rivets were measured according to the following protocol: Measures tissue thickness areas of 250 microns + / - 50 microns 1. Laser cut tissue with a thickness of 250 microns and a size of 6 mm × 40 mm with a sufficient amount of holes at 1 mm from the edge of the tensile model.

[0397] 2. Measure the tensile model in the middle of the 6 mm width and measure the tensile model at the ends and along the middle 40 mm; 3. Next, the tissue sample is placed in a freeze dryer in a vacuum at a temperature range of -70 degrees Celsius to 0 degrees Celsius for a period of 7-24 hours; 4. Next, a dried tissue polymer membrane is prepared by attaching the dried tissues together and injecting polymer rivets into the prepared holes (or injecting polymer into the prepared holes); 5. The construct (tissue-polymer-tissue) is hot pressed at 60 degrees Celsius at 2 bar in a thermal bond. That is, the solvent (e.g., THF) is evaporated for 4 days. Its main purpose is to melt the polymer components and connect them together to provide a joint.

[0398] 6. Then, the tissue samples were placed in saline for 12 hours for recovery; 7. Next, measure the thickness of the dried tissue again; 8. Finally, the tissue is tensile tested to determine the force at which it breaks (tears), and the resulting stress is calculated.

[0399]

[0400] As seen above, poorer results are obtained when evaporation is used. Furthermore, the more holes there are in the tissue and the smaller the diameter of each hole, the better the results obtained.

[0401] d) Hole size and shape and different hole arrangements

[0402] e) Different polymers: In the table below a summary of the stress results measured for the different polymers used is given.

[0403]

[0404] As can be seen in the table above, the Chronoflex C 75D provided the best results.

[0405] f) Alignment of holes, offset test between pre-created holes in two tissues: In this test, the holes in the tissue are not aligned. They have some overlap, but they are not concentric.

[0406]

[0407] It should be noted that according to another embodiment of the present invention, tissue-polymer-tissue connection can be achieved by at least partially coating 2 tissues with a polymer / making 2 tissues physically contact with the polymer (no need to create a hole). According to this embodiment, each tissue is at least partially coated (or contacted with a polymer), and then thermal bonding occurs to join the two tissues together.

[0408] In the mechanical property test, two 6 mm * 6 mm tissue samples were coated with polymer (coating thickness was 0.25-0.26 mm).

[0409]

[0410] As seen in the table above, such tissue-tissue connection (engagement) was achieved to withstand a force of 4.5 MPa.

[0411] Thus, a construct according to one embodiment of the invention comprises: At least two decellularized tissues and at least one polymer component, wherein the polymer component at least partially coats at least one surface region of each of the two decellularized tissues. The bonding between the two decellularized tissues is achieved by thermally bonding the layers together.

[0412] As mentioned above, thermal energy is applied to raise the temperature of the polymer to 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. When two sheets or sections of polymer components are in close contact under these conditions, polymer chain entanglement will occur, resulting in a joint. According to aspects of the present invention, the joint does not need to be achieved on the entire surface of the sheet or section. The site joint at one or more regions of the polymer component can be sufficient to provide a robust association of more than one component or any two polymer sheets or sections.

[0413] According to another embodiment, a tissue-to-metal connection / joining is provided. According to this embodiment, a single attachment mechanism is provided. According to this single attachment mechanism, the tissue (which is in physical contact with the polymer) is in contact with the metal coated with the polymer to promote joining. In this embodiment, the construct is an assembly of (a) n numbers of decellularized tissues and m numbers of metal elements, at least one of the decellularized tissues being at least partially associated with at least one polymer, and at least one of the metal elements being at least partially coated with a polymer. n and m are integers greater than 0. According to one embodiment, m is equal to n; according to another embodiment, m is substantially different from n.

[0414] According to one embodiment, the association of the at least one decellularized tissue with the polymer refers to physical contact / interaction, coating, adhesion, bonding, robust anchoring and any combination thereof. Alternatively or additionally, the association refers to:

[0415] (i) embedding a polymer sheet or segment at least partially into a surface region of a layer of tissue, whereby the embedding may be through a single site, or through two or more sites, on the surface region of the layer with or without penetrating the tissue; or (ii) anchoring the polymer sheet or segment into tissue to a depth of tissue that allows for secure association, whereby the anchoring may be through a single anchoring site, or through two or more anchoring sites, whereby the depth of anchoring or penetration of the polymer sheet or segment may vary, excluding puncture of the tissue; or (iii) completely anchoring the polymer sheet or segment into the tissue to completely penetrate the tissue from one face of the tissue to the other face, wherein the anchoring can be through a single anchoring site, or through two or more anchoring sites. Typically, such penetration will involve surface features that are configured to extend from one face of the decellularized tissue to another face through at least one hole formed in the tissue. As will be described in detail below, such features can be formed in situ after the hole is formed in the tissue, or can be provided on the polymer sheet or segment in a form selected and configured to puncture or penetrate the tissue.

[0416] The polymer sheets or segments are said to be associated with the tissue in a manner that ensures association with the decellularized tissue. One type of association or interaction present in the constructs of the present invention is via anchoring or piercing the decellularized tissue surface as detailed herein, or by forming holes in the tissue through which two polymer sheets or segments can be associated. In some embodiments of the present invention, where the construct is made of more than one construct component, each component includes, for example, a decellularized tissue confined between two sheets or segments of a polymer component, the association of more than one component can be achieved by polymer-to-polymer bonding.

[0417] According to this embodiment, bonding between tissue (already associated with the polymer) and the polymer coated metal is achieved by associating (e.g., physically contacting) the tissue (already associated with the polymer) and the at least partially polymer coated metal element together to promote bonding.

[0418] According to another embodiment, at least one solvent (e.g., THF, DMF) is dripped into the contact area between the tissue (associated with the polymer) and the at least partially coated metal element. According to another embodiment, the solvent partially dissolves the coating polymer (both the polymer associated with the tissue and the polymer-coated metal at the junction area) and fuses them together to provide a connection / joining of the tissue and the metal.

[0419] According to another embodiment, the use of a solvent at least partially dissolves the uppermost surface of the metal-coating polymer and the uppermost surface of the polymer associated with the tissue; thus, the polymer chains at the uppermost surface can move sufficiently to entangle with chains in another sheet or segment (and thereby fuse the tissue and metal together). Such an effect is a reflow effect. Within the scope of the present invention, the term "reflow" refers to a process in which at least partial dissolution of the polymer occurs and a rearrangement of the polymer chains produces bonding.

[0420] According to another embodiment, the polymer is first dissolved in at least one solvent and then applied only to the covering of the tissue / metal element.

[0421] 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.

[0422] According to another embodiment, when applied to thermal heating, the use of a solvent at least partially melts / dissolves the uppermost surface of the coating polymer of the metal and the uppermost surface of the polymer associated with the tissue. Within the scope of the present invention, the term "thermal reflow" or "thermal bonding" refers to a process in which dissolution occurs, after which the entire assembly is subjected to controlled heating (which results in a molten state of at least a portion of the polymer) to produce a permanent casting (and bonding) thereof. The coating of the metal element can be provided by dipping and / or brushing and / or spraying.

[0423] Reference is now made to FIG. 34c which illustrates an encapsulation frame 40 with a polymer (either by dip coating or spray coating) and tissue 10 associated with the polymer at specific connection areas to the metal frame.

[0424] Thus, according to this embodiment, the metal frame is coated with a polymer and the tissue is coated with the same polymer at the connection / joining regions.

[0425] 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, a hernia mesh or hernia repair device, a pelvic floor reconstruction device, a wound or burn dressing, a dura mater closure, and a cardiac patch.

[0426] According to another embodiment, the tissue is freeze-dried prior to applying the polymer to the tissue.

[0427] It should be noted that although the above and below disclose the connection of tissue to metal, it can be applied to any other material (eg, polymer material) having substantially the same mechanical properties.

[0428] The following measurements provide a test of the mechanical strength of the polymer and the bonding of the polymer to the tissue / metal framework: 1. Strength of polymer thickness when bonded to each other on top of metal bracket:

[0429] As can be seen in the table, the best mechanical properties are obtained by Chronoflex 75A and Chronoflex 75A; and the combination of Chronoflex 75A and Chronoflex 75D.

[0430] Medical Device Example - A needle-free heart valve using tissue-to-tissue bonding and / or tissue-to-metal bonding as described above is provided as follows: According to this example, the heart valve consists of 4 components: A metal stent (or frame) 11 made of nitinol, cobalt-chromium alloy or stainless steel; typically, three leaflets 12, an inner skirt 13 and an outer skirt 14 (see FIG. 35 a), in addition, as disclosed above and below, the heart valve comprises a polymer-based component. As disclosed below, the polymer component is used in its liquid form or as a sheet or film.

[0431] According to one embodiment of the present invention, the whole tissue portion is first laser cut and holes are created on the leaflets, inner skirt and outer skirt, which are made in 2D / 3D laser cutting (see Figure 36a).

[0432] Next, the polymer is dissolved in at least one solvent (eg, THF) or taken out as is, cast as a thin film (10-100 microns thick), and then laser cut to produce polymer sheets and rivets (for future connection of different parts of the tissue).

[0433] According to one embodiment, a polymer sheet is cast and laser cut into the shape of a 2D projection of the assembled heart valve (see FIG. 35b ).

[0434] According to one embodiment, the polymer sheet is punctured, and according to another embodiment, the polymer sheet is kept less punctured.

[0435] Next, freeze drying (lyophilization) of the tissue occurred at a temperature of -54°C, under a vacuum pressure reduced from 110 KPa to 2 Pa-3 Pa overnight.

[0436] Then, the coating of the stent (frame) is started. Chronoflex in tetrahydrofuran THF is used to provide at least one coating layer on the stent, preferably more than 10 coating layers are set on the frame. Preferably, the metal stent (frame) is coated by a method selected from brushing, spraying or dipping the metal stent (frame). According to one embodiment of the present invention, between at least one coating layer and another coating layer, heating is applied to dry the coating layer.

[0437] Next, the different tissue components are joined to each other by joining the polymer sheets to the tissues on both sides. The joining is achieved by injecting a Chronoflex solution dissolved in THF through the holes in the tissue. The Chronoflex solution dissolved in THF causes the tissues to adhere to each other. To further strengthen the adhesion, thermal bonding heating is applied. For all joined parts, thermal bonding is performed at a heating temperature of at least 6 bar and 80°C to strengthen and reinforce the joint.

[0438] It should be noted that at all times, all holes of, for example, the leaflets and skirt must be substantially aligned to result in improved coaptation.

[0439] Thermal bonding can be used to strengthen and reinforce the joint.

[0440] Next, after anastomosis, the assembly may be placed in saline for tissue recovery for at least 14 hours.

[0441] Next, the tissue-to-metal process begins to provide the final heart valve. It should be noted that while tissue-to-tissue bonding is performed while the tissue is dry, tissue-to-metal bonding is achieved while the tissue is wet (after recovery in saline).

[0442] In the tissue-to-metal bonding process, the tissue-polymer-tissue assembly is placed in a stent / frame, and the bonding of the tissue-polymer-tissue and the polymer-coated stent is obtained again by using a polymer. That is, the polymer is used to coat the tissue-polymer-tissue assembly and the relevant area of ​​the polymer-coated stent. Next, thermal bonding is applied. As mentioned above, when the polymer chains at the surface of a sheet or section are movable enough to entangle with the chains in another sheet or section, the bonding of the polymer sheet occurs. In order to achieve bonding, thermal energy can be applied to increase the temperature. When two sheets or sections of the polymer component are in close contact under these conditions, polymer chain entanglement will occur, resulting in a joint. According to aspects of the present invention, bonding does not need to be realized on the entire surface of the sheet or section. The site bonding at one or more regions of the polymer component can be sufficient to provide more than one component or the robust association of any two polymer sheets or sections.

[0443] It should be noted that engagement may be achieved in multiple placements along the perimeter of the stent.

[0444] According to another embodiment, the coated metal (i.e. polymer-metal-polymer) is encapsulated in the assembly of tissue-polymer-tissue; That is, a construct of tissue-polymer-tissue-polymer-metal-polymer-tissue-polymer-tissue is provided (see Figure 36 b-Figure 36 c). As seen in Figure 36 c, a part of the metal framework is encapsulated in the tissue-polymer-tissue assembly.

[0445] It should be noted that according to one embodiment, at least one type of polymer component may be used. Alternatively, two different polymers are used, each having a different viscosity.

[0446] The fully articulated sutureless valve assembly was tested for hemodynamics and durability. Fig.37 , Fig.37 The figure shows the hemodynamic behavior of a valve according to the invention.

[0447] In the test, 2 main parameters were presented: EOA - Effective Orifice Area - this provides a number for how well the valve is open and allows blood to flow through it (the higher the number, the better it functions); and RF - Regurgitant Fraction - This provides a % of how much blood leaks out of the valve in each pulse (ie, how much the valve is leaking).

[0448] As can be seen in the figure, the valve according to the present invention achieves 130 M cycles beyond the required 200 M cycles.

[0449] The terms "comprises," "comprising," "includes," "including," "having" and their cognates mean "including but not limited to."

[0450] The term "consisting of" means "including and limited to."

[0451] The term "consisting essentially of" means that the composition, method or structure may include additional ingredients, steps and / or parts, but only if the additional ingredients, steps and / or parts do not materially change the basic and novel characteristics of the claimed composition, method or structure.

[0452] As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, the term "a compound" or "at least one compound" includes more than one compound, including mixtures thereof.

[0453] Throughout this application, various embodiments of the present invention can be presented in range format.It should be understood that the description of range format is only for convenience and brevity, and should not be interpreted as a rigid limitation to the scope of the present invention.Therefore, the description of the scope should be considered to have specifically disclosed all possible sub-ranges and the individual numerical values ​​within the scope.For example, the description of the scope such as from 1 to 6 should be considered to have specifically disclosed sub-ranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc. and the individual numerals within the scope such as 1, 2, 3, 4, 5 and 6.No matter how the width of the scope, this applies.

[0454] Whenever a numerical range is indicated herein, it is intended to include any cited numerical value (fractional or integer) within the indicated range. The phrases "within the range of / between the first indicated number and the second indicated number" and "within the range from / between the first indicated number" to the second indicated number" are used interchangeably herein and are intended to include the first indicated number and the second indicated number and all fractional and integer numerical values ​​therebetween. As used herein, the term "method" refers to ways, means, techniques, and procedures for accomplishing a given task, including but not limited to those ways, means, techniques, and procedures known to or that have been developed from known ways, means, techniques, and procedures by practitioners in the fields of chemistry, pharmacology, biology, biochemistry, and medicine. As used herein, the term "treatment" includes abolishing, substantially inhibiting, slowing down, or reversing the development of a condition; substantially improving the clinical symptoms or aesthetic symptoms of a condition or substantially preventing the appearance of the clinical symptoms or aesthetic symptoms of a condition.

[0455] It should be understood that certain features of the invention described in the context of a separate embodiment for clarity are also provided in combination in a single embodiment. Conversely, various features of the invention described in the context of a single embodiment for brevity are also provided individually or in any suitable sub-combination or appropriately in any other described embodiment of the invention. Certain features described in the context of various embodiments are not considered to be essential features of those embodiments unless the embodiments do not work without those elements. Various embodiments and aspects of the invention as described above and as claimed in the claims section below find experimental support in the following examples.

Claims

1. A construct comprising at least one decellularized tissue, at least one element, and at least one polymer component; the at least one polymer component being in physical contact with at least a portion of at least one surface area of ​​the at least one decellularized tissue; wherein at least one polymer component at least partially coats at least one surface region of the element; Furthermore, the at least one surface region of the decellularized tissue that is in physical contact with the polymer component is in contact with the at least one surface region of the element that is coated with the polymer component.

2. A decellularized tissue, the decellularized tissue being physically associated with at least one element via at least one polymer component, the association comprising or consisting of: the polymer component being in at least partial physical contact with at least a portion of a surface area of ​​the at least one decellularized tissue; and at least a portion of the at least one element is coated by the at least one polymer component; wherein at least one surface region of the decellularized tissue associated with the polymer component is in contact with at least one surface region of the element coated by the polymer component.

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 is at least partially associated with at least one face of the decellularized tissue and at least partially associated with at least one surface region of the element; wherein the at least one surface region of the decellularized tissue in physical contact with the polymer component is in contact with the at least one surface region of the element coated by the polymer component.

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 metallic element, a polymeric material, any material having mechanical properties substantially different from the at least one decellularized tissue, and any combination thereof.

5. The construct of claim 4, wherein the at least one decellularized tissue at least partially in contact with the at least one polymer component is thermally bonded to provide the association.

6. A construct according to claim 5, wherein the thermal bonding is achieved by raising the temperature of the polymer components to above (a) the transition temperature of the polymer components for amorphous thermoplastic polymers; or (b) the melting temperature of the polymer components for semi-crystalline polymers to at least partially cause entanglement of polymer chains along at least one region of the polymer components.

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

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

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

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

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

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

13. The construct of any one of claims 1 to 3, comprising two or more components having decellularized tissue confined between two sheets or sections of a polymer component, wherein the components are associated with each other.

14. A construct according to claim 13, wherein at least two components are oppositely oriented.

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

16. The construct of any one of claims 1 to 15, further comprising at least one solvent; wherein the solvent is added to an area of ​​the decellularized tissue in physical contact with the polymer component and in contact with the at least one surface area of ​​the element coated with the polymer component.

17. A 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 coating at least one surface area of ​​the element.

18. A construct according to claim 17, wherein the construct is obtained by a backflow effect in which (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 coating at least one surface area of ​​the element are entangled along at least one regional chain of the polymer component.

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

20. The construct of any one of claims 16-19, 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. according to the construct described in any one in claim 1 to 20, wherein said polymer component is in the form of a granular layer or a granular coating, a polymer sheet, a polymer film, a polymer fiber or a polymer net, a gel, a hydrogel or a liquid film or a fluid film and any combination thereof.

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

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

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

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

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

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

28. A construct according to any one of claims 1 to 23, wherein the polymer component is or comprises an organosilicon polymer.

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

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

31. according to the construct of any one of claims 1 to 23, wherein the polymer component is a polymer selected from the following items or comprises a polymer selected from the following items: 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 alkyl (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 terephthalate / butylene terephthalate copolymers and combinations and copolymers thereof.

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

33. A construct according to any one of claims 1 to 23, wherein the polymer component is or comprises a polyether urethane selected from Pellethane, Elastane, Elastolan, Tecoflex, Biomer.

34. A construct according to any one of claims 1 to 23, wherein the polymer component is or comprises a polycarbonate urethane selected from the group consisting of Chronoflex, Biospan and Bionate.

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

36. A construct according to any one of claims 1 to 23, wherein the polymer component is or comprises Chronoflex, Tecoflex and any combination thereof.

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

38. The construct of claim 37, wherein the active material is selected from analgesics; antianxiety drugs; antiarrhythmics; antibacterials; antibiotics; anticoagulants and thrombolytics; anticonvulsants; antidepressants; antidiarrheals; antiemetics; antifungals; antihistamines; antihypertensives; anti-inflammatory drugs; antineoplastics; antipsychotics; antipyretics; antivirals; beta-blockers; corticosteroids; cytotoxins; hormones and sex hormones; enzymes; and vitamins.

39. The construct of claim 4, wherein the metal element is selected from the group consisting of a stent, a metal stent, a vascular graft, a heart valve, a membrane, a sealing device, a suture or staple, a hernia mesh or hernia repair device, a pelvic floor reconstruction device, a wound or burn dressing, a dura mater closure, and a cardiac patch.

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

41. A construct according to any one of claims 1 to 40, wherein at least one second decellularized tissue, at least a portion of at least one surface area of ​​the at least one second decellularized tissue is (a) at least partially coated with the at least one polymer component, and (b) is in physical contact with the at least one polymer component that is in physical contact with the at least one decellularized tissue, such that application of thermal bonding connects the at least one decellularized tissue and the at least one second decellularized tissue.

42. A construct according to any one of claims 1 to 41, wherein the polymer component that is in physical contact with at least a portion of at least one surface area of ​​the decellularized tissue (a) at least partially penetrates at least one surface area of ​​a second decellularized tissue; and / or (b) has at least one surface feature that extends out of one surface of the second decellularized tissue, and the surface feature passes through one surface of the second decellularized tissue to another surface through at least one hole formed in the tissue.

43. A construct according to claims 41-42 in the form of a multi-sheet construct.

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

45. The construct of claim 43, wherein the multi-sheet construct comprises a number of sheets of the second decellularized tissue and an equal number of sheets of the polymer component.

46. ​​The construct of any one of claims 41-42, wherein the at least one second decellularized tissue is dried prior to being coated with the polymer.

47. The construct of any one of claims 46, wherein the drying is freeze drying of the at least one decellularized tissue.

48. The construct of claim 49, wherein the polymer component is in the form of a particle layer or coating, a polymer sheet, a polymer film, a polymer fiber or a polymer net, a gel, a hydrogel, or a liquid film or a fluid film, and any combination thereof.

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

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

51. A kit comprising a construct according to any one of claims 1 to 50.

52. The apparatus of claim 51 configured as an implant.

53. The device of claim 51 or 52, wherein the device is selected from a stent, a metal stent, a vascular graft, a heart valve, a membrane, a sealing device, a suture or staple, a hernia mesh or hernia repair device, a pelvic floor reconstruction device, a wound or burn dressing, a dura mater closure, and a cardiac patch.

54. A process for producing a construct according to any one of claims 1 to 4, comprising contacting at least one surface region 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; Contacting at least one surface region of the at least one metal element having the at least one polymer with at least one surface region of the at least one decellularized tissue having the polymer; thereby forming the construct.

55. The process of claim 54, wherein the polymer coats at least one surface region of the at least one metal element; and / or coats at least one surface region of the at least one decellularized tissue.

56. The process of claim 55, wherein the polymer is cured.

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

58. A process 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 coating at least one surface area of ​​the element.

59. A process according to claim 58, wherein the construct is obtained by a reflow effect in which (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 coating at least one surface area of ​​the element are entangled along at least one regional chain of the polymer component.

60. The process of 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. A process 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. The process of claim 54, further comprising a. contacting the pierced surface area of ​​at least one decellularized tissue with at least one polymer, and b. allowing the at least one polymer to penetrate into the perforations (holes).

63. The process of claim 54, further comprising a. contacting at least one surface region of the decellularized tissue with at least one polymer, and b. allowing the at least one polymer to at least partially encapsulate the at least one decellularized tissue.

64. The process according to claim 60 further comprises at least one step selected from the following: (a) puncturing or forming holes in a surface area of ​​at least one decellularized tissue; (b) inserting the polymer into the perforations (holes); (c) solidifying the polymer; (d) allowing the at least one polymer to penetrate into the perforations (holes) and form a polymer sheet on the surface area; (e) any combination thereof.

65. The process of claim 55, further comprising at least one step selected from: (a) thermally bonding the at least one decellularized tissue and the at least one polymer by raising the temperature of the polymer component to above (i) the transition temperature of the polymer component for an amorphous thermoplastic polymer, or (ii) the melting temperature of the polymer component for a semi-crystalline polymer, so as to at least partially cause entanglement of polymer chains along at least one region of the polymer component to produce the construct; (b) selecting the polymer to be in the form of a particle layer or particle coating, a polymer sheet, a polymer film, a polymer fiber or a polymer mesh, a gel, a hydrogel, or as a liquid film or a fluid film, and any combination thereof; (c) any combination thereof.

66. A process according to claim 55, wherein at least one of the following is true: (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 area to form a construct component; and (d) any combination thereof.

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