Tissue-conductive scaffolding materials

Abstract

Disclosed herein is a bioactive polymer for forming a tissue scaffold, the polymer comprising a first monomer for binding water, a second monomer for imparting mechanical properties to the scaffold, and optionally a third monomer for binding to a natural or synthetic peptide or protein (NSPP), and a fourth monomer for imparting phase transition behavior, the scaffold forming a malleable structure upon hydration. Preferably, the first monomer is OEGMA, the second monomer is PLA / HEMA, the third monomer is NAS, and the fourth monomer is NIPAAm, the polymer comprising OEGMA in an amount of about 1 to about 15 mol%, PLA / HEMA in an amount of 5 to about 50 mol%, NAS in an amount of 0 to about 15 mol%, and NIPAAm in an amount of up to about 85 mol%.

Description

[Technical field]

[0001] Related Applications This application claims convention priority from Australian Provisional Patent Application 2022901056, filed April 21, 2022. The disclosure of AU'056 is incorporated herein by reference in its entirety.

[0002] The present invention relates to biocompatible polymers, particularly polymers useful in the formation of tissue scaffolds, which may be useful in the repair and regeneration of tissue.

[0003] The present invention relates to a prefabricated porous scaffold that can guide tissue integration, tissue interface regeneration and drug delivery applications. One embodiment of the present invention relates to the formation of a malleable scaffold that can be administered arthroscopically or by open surgical intervention. Upon contact with body tissue, the scaffold is adhesive and can be sutured, thereby allowing the immobilization of one or more tissues to support tissue regeneration and / or tissue interface integration.

[0004] In another embodiment, the present invention relates to a tissue-conductive medical filler. In a further embodiment, the polymer of the present invention can form a tissue scaffold. In yet another embodiment, the scaffold forms a malleable structure upon hydration. In yet another embodiment, the scaffold can form in situ when the temperature in the body increases after administration.

[0005] It is envisaged that the present invention will be useful for tissue engineering applications, including both cosmetic and therapeutic applications. The present invention will now be described with reference to preferred embodiments thereof, but those skilled in the art will appreciate that the spirit and scope of the invention may be embodied in many other forms. [Background technology]

[0006] Any discussion of prior art throughout this specification should in no way be taken as an admission that such prior art is widely known or forms part of the common general knowledge in the art.

[0007] Tissue engineering is a biomedical engineering field that uses a combination of cells, engineering materials, methods, and appropriate biochemical and physicochemical factors to restore, maintain, improve, or replace different types of living tissues. Tissue engineering often involves the use of cells arranged on tissue scaffolds in the formation of new living tissues for medical purposes, but is not limited to applications involving cells and tissue scaffolds. In practice, the term is closely related to applications that repair or replace tissues in part or in whole (e.g., bone, cartilage, blood vessels, bladder, skin, muscle, etc.). In many cases, the tissues involved require specific mechanical and structural properties for proper function. The term also relates to the use of cells within an artificially created support system (e.g., an artificial pancreas, or a bioartificial liver) to perform specific biochemical functions.

[0008] Scaffolds are materials engineered to induce desirable cellular interactions to contribute to the formation of new functional tissues for medical purposes. Cells are often seeded into these structures that can support three-dimensional tissue formation. Scaffolds mimic the extracellular matrix of native tissues, recapitulating the in vivo environment and allowing cells to influence their own microenvironment. They usually serve at least one of the following purposes: allow cell attachment and migration, deliver and retain cellular and biochemical factors, allow the diffusion of important cellular nutrients and expression products, and / or exert specific mechanical and biological influences to modify the behavior of the cellular phase.

[0009] Material selection is an essential aspect for creating scaffolds. The materials utilized can be natural or synthetic and can be biodegradable or non-biodegradable. Furthermore, they must be biocompatible, meaning that they do not cause any adverse effects to the cells. For example, silicone is a synthetic non-biodegradable material commonly used as a drug delivery material, while gelatin is a biodegradable natural material commonly used for cell culture scaffolds. The optimal material for each application will inevitably be different and will depend on the desired mechanical properties of the material. For example, tissue engineering of long bone defects requires a rigid scaffold with a compressive strength similar to cortical bone (100-150 MPa), which is much higher compared to scaffolds for skin regeneration.

[0010] There are several versatile synthetic materials that are used for many different scaffolding applications. One commonly used material is polylactic acid (PLA), a synthetic polymer. PLA is a polyester that degrades in the human body to form lactic acid, a naturally occurring chemical that is easily removed from the body. Similar materials are polyglycolic acid (PGA) and polycaprolactone (PCL). PLA is commonly combined with PGA to create polylactic-co-glycolic acid (PLGA). This is particularly useful because the degradation of PLGA can be tuned by changing the weight percentage of PLA and PGA. This tunability, along with its biocompatibility, makes PLGA an extremely useful material for scaffold fabrication.

[0011] Scaffolds may also be constructed from natural materials. Protein-based materials such as collagen or fibrin, and polysaccharide materials such as chitosan or glycosaminoglycans (GAGs) have all proven suitable in terms of cytocompatibility. Among the GAGs, hyaluronic acid, possibly combined with crosslinkers (e.g., glutaraldehyde, water-soluble carbodiimides, etc.), is a commonly used scaffold material. In addition, fragments of extracellular matrix proteins, such as RGD peptides, can be attached to non-bioactive materials to promote cell attachment. Another form of scaffold is decellularized tissue, which results from the chemical extraction of cells from tissue, leaving only the extracellular matrix. This has the advantage of a fully formed matrix specific to the desired tissue type. However, decellularized scaffolds may present immune problems to cells introduced in the future.

[0012] All patents and patent publications mentioned herein are incorporated by reference in their entirety.

[0013] International Publication No. 2013 / 091001 (WO'001) (PCT / AU2012 / 001566) relates to polymers, particularly polymers useful as hydrogels, and the use of hydrogels for the repair or restoration of tissue. In particular, the polymers and hydrogels of WO'001 can be used to repair or restore cartilage, particularly articular cartilage. The polymers include at least monomers for binding water, monomers for imparting mechanical properties, and monomers for binding extracellular proteins. Hydrogels include polymers that include at least monomers for binding water and monomers for binding extracellular proteins. Crosslinking polymers by binding extracellular matrix proteins forms hydrogels.

[0014] International Publication No. 2017 / 035587 (WO'587) (PCT / AU2016 / 050817) discloses biocompatible materials useful for tissue regeneration and repair, where the bioactive polymer can be in the form of a hydrogel, e.g., a thermoresponsive hydrogel. The bioactive polymers and resulting hydrogels of WO'587 can be used to regenerate bone tissue. Thus, this reference teaches a method of treating a bone defect in a mammal, comprising administering to the mammal a therapeutically effective amount of a hydrogel formed by a bioactive polymer to treat the bone defect.

[0015] International Publication No. 2017 / 015703 (WO'703) (PCT / AU2016 / 050653) discloses polymers comprising at least one antiseptic / analgesic / anti-inflammatory monomeric unit together with at least three additional monomeric units, the three additional monomeric units inducing a property selected from the group consisting of temperature activation, water solubility, mechanical strength, protein / polysaccharide binding ability, and combinations thereof. In particular, WO'703 discloses polymers in which the water-soluble monomeric unit is a hydrophilic ethylene glycol (OEGMA) moiety, the mechanical strength-imparting monomeric unit is a polylactide-co-2-hydroxy-ethylmethylacrylate (PLA / HEMA), the protein-reactive monomeric unit is an N-acryloxysuccinimide (NAS) moiety, and the thermosetting monomeric unit is an N-isopropylacrylamide (NIPAAm) moiety. The antiseptic / analgesic / anti-inflammatory monomeric unit comprises a methacrylate ester derivative of salicylic acid (5-HMA or 4-HMA, or a combination thereof).

[0016] International Publication No. 2021 / 119727 (WO'727) (PCT / AU2020 / 051332) teaches a composition comprising a polymer and a natural or synthetic peptide or protein (NSPP) as thymosin beta-4. The polymer comprises a first monomer for binding water, a second monomer for imparting mechanical properties, a third monomer for binding to the NSPP, and a fourth monomer for imparting phase transition behavior. In particular, the composition forms an adhesive and flowable hydrogel upon administration into or onto the body, thereby aiding in tissue repair and regeneration. Thus, WO'727 discloses a method of tissue repair and / or regeneration, which comprises administering the composition by injection or administration of an aerosol, thereby forming a hydrogel at the body temperature of the mammal.

[0017] All four applications referred to above are assigned to the applicant, Trimph IP Pty Ltd, Sydney, Australia.

[0018] It is an object of the present invention to overcome or ameliorate at least one of the disadvantages of the prior art, or to provide a useful alternative.

[0019] There is a general need in the art for new scaffolds for tissue repair and / or regeneration. There is a general need in the art for new scaffolds that allow for regeneration and integration of tissue interfaces. There is a general need in the art for new scaffolds that allow for tissue ingrowth as opposed to growing surface tissue. There is a general need for scaffolds for tissue repair and / or regeneration that negate the criticality of water solubility. There is a general need for tissue scaffolds that can be manufactured by using different processing methods. There is a general need for tissue scaffolds that can be tailored to modify their microenvironment to address requirements. There is a general need for tissue scaffolds that can be bound to peptides and / or proteins. There is a general need for tissue scaffolds that can be bound to different hydrophilic and hydrophobic drugs. There is a general need for glove-non-adherent tissue scaffolds for effective manipulation. There is a general need for malleable tissue scaffolds that allow for effective delivery to the site via arthroscopic or open surgical intervention. There is a general need for tissue scaffolds that adhere to the treatment site. There is a general need for sutureable (or stapled) tissue scaffolds for internal fixation.

[0020] It is against this background that the present invention has been developed, the various embodiments of which may find utility in relation to one or more of the general needs identified above.

[0021] In particular, the present invention is useful in soft tissue applications, such as skin grafting to secure graft tissue, preparing sites for future skin grafts, healing and / or strengthening and / or reinforcing and / or stabilizing tendons and / or ligaments, shoulder, knee and hip arthroplasty, repair of partial or full thickness rotator cuff tears (including but not limited to superior capsular reconstruction (SCR)), in cruciate ligaments (including anterior cruciate ligament, posterior cruciate ligament and medial cruciate ligament, ACL, PCL, MCL) and / or other ligament / tendon ruptures (including but not limited to Achilles ligament, biceps tendon, patellar tendon), and hard tissue applications. The present invention may also be useful in drug delivery applications.

[0022] Although the invention will be described with reference to specific examples, those skilled in the art will appreciate that the invention can be embodied in many other forms. Summary of the Invention

[0023] In a broad form, the invention is embodied as a filler material, which upon hydration forms a malleable scaffold. The scaffold is well tolerated in the body with minimal inflammatory response and can be used as a general matrix to support tissue healing and integration of two or more tissue interfaces. The scaffold is host tissue conductive but not inductive. The filler material can be used during surgery to hydrate with saline, the patient's own blood, platelet reach plasma (PRP), platelet reach fibrin, bone marrow aspirate, and / or other blood / cell / tissue products and / or extracts (from autologous and allogeneic sources) to form a malleable scaffold. The resulting scaffold can be administered arthroscopically or by open surgical intervention. Upon contact with body tissue, the scaffold is adhesive and suitable for suturing and surgical stapling, thereby allowing fixation of one or more tissues to support tissue regeneration and / or tissue interface integration.

[0024] According to a first aspect of the present invention there is provided a polymer for forming a tissue scaffold, comprising:

[0025] a first monomer for binding water;

[0026] a second monomer to impart mechanical properties to the scaffold;

[0027] Optionally, a third monomer for attachment to a natural or synthetic peptide or protein (NSPP);

[0028] a fourth monomer to impart phase transition behavior;

[0029] The scaffold is provided with a polymer that upon hydration forms a malleable structure.

[0030] In one embodiment, the first monomer is selected from polyethers, polyvinyl alcohol (PVA), poly(vinylpyrrolidone) (PVP), poly(amino acids), and dextran.

[0031] In one embodiment, the polyether is selected from polyethylene glycol (PEG), oligo(ethylene glycol) (OEG), polyethylene oxide (PEO), polyethylene oxide-co-propylene oxide (PPO), co-polyethylene oxide block copolymers or random copolymers thereof.

[0032] In one embodiment, the first monomer is oligo(ethylene)glycol monomethyl ether methacrylate (OEGMA).

[0033] In one embodiment, the second monomer is a methacrylate or a random copolymer that includes a methacrylate.

[0034] In one embodiment, the second monomer is selected from hydroxyethyl methacrylate (HEMA), hydroxyethyl methacrylate poly(lactic acid) copolymer (PLA / HEMA), poly(lactic acid), poly(caprolactone), poly(glycolide), poly(glycolide-co-lactide), or poly(glycolide-co-caprolactone).

[0035] In one embodiment, the second monomer is hydroxyethyl methacrylate poly(lactic acid) (PLA / HEMA).

[0036] In one embodiment, the third monomer has an electrophilic functional group for attachment to the NSPP.

[0037] In one embodiment, the third monomer is selected from N-hydroxysulfosuccinimide (SNHS), N-hydroxyethoxylated succinimide (ENHS) and N-acryloxysuccinimide (NAS).

[0038] In one embodiment, the third monomer is N-acryloxysuccinimide (NAS).

[0039] In one embodiment, the fourth monomer has a lower critical solution temperature (LCST) of less than about 37°C.

[0040] In one embodiment, the fourth monomer is selected from poly(ethylene oxide) / poly(propylene oxide) and poly(N-isopropylacrylamide) (PNIPAAm) homopolymers and copolymers.

[0041] In one embodiment, the fourth monomer is (N-isopropylacrylamide) (NIPAAm).

[0042] In one embodiment, the polymer comprises the first monomer in an amount of about 1 to about 15 mole percent.

[0043] In one embodiment, the polymer comprises the second monomer in an amount of about 5 to about 50 mole percent.

[0044] In one embodiment, the polymer comprises a third monomer in an amount of about 0 to about 15 mole %.

[0045] In one embodiment, the polymer comprises the fourth monomer in an amount of about 50 to about 85 mole percent.

[0046] In one embodiment, the polymer comprises a first monomer in an amount of about 1 to about 15 mol %, a second monomer in an amount of about 5 to about 50 mol %, a third monomer in an amount of 0 to about 15 mol %, and a fourth monomer in an amount that makes up 100% of the remainder of the polymer.

[0047] In one embodiment, the first monomer is OEGMA, the second monomer is PLA / HEMA, the third monomer is NAS, and the fourth monomer is NIPAAm, and the polymer comprises OEGMA in an amount from about 1 to about 15 mol %, PLA / HEMA in an amount from 5 to about 50 mol %, NAS in an amount from 0 to about 15 mol %, and NIPAAm in an amount up to about 85 mol %.

[0048] According to a second aspect of the present invention there is provided a tissue scaffold comprising a polymer defined according to the first aspect of the present invention, the scaffold forming a malleable structure upon hydration.

[0049] In one embodiment, the scaffold is hydrated during surgery, hi another embodiment, the scaffold is hydrated with saline, an aqueous solution, autologous or allogeneic blood, cell or tissue products, or a combination thereof.

[0050] In one embodiment, the scaffold is hydrated with the patient's own blood, platelet reach plasma (PRP), bone marrow aspirate, platelet reach fibrin, or other blood or tissue derived products or combinations thereof.

[0051] In one embodiment, the scaffold is hydrated with allogeneic blood, platelet reach plasma (PRP), platelet reach fibrin, or other blood or tissue derived products, or combinations thereof.

[0052] In one embodiment, the scaffold is non-adherent to surgical gloves, hi one embodiment, the scaffold is adhesive to the treatment site.

[0053] In one embodiment, the scaffold does not have tissue inductive properties.

[0054] In one embodiment, the scaffold is formed following administration to a mammal.

[0055] In one embodiment, the scaffold is formed at body temperature.

[0056] In one embodiment, administration is by injection or spray.

[0057] According to a third aspect of the present invention there is provided a malleable structure formed from a tissue scaffold as defined by the second aspect of the present invention.

[0058] According to a fourth aspect of the present invention there is provided a method of producing a tissue scaffold, the method comprising the steps of:

[0059] dissolving an aqueous solution of a polymer according to the first aspect of the invention in a buffer solution to obtain a resulting solution;

[0060] and freeze-drying the resulting solution.

[0061] The scaffolds have tunable morphology by adjusting the lyophilization parameters and concentration of the polymer.

[0062] In one embodiment, at least one drug or biological moiety is added to the resulting solution between the dissolving and lyophilization steps.

[0063] In one embodiment, the drug or biological moiety is selected from an antibiotic, a growth factor, a live virus, or a combination thereof.

[0064] According to a fifth aspect of the present invention there is provided a method of producing a tissue scaffold, the method comprising the steps of: - dissolving the polymer according to the first aspect of the invention in an organic solvent to obtain a solution; Precipitating the solution in water as an antisolvent to obtain a gel; and freeze-drying the gel, The scaffolds have alterable morphology by adjusting the freeze-drying parameters.

[0065] In one embodiment, the organic solvent is miscible in water.

[0066] According to a sixth aspect of the present invention there is provided a method of producing a tissue scaffold, the method comprising the steps of:

[0067] - dissolving the polymer according to the first aspect of the invention in an organic solvent to obtain a solution;

[0068] and electrospinning the solution.

[0069] According to a seventh aspect of the present invention there is provided a method of producing a tissue scaffold, the method comprising the steps of:

[0070] comprising dissolving a polymer as defined according to the first aspect of the present invention in an aqueous solution, autologous blood, a cell or tissue product, or a combination thereof;

[0071] The scaffold is injectable or sprayable.

[0072] According to an eighth aspect of the present invention, there is provided a method for treating a rotator cuff injury, comprising the steps of: skin grafting for immobilizing graft tissue; preparing the site for future skin grafts; healing and / or strengthening and / or reinforcing and / or stabilizing tendons and / or ligaments; repairing partial or full thickness rotator cuff tears; There is provided the use of a polymer according to the first aspect of the invention in the manufacture of a tissue scaffold for soft tissue healing in repair, shoulder, knee and hip arthroplasty, cruciate ligament and / or other ligament / tendon ruptures.

[0073] According to a ninth aspect of the present invention there is provided the use of a tissue scaffold according to the second aspect of the present invention for the repair and / or regeneration of tissue.

[0074] According to a tenth aspect of the present invention there is provided a method for tissue repair and / or regeneration; supporting skin grafts for anchoring graft tissue; preparing the site for future skin grafts; healing and / or strengthening and / or reinforcing and / or stabilising tendons and / or ligaments; repair of partial or full thickness rotator cuff tears; soft tissue healing in shoulder, knee and hip arthroplasties, cruciate ligaments and / or other ligaments / tendons, the method comprising administering to a mammal a tissue scaffold according to the second aspect of the present invention.

[0075] In one embodiment, administration is by injection or spray. In one embodiment, the scaffold is administered arthroscopically or by open surgical intervention. In one embodiment, the scaffold hydrates to form a malleable structure upon administration during surgery.

[0076] According to an eleventh aspect of the present invention there is provided a tissue scaffold according to the second aspect of the present invention for use in soft tissue healing in tissue repair and / or regeneration; supporting skin grafts for anchoring graft tissue; preparing the site for future skin grafts; healing and / or strengthening and / or reinforcing and / or stabilising tendons and / or ligaments; repair of partial or full thickness rotator cuff tears; shoulder, knee and hip arthroplasty, cruciate and / or other ligament / tendon ruptures.

[0077] In one embodiment, the scaffold is administered by injection or spray. In one embodiment, the scaffold is administered arthroscopically or by open surgical intervention. In one embodiment, the scaffold hydrates to form a malleable structure upon administration during surgery.

[0078] According to a twelfth aspect of the present invention there is provided a kit for forming a tissue scaffold comprising a polymer, the polymer comprising a first monomer for binding water, a second monomer for imparting mechanical properties to the scaffold, optionally a third monomer for binding to a natural or synthetic peptide or protein (NSPP) and a fourth monomer for imparting phase transition behaviour, wherein the scaffold forms a malleable structure upon hydration.

[0079] Definitions and Nomenclature In describing and claiming the present invention, the following terms are used in accordance with the definitions set forth below. It should also be understood that the terms used herein are for the purpose of describing particular embodiments of the present invention only, and are not intended to be limiting. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which the present invention pertains.

[0080] Unless the context clearly requires otherwise, throughout the description and claims, the words "comprise," "comprising," and the like are intended to be construed in their inclusive sense, i.e., "including, but not limited to," rather than in their exclusive or exhaustive sense.

[0081] The terms "preferred" and "preferably" refer to embodiments of the invention that may provide certain benefits, under certain circumstances. However, other embodiments may also be preferred, under the same or other circumstances. Furthermore, the recitation of one or more preferred embodiments does not imply that other embodiments are not useful, and is not intended to exclude other embodiments from the scope of the invention.

[0082] As used herein, phrases defining ranges or length limits, such as, for example, "1-5," refer to any integer between 1 and 5, i.e., 1, 2, 3, 4, and 5. In other words, any range defined by two integers explicitly recited is meant to include and disclose any integers defining the limits and any integers falling within that range.

[0083] Except in the operating examples or where otherwise indicated, all numbers expressing amounts of ingredients or reaction conditions used herein should be understood in all cases to be modified by the term "about". The examples are not intended to limit the scope of the invention. Hereinafter, or where otherwise indicated, "%" means "% by weight", "ratio" means "ratio by weight", and "parts" means "parts by weight".

[0084] Notwithstanding that the numerical ranges and parameters setting forth the broad scope of the invention are approximations, the numerical values ​​set forth in the specific examples are reported as precisely as possible, however, any numerical values ​​inherently contain certain errors necessarily resulting from the standard variation found in their respective testing measurements.

[0085] The following abbreviations are used herein: ECM Extracellular Matrix EHNS N-Hydroxyethoxylated Succinimide HEMA Hydroxyethyl Methacrylate LA Lactic Acid NAS N-acryloxysuccinimide NIPAAm N-isopropylacrylamide NSPP Natural or synthetic peptides or proteins OEG Oligo(ethylene glycol) OEGMA Oligo(ethylene)glycol monomethyl ether methacrylate PBS Phosphate Buffered Saline PEG Polyethylene glycol PEO Polyethylene Oxide PLA Poly(lactic acid) PLA / HEMA Hydroxyethyl Methacrylate Poly(lactic acid) PPO Polyethylene oxide-co-propylene oxide PVA Polyvinyl Alcohol PVP Poly(vinylpyrrolidone) PNPHO Poly(N-isopropylacrylamide-co-(N-acryloxysuccinimide)-co-(polylactide / 2-hydroxymethacrylate)-co-(oligo(ethylene glycol) / poly(NIPAAm-co-NAS-co-(PLA / HEMA)-co-OEGMA) PPHO Poly(N-isopropylacrylamide-co-(polylactide / 2-hydroxymethacrylate)-co-(oligo(ethylene glycol) / poly(NIPAAm-co-(PLA / HEMA)-co-OEGMA) SNHS N-Hydroxysulfosuccinimide TB4 Thymosin beta-4 or thymosin beta-4

[0086] Preferred embodiments of the present invention will now be described, by way of example only, with reference to the accompanying drawings, in which: [Brief description of the drawings]

[0087] [Figure 1(a)] 1 shows the surface of a PNPHO scaffold formed from a polymer of the present invention. [Figure 1(b)] The morphology is shown at a resolution of 100 microns. [Figure 1(c)] The morphology is shown at a resolution of 50 microns. [Figure 1(d)] The morphology is shown at a resolution of 20 microns. [Diagram 2] Handling of the PNPHO scaffold after hydration is shown, demonstrating the structural stability of the product during handling for effective manipulation and surgical administration. Sequential steps in defolding / unfolding the scaffold are shown. [Figure 3(a)]Figure 2 shows adhesion test of PNPHO scaffold onto soft tissue (epidermis). [Figure 3(b)] 1 shows a peel test of a PNPHO scaffold from soft tissue (epidermis). [Figure 4(a)] FIG. 1 shows adhesion testing of PNPHO scaffolds onto hard surfaces (simulating hard tissue, e.g. bone). [Figure 4(b)] 1 shows peel testing of PNPHO scaffolds onto hard surfaces (simulating hard tissue, e.g. bone). [Figure 5(a)] Figure 1 shows the H NMR spectrum of PPHO (NAS = 0 mol %) in CD3CN. The presence of characteristic monomer peaks confirmed the incorporation of PPHO. [Figure 5(b)] Comparative 1H NMR spectra of PNPHO and PPHO are shown. (a) 1H NMR spectrum of PPHO (NAS = 0 mol%), the presence of characteristic peaks confirms the incorporation of PPHO (NIPAAm = ~90 mol%, PLA / HEMA = ~10 mol% and OEGMA = ~1 mol%), and (b) 1H NMR spectrum of PNPHO (NAS = 10 mol%, OEGMA = 5 mol%). PPHO 1H NMR spectra were collected using CD3CN and PNPHO 1H NMR spectra were collected using CDCl3. The change in solvent required for NMR data acquisition was due to the hydrophobicity of PPHO preventing its dissolution in CDCl3. [Figure 5(c)] HPLC chromatograms at 230 nm showing PNPHO eluting at 6.2 minutes and PPHO eluting at 7.2 minutes. The difference in elution times between them confirms that PPHO elutes at a higher percentage of acetonitrile compared to PNPHO, further confirming the hydrophobicity of PPHO compared to PNPHO. [Figure 6(a)] Despite its inherent hydrophobicity, PPHO adheres to and exhibits adhesive properties on soft tissue. [Figure 6(b)] PPHO adhesion on hard surfaces to demonstrate adhesion of the product on simulated hard tissue. [Figure 7] Body weight changes over 42 days in mice implanted with PNPHO and / or PPHO scaffolds are shown (time point = days after implantation). [Figure 8] Digital images of macroscopic observations at various time points over a 42 day period are shown. Note that hair regrowth on the back of the mice does not correlate with the presence of the scaffold. This is a normal regrowth pattern and was generally consistent across all mice. [Figure 9] Digital images of mice sacrificed on days 7, 14, 21 and 42 after surgery, respectively, are shown. [Figure 10] Shown is an H&E image of skin histology of mouse ID "S-4" at 1 week. [Figure 11] H&E images of the skin implantation sites 1, 2, 3 and 6 weeks after surgery are shown. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0088] Detailed Description of the Preferred Embodiments The present invention will now be described more fully with reference to the accompanying examples and drawings, however it should be understood that the following description is illustrative only and should not be construed in any way as a restriction on the generality of the invention described above.

[0089] Reference will now be made in detail to certain specific embodiments of the invention. While the invention will be described in conjunction with the embodiments, it should be understood that the invention is not limited to these embodiments. On the contrary, the invention is intended to cover all alternatives, modifications, and equivalents which may be included within the scope of the present invention as defined by the claims.

[0090] One skilled in the art will recognize many methods and materials similar or equivalent to those described herein, which could be used in the practice of the present invention, and the present invention is in no way limited to the methods and materials described.

[0091] It will be understood that the invention disclosed and defined herein extends to all alternative combinations of two or more of the individual features mentioned or apparent from the text or drawings, all of which different combinations constitute various alternative embodiments of the invention.

[0092] Broadly disclosed herein is the use of tissue tailored polymers, preferably scaffolds formed from polymers that form malleable structures upon hydration.

[0093] polymer As used herein, the term "polymer" refers to a large molecule (macromolecule) composed of repeating structural units (monomers). These subunits are typically linked by covalent chemical bonds. The polymer may be a linear or branched polymer. Preferably, the polymer of the present invention is a copolymer comprising three or more different monomers. For example, in one embodiment, the polymer of the present invention comprises a first water-binding monomer, a second monomer capable of imparting mechanical properties to the tissue scaffold, and a third monomer having a functional group for binding to the NSPP.

[0094] As used herein, the term "monomer" refers to a structural unit that can combine to form a polymer, but may itself also be a polymer, or a derivative of a monomer or polymer. This type of monomer is also referred to herein as a "macromonomer." As used herein, a "macromonomer" is a polymer or oligomer, each of which has one end group that acts as a monomer molecule, such that each polymer or oligomer molecule contributes only a single monomer unit to the product polymer chain.

[0095] The polymers of the invention comprise a first monomer for binding water, a second monomer for imparting mechanical properties to the tissue scaffold, an optional third monomer for binding to a natural or synthetic peptide or protein (NSPP), and a fourth monomer for imparting phase transition behavior.

[0096] First monomer: Water-binding monomer As discussed above, the advantages of the tissue scaffold of the present invention may be at least partially attributed to the specific components that make up the polymer of the present invention.The particularly advantageous property of the polymer of the present invention is their water binding capacity.The presence of water within the scaffold of the present invention provides an environment that resembles both the natural environment of damaged tissue (which aids in tissue regeneration) and the necessary compression resistance for the scaffold.

[0097] Thus, preferred polymers for use herein should contain monomers or units capable of binding water in such a capacity that the polymer can form malleable structures when hydrated. In addition, the structures thus formed should have the necessary compression resistance and elasticity.

[0098] Those skilled in the art will understand that the water-binding monomer must be present in the polymer of the present invention in a sufficient proportion to produce a polymer that meets these requirements. In general, the proportion of water-binding monomer in the polymer is about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:10, about 1:20, about 1:30, about 1:40, about 1:50 molar ratio of water-binding monomer:mechanical strength monomer. In fact, the water-binding monomer should not only make the polymer hydrophilic, but also give the polymer a much greater water-binding capacity. Thus, the polymer according to the present invention has a water-binding capacity of about 70% to about 500%, about 80% to about 400%, about 90% to about 300% or about 100% to about 200%. For example, the water binding capacity of the polymers of the present invention may be about 70%, about 80%, about 90%, about 100%, about 110%, about 120%, about 130%, about 140%, about 150%, about 160%, about 170%, about 180%, about 190%, about 200%, about 210%, about 220%, about 230%, about 240%, about 250%, about 260%, about 270%, about 280%, about 290%, about 300%, about 310%, about 320%, about 330%, about 340%, about 350%, about 360%, about 370%, about 380%, about 390%, about 400%, about 410%, about 420%, about 430%, about 440%, about 450%, about 460%, about 470%, about 480%, about 490%, about 500%, about 510%, about 520%, about 530%, about 540%, about 550%, about 560%, about 570%, about 580%, about 590%, about 600%, about 610%, about 620%, about 630%, about 640%, about 650%, about 660%, about 670%, about 680%, about 690%, about 700%, about 710%, about 720%, about 730%, about 740%, about 750%, about 760%, about 770%, about 780%, about 790%, about 800%, about 810%, about 820%, about 830%, about 840%, about 850%, about 860%, about 870%, about 880%, about 890 about 280%, about 290%, about 300%, about 310%, about 320%, about 330%, about 340%, about 350%, about 360%, about 370%, about 380%, about 390%, about 400%, about 410%, about 420%, about 430%, about 440%, about 450%, about 460%, about 470%, about 480%, about 490%, or about 500%.

[0099] Suitable examples of water-binding monomers include those that can be synthesized into polymers such as polyethers (e.g., alkylene polyoxides such as polyethylene glycol (PEG), oligo(ethylene glycol) (OEG), polyethylene oxide (PEO), polyethylene oxide-co-propylene oxide (PPO), co-polyethylene oxide block or random copolymers, polyvinyl alcohol (PVA)), poly(vinylpyrrolidinone) (PVP), poly(amino acids) and dextran. Polyethers, and more particularly oligo(oxyalkylenes) (e.g., OEG), are particularly preferred because they have the requisite water-binding capacity, are simple to synthesize and / or purchase, and are inert, in the sense that they elicit minimal or no immune response from tissues in which they are placed.

[0100] Additionally, any of a variety of hydrophilic functional groups can be used to render the monomers (and thus the polymers formed from such monomers) water soluble. For example, functional groups such as phosphate, sulfate, quaternary amine, hydroxyl, amine, sulfonate, and carboxylate, which are water soluble, may be incorporated into the monomers to render them water soluble.

[0101] Monomers may also react with other compounds to form "macromonomers." Thus, the first monomer may optionally be a macromonomer.

[0102] A preferred first monomer, which is a macromonomer, is oligo(ethylene glycol) monomethyl ether methacrylate (OEGMA), which is a hydrophilic monomer composed of two hydrophilic monomers: ethylene glycol and methacrylate.

[0103] Preferably, the polymer comprises the first monomer in an amount of about 1 to about 15 mol %. In various embodiments, the first monomer may be present at 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, or about 15 mol %. In various embodiments, the first monomer may be present at about 1 to about 15 mol %, about 2 to about 14 mol %, about 3 to about 13 mol %, about 4 to about 12 mol %, about 5 to about 11 mol %, about 6 to about 10, about 7 to about 9 mol %, or about 8 mol %.

[0104] Second monomer: Monomer that imparts mechanical properties As discussed above, the advantageous properties of the tissue scaffolds of the invention may be due, in part, to the particular components that make up the polymers of the invention, which in some embodiments can contribute additional mechanical and adhesive properties to the scaffolds of the invention.

[0105] One skilled in the art will appreciate that monomers capable of imparting mechanical properties to the tissue scaffold should be present in the polymer of the invention in a proportion sufficient to produce a tissue scaffold having the desired mechanical properties. In general, the proportion of "mechanical" monomers in the polymer is about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:10, about 1:20, about 1:30, about 1:40, about 1:50 molar ratio of water binding monomer to "mechanical strength" monomer. Suitable examples of monomers capable of imparting mechanical properties (e.g., compression resistance) to the scaffold include acrylates such as hydroxyethyl methacrylate (HEMA), polyesters such as poly(lactic acid), poly(caprolactone), poly(glycolide), and random copolymers thereof (e.g., poly(glycolide-co-lactide) and poly(glycolide-co-caprolactone)).

[0106] Monomers may also react with other compounds to form “macromonomers.” A preferred second monomer that is a macromonomer is hydroxyethyl methacrylate poly(lactic acid) (PLA / HEMA).

[0107] Preferably, the polymer comprises the second monomer in an amount of about 1 to about 50 mole %. In various embodiments, the second monomer can be present at 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 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, or about 50 mole %. In various embodiments, the second monomer may be present at about 1 to about 15, about 2 to about 49, about 3 to about 48, about 4 to about 47, about 5 to about 46, about 6 to about 45, about 7 to about 44, about 8 to about 43, about 9 to about 42, about 10 to about 41, about 11 to about 40, about 12 to about 39, about 13 to about 38, about 14 to about 37, about 15 to about 36, about 16 to about 35, about 17 to about 34, about 18 to about 33, about 19 to about 34, about 20 to about 33, about 21 to about 30, about 22 to about 29, about 23 to about 28, about 24 to about 27, or about 25 to about 26 mol %.

[0108] Those skilled in the art will appreciate that since mechanical strength and adhesion are important factors in the present invention, the amount of the second monomer will occupy a wider range than the other monomers.

[0109] Third monomer: NSPP-linked monomer As discussed above, the tissue scaffolds used in the present invention can be optionally formed by combining a polymer with an NSPP. To effectively combine the polymer with the NSPP, the polymer preferably includes a monomer or unit that has crosslinking capability.

[0110] This cross-linking ability means that the polymer can bind to the NSPP and thereby cross-link the NSPP to form a scaffold that includes the NSPP, or, through a similar mechanism, the NSPP can act as a cross-linker, thereby cross-linking the polymer to form a scaffold.

[0111] To prepare a polymer capable of binding to an NSPP, one of skill in the art will understand that the monomer capable of binding to the NSPP must be present in the polymer of the invention in a sufficient proportion to crosslink with the NSPP, such that a tissue scaffold can be formed in the presence of water. Generally, the ratio of "crosslinking" monomers in the polymer is about 15:1, about 10:1, about 5:1, about 4:1, about 3:1, about 2:1, about 1:1, about 1:2, about 1:3, about 1:4, about 1:5, about 1:10, about 1:15 of crosslinking monomers:water binding monomers.

[0112] Monomers that can be attached to an NSPP generally have either electrophilic or nucleophilic functional groups, such that, for example, a nucleophilic functional group on the NSPP can react with an electrophilic functional group on the monomer to form a covalent bond.

[0113] Thus, for example, if the NSPP has a nucleophilic functional group such as an amine, the polymer may have an electrophilic functional group such as N-hydroxysuccinimide (NHS). Other electrophilic functional groups suitable for use in the present invention are N-hydroxysulfosuccinimide (SNHS) and N-hydroxyethoxylated succinimide (ENHS). An example of this type of monomer is N-acryloxysuccinimide (NAS). On the other hand, if the NSPP has an electrophilic functional group, the polymer may have a nucleophilic functional group such as an amine or a thiol.

[0114] Preferably, the polymer comprises a third monomer in an amount of up to 15 mol %. In various embodiments, the third monomer may be present at about 0, 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, or about 15 mol %. In various embodiments, the third monomer may be present at about 0 to about 1 mol %, about 1 to about 15 mol %, about 2 to about 14 mol %, about 3 to about 13 mol %, about 4 to about 12 mol %, about 5 to about 11 mol %, about 6 to about 10, about 7 to about 9 mol %, or about 8 mol %.

[0115] One of ordinary skill in the art will appreciate that the polymer may be formed from a hydrophobic composition and thus the third monomer is optional in the polymer.

[0116] The fourth monomer: phase transition monomer In another embodiment of the present invention, the polymer may further comprise a fourth monomer that can impart phase transition properties to the scaffold, thereby ensuring post-administration stability of the scaffold. Furthermore, these phase transition properties allow the polymer of the present invention to form a scaffold whose various properties (such as viscosity) can be altered by modifying factors such as pH and temperature. The scaffold is designed to have a lower critical solution temperature (LCST) below body temperature. A variety of thermoresponsive and injectable polymers are suitable for use in the present invention, including poly(ethylene oxide) / poly(propylene oxide) and poly(N-isopropylacrylamide) (PNIPAAm) copolymers.

[0117] Generally, the proportion of phase change monomer in the polymer is at least about a 3:1 molar ratio of phase change monomer:water binding monomer, which can be increased to, for example, about 10:1, about 11:1, about 12:1, about 13:1, about 14:1, about 15:1, about 16:1, about 17:1, about 18:1, about 19:1, about 20:1, about 25:1, about 30:1, about 35:1, about 40:1, about 45:1, about 50:1, about 55:1, about 60:1, about 65:1, about 70:1 molar ratios of phase change monomer:water binding monomer, up to about 75:1, about 80:1 and about 85:1.

[0118] Preferably, the polymer comprises a fourth monomer in an amount that comprises 100% of the remainder of the polymer composition. In one embodiment, the mole percent of the fourth monomer can be up to about 85%, preferably about 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, or 85 mole percent.

[0119] Other polymer properties It will be understood by those skilled in the art that by combining different types of monomers, polymers with a variety of different properties can be produced. Additionally, the properties of a polymer can be modified by incorporating certain monomers or functional groups into an existing polymer. For example, copolymerization of HEMA monomers with other monomers (such as methyl methacrylate) can be used to modify properties such as swelling and mechanical properties. Monomers can also be reacted with other compounds to form macromonomers (as defined above), which are then included in the polymers of the present invention. For example, HEMA can be reacted with lactide to form a HEMA-polylactic acid polymer (PLA / HEMA), which itself can be used as a monomer in the polymers of the present invention. Additionally, the monomers themselves can be combinations of monomer units that are then incorporated into the polymer. An example of this type of monomer is oligo(ethylene glycol) monomethyl ether methacrylate (OEGMA), which is a hydrophilic monomer composed of two hydrophilic monomers: ethylene glycol and methacrylate.

[0120] The preferred polymers of the present invention may be further modified with one or more moieties and / or functional groups. Any moiety or functional group may be used in accordance with the present invention. In some embodiments, the polymers may be modified with acyclic polyacetals derived from polyethylene glycol (PEG), carbohydrates, and / or polysaccharides. Additionally, as discussed above, hydrophilic groups may be incorporated into the monomers (and thus the polymer) to increase the water binding capacity of the polymer.

[0121] In terms of sequence, the copolymer can be a block copolymer, a graft copolymer, a random copolymer, a blend, a mixture and / or an adduct of any of the aforementioned and other polymers. Typically, the polymer according to the present invention is an organic polymer. Preferably, the polymer of the present invention is biocompatible. In some embodiments, the polymer is biodegradable. In other embodiments, the polymer is both biocompatible and biodegradable.

[0122] The preferred polymers of the present invention may also include other monomers in their structure, for example, the monomers may be polymers such as poly(vinyl alcohol) (PVA), polyesters, acrylic polymers, and ionic polymers, or monomers thereof.

[0123] If it is desired that the polymer be biodegradable or absorbable, one or more monomers having biodegradable bonds may be used. Alternatively, or in addition, the monomers may be selected such that the product of the reaction between them results in a biodegradable bond. For each approach, the monomers and / or bonds may be selected such that the resulting biodegradable polymer will degrade or be absorbed in a desired period of time, for example, from about 6 hours to about 6 months. Preferably, the monomers and / or bonds are selected such that the resulting product is non-toxic when the polymer degrades under physiological conditions.

[0124] Biodegradable bonds can be chemically or enzymatically hydrolyzable or absorbable. Exemplary chemically hydrolyzable biodegradable bonds include polymers, copolymers and oligomers of glycolide, lactide, caprolactone, dioxanone and trimethylene carbonate. Exemplary enzymatically hydrolyzable biodegradable bonds include peptidic bonds that can be cleaved by metalloproteinases and collagenases. Further exemplary biodegradable bonds include polymers and copolymers of poly(hydroxy acids), poly(orthocarbonates), poly(anhydrides), poly(lactones), poly(amino acids), poly(carbonates) and poly(phosphonates).

[0125] The chemical hydrolysis of lactide in the present invention results in an increase in the polymer's lower critical solution temperature (LCST) (by decreasing the overall hydrophobicity of the polymer) and therefore its bioabsorption capacity.

[0126] Preferred Polymers The polymer preferably comprises the first monomer in an amount of about 1 to about 15 mol %. In various embodiments, the first monomer may be present at 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, or about 15 mol %. Preferably, the first monomer is OEGMA.

[0127] The polymer preferably comprises the second monomer in an amount of about 5 to about 50 mole %. In various embodiments, the second monomer may be present at 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 21, about 22, about 23, about 24, about 25, about 26, about 27, about 28, about 29, about 30, about 31, about 32, about 33, about 34, about 35, about 36, about 37, about 38, about 39, about 40, about 41, about 42, about 43, about 44, about 45, about 46, about 47, about 48, about 49, or about 50 mole %. Preferably, the second monomer is PLA / HEMA.

[0128] The polymer preferably comprises a third monomer in an amount of up to 15 mol%. In various embodiments, the third monomer may be present at about 0, 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, or about 15 mol%. Preferably, the third monomer is NAS.

[0129] The polymer preferably comprises a fourth monomer in an amount that constitutes 100% of the remainder of the polymer composition, for example, between about 50 and about 85 mol %. In one embodiment, the mol % of the fourth monomer can be up to about 85%, preferably about 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84 or 85 mol %. Preferably, the fourth monomer is NIPAAm.

[0130] The percentages given herein relate to the composition of the final polymer and not to the amounts of feed utilized in forming the polymer.

[0131] In one embodiment, the polymer preferably comprises a first monomer in an amount of about 1 to about 15 mol %, a second monomer in an amount of about 5 to about 50 mol %, a third monomer in an amount up to 15 mol %, and a fourth monomer in an amount up to about 85 mol %.

[0132] Preferably, the first monomer is OEGMA, the second monomer is PLA / HEMA, the third monomer is NAS, and the fourth monomer is NIPAAm.

[0133] In another embodiment, the polymer preferably comprises a first monomer in an amount of about 7 mol %, a second monomer in an amount of about 30 mol %, a third monomer in an amount of about 7 mol %, and a fourth monomer in an amount of about 53 mol %.

[0134] Preferably, the first monomer is OEGMA, the second monomer is PLA / HEMA, the third monomer is NAS, and the fourth monomer is NIPAAm.

[0135] In one embodiment, the polymer of the present invention has formula (I): [ka] is a polymer of During the ceremony, A is the first monomer (water-binding monomer), e.g., OEGMA; B is a second monomer (a monomer capable of imparting mechanical properties to the tissue scaffold), e.g., PLA / HEMA; C is a third monomer (a monomer having a functional group for attachment to an NSPP), e.g., NAS; D is a fourth monomer (a monomer capable of imparting phase transition properties to the scaffold), for example, NIPAAm).

[0136] In various embodiments, m is an integer from 1-20, n is an integer from 1-20, p is an integer from 0-20, and q is an integer from 1-20.

[0137] Exemplary polymers of the present invention are represented by the following formula (Ia): [ka] is represented by where A is the water binding monomer OEGMA, B is the reinforcing monomer PLA / HEMA, C is the crosslinker NAS, D is the phase transfer monomer NIPAAm, and m, n and p, q, x and y are as defined above.

[0138] One skilled in the art will recognize that monomers A, B, C and D can be present in the polymer in any order, provided that the required water binding, strengthening and / or crosslinking capabilities are achieved.

[0139] It has also been discovered that some monomers, such as PLA / HEMA, polyesters, such as poly(lactic acid), poly(caprolactone), poly(glycolide), and random copolymers thereof (e.g., poly(glycolide-co-lactide) and poly(glycolide-co-caprolactone), as well as other biodegradable and biocompatible polymers, can increase the LCST of the preferred polymers used in the present invention during degradation of the biodegradable segment (e.g., PLA) in vivo, resulting in bioresorption of the polymer. This provides the additional advantage that the polymers used in the present invention can be designed to be biodegradable in vivo.

[0140] The overall size of preferred polymers for use in the present invention may vary depending on factors such as the type of monomer incorporated into the polymer, the type of NSPP sought to be used to form the scaffold, and the conditions under which the protein is attached to the polymer. In general, however, preferred polymers for use in the present invention may be molecules of about 1 to about 100 kDa, about 5 to about 60 kDa, or about 30 kDa. In various embodiments, the polymers of the present invention may be molecules of about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99 or about 100 kDa molecule.

[0141] PNPHO A preferred polymer of the present invention is poly(NIPAAm-co-NAS-co-(PLA / HEMA)-co-OEGMA), or "PNPHO." The polymer PNPHO preferably comprises OEGMA in an amount of about 1 to about 15 mol %, PLA / HEMA in an amount of about 5 to about 50 mol %, NAS in an amount up to 15 mol %, and NIPAAm in an amount that constitutes 100%, for example about 50 to about 85 mol %, of the remainder of the polymer composition.

[0142] The percentages given herein relate to the composition of the final polymer and not to the amounts of feed utilized in forming the polymer.

[0143] A preferred form of polymer PNPHO for use in the present application is a polymer of formula (Ia), as shown above, wherein x is in the range of 1 to 1000, y is in the range of 1 to 1000, and m, n, p, and q are in the range of 1 to 20.

[0144] One skilled in the art will recognize that monomers A, B, C and D can be present in the polymer in any order, provided that the required water binding, strengthening and / or crosslinking capabilities are achieved.

[0145] PPHO Another preferred polymer of the present invention is poly(NIPAAm-co-(PLA / HEMA)-co-OEGMA), or "PPHO." The polymer PPHO preferably comprises OEGMA in an amount of about 1 to about 15 mol %, PLA / HEMA in an amount of about 5 to about 50 mol %, and NIPAAm in an amount that comprises 100%, for example about 50 to about 85 mol %, of the remainder of the polymer composition. In a preferred embodiment, the PPHO is about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or about 15 mol % of OEGMA, and / or about 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45 , 46, 47, 48, 49 or about 50 mol% PLA / HEMA, and / or NIPAAM in an amount of about 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 78, 79, 80, 81, 82, 83, 84 or about 85 mol%.

[0146] The percentages given herein relate to the composition of the final polymer and not to the amounts of feed utilized in forming the polymer.

[0147] A preferred form of polymeric PPHO for use in the present application is a polymer of formula (II), as shown below: wherein x is in the range of 1 to 1000, y is in the range of 1 to 1000, and m, n, and q are in the range of 1 to 20.

[0148] One skilled in the art will recognize that monomers A, B, and D can be present in the polymer in any order, provided that the required water binding, strengthening, and / or crosslinking capabilities are achieved.

[0149] Polymer synthesis Those skilled in the art will be aware of suitable methods for synthesizing the preferred polymers used in the present invention, including methods such as ring-opening polymerization, addition polymerization (including free radical polymerization), and condensation polymerization.

[0150] The formation of the preferred polymers, PNPHO and PPHO, is described in the Examples below.

[0151] Composition for forming a scaffold The present invention also relates to polymers for forming tissue scaffolds, comprising a first water-binding monomer, a second monomer that confers mechanical properties, an optional third monomer that is an NSPP-binding monomer, and a fourth monomer that can impart phase transition properties to the scaffold.

[0152] As used herein, the term "composition" refers to a solid or liquid composition that contains the above-mentioned components. In some embodiments, other components such as pharma- ceutically acceptable excipients and biologically active agents (e.g., drugs, vitamins and minerals) can also be included in the composition of the present invention to aid in the repair and / or regeneration of target tissue and / or provide a method for achieving targeted delivery of biologically active compounds.

[0153] Excipients and Bioactive Agents Pharmaceutically acceptable excipients may be included in the compositions and / or scaffolds of the present invention and may include any and all solvents, dispersion media, inert diluents, or other liquid vehicles, dispersion or suspension aids, granulating agents, surfactants, disintegrants, isotonicity agents, thickening or emulsifying agents, preservatives, binders, lubricants, buffers, oils, etc., suitable for the particular dosage form desired. Remington (Gennaro, AR, Remington: The Science and Practice of Pharmacy, 21st Ed (2006) Lippincott Williams & Wilkins) discloses various excipients used in the formulation of pharmaceutical compositions and known techniques for their preparation. The use of any conventional excipient is considered within the scope of the present invention, except insofar as it is incompatible with the substance or its derivatives, such as by producing any undesirable biological effects or otherwise interacting in a deleterious manner with any other component(s) of the pharmaceutical composition.

[0154] Excipients such as coloring agents, coating agents, sweetening, flavoring, and perfuming agents can be present in the composition, according to the judgment of the formulator.

[0155] Biologically active agents or drug compounds that may be added to the compositions and / or scaffolds of the invention include proteins, glycosaminoglycans, carbohydrates, nucleic acids, as well as inorganic and organic bioactive compounds such as enzymes, antibiotics, antineoplastic agents, local anesthetics, hormones, angiogenic agents, anti-angiogenic agents, growth factors (e.g., insulin-like growth factor-1 (IGF-1), basic fibroblast growth factor (bFGF) and transforming growth factor-b (TGFb)), antibodies, neurotransmitters, psychoactive drugs, anti-cancer drugs, chemotherapeutic drugs, drugs affecting the reproductive organs, genes and oligonucleotides.

[0156] Compositions containing ingredients such as excipients and / or bioactive agents can be prepared by combining the polymers of the invention with NSPPs, combining it with one or more other ingredients, and then lyophilizing the resulting composition to provide a ready-to-use tissue scaffold.

[0157] The amounts of polymer, NSPP and bioactive agent present in the composition will necessarily depend on the particular drug and condition being treated. One of ordinary skill in the art will know the appropriate agents and amounts to use to treat a condition.

[0158] Natural or synthetic peptides or proteins NSPPs can be important because they provide additional mechanical properties to the scaffold (such as strength and elasticity), as well as provide an environment at the repair site that mimics the natural environment, thereby aiding in tissue repair and regeneration.

[0159] It is important that the NSPP contains a side chain or other functional group that is exposed to allow reaction with a functional group of the NSPP-binding monomer(s) thereby attaching the NSPP to the polymer via the NSPP-binding monomer(s). Examples of suitable side chains include glutamic acid or lysyl side chains.

[0160] The present invention also contemplates the use of variants of NSPP, such as species variants or polymorphic variants. The present invention is intended to cover all functionally active variants of NSPP that exhibit the same activity. This includes apo- and halo-forms of NSPP, post-translationally modified forms, and glycosylated or deglycosylated derivatives. Such functionally active fragments and variants include, for example, those with conservative amino acid substitutions.

[0161] Preferably, the NSPP(s) for use in the present invention are obtained from recombinant sources, but may also be extracted from natural sources or synthesized.

[0162] scaffold The present invention also relates to a tissue scaffold comprising a polymer according to the present invention and an optional NSPP, the polymer comprising a first water-binding monomer, a second monomer imparting mechanical properties, an optional third monomer being an NSPP-binding monomer comprising a functional group capable of binding to the NSPP, and a fourth monomer capable of imparting phase transition properties to the scaffold, the scaffold forming a malleable structure upon hydration. Preferably, the first monomer is OEGMA, the second monomer is PLA / HEMA, the third monomer is NAS, and the fourth monomer is NIPAAm.

[0163] In one embodiment, the scaffold comprises a polymer having the above monomer to ensure the post-administration stability of the scaffold.Specifically, the LCST of the scaffold is below body temperature or about 37°C to avoid the formation or dissolution of unstable complexes in vivo.One example of the monomer useful for this purpose is NIPAAm.

[0164] In another embodiment, the amount of the first monomer in the polymer does not exceed about 15 mol % to ensure that the LCST of the scaffold is below body temperature or about 37° C. One example of a monomer useful for this purpose is OEGMA.

[0165] In yet another embodiment, the amount of the second monomer in the polymer can be as high as 50 mole % to act as the backbone of the polymer.

[0166] cell The scaffolds of the present invention may also include cells to assist in the repair and / or regeneration of the target tissue.

[0167] In general, the cells used in accordance with the present invention can be any type of cell. The cells must be viable when embedded in the scaffold of the present invention.

[0168] In some embodiments, cells that can be embedded in the scaffold according to the present invention include, but are not limited to, mammalian cells (e.g., human cells, primate cells, mammalian cells, rodent cells, etc.), avian cells, fish cells, insect cells, plant cells, fungal cells, bacterial cells, and hybrid cells. In some embodiments, exemplary cells that can be embedded in the scaffold include stem cells, totipotent cells, pluripotent cells, and / or embryonic stem cells.

[0169] In some embodiments, exemplary cells that can be embedded in the scaffold according to the present invention include, but are not limited to, primary cells and / or cell lines from any tissue. For example, cardiomyocytes, muscle cells, hepatocytes, keratinocytes, melanocytes, neurons, astrocytes, embryonic stem cells, adult stem cells, hematopoietic stem cells, hematopoietic cells (e.g., monocytes, neutrophils, macrophages, etc.), ameloblasts, fibroblasts, chondrocytes, osteoblasts, osteoclasts, neurons, sperm cells, egg cells, liver cells, epithelial cells from the lung, epithelial cells from the digestive tract, epithelial cells from the intestine, liver, epithelial cells from the skin, etc., and / or hybrids thereof may be embedded in the scaffold according to the present invention.

[0170] Exemplary mammalian cells that can be embedded in a scaffold according to the invention include, but are not limited to, Chinese hamster ovary (CHO) cells, HeLa cells, Madin-Darby canine kidney (MDCK) cells, baby hamster kidney (BHK cells), NSO cells, MCF-7 cells, MDA-MB-438 cells, U87 cells, A172 cells, HL60 cells, A549 cells, SP10 cells, DOX cells, DG44 cells, HEK293 cells, SHSY5Y, Jurkat cells, BCP-1 cells, COS cells, Vero cells, GH3 cells, 9L cells, 3T3 cells, MC3T3 cells, C3H-10T1 / 2 cells, NIH-3T3 cells, and C6 / 36 cells.

[0171] In some embodiments, it is desirable for the cells to be distributed evenly throughout the scaffold. An even distribution can help provide a more uniform tissue-like scaffold that provides a more uniform environment for the encapsulated cells. In some embodiments, the cells are located on the surface of the scaffold. In some embodiments, the cells are located in the interior of the scaffold. In some embodiments, the cells are layered within the scaffold. In some embodiments, the scaffold comprises different cell types.

[0172] In some embodiments, the conditions under which cells are embedded in the scaffold are altered to maximize cell viability. In some embodiments, for example, cell viability increases at lower polymer concentrations. In some embodiments, cells located at the periphery of the hydrogel tend to have reduced viability compared to cells fully encapsulated within the hydrogel. In some embodiments, the conditions of the surrounding environment (e.g., pH, ionic strength, nutrient availability, temperature, oxygen availability, osmolality, etc.) may need to be adjusted and / or altered to maximize cell viability.

[0173] In some embodiments, cell viability can be measured by monitoring one of many indicators of cell viability. In some embodiments, indicators of cell viability include, but are not limited to, intracellular esterase activity, plasma membrane integrity, metabolic activity, gene expression, and protein expression. By way of example only, when cells are exposed to a fluorogenic esterase substrate (e.g., calcein AM), live cells fluoresce green as a result of intracellular esterase activity that hydrolyzes the esterase substrate to a green fluorescent product. By way of another example, when cells are exposed to a fluorescent nucleic acid stain (e.g., ethidium homodimer-1), dead cells fluoresce red because their plasma membranes are compromised and therefore permeable to the high affinity nucleic acid stain.

[0174] Generally, the number of cells in the composition is an amount that allows for the formation of a hydrogel according to the present invention. In some embodiments, the amount of cells suitable for forming a hydrogel according to the present invention ranges from about 0.1% to about 80% by weight, from about 1.0% to about 50% by weight, from about 1.0% to about 40% by weight, from about 1.0% to about 30% by weight, from about 1.0% to about 20% by weight, from about 1.0% to about 10% by weight, from about 5.0% to about 20% by weight, or between about 5.0% and about 10% by weight.

[0175] In some embodiments, the amount of cells in a composition suitable for forming a hydrogel according to the invention is about 5% by weight. In some embodiments, the concentration of cells in a precursor solution suitable for forming a hydrogel according to the invention is about 10 to about 1 x 10 8 cells / mL, approximately 100 to approximately 1×10 7 cells / mL, approximately 1×10 3 ~Approx. 1×10 6 cells / mL, or approximately 1 x 10 4 ~Approx. 1×10 5 In some embodiments, a single hydrogel comprises a population of identical cells and / or cell types. In some embodiments, a single hydrogel comprises a population of non-identical cells and / or cell types.

[0176] In some embodiments, a single hydrogel may contain at least two different types of cells. In some embodiments, a single hydrogel may contain 3, 4, 5, 10 or more types of cells. By way of example only, in some embodiments, a single hydrogel may contain only embryonic stem cells. In some embodiments, a single hydrogel may contain both embryonic stem cells and hematopoietic stem cells.

[0177] Culture medium Cells may be grown and / or maintained using any of a variety of cell culture media, including complex media and / or serum-free media, which can support the growth of one or more cell types or cell lines. Typically, cell culture media contain buffers, salts, energy sources, amino acids (e.g., natural amino acids, unnatural amino acids, etc.), vitamins, and / or trace elements. Cell culture media may optionally contain a variety of other components, including, but not limited to, carbon sources (e.g., natural sugars, unnatural sugars, etc.), cofactors, lipids, sugars, nucleosides, animal-derived components, hydrolysates, hormones, growth factors, surfactants, indicators, minerals, activators of specific enzymes, activator inhibitors of specific enzymes, enzymes, organics, and / or small molecule metabolites.

[0178] Cell culture media suitable for use in accordance with the present invention are commercially available from a variety of sources, such as ATCC (Manassas, Va.). In certain embodiments, one or more of the following media are used to grow cells: RPMI-1640 medium, Dulbecco's Modified Eagle's Medium, Minimal Essential Medium Eagle, F-12K medium, Iscove's Modified Dulbecco's Medium.

[0179] Those skilled in the art will recognize that the cells listed herein represent an exemplary, non-exhaustive list of cells that may be encapsulated within a precursor solution according to the present invention.

[0180] Purpose The present invention aims to provide a scaffold that supports the natural healing of damaged tissue without inducing any specific tissue formation. The present invention aims to enable immobilization of one / two or more tissues to assist tissue regeneration and / or tissue interface integration with minimal foreign body reaction.

[0181] Preferably, the tissue scaffold of the present invention is hydrated intraoperatively with saline, the patient's own blood, platelet reach plasma (PRP), platelet reach fibrin and / or other autologous blood / cell / tissue products to form a malleable scaffold. The resulting scaffold can be administered arthroscopically or by open surgical intervention. Upon contact with body tissue, the scaffold is adhesive and can be sutured, thereby allowing fixation of one or more tissues to support tissue regeneration and / or tissue interface integration.

[0182] The scaffold of the present invention can preferably be manufactured using different processing methods.

[0183] The scaffolds of the present invention preferably negate the criticality of water solubility.

[0184] The scaffolds of the present invention can preferably be tailored to modify their microenvironments to address different requirements.

[0185] The scaffolds of the present invention preferably bind a variety of hydrophilic and hydrophobic drugs.

[0186] The scaffolds of the present invention are preferably non-adherent to gloves for effective manipulation.

[0187] The scaffolds of the present invention are preferably malleable, allowing for effective delivery to the site via arthroscopic or open surgical intervention.

[0188] The scaffolds of the present invention are preferably adhesive to the treatment site, and the scaffolds of the present invention are preferably sutureable for internal fixation.

[0189] The present invention has been developed for use in skin grafts for fixation of graft tissue, repair of partial or full thickness rotator cuff tears, total shoulder arthroplasty, tendon healing in anterior cruciate ligament (ACL) and / or other ligament / tendon ruptures.

[0190] kit The present invention provides various kits that include one or more scaffolds and / or polymers of the present invention. For example, in one embodiment, the present invention provides a kit that includes a scaffold and / or polymer and instructions for use. The kit may include multiple different scaffolds and / or polymers. The kit may optionally include cells, NSPPs, biologically active compounds, etc. The kit may include any of several additional components or reagents in any combination. Not all of the various combinations are explicitly described, and each combination is within the scope of the present invention.

[0191] PNPHO / PPHO composition PNPHO was synthesized according to the synthetic procedures generally described in the applicant's previous publication, WO 2013 / 091001. PPHO was synthesized in a similar manner, except for the addition of NAS.

[0192] The synthesis of PNPHO copolymers was performed in accordance with that shown in WO 2013 / 091001, as evidenced by the proton peaks of each monomer. 1 The synthesis of PPHO was confirmed by H NMR spectroscopy. Similarly, the synthesis of PPHO was confirmed by evidence of the proton peaks of each monomer, as shown in Figure 5(a). 1 The hydrophobicity of PPHO was confirmed by H NMR spectroscopy. Comparative NMR spectra of PNPHO and PPHO are shown in Figure 5(b). As shown in Figure 5(c), the HPLC chromatogram at 230 nm shows PNPHO eluting at 6.2 min and PPHO eluting at 7.2 min. The difference in elution times indicates that PPHO elutes at a higher percentage of acetonitrile compared to PNPHO, thus confirming the hydrophobicity of PPHO compared to PNPHO.

[0193] Characteristic proton peaks were detected for NIPAAm (a and b), NAS (e), PLA / HEMA (f, h, k) and OEGMA (m and n). The final composition of the copolymer was calculated based on the integral of these peaks from each monomer for NIPAAm (a), NAS (e / 2-f), PLA / HEMA (h) and OEGMA (n / 2). In this study, the copolymers are denoted as PNPHO, with the subscript added corresponding to the PLA / HEMA (lactate length) to OEGMA molar ratio. For example, PNPHO 8(6)3 represents a copolymer synthesized with 8 mol % PLA / HEMA and 3 mol % OEGMA with lactate length 6. A variety of copolymers were produced.

[0194] Fabrication of tissue scaffolds from PNPHO / PPHO The tissue scaffolds of the present invention can be manufactured from PNPHO / PPHO solutions using different methods. The resulting PNPHO / PPHO scaffolds and their morphology are shown in Figure 1. Upon hydration, the scaffolds can be shaped and optionally delivered arthroscopically, as shown in Figure 2. The product is suitable for suturing and surgical stapling. The product is not adhesive to gloves, allowing for effective manipulation.

[0195] Upon contact with different tissues (as shown in Figures 3, 4 and 6), the product is adhesive and allows for in vivo immobilization of one or more tissues with or without suturing / internal stabilization.

[0196] In general, the PHPHO / PPHO scaffolds of the present invention can be prepared by lyophilization of an aqueous composition, where a PHPHO / PPHO solution with or without peptide / protein components is dissolved in a buffer solution, different drugs / moieties (antibiotics, growth factors, live viruses) can be added to the solution for final product embedding, the resulting solution is lyophilized, the morphology of the complex can be modified by adjusting the lyophilization parameters and the concentration of the PHPHO polymer, and the final product is in the form of a ready-to-use scaffold with or without embedded biological moieties.

[0197] Exemplary Method 1 The first method is used, in which the polymer is water-soluble: it involves dissolving an aqueous solution of the polymer in a buffer solution and freeze-drying the resulting solution, and the morphology of the scaffold is modified by adjusting the freeze-drying parameters and the concentration of the polymer.

[0198] Exemplary Method 2 A second method is used where the polymer is hydrophobic (e.g., PPHO), which involves dissolving the polymer in an organic solvent to obtain a solution, precipitating the solution in water as a poor solvent to obtain a gel, and freeze-drying the gel, where the morphology of the scaffold is modified by adjusting the freeze-drying parameters.

[0199] Other preparation methods may include porogen leaching and / or gas foaming.

[0200] Exemplary Method 3 The method is used to create an injectable or sprayable scaffold, which involves dissolving a PPHO polymer in an aqueous solution, autologous blood, cell or tissue products, or a combination thereof.

[0201] Scaffolding Manufacturing According to the present invention, scaffolds are made by either freeze-drying PNPHO and / or PPHO to form a porous membrane, as outlined above and shown in Figures 7 and 10. Additionally, scaffolds can be fabricated by press compaction of PNPHO and / or PPHO powders. Although SEM imaging of both configurations showed similar microstructures and topographies, the scaffolds made using press compaction were used for in vivo biocompatibility analysis.

[0202] The aim of this study was to investigate the biodegradability of freeze-dried TM scaffolds in a mouse model. Twenty-four male Balb / c mice, approximately 10 weeks of age, were subcutaneously implanted with freeze-dried PNPHO and PPHO scaffolds. Ultrasound measurements were performed weekly to determine in situ presence and volume. Animals were sacrificed 7, 14, 21 and 42 days after implantation. Biopsies of the implantation sites were taken, fixed in 10% formalin, embedded in paraffin, sectioned and stained with H&E for histological analysis.

[0203] Surgical procedure and product preparation PNPHO and PPHO scaffolds were used. Prior to surgery, mice were anesthetized with 3% isoflurane. General anesthesia was noted by lack of response to toe pinch. A 1 cm incision was made on the mouse dorsal skin and the scaffolds were inserted subcutaneously. The incision was then closed with sutures and cleaned as necessary.

[0204] Material Handling Prior to implantation, the scaffolds were hydrated in 2 mL of warm water at approximately 37° C. for at least 3 minutes. The scaffolds were found to be cohesive and moldable. They could be easily picked up with forceps and easily and conveniently delivered into a pouch created on the back of the mouse.

[0205] recovery All experimental mice recovered from anesthesia within 5 minutes. The mice were recorded as healthy with no signs of discomfort or stress. The procedure did not affect the mobility of the mice, and no scaffolds were detected emerging from the incision wound after it was closed by suturing. The scaffolds appeared to remain solid for some time inside the mouse.

[0206] Animal health and weight No abnormalities in health (and significant weight loss) were noted in all mice over the experimental period, except for mouse ID "S-21." This mouse showed significant weight loss on the fourth day after surgery, with no other abnormal signs or signs of distress, the mouse was behaving normally, had no wounds or bites, or exhibited hair loss. As all other parameters were normal, the mouse was kept for further observation. The weight of this mouse steadily increased on the seventh day and returned to normal levels on the eleventh day.

[0207] Macroscopic observation and ultrasonic analysis Mice were scanned with high frequency ultrasound to determine the volume of the scaffold after implantation. Ultrasound was performed immediately after implantation and on days 0, 4, 7, 11, and 14. Immediately after treatment, the scaffold was visually detected and palpable, but ultrasound imaging did not detect the scaffold, likely due to its solid structure.

[0208] Biopsy and autopsy At days 7, 14, 21 and 42, groups of mice were sacrificed and scored macroscopically for the presence of scaffolds. Skin biopsies were collected and preserved in 10% formalin for histological analysis. The scaffolds were integrated within the host environment at all time points of harvest. The panniculus carnosus layer was found to be more adherent to the underlying structures compared to historical control groups and the experience of this anatomical layer in mice. The skin was more adherent to the subcutaneous fascia and muscle tissue. The scaffolds appear to be integrated within the tissue layers of the underlying tissues.

[0209] Textile analysis Skin biopsies collected from the implantation sites were processed and stained with H&E to analyze the presence of the TM scaffold, cell infiltration, inflammatory response, and angiogenesis.

[0210] Histological evaluation of the site after 1 week in Figure 10 showed scaffold structure with cellular infiltration (indicated by blue arrow). The scaffold implanted in the back of the mouse should seat beneath the panniculus carnosus layer (indicated by orange arrow). The underlying area shows large spaces with scaffold with cellular infiltration (indicated by yellow arrow).

[0211] The results in Figure 11 showed that at week 1, the scaffold was histologically present and infiltrated with numerous cells (indicated by red arrows). At weeks 2, 3 and 6, remaining scaffolds could be detected histologically, while the host tissue had a loose connective tissue structure, possibly fascia (indicated by green arrows). It is important to note that both PNPHO and PPHO are highly soluble in ethanol and therefore removed during the processing steps. Nevertheless, there was no sign of scaffold encapsulation, fibrosis and fibrous tissue formation around the scaffold, confirming the biocompatibility of the product.

[0212] The mild cellular infiltration observed 1 week after implantation was consistent with the early stages of wound healing and foreign body reaction. No obvious multinucleated giant cells were detected in the area where the scaffold was present. Large areas of cellular infiltration were observed around the sutures, which is expected and considered a typical foreign body reaction in the biodegradation of sutures. No cellular inflammatory reaction was observed at weeks 2, 3, and 6.

[0213] Other notes The scaffolds used in the experiments could be handled in a way that allowed for easy implantation and convenient administration after hydration. The healthy recovery of all experimental mice and the absence of any abnormal signs of discomfort or stress immediately after the procedure also indicate their biocompatibility with in vivo life.

[0214] The detection of solid scaffolds in the mouse body immediately after implantation suggests that the scaffolds remained in situ and were not easily removed or degraded in the body. The lack of significant weight loss in all mice also indicates the overall health and stability of the animals during the experimental period. The results of the procedure suggest that the scaffolds used were effective in their intended purpose and the implantation procedure was overall successful. Macroscopically, the researchers could easily identify the tablet-shaped scaffolds subcutaneously immediately after the implantation procedure. The scaffolds did not turn into a liquid state and remained solid. [Industrial Applicability]

[0215] It will be understood that the present invention is easily applicable in the surgical and biomedical fields. The PNPHO or PPHO scaffold of the present invention can be used in any anatomical site where internal fixation of one or more tissues is required to aid healing. These include, but are not limited to, skin grafts for fixing graft tissue, repair of partial or full thickness rotator cuff tears, total shoulder arthroplasty, tendon healing in anterior cruciate ligament (ACL) and / or other ligament / tendon tears, etc.

Claims

1. A polymer for forming a tissue scaffold in mammals, wherein the polymer is A first monomer for binding water, which is a polyether selected from polyethylene glycol (PEG), oligo(ethylene glycol) (OEG), polyethylene oxide (PEO), polyethylene oxide-copropylene oxide (PPO), co-polyethylene oxide block, and random copolymers thereof, A second monomer for imparting mechanical properties to the scaffold, the second monomer being a polyether selected from hydroxyethyl methacrylate (HEMA), hydroxyethyl methacrylate poly(lactic acid) copolymer (PLA / HEMA), poly(lactic acid), poly(caprolactone), poly(glycolide), poly(glycolide-colactide), and poly(glycolide-cocaprolactone), Optionally, a third monomer for binding to a natural or synthetic peptide or protein (NSPP), selected from N-hydroxysulfosuccinimide (SNHS), N-hydroxyethoxylated succinimide (ENHS), and N-acrylooxysuccinimide (NAS), The scaffold comprises a fourth monomer for imparting liquid-solid phase transition behavior to the scaffold, The third monomer has a lower critical solution temperature (LCST) of less than approximately 37°C, and the third monomer is selected from poly(ethylene oxide) / poly(propylene oxide), and poly(N-isopropylacrylamide) (PNIPAAm) homopolymers and copolymers. The aforementioned scaffold is a polymer that forms a malleable structure upon hydration.

2. The polymer according to claim 1, wherein the first monomer is oligo(ethylene) glycol monomethyl ether methacrylate (OEGMA).

3. The polymer according to claim 1, wherein the second monomer is hydroxyethyl methacrylate poly(lactic acid) (PLA / HEMA).

4. The polymer according to claim 1, wherein the third monomer is N-acrylooxysuccinimide (NAS).

5. The polymer according to claim 1, wherein the fourth monomer is (N-isopropylacrylamide) (NIPAAm).

6. The first monomer is OEGMA, the second monomer is PLA / HEMA, the third monomer is NAS if present, and the fourth monomer is NIPAAm. The polymer according to claim 1, wherein the polymer comprises about 1 to about 15 mol% of OEGMA, 5 to about 50 mol% of PLA / HEMA, 0 to about 15 mol% of NAS if present, and up to about 85 mol% of NIPAAm.

7. A tissue scaffold comprising the polymer described in claim 1, wherein the scaffold forms a malleable structure upon hydration.

8. The tissue scaffold according to claim 7, wherein the scaffold can be hydrated during surgery.

9. The tissue scaffold according to claim 8, wherein the scaffold is formed by body heat.

10. The tissue scaffold according to claim 9, wherein the scaffold is formed after administration to a mammal, and the administration is by injection or spray.

11. A malleable structure formed from a tissue scaffold according to any one of claims 7 to 10.

12. A method for creating a tissue scaffold, The steps of dissolving an aqueous solution of the polymer described in claim 1 in a buffer solution to obtain the resulting solution, The step includes freeze-drying the resulting solution, A method wherein the scaffold has a form that can be modified by adjusting the freeze-drying parameters and concentration of the polymer.

13. A method for creating a tissue scaffold, The steps of dissolving the polymer described in claim 1 in an organic solvent to obtain a solution, The steps include: precipitation of the above solution in water as a poor solvent to obtain a gel, The step of freeze-drying the gel is included, A method wherein the scaffold has a form that can be modified by adjusting the freeze-drying parameters.

14. A method for creating a tissue scaffold, A step of dissolving the polymer described in claim 1 in an organic solvent to obtain a solution, A method comprising the step of electrospinning the aforementioned solution.

15. A method for creating a tissue scaffold, The polymer described in claim 1 is dissolved in an aqueous solution, autologous blood, cell or tissue product, or a combination thereof. A method wherein the scaffold is injectable or sprayable.

16. Use of the tissue scaffold according to claim 7 for tissue repair and / or regeneration.

17. The tissue scaffold according to claim 7 for use in soft tissue healing in shoulder arthroplasty, knee arthroplasty and hip arthroplasty, cruciate ligament and / or ligament ruptures.

18. The tissue scaffold according to claim 17, wherein the scaffold is administered by injection or spray, or by arthroscopic or open surgical intervention.

19. The tissue scaffold according to claim 18, wherein the scaffold is hydrated during administration in surgery to form a malleable structure.

20. A kit for use in forming a tissue scaffold, wherein the kit comprises a polymer as described in claim 1 and means for administering a surgically effective amount of the polymer to tissue requiring repair or regeneration, the scaffold forming a malleable structure upon hydration.

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