RU2795561C1 - Tissue and organ substitutes and methods for their manufacture - Google Patents

Abstract

FIELD: biotechnology.

SUBSTANCE: biological tissue or organ substitutes for transplants and implants, as well as to methods for their production. The composition of the tissue or organ substitute contains a tissue engineering construct containing one or more bioink compositions, where one or more bioink compositions contain from 5 to 200 mg/ml of collagen; as well as a biocompatible support structure containing one or more external supports, internal supports or a combination thereof from a biocompatible material; wherein the composition of the tissue or organ substitute has a three-dimensional 3D shape, where the tissue-engineering construct has the shape of an organ or tissue; and the biomaterial is present in an amount of 1 to 100 wt.% of the biocompatible support structure. A method for manufacturing a tissue or organ substitute composition includes: manufacturing a biocompatible support structure; making a tissue engineering construct comprising applying bio-colors to or around a biocompatible support structure, wherein one or more bioink compositions contain 5 to 200 mg/mL of collagen; crosslinking the bioinks and optionally repeating the application and crosslinking steps to form a tissue engineering construct; and curing the tissue or organ substitute composition; wherein the composition of the substitute has a 3D shape, where the tissue engineering structure has the shape of an organ or tissue; the biocompatible support structure comprises one or more external supports, internal supports, or combinations thereof of a biocompatible material; and the biocompatible material is present in an amount of 1 to 100 wt.% of the biocompatible support structure.

EFFECT: possibility to obtain tissue or organ substitute compositions.

57 cl, 13 dwg

Description

Cross-reference to related applications

This application claims priority over U.S. Provisional Application No. 62/826,745, filed March 29, 2019, the disclosure of which is incorporated herein by reference in its entirety.

Technical field

This technology generally refers to biological tissue or organ substitutes for transplants and implants, as well as methods for their production. More specifically, this technology relates to tissue and organ substitutes having a biocompatible support structure and tissue-engineered design, as well as methods for their manufacture.

Brief summary of the invention

In one aspect, this technology provides the use of a tissue or organ substitute that includes a tissue engineered construct containing one or more biostain compositions and a biocompatible support structure that includes one or more external supports, internal supports, or combinations thereof of a biocompatible material, wherein said the composition has a three-dimensional 3D shape, and the biocompatible material is present in an amount of from about 1% to about 100% by weight of the biocompatible support structure.

In a related aspect, this technology provides a method of making a composition for a tissue or organ substitute, which includes making a biocompatible support structure, as described herein, in any embodiment; making a tissue engineered construct as described herein, in any embodiment, comprising applying one or more biostain compositions, as described herein, in any embodiment, to or around a biocompatible support structure; crosslinking one or more bioink compositions and optionally repeating the application and crosslinking steps to form a tissue engineered construct; and curing the tissue or organ substitute composition; wherein the biocompatible support structure includes one or more of external supports, internal supports, or combinations thereof of a biocompatible material, and wherein the biocompatible material is present in an amount of from about 1% to about 100% by weight of the biocompatible support structure.

Brief description of graphic materials

Fig. 1A shows a perspective view of a tissue/organ substitute composition according to an embodiment of this technology that includes a biocompatible support structure, wherein the biocompatible support structure has an external support partially covering the outer two-dimensional (2D) surfaces of the tissue engineered construct and an internal support partially encapsulated by the tissue engineered structure. design.

Fig. 1B shows a perspective view of a biocompatible support structure according to an embodiment of this technology having an internal support coupled to one or more 2D surfaces of the external structure.

Fig. 1C shows a perspective view of an outer support having one or more holes extending through it from one 2D surface to an opposite parallel 2D surface.

Fig. 1D shows a perspective view of an internal support according to an embodiment of this technology having one or more holes extending through them from one 2D surface to an opposite parallel 2D surface of the internal support, and also having a cavity.

Fig. 2A shows a 2D sheet having one or more holes in accordance with an embodiment of this technology.

Fig. 2B shows a dissolvable mold for forming the inner support of an ear-shaped tissue engineered construct in accordance with an embodiment of the present technology.

Fig. 2C shows a 2D porous sheet thermoformed onto an internal support mold in accordance with an embodiment of this technology.

Fig. 3A shows an internal view of a mold configured and contoured for injection molding an ear-shaped tissue engineered structure in accordance with an embodiment of the present technology.

Fig. 3B shows the appearance of a mold configured and contoured for injection molding an ear-shaped tissue engineered structure in accordance with an embodiment of this technology.

As discussed below, FIG. 4A-4D show block diagrams depicting exemplary methods for fabricating a biocompatible support structure and a tissue engineered construct for a tissue or organ substitute composition.

Fig. 4A shows a general exemplary method for making a tissue or organ substitute composition in accordance with an embodiment of the present technology.

Fig. 4B shows a typical manufacturing process for biocompatible support structures (external supports and/or internal supports).

Fig. 4C shows an exemplary fabrication process for a tissue engineered construct in accordance with at least one embodiment.

Fig. 4D shows a flowchart of an exemplary method for curing a structure in accordance with at least one embodiment.

Detailed description of the invention

This document describes various embodiments. It should be noted that specific embodiments are not intended to exhaustively describe or limit the broader aspects discussed herein. One aspect described in connection with a particular embodiment is not necessarily limited to that embodiment and may be practiced with any other embodiment(s).

As used herein, the term "about" will be understood by those skilled in the art and will vary to some extent depending on the context in which it is used. If there are uses of a term that are not clear to those skilled in the art, given the context in which it is used, "about" will mean up to plus or minus 10% of the particular term.

The use of terms in the singular and similar references to the singular in the context of the description of the claimed invention (especially in the context of the claims) should be construed as embracing the terms in both the singular and the plural, unless otherwise specified in this document or clearly contrary to the context. The listing of ranges of values in this document is merely intended to serve as a concise way of individually referring to each individual value falling within that range, unless otherwise noted herein, and each individual value is included in the description as if it were separately stated. in this document. All of the methods described herein may be performed in any suitable order, unless otherwise indicated herein or otherwise clearly contradicts the context. The use of any and all examples or exemplary language (eg, "such as") provided herein is only intended to better illustrate embodiments of the invention and is not intended to limit the scope of the claims unless otherwise stated. No wording in the specification should be construed as indicating any non-declared element as essential.

The term "biocompatible material" refers to a material derived from a natural or synthetic source that has the ability to perform its desired function in the subject's body without causing any undesirable local or systemic effects. As used herein, the term "biocompatible materials" may also refer to materials that are biodegradable, bioabsorbable, bioresorbable, or a combination of two or more of these under physiological conditions. The term "biodegradable" refers to a material that can be broken down into basic substances by normal environmental processes and/or by the action of living beings such as microorganisms. The term "bioabsorbable" refers to a material capable of being absorbed into living tissue. The term "bioresorbable" refers to a material that, when placed in the human body, begins to dissolve (resorb) and slowly be replaced by growing tissue. In the context of this document, in any embodiment, the terms "biodegradable", "bioabsorbable" and "bioresorbable" are used interchangeably.

As used herein, a "subject" or "patient" is a mammal as described herein. The terms "subject" and "patient" may be used interchangeably. As used herein, the term "mammal" includes, but is not limited to, a cat, dog, rodent, or primate. For example, in any embodiment of the present invention, the mammal is a human.

The term "hydrogel" or "gel" refers to the substance formed when an organic polymer (natural or synthetic) solidifies or solidifies to create an open lattice three-dimensional structure that traps water or other solution molecules to form a gel. Curing may occur, for example, by aggregation, coagulation, hydrophobic interactions or crosslinking. Hydrogels can be cellular (ie containing cells) or acellular (ie without cells). Hydrogels containing cells can solidify rapidly, causing the cells to remain uniformly suspended in the mold (or around or within another solidified gel) until the gel has solidified. Hydrogels can also be biocompatible, eg non-toxic to the cells suspended in the hydrogel.

In the context of this document, the term "2D thickness" refers to the thickness of the cross-section of an external support or an internal support or support element (for example, a truss element or a support like a truss) from one surface to the opposite parallel surface of the supports or support elements.

In the context of this document, the term "collagen" refers to a major connective tissue protein that has high tensile strength and is found in most multicellular organisms. Collagen is the main fibrous protein and also the non-fibrillar basement membrane protein. It contains large amounts of glycine, proline, hydroxyproline and hydroxylysine. Collagen is found throughout the body and comes in at least 12 types (type I-XII).

Tissue/organ substitutes according to this technology

In one aspect, this technology provides the use of a tissue or organ substitute that includes a tissue engineered construct containing one or more biostain compositions and a biocompatible support structure that includes one or more external supports, internal supports, or combinations thereof of a biocompatible material, wherein said the composition has a three-dimensional 3D shape, and the biocompatible material is present in an amount of from about 1% to about 100% by weight of the biocompatible support structure.

Fig. 1A shows a perspective view of a tissue/organ substitute composition 100 according to an embodiment of this technology that includes a biocompatible support structure 110, wherein the biocompatible support structure has an external support 120 partially covering the outer two-dimensional 2D surfaces of the tissue engineered structure 140 and an internal support. 130 partially encapsulated with tissue engineered construct 140.

Fig. 1B shows a perspective view of a biocompatible support structure 110 in accordance with an embodiment of the present technology having an internal support 140 coupled to one or more 2D surfaces of the external structure 120. FIG. 1C shows a perspective view of an outer support 120 having one or more holes 150 extending through from one 2D surface to an opposite parallel 2D surface. Fig. 1D shows a perspective view of an internal support 130 in accordance with an embodiment of the present technology, having one or more holes 160 extending through from one 2D surface to the opposite parallel 2D surface of the internal support, and having cavities 170.

Tissue engineering design

The tissue/organ substitute of this technology includes a tissue engineered construct that includes one or more biopaint compositions. In any embodiment of the present invention, the tissue engineered construct may be of any suitable shape or size. For example, in any embodiment of the present invention, the tissue engineered construct may have a customized 3D structure of various shapes and sizes for a range of clinical applications, such as facial or craniofacial applications, artificial cartilage, or combinations thereof. In any embodiment of the present invention, the tissue engineered construct may be in the form of an organ or tissue of the subject. For example, in any embodiment of the present invention, the organ or tissue is selected from the ear, nose, meniscus, articular cartilage, or heart valve. In one embodiment of the invention, said organ is the ear. Fig. 1A shows an embodiment of a human ear-shaped tissue engineered structure 140 in which the auricle is partially covered by an outer support 120 in accordance with an embodiment of the present technology, and "ear-shaped" tissue engineered portions partially encapsulate the inner support 130.

In any embodiment of the present invention, the tissue engineering system may have a 3D structure based on a 3D computer model. For example, in any embodiment of the present invention, the computer model may include a 3D contralateral organ or tissue of the subject. In any embodiment of the invention, the subject is a mammal (eg, a cat, dog, rodent, or primate). In any embodiment of the invention, the subject is a human. A 3D computer model can be generated using 3D imaging tools. Suitable 3D imaging tools include, but are not limited to, clinical computed tomography (CT), magnetic resonance imaging (MRI), laser scanning, 3D cameras, or combinations of any two or more of these.

The tissue engineered structure may be configured such that the size of said tissue engineered structure is larger than the size of the biocompatible support structure, with the tissue engineered structure partially or completely in contact with the biocompatible support structure. For example, in any embodiment of the present invention, the tissue engineered construct may be configured such that the internal supports of the biocompatible support structure are wholly or partially encapsulated within said tissue engineered construct.

In any embodiment of the present invention, the tissue engineered construct may be configured such that said tissue engineered structure is smaller than the biocompatible support structure, wherein said tissue engineered construct is wholly or partially covered and/or enclosed within the outer supports of the biocompatible support structure. For example, in any embodiment of the present invention, one or more of the 2D external surfaces of the tissue engineered construct may be covered and/or enclosed by the outer support of a biocompatible support structure. In any embodiment of the present invention, the tissue engineered construct may be sized beyond the capabilities of the outer support of the biocompatible support structure such that the tissue construct protrudes (or protrudes) beyond the outer 2D surface of the outer support. For example, in any embodiment of the present invention, the tissue engineered construct may protrude (or protrude) into one or more of the holes in the outer support of the biocompatible support structure.

In any embodiment of the present invention, the tissue engineered construct may include a unique set of mechanical properties to enable it to function properly. For example, in any embodiment of the present invention, the tissue engineered construct may have one or more of the following characteristics: an equilibrium modulus of about 2 kPa to about 2 MPa, an instantaneous modulus of about 20 kPa to about 20 MPa, and a hydraulic permeability of about 1x10 ^-16 m ² /Pa s to about 1x10 ^-8 m ² /Pa s. In any embodiment of the invention, the tissue engineered construct may have an equilibrium modulus of about 2 kPa to about 2 MPa, about 2 kPa to about 1.5 MPa, about 2 kPa to about 1 MPa, about 2 kPa to about 800 kPa, from about 2 kPa to about 600 kPa, from about 2 kPa to about 400 kPa, or any range including and/or within any two of the previous values. For example, in any embodiment of the present invention, the equilibrium modulus may be 2 kPa, about 10 kPa, about 25 kPa, about 50 kPa, about 100 kPa, about 200 kPa, about 300 kPa, about 400 kPa, about 500 kPa, about 600 kPa , about 700 kPa, about 800 kPa, about 900 kPa, about 1 MPa, about 1.1 MPa, about 1.2 MPa, about 1.3 MPa, about 1.4 MPa, about 1.5 MPa, about 1, 6 MPa, about 1.7 MPa, about 1.8 MPa, about 1.9 MPa, about 2 MPa, or any range including and/or within any two of the previous values.

In any embodiment of the present invention, the tissue engineered construct may have an instantaneous modulus of about 20 kPa to about 20 MPa, 50 kPa to about 15 MPa, about 100 kPa to about 10 MPa, about 500 kPa to about 8 MPa, about 1 MPa up to about 5 MPa, or in any range including and/or within any two of the previous values. For example, in any embodiment of the present invention, the tissue engineered construct may have an instantaneous modulus of about 20 kPa, about 40 kPa, about 60 kPa, about 80 kPa, about 100 kPa, about 200 kPa, about 300 kPa, about 400 kPa, about 500 kPa, about 600 kPa, about 700 kPa, about 800 kPa, about 900 kPa, about 1 MPa, about 2 MPa, about 3 MPa, about 4 MPa, about 5 MPa, about 6 MPa, about 7 MPa, about 8 MPa, about 9 MPa, approx. 10 MPa, approx. 11 MPa, approx. 12 MPa, approx. 13 MPa, approx. 14 MPa, approx. 15 MPa, approx. 16 MPa, approx. 17 MPa, approx. 18 MPa, approx. including any two of the previous values, and/or within them. In any embodiment of the present invention, the tissue engineered construct may have a hydraulic permeability from about 11 x 10 ^-16 m ² /PA s to about 1 x 10 ^-8 m ² /PA s, from about 1 x 10 ^-13 m ² /PA s to about 9 x 10 ^-10 m ² /PA s, from about 1 x 10 ^-12 m ² /PA s to about 6 x 10 ^-10 , from about 1 x 10 ^-11 m ² /PA s up to about 3 x 10 ^-10 m ² /Pa·s or in any range including and/or within any two of the previous values.

The tissue engineered construct may include one or more biopaint compositions. The term "bio-paint" or "bio-paint composition" refers to a paint derived from a biomaterial. For example, in any embodiment of the present invention, biopaints may include, but are not limited to, a hydrogel (e.g., alginate hydrogel), hyaluronic acid, elastin, laminin, agarose, collagen, chitosan, fibrin, hyaluronic acid, carrageenan, polyethylene oxide, polypropylene oxide, polyethylene oxide copolymer, and polypropylene oxide, hydroxypropylmethylcellulose, propylene fumarate-ethylene glycol copolymer, poly(ethylene glycol)-co-poly(lactic acid), poly(vinyl alcohol), KDL12 oligopeptides, poly(n-isopropylacrylamide), or combinations of two or more of them. The bioink can have a controlled crosslinking rate by adjusting environmental variables including but not limited to temperature, pH, ionic strength, heat, UV light, or the addition of chemical crosslinkers such as calcium, magnesium, barium, chondroitin, sulfate, carbodiimides, ribose, riboflavin and thrombin. Suitable hydrogels for use in biopaints for tissue engineering are described in US Pat. No. 9,044,335 entitled "COMPOSITE TISSUE-ENGINEERED INTERVERTEBRAL DISC WITH SELF-ASSEMBLED ANNULAR ALIGNMENT" filed May 5, 2010, the entire contents of which are incorporated herein by reference.

In any embodiment of the present invention, one or more tissue engineered biopaint compositions may include an alginate hydrogel. For example, in any embodiment of the present invention, the alginate hydrogel may be present in an amount of from about 0.5% (w/v) to about 10% (w/v). For example, in any embodiment of the present invention, the alginate hydrogel may be present in an amount of about 0.5% (w/v), about 0.6% (w/v), about 0.7% (w/v .), about 0.8% (w/v), about 0.9% (w/v), about 1.0% (w/v), about 1.5% (w/v). /v), about 2.0% (w/v), about 2.5% (w/v), about 3.0% (w/v), about 3.5% ( w/v), about 4.0% (w/v), about 4.5% (w/v), about 5.0% (w/v), about 5.0 % (w/v), about 5.5% (w/v), about 6.0% (w/v), about 6.5% (w/v), about 7 .0% (w/v), about 7.5% (w/v), about 8.0% (w/v), about 8.5% (w/v), about 9.0% (w/v), about 9.5% (w/v), about 10.0% (w/v), or any range including any two of the previous values, and/or within them.

In any embodiment of the present invention, one or more tissue engineered biostain compositions may include collagen. Collagen-based materials are preferred for use in tissue engineering structures. Methods for harvesting collagen for use in biogel compositions, methods for generating 3D structures using biogel compositions, and methods for preparing biogel compositions for use in 3D printing systems are described in PCT application serial number PCT/US2017/034582 titled "3D PRINTABLE BIO GEL AND METHOD OF USE, filed May 25, 2017, with Patent Registry Number 113066-0104, the entire content of which is incorporated herein by reference to reference information and methods set forth therein.

In any embodiment of the present invention, the one or more tissue engineered biostain compositions may include collagen in an amount greater than about 5 mg/mL. Suitable amounts of collagen in biopaints may include, but are not limited to, from about 5 mg/mL to about 200 mg/mL. For example, in any embodiment of the present invention, collagen may be present in an amount of about 5 mg/ml, about 10 mg/ml, about 15 mg/ml, about 20 mg/ml, about 25 mg/ml, about 30 mg/ml, about 35 mg/ml, about 40 mg/ml, about 45 mg/ml, about 50 mg/ml, about 55 mg/ml, about 60 mg/ml, about 65 mg/ml, about 70 mg/ml, about 75 mg /ml, about 80 mg/ml, about 85 mg/ml, about 90 mg/ml, about 95 mg/ml, about 100 mg/ml, about 105 mg/ml, about 110 mg/ml, about 115 mg/ml , about 120 mg/ml, about 125 mg/ml, about 130 mg/ml, about 135 mg/ml, about 140 mg/ml, about 145 mg/ml, about 150 mg/ml, about 155 mg/ml, about 160 mg/ml, about 165 mg/ml, about 170 mg/ml, about 175 mg/ml, about 180 mg/ml, about 185 mg/ml, about 190 mg/ml, about 195 mg/ml, about 200 mg /ml or in any range, including any two of the previous values, and/or within them.

In any embodiment of the present invention, the tissue engineered construct may include one or more biostain compositions containing type I collagen, type III collagen, type IV collagen, other fibrillar collagens, or a combination thereof.

In any embodiment of the present invention, one or more tissue engineered biopaint compositions may further comprise a neutralizer and a cell medium as described herein, in any embodiment. Suitable neutralizers may include, but are not limited to, formulations containing, for example, a weak acid. In any embodiment of the invention, said tissue engineered construct may be cellular or acellular. For example, in any embodiment of the present invention, the tissue-engineered biopaints may include a cell medium containing a population of cells, which may include, but is not limited to, living cells obtained from and/or isolated from a subject's tissue. The cell environment may further include other living cells; for example, in any embodiment of the present invention, biopaints may include epidermal cells, chondrocytes and other cells that form mesenchymal stem cells, adipose tissue-derived stem cells, cartilage, macrophages, adipocytes, dermal cells, muscle cells, hair follicles, fibroblasts, organ cells, osteoblasts, osteocytes and other cells that form bones, endothelial cells, mucosal cells, pleural cells, ear canal cells, tympanic membrane cells, peritoneal cells, Schwann cells, corneal epithelial cells, gingival cells, neuronal stem cells of the central nervous system systems, tracheal epithelial cells, or combinations of two or more of these.

In any embodiment of the present invention, the cell population may be present in an amount from about 1.0 x 10 ⁵ cells/ml to about 5.0 x 10 ⁷ cells/ml. For example, in any embodiment of the present invention, the number of cells present in the tissue engineered biostains may include about 1.0 x 10 ⁵ cells/ml, about 2.0 x 10 ⁵ cells/ml, about 3.0 x 10 ⁵ cells/ml. ml, approx. 4.0 x 10 ⁵ cells/ml, approx. 5.0 x 10 ⁵ cells/ml, approx. 6.0 x 10 ⁵ cells/ml, approx. 7.0 x 10 ⁵ cells/ml, approx. 8.0 x 10 ⁵ cells/ml, approx. 9.0 x 10 ⁵ cells/ml, approx. 1.0 x 10 ⁶ cells/ml, approx. 2.0 x 10 ⁶ cells/ml, approx. 3.0 x 10 ⁶ cells/ml , about 4.0 x ¹⁰⁶ cells/ml, about 5.0 x ¹⁰⁶ cells/ml, about 6.0 x ¹⁰⁶ cells/ml, about 7.0 x ¹⁰⁶ cells/ml, about 8.0 x 10 ⁶ cells/ml, about 9.0 x 10 ⁶ cells/ml, about 1.0 x 10 ⁷ cells/ml, about 2.0 x 10 ⁷ cells/ml, about 3.0 x 10 ⁷ cells/ml, about 4.0 x 10 ⁷ cells/ml, about 5.0 x 10 ⁷ cells/ml, or any range including and/or within any two of the previous values. Suitable numbers of cells present in tissue engineered biostains include, but are not limited to, from about 1.0 x 10 ⁵ cells/mL to about 5.0 x 10 ⁷ cells/mL, from about 1.0 x 10 ⁵ cells/mL to about 1.0 x 10 ⁷ cells/ml, from about 1.0 x 10 ⁵ cells/ml to about 5.0 x 10 ⁶ cells/ml, from about 1.0 x 10 ⁵ cells/ml to about 5.0 x 10 ⁶ cells/ml or in any range including any two of the previous values, and/or within them.

In any embodiment of the present invention, one or more tissue engineered biostain compositions may be cell-free if the biostains do not contain living cells. In any embodiment of the present invention, the tissue engineered construct may be configured such that said tissue engineered construct has regions or components that may be acellular or cellular. For example, the tissue engineered construct may include a volume of the biopaint composition completely or partially encapsulated in an internal support separating the volume of the biopaint composition from other parts of the tissue engineered construct. In any embodiment of the present invention, the volume of the encapsulated biopaint composition may be different from the volume of non-encapsulated tissue-engineered biopaints. For example, in any embodiment of the present invention, said tissue engineered construct may include multiple volumes of the same or different biostain compositions encapsulated by two or more internal supports. In the present invention, the design, the tissue engineering design, includes a plurality of biocolor compositions.

In any embodiment of the present invention, the one or more tissue engineered biopaint compositions may further comprise carriers or additives. Suitable carriers may include, but are not limited to, water, aqueous solutions of ionic salts (e.g., physiological saline), phosphate buffered saline (PBS), cell medium, fetal bovine serum (FBS), Dulbecco's Minimum Essential Medium (DMEM), fibroblast growth factor (bFGF) and the like. or their combinations. Suitable additives include, among others, growth factors and cross-linking agents. Suitable crosslinking agents include, but are not limited to, riboflavin, ribose, polyethylene glycol (PEG), glutaraldehyde, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, genipin, chitosan, and the like. or their combinations.

Biocompatible support structure

The present technology provides a biocompatible support structure that includes one or more external supports, internal supports, or a combination thereof made from a biocompatible material. Fig. 1B shows an embodiment of a biocompatible support structure 110 having an external support 120 and an internal support 130, with the internal support coupled to one or more 2D surfaces of the external support. In the context of this document, the term "outer supports" refers to one or more structural components adapted to various 2D or 3D shapes and sizes and configured for use as an exoskeletal structure covering one or more surfaces of a tissue engineered structure. Fig. 1C shows an outer support 120 configured to cover a tissue-engineered ear-shaped shell. For example, in any embodiment of the present invention, the outer support may completely encapsulate the tissue engineered structure, such that the outer support will completely cover the outer surfaces of the tissue engineered structure. In any embodiment of the present invention, the outer support may partially cover one or more outer surfaces of the tissue engineered structure. Fig. 1C shows an embodiment of an external support 120 partially enclosing the shell of a tissue-engineered structure in the shape of a human ear 140. In any embodiment of the present invention, the external support may include one or more cavities (i.e., holes) to allow the flow of materials (e.g., nutrients, biological waste products, cellular and tissue materials, for example, growing vascular and nervous structures, or the like, or combinations thereof), while in the specified cavity there is a tissue engineering structure.

As used herein, the term "internal support" refers to one or more structural components adapted to various 2D or 3D shapes and sizes and configured to serve as an internal scaffold that is wholly or partially encapsulated by a tissue engineered construct. For example, in any embodiment of the present invention, the internal support may be completely encapsulated by the tissue engineered structure such that no portion of the internal support extends beyond the 2D outer surfaces of the tissue engineered structure. In any embodiment of the present invention, the internal support may be partially encapsulated by the tissue engineered structure such that one or more portions of the internal support extend beyond one or more two-dimensional outer surfaces of the tissue engineered structure. In any embodiment of the present invention, the inner support may be connected to one or more two-dimensional surfaces of the outer support and placed distally from them. In any embodiment of the present invention, the internal support may include one or more cavities (i.e., holes) to allow the flow of materials (e.g., nutrients, biological waste products, cellular and tissue materials, e.g., growing vascular and neural structures, or the like, or combinations thereof), wherein said cavity may include a volume of a bio-ink composition, the same as or different from the bio-paints, not present in the cavity. Fig. 1D shows an inner support 130 having a cavity 170.

In any embodiment of the present invention, the internal support may be 2D or 3D shaped such that it occupies a volume of space within the tissue engineered construct that is less than the total volume of the tissue engineered construct. For example, in any embodiment of the present invention, the outer support may partially cover one or more outer surfaces of the tissue engineered structure. In any embodiment of the present invention, the internal support may be wholly or partially ruptured, absorbed, or resorbed to form one or more cavities, channels, or combinations thereof in the tissue engineered construct. In any embodiment of the present invention, one or more cavities and/or channels may extend beyond one or more 2D outer surfaces of the tissue engineered construct. For example, in any embodiment of the present invention, the cavity and/or channel may extend through them such that one 2D outer surface of the tissue engineered structure is in fluid communication with another 2D outer surface of the tissue engineered structure. In any embodiment of the present invention, one or more cavities and/or channels may extend beyond one 2D outer surface of the tissue engineered structure such that there is no fluid communication with the other 2D outer surface of the tissue engineered construct. In any embodiment of the present invention, one or more cavities and/or channels may be filled or facilitate the flow of materials (eg, nutrients, biological waste products, cellular and tissue materials, and the like, or combinations thereof).

In any embodiment of the present invention, the internal support may be configured to encapsulate the volume of the biopaint composition completely or partially within the tissue engineered construct. In the present invention, the internal supports form one or more distinct pockets of a fully or partially encapsulated bioink composition within the tissue engineered construct. For example, the volume of the bioink composition encapsulated by the internal support may be the same or different from the volume of the bioink composition not encapsulated by the internal support. In any embodiment of the present invention, the volume of the encapsulated biopaint composition may be different from the volume of non-encapsulated tissue-engineered biopaints.

Without being limited by theory, it is believed that the outer support and/or the inner support increase the rigidity of the tissue engineered structure. In addition, the outer and/or inner support may further enhance the compressive modulus, elastic modulus, flexural stiffness, and shear strength. This can limit shape distortion during external compression by retaining the biomaterial. For example, supports add to the axial and flexural stiffness, as well as other mechanical properties described in this document, of tissue-engineered structures, providing in some cases a stiffer material inside, and for external supports, this is achieved with minimal use of material by increasing the moment cross-sectional inertia. In any embodiment of the present invention, one or more external supports, internal supports, or combinations thereof of a biocompatible support structure increase the value of one or more of the following characteristics: axial and bending stiffness, compression modulus, modulus of elasticity, bending stiffness, and tensile strength. shift of the tissue-engineered structure by about 5-10,000 times. For example, in any embodiment of the present invention, the axial and bending stiffness, compression modulus, elastic modulus, bending stiffness, and shear strength can be increased by about 5 times, about 10 times, about 20 times, about 30 times. , about 40 times, about 50 times, about 60 times, about 70 times, about 80 times, about 90 times, about 100 times, about 200 times, about 300 times, about 400 times , about 500 times, about 1000 times, about 5000 times, about 10000 times, or any range including and/or within any two of the previous values.

In any embodiment of the present invention, another external support, an internal support, or combinations thereof have a two-dimensional 2D thickness suitable to achieve the desired mechanical properties, such as the flexural modulus of the biocompatible support, or to impart the desired tensile, compressive, and flexural mechanical properties to the tissue-engineered structure. , as well as load power. For example, in any embodiment of the present invention, the outer supports or internal supports have a 2D thickness of about 10 microns to about 2000 microns. For example, in any embodiment of the present invention, the outer supports or the inner supports may have a 2D thickness of about 10 µm, about 20 µm, about 30 µm, about 40 µm, about 50 µm, about 60 µm, about 70 µm, about 80 µm, approx. 90 µm, approx. 100 µm, approx. 150 µm, approx. 200 µm, approx. 250 µm, approx. 300 µm, approx. 350 µm, approx. µm, approx. 700 µm, approx. 750 µm, approx. 800 µm, approx. 850 µm, approx. 900 µm, approx. 950 µm, approx. approx. 1300 µm, approx. 1350 µm, approx. 1400 µm, approx. 1450 µm, approx. 1500 µm, approx. 1550 µm, approx. µm, about 1950 µm, about 2000 µm, or any range including and/or within any of the previous values. Suitable 2D thicknesses include, but are not limited to, values from about 10 µm to about 2000 µm, from about 10 µm to about 1500 µm, from about 10 µm to about 1000 µm, from about 10 µm to about 500 µm, or are in any range, including any of the previous values and/or values in between.

In any embodiment of the present invention, the external supports or internal supports may include one or more holes passing through them from one 2D surface to the opposite parallel surface of the external support and / or internal support, allowing the flow of materials (for example, nutrients, biological products life activity, cellular and tissue materials, etc. or combinations thereof). Fig. 1C shows an outer support 120 having one or more holes 150, and FIG. 1D shows an internal support having one or more holes 160. The one or more holes may have an average size in the range of about 1 micron to about 10,000 microns. In any embodiment of the present invention, the average size of one or more holes may include, but is not limited to, about 1 µm, about 5 µm, about 10 µm, about 20 µm, about 30 µm, about 40 µm, about 50 µm, about 1 µm , approx. 5 µm, approx. 10 µm, approx. 20 µm, approx. 30 µm, approx. 40 µm, approx. 50 µm, approx. 60 µm, approx. 300 µm, approx. 400 µm, approx. 500 µm, approx. 600 µm, approx. 700 µm, approx. 800 µm, approx. 900 µm, approx. , approx. 4000 µm, approx. 4500 µm, approx. 5000 µm, approx. 5500 µm, approx. 6000 µm, approx. 6500 µm, approx. 10,000 µm, or any range including and/or within any two of the previous values. Suitable ranges include about 1 µm to about 10,000 µm, about 5 µm to about 7500 µm, about 10 µm to about 7000 µm, about 20 µm to about 5000 µm, about 35 µm to about 2500 µm, or are in any range, including any of the previous values and/or values in between. One or more holes may be of any shape; for example, the shape of one or more holes may be round, oval, elliptical, polygonal, and the like. or their combinations.

In any embodiment of the present invention, one or more external support surfaces and/or internal support surfaces may include one or more protrusions connected to and extending distally from one or more 2D external support and/or internal support surfaces. . For example, in any embodiment of the present invention, the protrusions may include, but are not limited to, spikes, knots, knots, struts, ridges, and the like. or their combinations.

In any embodiment of the invention, the biocompatible support structure may include any combination of external supports and/or internal supports. For example, a biocompatible support structure may include only one or more external supports, as described herein, in any embodiment. A biocompatible support in any embodiment of the present invention may include only one or more internal supports. In any embodiment of the invention, the biocompatible support structure may include one or more external supports and one or more internal supports. For example, a portion of the inner leg may be connected to the 2D surface of the outer leg. In any embodiment of the present invention, the biocompatible support structure may include an external support and an internal support, with one or more of the external supports not connected to or in contact with one or more of the internal supports.

The biocompatible support structure may include a biocompatible material. For example, in any embodiment of this invention, the biocompatible material may include, but is not limited to, polysaccharides, biocompatible polymers, rubber, silicon, biocompatible metals (e.g., steel, cobalt-chromium alloys, titanium, titanium alloys, magnesium, magnesium alloys, zinc, zinc alloys, iron alloys, etc. or combinations thereof), biocompatible ceramics, polyethylene glycol, polypropylene glycol, polyamino acids, natural and biopolymers (e.g. glycosaminoglycans, cellulose, chitosan, chitin, dextrans, gelatin, collagen, lignins, polyamino acids, glycoproteins, elastine , laminins) or combinations of two or more of them. In any embodiment of this invention, the biocompatible material may include biocompatible polymers; for example, in any embodiment of the present invention, biocompatible polymers may include polylactides (PLA), polyglycolic acid (PGA), lactide-glycolide copolymer (PLGA), polydioxanone (PDO), polycaprolactones, and combinations thereof.

In any embodiment of this invention, the biocompatible polymer may have a melting point from about 50°C to about 290°C. For example, in any embodiment of this invention, the biocompatible polymer may have a melting point of about 50°C, about 55°C, about 60°C, about 65°C, about 70°C, about 75°C, about 80°C. C, about 85°C, about 90°C, about 95°C, about 100°C, about 110°C, about 120°C, about 130°C, about 140°C, about 150°C, about 160° C, about 170°C, about 180°C, about 190°C, about 200°C, about 210°C, about 220°C, about 230°C, about 240°C, about 250°C, about 260° C, about 270°C, about 280°C, about 290°C, or any range including and/or within any two of the previous values.

In any embodiment of the invention, the biocompatible material may further comprise one or more additives. For example, one or more additives may include, among other things, cross-linking agents.

The biocompatible material may be biodegradable, bioabsorbable, bioresorbable, or a combination thereof. For example, in any embodiment of the present invention, the biocompatible material may be degraded, absorbed, or resorbed under physiological conditions at a rate of from about 1 week to about 7 years. Suitable degradation, absorption or resorption rates may include, but are not limited to, about 1 week, about 2 weeks, about 3 weeks, about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 1 year, about 1.5 years, about 2 years, about 2.5 years, about 3 years, about 3.5 years, about 4 years, about 4.5 years, about 5 years, about 5.5 years, about 6 years, about 6.5 years, about 7 years, or be in ranges including and/or between any two of the previous values. For example, in any embodiment of the present invention, the rate of degradation, absorption, or resorption may include from about 1 week to about 7 years, from about 1 week to about 5 years, from about 1 week to about 3 years, from about 1 week to about 1 of the year. from about 1 week to about 6 months, from about 1 week to about 3 months. Suitable biodegradable, bioabsorbable and/or bioresorbable materials include, but are not limited to, PLA, PGA, PLGA, PDO, polycaprolactones, bioresorbable metal alloys, and combinations thereof.

In any embodiment of the present invention, the inner support may be a different biomaterial from the material of the outer support. For example, the inner support may be a biomaterial whose rate of degradation, absorption, or resorption is different from that of the outer support material. In any embodiment of the present invention, the inner support biomaterial has a longer rate of degradation, absorption, or resorption than the outer support material. In any embodiment of the present invention, the inner support biomaterial has a lower rate of degradation, absorption, or resorption than the outer support material.

When the material is or includes a biopolymer, protein, glycoprotein (eg as defined above), the concentration of this component may be from about 1000 mg/mL to about 2500 mg/mL. For example, in any embodiment of the present invention, the biocompatible material may be present at a concentration of about 1000 mg/ml, about 1100 mg/ml, about 1200 mg/ml, about 1300 mg/ml, about 1400 mg/ml, about 1500 mg/ml. mg/ml, about 1600 mg/ml, about 1700 mg/ml, about 1800 mg/ml, about 1900 mg/ml, about 2000 mg/ml, about 2100 mg/ml, about 2200 mg/ml, about 2300 mg/ml ml. ml, about 2400 mg/ml, about 2500 mg/ml, or in any range including and/or within any two of the previous values. In any embodiment of the present invention, the biocompatible material may be present at a concentration of from about 1500 mg/ml to about 2500 mg/ml, from about 1750 mg/ml to about 2200 mg/ml, from about 2000 mg/ml to about 2300 mg/ml or in any range including and/or within any two of the previous values.

The biocompatible material may be present in the biocompatible support structure in an amount of from about 1% to 100% by weight of the biocompatible support structure. For example, in any embodiment of the present invention, the amount of biocompatible material present by weight in the biocompatible support structure may be about 1%, about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, about 95 % or be in any range including and/or within any two of the previous values. Suitable amounts of biocompatible material in the biocompatible support structure include, but are not limited to, about 5% to about 100%, 10% to about 100%, about 50% to about 100%, about 5% to about 20%, about 90% to about 100%, or within any range including and/or within any two of the previous values.

The biocompatible support structure has mechanical properties to maintain the mechanical properties of the tissue engineered structure. In any embodiment of the present invention, the biocompatible support structure may have one or more of the following characteristics: a flexural modulus of about 0.2 GPa to about 100 GPa, a modulus of elasticity of about 0.02 GPa to about 100 GPa, or a tensile strength of gap from about 1 MPa to about 1000 MPa.

In any embodiment of the invention, the biocompatible support structure may have a flexural modulus of about 0.2 GPa to about 100 GPa. For example, in any embodiment of the present invention, the biocompatible support structure may have a flexural modulus of about 0.2 GPa, about 0.3 GPa, about 0.4 GPa, about 0.5 GPa, about 0.6 GPa, about 0. 7 GPa, about 0.8 GPa, about 0.9 GPa, about 1 GPa, about 2 GPa, about 3 GPa, about 4 GPa, about 5 GPa, about 6 GPa, about 7 GPa, about 8 GPa, about 9 GPa , about 10 GPa, about 11 GPa, about 12 GPa, about 13 GPa, about 14 GPa, about 15 GPa, about 16 GPa, about 17 GPa, about 18 GPa, about 19 GPa, about 20 GPa, about 25 GPa, about 30 GPa, about 35 GPa, about 40 GPa, about 45 GPa, about 50 GPa, about 55 GPa, about 60 GPa, about 65 GPa, about 70 GPa, about 75 GPa, about 80 GPa, about 85 GPa, about 90 GPa , about 95 GPa, about 100 GPa, or in any range including and/or within any two of the previous values. Suitable flexural modulus values may include, but are not limited to, about 0.2 GPa to about 100 GPa, about 0.2 GPa to about 50 GPa, about 0.2 GPa to about 25 GPa, about 0.2 GPa to about 15 GPa, from about 1 GPa to about 15 GPa. GPa, from about 5 to about 15 GPa, or ranges including and/or within any two of the previous values.

In any embodiment of the present invention, the biocompatible support structure may have a modulus of elasticity from about 0.02 GPa to about 100 GPa. For example, in any embodiment of the present invention, the biocompatible support structure may have an elastic modulus of about 0.02 GPa, 0.04 GPa, about 0.06 GPa, about 0.08 GPa, about 0.1 GPa, about 0.2 GPa, about 0.3 GPa, about 0.4 GPa, about 0.5 GPa, about 0.6 GPa, about 0.7 GPa, about 0.8 GPa, about 0.9 GPa, about 1 GPa, about 2 GPa, about 3 GPa, about 4 GPa, about 5 GPa, about 6 GPa, about 7 GPa, about 8 GPa, about 9 GPa, about 10 GPa, about 11 GPa, about 12 GPa, about 13 GPa, about 14 GPa, about 15 GPa, about 16 GPa, about 17 GPa, about 18 GPa, about 19 GPa, about 20 GPa, about 25 GPa, about 30 GPa, about 35 GPa, about 40 GPa, about 45 GPa, about 50 GPa, about 55 GPa, about 60 GPa, about 65 GPa, about 70 GPa, about 75 GPa, about 80 GPa, about 85 GPa, about 90 GPa, about 95 GPa, about 100 GPa, or any range including any two of the previous values, and/or within their limits. Suitable modulus values may include, but are not limited to, about 0.2 GPa to about 100 GPa, about 0.2 GPa to about 50 GPa, about 0.2 GPa to about 25 GPa, about 0.2 GPa to about 15 GPa, from about 1 GPa to about 15 GPa. GPa, from about 5 to about 15 GPa, or ranges including and/or may be within any two of the previous values.

In any embodiment of the present invention, the biocompatible support structure may have a tensile strength of from about 1 MPa to about 1000 MPa. For example, in any embodiment of the present invention, the biocompatible support structure may have a tensile strength of about 1 MPa, about 2 MPa, about 3 MPa, about 4 MPa, about 5 MPa, about 6 MPa, about 7 MPa, about 8 MPa, about 9 MPa, about 10 MPa, about 20 MPa, about 25 MPa, about 30 MPa, about 35 MPa, about 40 MPa, about 45 MPa, about 50 MPa, about 55 MPa, about 60 MPa, about 65 MPa, about 70 MPa , about 75 MPa, about 80 MPa, about 85 MPa, about 90 MPa, about 95 MPa, about 100 MPa, about 150 MPa, about 200 MPa, about 250 MPa, about 300 MPa, about 350 MPa, about 400 MPa, about 450 MPa, approx. 500 MPa, approx. 550 MPa, approx. 600 MPa, approx. 650 MPa, approx. 700 MPa, approx. 750 MPa, approx. 800 MPa, approx. 850 MPa, approx. , including any two of the previous values, and/or within them.

The tissue or organ substitute composition includes a tissue engineered construct and a biocompatible support structure taken together in any configuration as described herein and has improved mechanical properties (eg, strength).

In any embodiment of the present invention, one or more external supports, internal supports, or combinations thereof of a biocompatible support structure can increase the load capacity of the tissue engineered structure from about 1% to about 10,000%. For example, in any embodiment of the present invention, the load capacity of the tissue engineered structure can be increased from about 1% to about 10000%, from about 1% to about 1000%, from about 1% to about 100%, from about 1% to about 10%. or in any range including and/or within any two of the previous values. Suitable load power increases may include, but are not limited to, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, about 10% , about 11%, about 12%, about 13%, about 14%, about 15%, about 16%, about 17%, about 18%, about 19%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 200%, about 300%, about 400%, about 500%, about 600%, about 700%, about 800% , about 900%, about 1000%, about 2000%, about 3000%, about 4000%, about 5000%, about 6000%, about 7000%, about 8000%, about 9000%, about 10000% or can be in any range , including any two of the previous values, and/or within them.

The tissue or organ substitute composition, in any embodiment of the present invention, can withstand compression from about 1 kN to about 10,000 kN and a shear force from about 0.2 kN to about 1,000 kN. For example, in any embodiment of the present invention, the tissue or organ substitute composition of the present technology can withstand a compression of about 1 kN, 2 kN, about 3 kN, about 4 kN, about 5 kN, about 6 kN, about 7 kN, about 8 kN , approx. 9 kN, approx. 10 kN, approx. 20 kN, approx. 30 kN, approx. 40 kN, approx. 50 kN, approx. 60 kN, approx. 70 kN, approx. 80 kN, approx. 300 kN, approx. 400 kN, approx. 500 kN, approx. 600 kN, approx. 700 kN, approx. 800 kN, approx. 900 kN, approx. 1000 kN, approx. 1500 kN, approx. , approx. 4000 kN, approx. 4500 kN, approx. 5000 kN, approx. 5500 kN, approx. 6000 kN, approx. 6500 kN, approx. 7000 kN, approx. 10,000 kN, or in any range including and/or within any two of the previous values. Suitable compression ranges may include, but are not limited to, about 1 kN to about 10,000 kN, about 1 kN to about 5,000 kN, about 1 kN to about 1,000 kN, about 1 kN to about 500 kN, about 1 kN to about 15 kN, from about 4 kN to about 10 kN, or ranges including and/or within any two of the preceding values.

In any embodiment of the present invention, the tissue or organ substitute composition of the present technology can withstand a shear force of from about 0.2 kN to about 1000 kN. For example, in any embodiment of the present invention, the tissue or organ substitute composition of the present technology can withstand a shear force that can be about 0.2 kN, about 0.3 kN, about 0.4 kN, about 0.5 kN, about 0 .6 kN, about 0.7 kN, about 0.8 kN, about 0.9 kN, about 1.0 kN, about 10 kN, about 50 kN, about 100 kN, about 150 kN, about 200 kN, about 250 kN, approx. 300 kN, approx. 350 kN, approx. 400 kN, approx. 450 kN, approx. 500 kN, approx. 550 kN, approx. 600 kN, approx. 650 kN, approx. about 900 kN, about 950 kN, about 1000 kN, or may be in any range including and/or within any two of the previous values. Suitable shear force ranges include, but are not limited to, about 0.2 kN to about 1000 kN, about 1 kN to about 500 kN, about 10 kN to about 100 kN, about 0.2 kN to about 0.9 kN or are in any range including and/or within any two of the previous values.

Ways

This technology provides a method for making a tissue or organ substitute composition that includes making a biocompatible support structure as described herein, in any embodiment; making a tissue engineered construct as described herein, in any embodiment, comprising: applying one or more biostain compositions as described herein, in any embodiment, to or around a biocompatible support structure; crosslinking one or more bioink compositions and optionally repeating the application and crosslinking steps to form a tissue engineered construct; and curing the tissue or organ substitute composition; wherein the biocompatible support structure includes one or more of external supports, internal supports, or combinations thereof of a biocompatible material, and wherein the biocompatible material is present in an amount of from about 1% to about 100% by weight of the biocompatible support structure.

Fig. 4A is a flowchart 400 showing an embodiment of a method for making a tissue or organ substitute composition. The method includes fabricating biocompatible support structures (process 401), fabricating a tissue engineered construct (process 402), and curing the tissue engineered construct (process 403). Each of the processes (eg, sub-processes, subroutines, or steps) 401, 402, and 403 are discussed below with reference to FIG. 4B-4D.

A method for manufacturing a biocompatible support structure may include one or more manufacturing systems or methods including, but not limited to, thermoforming, injection molding, rotational molding, positive molds, negative molds, extrusion, subtractive manufacturing, cutting, laser cutting, drilling, water jet cutting, stamping, direct thermal cutting, chemical etching, selective thermal melting, dipping processes, spray coating, selective electrostatic coating, milling and machining, 3D 3D printing, or combinations of two or more of these methods. For example, in any embodiment of the present invention, the manufacturing method may include three-dimensional printing. Suitable 3D printing methods may include, but are not limited to, inkjet printing, layer-by-layer printing, extrusion printing, or bioprinting. For example, in any embodiment of the present invention, the application may include the application of one or more layers of biocompatible material. For example, in any embodiment of the present invention, 3D printing may include: applying one or more layers of a biocompatible material to a substrate; optionally crosslinking the deposited biocompatible material; and optionally repeating the application and optional crosslinking steps to obtain a biocompatible support structure; wherein the resulting biocompatible support structure includes one or more of external supports, internal supports, or combinations thereof of a biocompatible material, and wherein the biocompatible material is present in an amount of from about 1% to about 100% by weight of the biocompatible support structure.

In any embodiment of the present invention, a biocompatible support structure can be manufactured using biocompatible materials as described herein, in any embodiment, using 3D printing systems, as described in PCT application serial number PCT/2018/034457 called "ASEPTIC PRINTER SYSTEM INCLUDING DUAL ARM-MECHANISM", filed May 24, 2018, with Patent Registry Number 113066-0107, the entire contents of which are incorporated herein by reference for reference and description of the methods set forth therein.

In any embodiment of the present invention, a method for manufacturing a biocompatible support structure may include injection molding. For example, in any embodiment of the present invention, application may include applying a biocompatible material to a mold of a biocompatible support structure; optionally crosslinking the biocompatible material in the mold and optionally repeating the application and optional crosslinking steps; and removing the biocompatible support structure from the mold. In any embodiment of the present invention, the specified mold may be soluble or insoluble, for example, from a polymer, metal or biomaterial. For example, in any embodiment of the invention, said mold may be dissolvable such that the mold dissolves to form an internal cavity in one or more external supports, internal supports, or combinations thereof. In any embodiment of the invention, said mold may be a soluble material in granular form such as sand, powder, etc., or combinations thereof. Fig. 2B shows a dissolvable mold 190 for forming an internal support for an ear-shaped tissue engineered construct.

Fig. 4B is a flow diagram relating to an embodiment of a method for manufacturing a typical biocompatible support (process 401) for an internal and/or external support.

In at least one embodiment of the present invention, process 401 includes selecting one of the internal or external supports for manufacture. After determining the manufacture of the internal support (step 410), a typical manufacturing process includes forming a 2D sheet of biocompatible material (step 411), thermoforming the 2D sheet onto a mold (step 412), removing the mold (step 413), and stitching biocompatible material to obtain internal support (step 414).

As further shown in FIG. 4B, after determining the manufacture of the outer support (step 420), a typical manufacturing process includes applying the biocompatible material via 3D printing or injection molding (step 421), cross-linking the biocompatible material (step 422), and separating the outer support from the substrate or mold. (eg, injection molds) (step 423).

In at least one embodiment of the present invention, for the manufacture of an external support, a typical technique includes applying a biocompatible material through a fused deposition process, using a 3D printer or injection molding, and optionally crosslinking the biocompatible material to form an external support. To make an internal support, a typical technique involves applying a biocompatible material to form a 2D sheet, thermoforming the 2D sheet on a dissolvable mold, dissolving the mold, and optionally crosslinking the biocompatible material to form an internal support.

In addition, in at least one embodiment of the present invention, the fabrication technology of the tissue engineered construct may include applying bioinks using a 3D bioprinter and may further include placing a biocompatible support structure on a substrate (e.g., a bioprinting work surface) and applying the bioinks by printing. onto and/or around the support structure to obtain said tissue-engineered ear-shaped structure. As further discussed herein, said method may include applying the bioinks using a molding method and may include placing a biocompatible support structure within a mold of the tissue engineered construct (e.g., an ear-shaped mold) and applying one or more layers of the bioinks to the press. -shape.

In addition, the method for manufacturing a tissue engineered structure according to an embodiment of the present invention may include a curing process. The curing process may include placing the structure in a media bath (eg, at about 37°C) and curing overnight (eg, for about 12 hours) for 3D printed designs; or placing the mold in a media bath, removing the structure from the mold after a couple of hours (eg 2 hours) and curing the structure overnight for injection molded structures.

In any embodiment of the present invention, the method of manufacturing a biocompatible support structure may include thermoforming. The thermoforming method may include any suitable known thermoforming methods, such as methods suitable for thermoforming a porous and/or non-porous 2D sheet. Such thermoforming methods may include heating a 2D sheet and vacuuming on a mold, or heating a 2D sheet and forming using a positive and negative mold.

For example, in any embodiment of the present invention, thermoforming includes contacting a 2D sheet of biocompatible material with a substrate, wherein the substrate includes one or more molds formed to form one or more external supports, internal supports, or combinations thereof; heating the 2D sheet of biocompatible material to thermoform the 2D sheet according to the shape of one or more molds to form one or more external supports, internal supports, or combinations thereof; and removing one or more external supports, internal supports, or combinations thereof from the mold. In any embodiment of the present invention, thermoforming may further include repeating one or more of the following processes: contacting, heating, and removing one or more molds to obtain the desired shape of one or more outer supports, internal supports, or combinations thereof. Fig. 2C shows a 2D porous sheet thermoformed onto an inner support mold.

In any embodiment of the present invention, the 2D sheet may be made from 3D printed, spun, woven or knitted fibers. For example, in any embodiment of the present invention, the 2D sheet may be a 3D printed fiber using the 3D printing techniques described herein. In any embodiment of the present invention, the 2D sheet may be porous or non-porous. In any embodiment of the present invention, the porous 2D sheet may include one or more apertures having an average size in the range of about 1 micron to about 10,000 microns, as described herein in any embodiment of the present invention. One or more holes may be of any shape; for example, the shape of one or more holes may be round, oval, elliptical, polygonal, and the like. or their combinations. Fig. 2A shows a 2D sheet 180 having one or more holes in accordance with an embodiment of this technology.

In any embodiment of the present invention, the fibers may have a diameter of from about 10 microns to about 2000 microns. For example, in any embodiment of the present invention, the outer supports or the inner supports may have a 2D thickness of about 10 µm, about 20 µm, about 30 µm, about 40 µm, about 50 µm, about 60 µm, about 70 µm, about 80 µm, approx. 90 µm, approx. 100 µm, approx. 150 µm, approx. 200 µm, approx. 250 µm, approx. 300 µm, approx. 350 µm, approx. µm, approx. 700 µm, approx. 750 µm, approx. 800 µm, approx. 850 µm, approx. 900 µm, approx. 950 µm, approx. approx. 1300 µm, approx. 1350 µm, approx. 1400 µm, approx. 1450 µm, approx. 1500 µm, approx. 1550 µm, approx. 1600 µm, approx. µm, about 1950 µm, about 2000 µm, or may be in any range including and/or within any two of the previous values. Suitable 2D thickness values include, but are not limited to, values from about 10 µm to about 2000 µm, from about 10 µm to about 1500 µm, from about 10 µm to about 1000 µm, from about 10 µm to about 500 µm, or are in any range, including any of the previous values and / or within them.

In any embodiment of the present invention, the substrate may include a 3D printer surface (eg, a 3D printer bed). In any embodiment of the present invention, said substrate may include a mold suitable for use in one or more of the following methods: injection molding, rotational molding, positive molds, negative molds, and the like. In any embodiment of the present invention, said substrate may include the surface of a manufacturing device suitable for performing subtractive manufacturing, milling and machining, or the like. In any embodiment of the present invention, said substrate may be a mold having a 3D shape of a tissue engineered structure.

In any embodiment of the present invention, the biocompatible material is crosslinked. For example, in any embodiment of the present invention, the biocompatible material may be subjected to crosslinking conditions to produce a crosslinked or polymerized biocompatible material. Suitable crosslinking conditions may include, but are not limited to, UV irradiation. In any embodiment of the invention, the biocompatible material may not include crosslinking. For example, in any embodiment of the present invention, after deposition, the biomaterial may include a cooling step, a fusion step, and the like. or their combinations.

In the method according to this technology, the method of manufacturing a biocompatible support structure may include applying one or more layers of biocompatible material so that one or more external supports and/or internal supports have a 2D thickness of from about 10 μm to about 2000 μm, as described in this document, in any embodiment.

In the method according to this technology, the biocompatible material may include a biocompatible material, as described in this document, in any embodiment. In any embodiment of this invention, the biocompatible polymer may have a melting point from about 50°C to about 290°C, as described herein, in any embodiment.

In any embodiment of the present invention, the biocompatible support structure obtained by the manufacturing process may have one or more of the following characteristics: a flexural modulus of about 0.2 GPa to about 100 GPa, a modulus of elasticity of about 0.2 GPa to about 100 GPa or tensile strength from about 1 MPa to about 1000 MPa, as described herein, in any embodiment.

In any embodiment of the invention, said method may further comprise sterilizing the biocompatible support structure. For example, in any embodiment of the present invention, sterilization may include, but is not limited to, gamma irradiation, peracetic acid incubation, autoclaving, UV irradiation, peroxide sterilization, supercritical fluid treatment, and the like. or their combinations.

Fabrication of a tissue engineered construct involves applying one or more layers of one or more biopaint compositions as described herein, in any embodiment. In any embodiment of the present invention, the fabrication of a tissue engineered construct may include one or more fabrication systems or methods; for example, in any embodiment of this invention, one or more manufacturing systems or methods may include injection molding, rotational molding, positive dies, negative dies, subtractive manufacturing, milling and machining, and 3D printing. In any embodiment of the present invention, said manufacturing method may include 3D printing, as described herein, in any embodiment. For example, in any embodiment of the invention, application may include applying one or more layers of one or more bioink compositions to or around a biocompatible support structure using a 3D printer; cross-linking one or more bio-paint compositions and optionally repeating the application and cross-linking steps to obtain a tissue engineered construct; and curing the tissue or organ substitute composition.

Fig. 4C shows a fabrication process 402 for fabricating a tissue engineered construct, in accordance with at least one embodiment. Process 402 may include using at least one 3D printer to print a biocompatible support structure (block 430). The process further includes placing the biocompatible support structure on a substrate (eg, a work surface) (step 431) and applying bioinks to and/or around the biocompatible support structure (step 432).

In any embodiment of the invention, fabrication of the tissue engineered structure may include injection molding. For example, in any embodiment of the invention, application may include applying one or more bioink compositions to a mold containing a biocompatible support structure; cross-linking one or more bio-ink compositions in a mold, and optionally repeating the application and cross-linking steps to obtain a tissue-engineered construct; and curing the tissue or organ substitute composition, wherein the mold may be removed before or after the curing step. In any embodiment of the invention, said method may include removing the mold after the tissue or organ substitute composition has cured. In any embodiment of the invention, said method may include removing the mold prior to curing the tissue or organ substitute composition. Fig. 3A shows an internal view of a mold 191 configured and contoured for injection molding an ear-shaped tissue engineered structure and having halves 191, 192 as first and second mold parts. Mold part 192 includes first cavity 194 and mold part 193 includes second cavity 195. FIG. 3B shows the appearance of a mold 191 configured and contoured for injection molding an ear-shaped tissue engineered structure.

As discussed above, FIG. 4C depicts a fabrication process 402 of a tissue engineered construct, in accordance with at least one embodiment. In certain embodiments of the present invention, process 402 includes fabrication of a tissue engineered construct by injection molding (step 440). This process may further include placing the biocompatible support structure in the mold (step 441) and applying bioinks to the mold (step 442).

In any embodiment of the invention, the bioinks are crosslinked. For example, in any embodiment of the present invention, bioinks can be subjected to crosslinking conditions to produce crosslinked or polymerized bioinks. For example, in any embodiment of the present invention, crosslinking may include UV irradiation, addition of salts, pH neutralization, thermal crosslinking, and the like. or their combinations.

The tissue or organ substitute composition undergoes a curing step after crosslinking. In any embodiment of the present invention, said method comprises curing the tissue or organ substitute composition at a temperature of from about 4°C to about 50°C. Suitable cure temperatures may include, but are not limited to, about 4°C to about 50°C, about 15°C to about 45°C, about 30°C to about 40°C, about 34°C to about 37 °C, from about 35°C to about 37°C, from about 36°C to about 37°C, or may be in any range including and/or within any two of the previous values. In any embodiment of the present invention, the tissue or organ substitute composition may be placed in a buffer solution or cell medium during the curing step. In any embodiment of the present invention, suitable buffer solutions include, but are not limited to, phosphate buffered physiological saline, sodium chloride solution, phosphate-containing solutions, phosphate buffer solution, Hank's balanced salt solution, and MES buffer. Suitable cell media may include, but are not limited to, serum, serum-free medium, HEPES, DMEM, and the like. or combinations thereof, supplemented with bFGF, FBS, ascorbate, and the like. or their combinations. In any embodiment of the present invention, said method may include curing the tissue or organ substitute composition in a buffer solution and a cellular medium. In any embodiment of the present invention, curing may occur within from about 3 hours to about 24 hours. For example, in any embodiment of the present invention may occur for about 3 hours, about 4 hours, about 5 hours, about 6 m, about 7 hours, about 8 hours, about 9 hours, about 10 hours, about 11 hours, about 12 hours, about 13 hours, about 14 hours, about 15 hours, about 16 hours, about 17 hours, about 18 hours, about 19 hours, about 20 hours, about 21 hours, about 22 hours, about 23 hours, about 24 hours or in any range including and/or within any two of the previous values.

Fig. 4D shows a flowchart of a process in accordance with an embodiment. In particular, a process 403 for curing a tissue engineered construct is demonstrated. In accordance with an embodiment, process 403 includes placing the structure in a bath of medium (step 451) and allowing the structure to cure for a sufficient period of time (step 452).

In any embodiment of this invention, one or more biopaint compositions may include, but are not limited to, alginate, agarose, collagen, chitosan, fibrin, hyaluronic acid, laminin, elastin, carrageenan, polyethylene oxide, polyethylene oxide-polypropylene copolymer, hydroxypropyl methylcellulose, propylene fumarate copolymer, and ethylene glycol, poly(ethylene glycol)-co-poly(lactic acid), poly(vinyl alcohol), KDL12 oligopeptides, poly(n-isopropylacrylamide), or combinations of two or more of them. One or more biopaint compositions can have a controlled crosslinking rate by adjusting environmental variables including, but not limited to, temperature, pH, ionic strength, heat, light, or the addition of chemical crosslinkers such as calcium, magnesium, barium, chondroitin, sulfate and thrombin, riboflavin, cabodiimides and the like. Suitable hydrogels for use in biopaints for tissue engineering are described in US Pat. No. 9,044,335 entitled "COMPOSITE TISSUE-ENGINEERED INTERVERTEBRAL DISC WITH SELF-ASSEMBLED ANNULAR ALIGNMENT" filed May 5, 2010, the entire contents of which are incorporated herein by reference.

In any embodiment of this invention, one or more biopaint compositions may include an alginate hydrogel, as described herein, in any embodiment. For example, in any embodiment of the invention, the alginate hydrogel may be present in an amount of from about 0.5% (w/v) to about 10% (w/v). In any embodiment of the present invention, one or more biopaint compositions may include collagen, as described herein, in any embodiment. For example, in any embodiment of the present invention, one or more biopaint compositions may include more than about 5 mg/mL of collagen. In any embodiment of the present invention, one or more biopaint compositions may include type I collagen, type II collagen, or a combination thereof.

In any embodiment of the present invention, one or more biopaint compositions may additionally include a neutralizer and a cell medium as described herein, in any embodiment. Suitable neutralizers may include, but are not limited to, formulations containing, for example, a weak acid. In any embodiment of the present invention, one or more biopaint compositions may be cellular or acellular. For example, in any embodiment of the present invention, one or more biostain compositions may include cell media containing living cells. The cell environment may further include other living cells; for example, in any embodiment of the present invention, biostains may include epidermal cells, chondrocytes and other cells that form mesenchymal stem cells, adipose-derived stem cells, macrophages, adipocytes, dermal cells, muscle cells, hair follicles, fibroblasts, organ cells , osteoblasts, osteocytes and other cells that form bones, endothelial cells, mucosal cells, pleural cells, ear canal cells, tympanic membrane cells, peritoneal cells, Schwann cells, corneal epithelial cells, gingival cells, neuronal stem cells of the central nervous system, epithelial cells of the trachea, or combinations of two or more of them. In any embodiment of the present invention, one or more biopaint compositions may be cell-free if one or more biopaint compositions do not contain living cells.

In any embodiment of the present invention, one or more biopaint compositions may further include carriers or additives. Suitable carriers may include, but are not limited to, water, aqueous solutions of ionic salts (e.g., sodium hydroxide), phosphate buffered saline (PBS), cell medium, fetal bovine serum (FBS), Dulbecco's Minimum Essential Medium (DMEM), fibroblast growth factor ( bFGF), etc. or their combinations. Suitable additives include, among other things, cross-linking agents. Suitable crosslinking agents include, but are not limited to, riboflavin, ribose, polyethylene glycol (PEG), glutaraldehyde, 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide hydrochloride, genipin, chitosan, and the like. or their combinations.

In any embodiment of the present invention, the manufacture of the biocompatible support structure or the manufacture of the tissue engineered construct may occur sequentially or simultaneously.

In any embodiment of this method, the tissue engineered construct may be of any suitable shape or size. For example, in any embodiment of the present invention, the tissue engineered construct may have a customized 3D structure of various shapes and sizes for a range of clinical applications, such as facial or craniofacial applications, artificial cartilage, or combinations thereof. In any embodiment of the present invention, the tissue engineered construct may be in the form of an organ or tissue of the subject. For example, in any embodiment of the present invention, the organ or tissue is selected from the ear, nose, meniscus, articular cartilage, trachea, or heart valve. In one embodiment of the invention, said organ is the ear.

In any embodiment of the present invention, the tissue engineering system may have a 3D structure based on a 3D computer model. For example, in any embodiment of the present invention, the computer model may include a 3D contralateral organ or tissue of the subject. In any embodiment of the invention, the subject is a mammal (eg, a cat, dog, rodent, or primate). In any embodiment of the invention, the subject is a human. A 3D computer model can be generated using 3D imaging tools. Suitable 3D imaging tools include, but are not limited to, clinical computed tomography (CT), magnetic resonance imaging (MRI), laser scanning, 3D cameras, or combinations of any two or more of these.

In any embodiment of the present invention, the tissue or organ substitute composition can withstand one or more compressions from about 1 to about 10,000 kN or shear forces from about 0.2 to about 1,000 kN, as described herein in any embodiment.

In at least one embodiment of the present invention, methods for making a tissue or organ substitute composition according to the present technology are a method in accordance with the steps illustrated in FIG. 4A-4D.

The present invention, thus described in general terms, will be better understood by reference to the above examples, which are given by way of illustration and are not intended to limit the present invention.

Although certain embodiments of the present invention have been illustrated and described, it should be understood that changes and modifications may be made to them in accordance with ordinary skill in the art without deviating from the technology in its broader aspects, as defined in the following formula inventions.

The embodiments of the present invention illustratively described herein may be appropriately practiced in the absence of any element or elements, limitation or limitations not specifically described herein. Thus, for example, the terms "comprising", "including", "comprising", etc. should be understood in a broad sense and without limitation. In addition, the terms and expressions used herein have been used as terms of description and not limitation, and the use of such terms and expressions is not intended to exclude any equivalents of the features or parts shown and described, but it is recognized that various modifications are possible in within the scope of the claimed technology. In addition, the phrase "consisting essentially of" should be understood to include those elements that are specifically listed and those additional elements that do not significantly affect the main and new characteristics of the claimed technology. The phrase "consisting of" excludes any unspecified element.

The present invention should not be limited to the specific embodiments described in this application. Many modifications and variations can be made without departing from its spirit and scope, as will be apparent to those skilled in the art. Functionally equivalent methods and compositions within the scope of this invention, in addition to those listed herein, will be apparent to those skilled in the art from the foregoing descriptions. Such modifications and changes are intended to be within the scope of the appended claims. Therefore, the invention is to be limited only by the definitions of the appended claims together with the full scope of the equivalents that apply to these claims. It should be understood that this invention is not limited to specific methods, reagents, compounds or compositions, which, of course, may vary. It should also be understood that the terminology used herein is intended solely to describe specific embodiments of the present invention and is not intended to limit the scope of the invention.

Furthermore, if features or aspects of the present invention are described in terms of Markush groups, those skilled in the art will appreciate that the present invention is also thus described in terms of any individual member or subgroup of members of the Markush group.

As will be appreciated by one of ordinary skill in the art, for any and all purposes, especially in terms of providing a written description, all ranges described herein also cover any and all possible subranges and combinations thereof. Any listed range can be readily recognized as being sufficiently descriptive and allowing the same range to be broken down into at least equal halves, thirds, quarters, fifths, tenths, and so on. As a non-limiting example, each range discussed herein can be easily divided into lower third, middle third, and upper third, and so on. As will also be appreciated by one of ordinary skill in the art, all terms such as "up to", "at least", "greater than", "less than" and the like include the indicated number and refer to ranges that may subsequently be subdivided. into subbands as described above. Finally, as one of skill in the art will appreciate, the range includes each individual member.

Although this description contains many specific implementation details, they should not be construed as limitations on the scope of the claims made, but rather as descriptions of features specific to particular implementations. Some of the features described in this document in the context of individual implementations may also be implemented in combination in a single implementation. Conversely, various functions described in the context of one implementation may also be implemented in multiple implementations individually or in any suitable subcombination. Moreover, although the features may be described above as operating in certain combinations and even originally claimed as such, one or more features of the claimed combination may in some cases be excluded from the combination, and the claimed combination may be directed to a subcombination or variation of the subcombination.

The terms "paired", "connected" and the like, which can be used in this document, means the connection of two components, directly or indirectly, with each other. Such a connection may be fixed (for example, permanent) or movable (for example, removable or detachable). Such a connection can be achieved with two components or two components and any additional intermediate components integrally formed with each other, or with two components or two components and any additional intermediate components attached to each other.

All publications, patent applications, granted patents and other documents mentioned in this specification are incorporated herein by reference, as if each individual publication, patent application, patent granted or other document is specifically and individually indicated for inclusion as links in full. Definitions contained in text incorporated by reference are excluded to the extent that they conflict with the definitions.

Claims (84)

1. Composition of a tissue or organ substitute, containing: a tissue engineered construct containing one or more biocolor compositions, where one or more biocolor compositions contain from 5 to 200 mg/ml of collagen; and a biocompatible support structure comprising one or more external supports, internal supports, or a combination thereof of a biocompatible material; wherein: the composition of the tissue or organ substitute has a three-dimensional 3D shape, where the tissue-engineered structure has the shape of an organ or tissue; And the biomaterial material is present in an amount of 1 to 100% by weight of the biocompatible support structure. 2. Composition according to claim 1, characterized in that one or more bio-paint compositions contain a hydrogel. 3. The composition according to p. 2, characterized in that one or more compositions of biocolors contain hyaluronic acid present in an amount of from 0.5 to 10% wt./about. 4. Composition according to claim 2, characterized in that one or more bio-paint compositions contain an alginate hydrogel present in an amount of 0.5. up to 10% w/v 5. Composition according to claim 1, characterized in that the collagen is selected from type I collagen, type III collagen, type IV collagen, other fibrillar collagens, or a combination of two or more of them. 6. The composition according to any one of paragraphs. 1-5, characterized in that the tissue engineering construct additionally includes a population of cells present at a concentration of 1.0x10 ⁵ to 5.0x10 ⁷ cells/ml. 7. The composition according to any one of paragraphs. 1-6, characterized in that one or more compositions of bioinks additionally contain a carrier, a crosslinking agent, or a combination thereof. 8. The composition according to any one of paragraphs. 1-7, characterized in that the tissue-engineering construct has the shape of an organ or tissue of the subject. 9. The composition according to any one of paragraphs. 1-8, characterized in that the organ or tissue is selected from the ear, nose, meniscus, articular cartilage or heart valve. 10. Composition according to claim 9, characterized in that said organ is an ear. 11. The composition according to any one of paragraphs. 1-10, characterized in that the biocompatible material is selected from the group consisting of polysaccharides, biocompatible polymers, rubber, silicon, biocompatible metals, biocompatible ceramics, polyethylene glycol, polypropylene glycol, polyamino acids, natural and biopolymers, or a combination of two or more of them. 12. Composition according to claim 11, characterized in that the biocompatible material is selected from the group consisting of PLA, PGA, PLGA, PDO, polycaprolactones, bioresorbable metal alloys, and combinations thereof. 13. Composition according to claim 12, characterized in that the biocompatible material is decomposed, absorbed or resorbed at a rate of 1 week to 7 years. 14. The composition according to any one of paragraphs. 11-13, characterized in that the biocompatible support structure contains one or more external supports and one or more internal supports. 15. The composition according to claim 14, characterized in that the outer support of one or more external supports contains a biocompatible material that is different from or is the same as the internal support of one or more internal supports. 16. The composition according to claim 15, characterized in that the inner support is decomposed, absorbed or resorbed at a rate that is the same as or different from the rate of the outer support. 17. The composition according to any one of paragraphs. 1-16, characterized in that the outer support has a 3D configuration to fully or partially cover one or more surfaces of the tissue engineered structure. 18. Composition according to any one of paragraphs. 1-17, characterized in that the outer support has one or more holes passing through one or more 2D surfaces of the outer support. 19. The composition according to any one of paragraphs. 1-18, characterized in that the internal support is fully or partially encapsulated in a tissue-engineered structure. 20. The composition according to any one of paragraphs. 1-19, characterized in that the inner support is in contact with one or more 2D surfaces of the outer support. 21. The composition according to any one of paragraphs. 1-20, characterized in that the inner support has a 2D or 3D shape. 22. The composition according to claim 21, characterized in that the internal support is completely or partially decomposed, absorbed or resorbed with the formation of one or more cavities, channels or combinations thereof in the tissue engineering structure. 23. The composition according to any one of paragraphs. 1-22, characterized in that the internal support is configured to fully or partially encapsulate the amount of bio-colors in the tissue engineering construct. 24. The composition according to any one of paragraphs. 1-23, characterized in that the outer support and the inner support have a 2D thickness of 10 to 2000 µm. 25. The composition according to any one of paragraphs. 1-24, characterized in that the biocompatible support structure has one or more of the following characteristics: a flexural modulus of 0.2 to 100 GPa, an elastic modulus of 0.02 to 100 GPa, or a tensile strength of 1 to 1000 MPa. 26. The composition according to any one of paragraphs. 1-25, characterized in that one or more external supports, one or more internal supports, or combinations thereof increase the rigidity of the fabric structure from 5 to 10,000 times. 27. The composition according to any one of paragraphs. 1-25, characterized in that one or more external supports, one or more internal supports, or combinations thereof increase the load capacity of the tissue engineering structure from 1 to 10,000%. 28. The composition according to any one of paragraphs. 1-27, characterized in that said composition withstands compression from 1 to 10,000 kN, shear force from 0.2 to 1,000 kN, or a combination thereof. 29. A method for manufacturing a composition of a tissue or organ substitute, including: production of a biocompatible support structure; fabrication of a tissue engineering structure containing: applying biocolors to or around a biocompatible support structure, wherein one or more biocolor compositions contain 5 to 200 mg/mL of collagen; crosslinking the bioinks and optionally repeating the application and crosslinking steps to form a tissue engineered construct; and curing the tissue or organ substitute composition; wherein: the composition of the substitute has a 3D shape, where the tissue engineering structure has the shape of an organ or tissue; the biocompatible support structure comprises one or more external supports, internal supports, or combinations thereof of a biocompatible material; and the biocompatible material is present in an amount of 1 to 100% by weight of the biocompatible support structure. 30. The method according to claim 29, characterized in that said method for manufacturing a biocompatible support structure includes one or more of the following processes: thermoforming, injection molding, rotational molding, positive molds, negative molds, subtractive manufacturing, milling and machining, 3D printing, and combinations thereof. 31. The method according to claim 29 or 30, characterized in that said method for manufacturing a biocompatible support structure includes: applying one or more layers of biocompatible material to the substrate; cross-linking of biocompatible material; optionally repeating the application and crosslinking steps to obtain one or more external supports, one or more internal supports, or combinations thereof; and separating the substrate from one or more external supports, one or more internal supports, or combinations thereof. 32. The method of claim. 31, characterized in that the application includes 3D printing one or more layers of biocompatible material. 33. The method of claim 32, wherein the 3D printing is selected from the group consisting of inkjet printing, layer printing, extrusion printing, and bioprinting. 34. The method according to claim 29 or 30, characterized in that said method for manufacturing a biocompatible support structure includes injection molding, while injection molding includes: applying one or more layers of biocompatible material to a substrate, wherein the substrate is a form of one or more external supports, internal supports, or combinations thereof; cross-linking the biocompatible material in the mold; and removing one or more external supports, internal supports, or combinations thereof from the mold. 35. The method according to claim 29 or 30, characterized in that the biocompatible material is in the form of a 2D sheet. 36. The method according to claim 35, characterized in that the 2D sheet is made from woven or knitted fibers. 37. The method according to claim 35 or 36, characterized in that the fibers have a diameter of from 10 to 2000 microns. 38. The method according to any one of paragraphs. 35-37, characterized in that the 2D sheet is a porous sheet. 39. The method according to any one of paragraphs. 35-37, characterized in that said method for manufacturing a biocompatible support structure includes thermoforming, wherein thermoforming includes: contacting a 2D sheet of biocompatible material with a substrate, wherein the substrate is in the form of one or more external supports, internal supports, or combinations thereof; heating the 2D sheet of biocompatible material to thermoform the 2D sheet according to the shape of the mold to form one or more external supports, internal supports, or combinations thereof; removing one or more external supports, internal supports, or combinations thereof from the mold. 40. The method according to any one of paragraphs. 29-39, characterized in that the outer support has one or more holes passing through one or more 2D surfaces of one or more external supports, internal supports, or combinations thereof. 41. The method according to any one of paragraphs. 29-40, characterized in that the biocompatible material is selected from the group consisting of polysaccharides, biocompatible polymers, rubber, silicon, biocompatible metals, biocompatible ceramics, polyethylene glycol, polypropylene glycol, polyamino acids, natural and biopolymers, or a combination of two or more of them. 42. The method of claim 41, wherein the biocompatible material is a biocompatible polymer selected from the group consisting of PLA, PGA, PLGA, PDO, polycaprolactones, and combinations thereof. 43. The method according to p. 42, characterized in that the biocompatible polymer has a melting point from 50 to 290°C. 44. The method according to any one of paragraphs. 29-43, characterized in that the outer support and the inner support have a 2D thickness of 100 to 1200 µm. 45. The method according to any one of paragraphs. 29-43, characterized in that said method for manufacturing a tissue engineered structure includes one or more of the following processes: injection molding, rotational molding, positive molds, negative molds, subtractive manufacturing, milling and machining, and three-dimensional 3D printing. 46. The method according to claim 45, characterized in that the application includes 3D printing of one or more layers of bioinks. 47. The method of claim 46, wherein the 3D printing is selected from the group consisting of inkjet printing, layer printing, extrusion printing, and bioprinting. 48. The method according to claim 45, characterized in that said method for manufacturing a tissue engineering structure includes injection molding, while injection molding includes: applying bio-colors to a mold, said mold containing a biocompatible support structure; crosslinking the bioinks in a mold and optionally repeating the application and crosslinking steps to obtain a tissue or organ substitute composition; removing the tissue or organ substitute composition from the mold, wherein the curing step occurs before removal from the mold, after removal from the mold, or a combination thereof. 49. The method according to any one of paragraphs. 29-48, characterized in that this method provides for the manufacture of a biocompatible base and tissue engineering construct sequentially or simultaneously. 50. The method according to any one of paragraphs. 29-49, characterized in that the composition of the tissue or organ substitute is cured at a temperature of 4 to 50°C. 51. The method according to any one of paragraphs. 29-50, characterized in that the bio-paint contains a hydrogel. 52. The method according to p. 51, characterized in that biopaint contains alginate hydrogel present in an amount of from 0.5 to 10% wt./about. 53. The method according to any one of paragraphs. 29-52, characterized in that said collagen is selected from type I collagen, type II collagen, or a combination thereof. 54. The method according to any one of paragraphs. 29-53, characterized in that the biostain additionally contains a population of cells present at a concentration of 1.0x10 ⁵ to 5.0x10 ⁷ cells/ml. 55. The method according to any one of paragraphs. 29-53, characterized in that the tissue engineering construct has the shape of an organ or tissue of the subject. 56. The method according to any one of paragraphs. 29-55, characterized in that the organ or tissue is selected from the ear, nose, meniscus, articular cartilage or heart valve. 57. The method according to any one of paragraphs. 29-56, characterized in that said organ is an ear.

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