EP4577255A1 - Compositions céramiques imprimées en 3d et procédés d'utilisation - Google Patents

Compositions céramiques imprimées en 3d et procédés d'utilisation

Info

Publication number
EP4577255A1
EP4577255A1 EP23858242.3A EP23858242A EP4577255A1 EP 4577255 A1 EP4577255 A1 EP 4577255A1 EP 23858242 A EP23858242 A EP 23858242A EP 4577255 A1 EP4577255 A1 EP 4577255A1
Authority
EP
European Patent Office
Prior art keywords
weight
lactide
glycolide
ink
formulation
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Pending
Application number
EP23858242.3A
Other languages
German (de)
English (en)
Inventor
Luis Alvarez
Todd HEIL
David Long
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Theradaptive Inc
Original Assignee
Theradaptive Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Theradaptive Inc filed Critical Theradaptive Inc
Publication of EP4577255A1 publication Critical patent/EP4577255A1/fr
Pending legal-status Critical Current

Links

Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/40—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material
    • A61L27/44—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix
    • A61L27/46—Composite materials, i.e. containing one material dispersed in a matrix of the same or different material having a macromolecular matrix with phosphorus-containing inorganic fillers
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/54—Biologically active materials, e.g. therapeutic substances
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L27/00—Materials for grafts or prostheses or for coating grafts or prostheses
    • A61L27/50—Materials characterised by their function or physical properties, e.g. injectable or lubricating compositions, shape-memory materials, surface modified materials
    • A61L27/58—Materials at least partially resorbable by the body
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
    • B29C64/00—Additive manufacturing, i.e. manufacturing of three-dimensional [3D] objects by additive deposition, additive agglomeration or additive layering, e.g. by 3D printing, stereolithography or selective laser sintering
    • B29C64/10—Processes of additive manufacturing
    • B29C64/106—Processes of additive manufacturing using only liquids or viscous materials, e.g. depositing a continuous bead of viscous material
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y70/00—Materials specially adapted for additive manufacturing
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B33—ADDITIVE MANUFACTURING TECHNOLOGY
    • B33Y—ADDITIVE MANUFACTURING, i.e. MANUFACTURING OF THREE-DIMENSIONAL [3D] OBJECTS BY ADDITIVE DEPOSITION, ADDITIVE AGGLOMERATION OR ADDITIVE LAYERING, e.g. BY 3D PRINTING, STEREOLITHOGRAPHY OR SELECTIVE LASER SINTERING
    • B33Y80/00—Products made by additive manufacturing
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2300/00—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices
    • A61L2300/40—Biologically active materials used in bandages, wound dressings, absorbent pads or medical devices characterised by a specific therapeutic activity or mode of action
    • A61L2300/412—Tissue-regenerating or healing or proliferative agents
    • A61L2300/414—Growth factors
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61L—METHODS OR APPARATUS FOR STERILISING MATERIALS OR OBJECTS IN GENERAL; DISINFECTION, STERILISATION OR DEODORISATION OF AIR; CHEMICAL ASPECTS OF BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES; MATERIALS FOR BANDAGES, DRESSINGS, ABSORBENT PADS OR SURGICAL ARTICLES
    • A61L2430/00—Materials or treatment for tissue regeneration
    • A61L2430/02—Materials or treatment for tissue regeneration for reconstruction of bones; weight-bearing implants

Definitions

  • the caprolactone/glycolide copolymer is caprolactone/glycolide copolymer (95:5). In some embodiments, the caprolactone/glycolide copolymer is caprolactone/glycolide copolymer (90:10). Further provided is a formulation comprising about 55% to about 65% by weight ⁇ TCP, about 15% to about 25% by weight poly(D,L-lactide-co-glycolide) copolymer, about 5% to about 15% PEG having a molecular weight from about 500 g/mol to about 15,000 g/mol, and about 5% to about 15% PEG having a molecular weight from about 25,000 g/mol to about 50,000 g/mol.
  • the formulation comprises about 60% by weight ⁇ TCP, about 20% by weight glycolide/L-lactide copolymer, about 10% by weight PEG having a molecular weight from about 500 g/mol to about 15,000 g/mol, and about 10% PEG having a molecular weight from about 25,000 g/mol to about 50,000 g/mol.
  • the formulation comprises about 60% by weight ⁇ TCP, about 20% by weight glycolide/L-lactide copolymer, about 10% by weight PEG Attorney Docket No: 50222-711.601 having a molecular weight of about 8,000 g/mol, and about 10% PEG having a molecular weight of about 35,000 g/mol.
  • the therapeutic agent comprises one or more polypeptides selected from Table 1, or a functional portion thereof.
  • the therapeutic agent comprises a bone morphogenetic protein (BMP).
  • BMP bone morphogenetic protein
  • the therapeutic agent comprises a targeting moiety that non-covalently binds to the structure.
  • the targeting moiety binds to the printed three-dimensional structure with an affinity of about 1 pM to about 100 ⁇ m.
  • the targeting moiety comprises a polypeptide at least about 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to any one of the sequences of Tables 5-6.
  • the targeting moiety comprises about 2, 3, 4, 5, 6, 7, 8, 9, or 10 sequences selected from the sequences of Tables 5-6.
  • the therapeutic agent comprises or is part of a chimeric polypeptide comprising a sequence at least about 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to any one of SEQ ID NOS: 433-441.
  • the structure has a density of about 1 g/cm3 to about 1.5 g/cm3.
  • the structure has an open porosity of about 25% to about 40%.
  • FIGS.1A-1C are images from scanning electron microscopy (SEM images) of an example 3D printed object made with ink formulation #1 as outlined in Example 2.
  • FIG.1A and FIG.1C Attorney Docket No: 50222-711.601 are SEM images of the surface of the object at increasing magnifications.
  • FIG. 1B is an SEM image of the side of the object.
  • FIGS.2A-2C are images from scanning electron microscopy (SEM images) of an example 3D printed object made with ink formulation #2 as outlined in Example 2.
  • FIG.2A and FIG.2C are SEM images of the surface of the object at increasing magnifications.
  • FIG. 2B is an SEM image of the side of the object.
  • FIGS.3A-3C are images from scanning electron microscopy (SEM images) of an example 3D printed object made with ink formulation #3 as outlined in Example 2.
  • FIG.3A and FIG.3C are SEM images of the surface of the object at increasing magnifications.
  • FIG. 3B is an SEM image of the side of the object.
  • FIGS.4A-4C are images from scanning electron microscopy (SEM images) of an example 3D printed object made with ink formulation #4 as outlined in Example 2.
  • FIG.4A and FIG.4C are SEM images of the surface of the object at increasing magnifications.
  • FIG. 4B is an SEM image of the side of the object.
  • FIGS.5A-5C are images from scanning electron microscopy (SEM images) of an example 3D printed object made with ink formulation #5 as outlined in Example 2.
  • FIG.5A and FIG.5C are SEM images of the surface of the object at increasing magnifications.
  • FIG. 5B is an SEM image of the side of the object.
  • FIGS.6A-6C are images from scanning electron microscopy (SEM images) of an example 3D printed object made with ink formulation #6 as outlined in Example 2.
  • FIG.6A and FIG.6C are SEM images of the surface of the object at increasing magnifications.
  • FIG. 6B is an SEM image of the side of the object.
  • FIGS.7A-7C are images from scanning electron microscopy (SEM images) of an example 3D printed object made with ink formulation #7 as outlined in Example 2.
  • FIG.7A and FIG.7C are SEM images of the surface of the object at increasing magnifications.
  • FIG. 7B is an SEM image of the side of the object.
  • FIGS.8A-8C are images from scanning electron microscopy (SEM images) of an example 3D printed object made with ink formulation #8 as outlined in Example 2.
  • FIG.8A and FIG.8C are SEM images of the surface of the object at increasing magnifications.
  • FIG. 8B is an SEM image of the side of the object.
  • FIGS.9A-9C are images from scanning electron microscopy (SEM images) of an example 3D printed object made with ink formulation #9 as outlined in Example 2.
  • FIG.9A and FIG.9C are SEM images of the surface of the object at increasing magnifications.
  • FIG. 9B is an SEM image of the side of the object.
  • FIGS. 10A-10C are images from scanning electron microscopy (SEM images) of an example 3D printed object made with ink formulation #10 as outlined in Example 2.
  • FIG.10A and FIG. 10C are SEM images of the surface of the object at increasing magnifications.
  • FIG. 10B is an SEM image of the side of the object.
  • FIGS. 11A-11C are images from scanning electron microscopy (SEM images) of an example 3D printed object made with ink formulation #11 as outlined in Example 2.
  • FIG.11A and FIG. 11C are SEM images of the surface of the object at increasing magnifications.
  • FIG. 11B is an SEM image of the side of the object.
  • FIG. 12A-12C are images from scanning electron microscopy (SEM images) of an example 3D printed object made with ink formulation #12 as outlined in Example 2.
  • FIG.12A and FIG. 12C are SEM images of the surface of the object at increasing magnifications.
  • FIG. 12B is an SEM image of the side of the object.
  • FIG. 13A is a photograph of an example flexible 3-layer membrane made with ink formulation #163D printed with a 400 micron nozzle as outlined in Example 2.
  • FIGS.13B-13D are images from scanning electron microscopy (SEM images) of the flexible 3-layer membrane at increasing magnifications.
  • FIG.14A is a photograph of an example gyroid scaffold made with ink formulation #16 3D printed with a 400 micron nozzle as outlined in Example 2.
  • FIGS.14B-14D are images from scanning electron microscopy (SEM images) of the gyroid scaffold at increasing magnifications.
  • FIG. 15A is a photograph of an example flexible 3-layer membrane made with ink formulation #183D printed with a 400 micron nozzle as outlined in Example 2.
  • FIG.15B is a photograph of a hollow cylinder made with ink #183D printed with a 400 micron nozzle as outline in Example 2.
  • FIG. 16A is a photograph of an example flexible 3-layer membrane made with ink formulation #193D printed with a 400 micron nozzle as outlined in Example 2.
  • FIG.16B is a photograph of a hollow cylinder made with ink #193D printed with a 400 micron nozzle as outline in Example 2.
  • FIG. 15A is a photograph of an example flexible 3-layer membrane made with ink formulation #193D printed with a 400 micron nozzle as outline in Example 2.
  • FIG.16B is a photograph of a hollow cylinder made with ink #193D printed with a
  • the formulations include a ceramic material such as calcium phosphate (e.g., tricalcium phosphate, beta tricalcium phosphate, alpha tricalcium phosphate), hydroxyapatite, fluorapatite, bone (e.g., demineralized bone), glasses (bioglasses) such as silicates, vanadates, and related ceramic minerals, or chelated divalent metal ions, or a combination thereof.
  • the ceramic material comprises beta-tricalcium phosphate ( ⁇ -TCP).
  • the formulation is about 30-70, 30-65, 30-60, 30-55, 30-50, 30-45, 30-40, 30-35, 35-70, 35-65, 35-60, 35-55, 35-50, 35-45, 35-40, 40-70, 40-65, 40- 60, 40-55, 40-50, 40-45, 45-70, 45-65, 45-60, 45-55, 45-50, 50-70, 50-65, 50-60, 50-55, 55-70, 55-65, 55-60, 60-70, 60-65, or 65-70 percent ceramic by weight of the formulation.
  • the formulation is about 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%, or 70% ceramic by weight.
  • the formulation is about 60% ceramic by weight.
  • the ceramic is ⁇ -TCP.
  • the ⁇ -TCP is introduced into the formulation as a powder.
  • the formulation comprises one or more additional components.
  • additional components include water, polymer (including copolymer), antifoaming agent, dispersing agent, solvent, particulate or sacrificial pore former, blowing agent, and plasticizer.
  • the formulation comprises one or more polymers, e.g., about 1, 2, 3, 4, or 5 polymers.
  • Non-limiting examples of polymers include poly(ethylene oxide), poly(propylene oxide), polyethylene glycol (PEG), and polyester.
  • the polymer is a water soluble polymer, e.g., PEG.
  • the formulation comprises a polymer that is about 5-30 percent by weight of the formulation.
  • the formulation comprises a polymer that is about 10-30 percent by weight of the formulation. In some embodiments, the formulation is about 20-60 percent total polymer by weight. For instance, the total polymer includes two or more polymers in the formulation, where the total percentage of polymers in the formulation is about 20-60 percent of the weight of the formulation. In an example embodiment, a first polymer is present at about 5-15% by weight of the formulation, and a second Attorney Docket No: 50222-711.601 polymer is present at about 5-15% by weight of the formulation. In some embodiments, the formulation is about 30 to about 50 percent total polymer by weight. As non-limiting examples, the formulation is about 35-45 percent total polymer by weight. In an example, the polymer comprises a poloxamer.
  • Poloxamers are block copolymers of poly(ethylene oxide) (PEO) and poly(propylene oxide) (PPO).
  • a non-limiting example of a poloxamer is poloxamer 407, such as Pluronic® F-127.
  • the formulation comprises about 5-20, 5-15, 5-10, 10-20, 10-15, or 15-20 percent poloxamer 407 by weight.
  • the polymer comprises polyethylene glycol (PEG).
  • the formulation comprises about 5-30, 5-25, 5-20, 5- 15, 5-10, 10-30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20-30, 20-25, or 25-30 percent by weight PEG.
  • the formulation comprises about 5-30, or 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 or 30 percent by weight PEG.
  • PEG can have a molecular weight from 500 g/mol to 35,000 g /mol.
  • the formulation comprises PEG having a molecular weight of 1,500 g/mol. In a further nonlimiting example embodiment, the formulation comprises PEG having a molecular weight of 8,000 g/mol. In a further nonlimiting example embodiment, the formulation comprises PEG having a molecular weight of 20,000 g/mol. In a further nonlimiting example embodiment, the formulation comprises PEG having a molecular weight of 35,000 g/mol. In a nonlimiting example embodiment, the formulation comprises a first PEG at about 5-15 percent by weight and a second PEG at about 5-15 percent by weight.
  • the polymer comprises polydioxanone (PDS).
  • PDS polydioxanone
  • the formulation comprises about 10-30, 10-25, 10-20, 10- 15, 15-30, 15-25, 15-20, 20-30, 20-25, or 25-30 percent by weight PDS.
  • the formulation comprises about 15-25, or 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 percent by weight PDS.
  • the polymer comprises poly-l-lactide.
  • the formulation comprises about 10-30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20-30, 20-25, or 25-30 percent by weight poly-l-lactide.
  • the formulation comprises about 15-25, or 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 percent by weight poly-l-lactide.
  • the formulation comprises about 15-25, or 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25 percent by weight glycolide/L-lactide copolymer.
  • the copolymer comprises glycolide and lactide (e.g., L-lactide).
  • the first polymer comprises poly-l-lactide and the second polymer comprises PEG.
  • the first polymer comprises a copolymer and the second polymer comprises PEG.
  • the copolymer may comprise PCL and polyglycolide (e.g., 95mol% polycaprolactone, 5mol% polyglycolide; 90mol% polycaprolactone, 10mol% polyglycolide).
  • the blowing agent comprises about 5-20, 5-18, 5-16, 5-14, 5-12, 5-10, 5-8, 5-6, 6- 20, 6-18, 6-16, 6-14, 6-12, 6-10, 6-8, 8-20, 8-18, 8-16, 8-14, 8-12, 8-10, 10-20, 10-18, 10-16, 10- 14, 10-12, 5-15, or about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 percent by weight blowing agent.
  • the blowing agent releases carbon dioxide base during printing to create a foamed structure that can increase porosity of the 3D printed structure.
  • blowing agents include baking powder (e.g., monocalcium phosphate, sodium bicarbonate, corn starch) and azodicarbonamide.
  • the blowing agent comprises sodium bicarbonate. In some cases, the formulation comprises about 5-15, or about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 percent by weight sodium bicarbonate. In some embodiments, the blowing agent provides for micropores in the structure having an average diameter of about 1 micron to about 500 microns, or about 50 microns to about 250 microns, or about 150 microns. In some embodiments, a formulation comprises a ceramic material (e.g., ⁇ -TCP) and a polymer. Polymers include PEO, PPO, PDS, PEG, polyester, copolymers, or a combination thereof.
  • the formulation is about 30-70, 30-65, 30-60, 30-55, 30-50, 30- 45, 30-40, 30-35, 35-70, 35-65, 35-60, 35-55, 35-50, 35-45, 35-40, 40-70, 40-65, 40-60, 40-55, 40-50, 40-45, 45-70, 45-65, 45-60, 45-55, 45-50, 50-70, 50-65, 50-60, 50-55, 55-70, 55-65, 55- 60, 60-70, 60-65, or 65-70 percent ceramic material (e.g., ⁇ -TCP) by weight, e.g., about 30%, Attorney Docket No: 50222-711.601 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%,
  • the formulation comprises about 5-20, 5-15, 5-10, 10-20, 10-15, or 15-20 percent poloxamer 407 by weight. In some cases, the formulation comprises about 10-30, 10-25, 10-20, 10-15, 15-30, 15- 25, 15-20, 20-30, 20-25, or 25-30 percent by weight PEG. In some cases, the formulation comprises about 5-15 percent by weight a first PEG and 5-15 percent by weight a second PEG. In some cases, the formulation comprises about 10-30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20- 30, 20-25, or 25-30 percent by weight PCL. In some cases, the formulation comprises about 10- 30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20-30, 20-25, or 25-30 percent by weight PDS.
  • the formulation comprises about 10-30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20- 30, 20-25, or 25-30 percent by weight caprolactone/glycolide copolymer (95:5). In some cases, the formulation comprises about 10-30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20-30, 20-25, or 25-30 percent by weight caprolactone/glycolide copolymer (90:10). In some cases, the formulation comprises about 10-30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20-30, 20-25, or 25-30 percent by weight poly(D,L-lactide-co-glycolide) copolymer (50:50).
  • the formulation comprises about 10-30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20-30, 20-25, or 25-30 percent by weight dioxanone/L-lactide copolymer (90:10). In some cases, the formulation comprises about 10-30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20-30, 20-25, or 25-30 percent by weight glycolide/L-lactide copolymer (95:5). In some cases, the formulation comprises about 10-30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20-30, 20-25, or 25-30 percent by weight poly- l-lactide. In some embodiments, the formulation further comprises an antifoaming agent.
  • the formulation further comprises a dispersing agent. In some embodiments, the formulation further comprises a solvent. In some embodiments, the formulation further comprises a plasticizer. In some embodiments, the formulation further comprises a particulate or sacrificial pore former. In some cases, the formulation further comprises a blowing agent. In some embodiments, a formulation comprises a ceramic material (e.g., ⁇ -TCP) and a particulate or sacrificial pore former. The particulate may be water soluble.
  • Non-limiting examples of particulates include salts and sugars, e.g., sodium chloride, calcium chloride, sucrose, trehalose (e.g., ⁇ , ⁇ trehalose dihydrate), and mannitol (e.g., D-mannitol).
  • the pore former may be a water soluble polymer such as PEG.
  • the formulation is about 30-70, 30-65, 30-60, 30-55, 30-50, 30-45, 30-40, 30-35, 35-70, 35-65, 35-60, 35-55, 35-50, 35-45, 35- 40, 40-70, 40-65, 40-60, 40-55, 40-50, 40-45, 45-70, 45-65, 45-60, 45-55, 45-50, 50-70, 50-65, 50-60, 50-55, 55-70, 55-65, 55-60, 60-70, 60-65, or 65-70 percent ceramic material (e.g., ⁇ -TCP) Attorney Docket No: 50222-711.601 by weight, e.g., about 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%,
  • the formulation comprises about 1-10, 1-9, 1-8, 1-7, 1-6, 1-5, 2-10, 2-9, 2-8, 2-7, 2-6, 2-5, 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-10, 5-9, 5-8, 5-7, or 5-6 percent by weight particulate.
  • the particulate comprises sucrose.
  • the formulation comprises about 10-30% or about 5-15% percent by weight sacrificial pore former, such as a polymer.
  • the formulation further comprises water.
  • the formulation further comprises a polymer.
  • the formulation is about 30-70, 30-65, 30-60, 30-55, 30-50, 30-45, 30-40, 30-35, 35-70, 35-65, 35-60, 35-55, 35-50, 35-45, 35- 40, 40-70, 40-65, 40-60, 40-55, 40-50, 40-45, 45-70, 45-65, 45-60, 45-55, 45-50, 50-70, 50-65, 50-60, 50-55, 55-70, 55-65, 55-60, 60-70, 60-65, or 65-70 percent ceramic material (e.g., ⁇ -TCP) by weight, e.g., about 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%, 6
  • a formulation further comprises a dispersing agent. In some embodiments, the formulation further comprises a solvent. In some embodiments, the formulation further comprises a plasticizer. In some embodiments, the formulation further comprises a particulate or sacrificial pore former. Attorney Docket No: 50222-711.601
  • a formulation comprises a ceramic material and one or more polymers. In some embodiments, the formulation comprises about 30% to about 70% a ceramic material (e.g., ⁇ -TCP).
  • the formulation comprises about 30-70, 30-65, 30-60, 30-55, 30-50, 30-45, 30-40, 30-35, 35-70, 35-65, 35-60, 35-55, 35-50, 35-45, 35-40, 40-70, 40-65, 40-60, 40- 55, 40-50, 40-45, 45-70, 45-65, 45-60, 45-55, 45-50, 50-70, 50-65, 50-60, 50-55, 60-70, 60-65, 65-70, 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, or 70 percent by weight a ceramic material (e.g., ⁇ -TCP).
  • a ceramic material e.g., ⁇ -TCP
  • the formulation comprises a first polymer, e.g., about 10-30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20-30, 20-25, or 25-30 percent by weight first polymer.
  • the first polymer may be polycaprolactone (PCL).
  • the first polymer may be polydioxanone (PDS).
  • the first polymer may be poly-l-lactide.
  • the first polymer may be a co- polymer, e.g., caprolactone/glycolide copolymer, poly(D,L-lactide-co-glycolide) copolymer, dioxanone/L-lactide copolymer, or glycolide/L-lactide copolymer.
  • the formulation comprises a second polymer, e.g., about 10-30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20-30, 20-25, or 25-30 percent by weight second polymer.
  • the first and/or second polymer may be water soluble or non-water soluble.
  • the formulation comprises a first polymer, a second polymer, and/or a third polymer.
  • the third polymer may be water soluble or non-water soluble.
  • the second polymer may be polyethylene glycol (PEG).
  • the second polymer may be PEG with a MW of about 8000g/mol.
  • the third polymer may be PEG.
  • the third polymer may be PEG with a MW of about 35,000g/mol.
  • the formulation comprises about 30-70% by weight ceramic, about 10-30% by weight a first polymer, and about 10-30% by weight a second polymer. In a non-limiting embodiment, the formulation comprises about 30-70% by weight ceramic, about 10-30% by weight a first polymer, and about 5-15% by weight a second polymer, and about 5-15% by weight of a third polymer.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight PCL, and about 10-30% by weight PEG.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight poly-l-lactide, and about 10-30% by weight PEG.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% Attorney Docket No: 50222-711.601 by weight poly-l-lactide, about 5-15% by weight PEG (8000 MW), and about 5-15% by weight PEG (35000 MW).
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight copolymer, and about 10-30% by weight PEG.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight copolymer, about 5-15% by weight PEG (8000 MW), and about 5-15% by weight PEG (35000 MW).
  • co-polymers include caprolactone/glycolide copolymer, poly(D,L- lactide-co-glycolide) copolymer, dioxanone/L-lactide copolymer, or glycolide/L-lactide copolymer.
  • the formulation comprises a particulate and/or pore former. The particulate may be water soluble. In some cases, the particulate comprises sucrose.
  • the formulation comprises about 1-10, 1-9, 1-8, 1-7, 1-6, 1-5, 2-10, 2-9, 2-8, 2-7, 2-6, 2-5, 3-10, 3-9, 3-8, 3-7, 3-6, 3-5, 4-10, 4-9, 4-8, 4-7, 4-6, 4-5, 5-10, 5-9, 5-8, 5-7, or 5-6 percent by weight particulate.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight PCL, about 10-30% by weight PEG, and about 1-10% by weight particulate.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight PCL, PDS, poly-l-lactide, caprolactone/glycolide copolymer, poly(D,L-lactide-co-glycolide) copolymer, dioxanone/L-lactide copolymer, or glycolide/L- lactide copolymer, about 5-15% by weight 8000 MW PEG, about 5-15% by weight 35,000 MW PEG, and about 1-10% by weight particulate.
  • the formulation comprises PEG, and the PEG is a pore former.
  • the PEG is present at about 10-30% by weight, or about 5-15% by weight 8000 MW PEG and about 5-15% by weight 35,000 MW PEG.
  • the formulation comprises a blowing agent.
  • the blowing agent comprises sodium bicarbonate.
  • the formulation comprises about 5-20, 5-18, 5-16, 5-14, 5-12, 5-10, 5-8, 5-6, 6-20, 6-18, 6-16, 6-14, 6-12, 6-10, 6-8, 8-20, 8-18, 8-16, 8-14, 8-12, 8-10, 10-20, 10-18, 10-16, 10-14, 10-12, 5-15, or about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, or 15 percent by weight blowing agent.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight PCL, about 10-30% by weight PEG, and about 5-20% by weight blowing agent.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight PCL, PDS, poly-l-lactide, caprolactone/glycolide copolymer, poly(D,L-lactide-co-glycolide) copolymer, dioxanone/L- lactide copolymer, or glycolide/L-lactide copolymer, about 5-15% by weight 8000 MW PEG, about 5-15% by weight 35,000 MW PEG, and about 5-20% by weight blowing agent.
  • the polymer of the formulation is a copolymer, such as a PCL and polyglycolide copolymer.
  • the copolymer comprises about 80-99, 80-98, 80-97, 80-96, 80-95, 80-94, 80-93, 80-92, 80-91, 80-90, 80-89, 80-88, 80-87, 80-86, 80-85, 85- 99, 85-98, 85-97, 85-96, 85-95, 85-94, 85-93, 85-92, 85-91, 85-90, 90-99, 90-98, 90-97, 90-96, 90-95, 90-94, 90-93, 90-92, 90-91, 95-99, 95-98, 95-97, or 95-96 percent molar PCL, and about 1-20, 1-18, 1-16, 1-14, 1-12, 1-10, 1-8, 1-6, 1-4, 1-2, 2-20, 2-18
  • the copolymer comprises about 90-95 percent by mole PCL and about 5-10 percent by mole polyglycolide.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight PCL and polyglycolide copolymer (e.g., 95mol% polycaprolactone, 5mol% polyglycolide), and about 10-30% by weight PEG.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight PCL and polyglycolide copolymer (e.g., 90mol% polycaprolactone, 10mol% polyglycolide), and about 10-30% by weight PEG.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight PCL and polyglycolide copolymer (e.g., 90mol% polycaprolactone, 10mol% polyglycolide, which may be referred to as caprolactone/glycolide copolymer (90:10)), and about 5-15% by weight 8000 MW PEG, about 5-15% by weight 35,000 MW PEG.
  • polyglycolide copolymer e.g., 90mol% polycaprolactone, 10mol% polyglycolide, which may be referred to as caprolactone/glycolide copolymer (90:10)
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight PCL and polyglycolide copolymer (e.g., 95mol% polycaprolactone, 5mol% polyglycolide), and about 5-15% by weight 8000 MW PEG, about 5-15% by weight 35,000 MW PEG.
  • the polymer of the formulation is a copolymer, such as a PDS and polyglycolide copolymer.
  • the copolymer comprises about 80-99, 80-98, 80-97, 80-96, 80-95, 80-94, 80-93, 80-92, 80-91, 80-90, 80-89, 80-88, 80-87, 80-86, 80-85, 85- 99, 85-98, 85-97, 85-96, 85-95, 85-94, 85-93, 85-92, 85-91, 85-90, 90-99, 90-98, 90-97, 90-96, 90-95, 90-94, 90-93, 90-92, 90-91, 95-99, 95-98, 95-97, or 95-96 percent molar PDS, and about 1-20, 1-18, 1-16, 1-14, 1-12, 1-10, 1-8, 1-6, 1-4, 1-2, 2-20, 2-18, 2-16, 2-14, 2-12, 2-10, 2-8, 2-6, 2-4, 3-20, 3-18, 3-16, 3-14, 3-12, 3-10, 3-8, 3-6, 3-4,
  • the copolymer comprises about 90-95 percent by mole PDS and about 5-10 percent by mole polyglycolide.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight PDS and polyglycolide copolymer (e.g., 90mol% PDS, 10mol% polyglycolide), and about 10-30% by weight PEG.
  • the Attorney Docket No: 50222-711.601 formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight PDS and polyglycolide copolymer (e.g., 90mol% PDS, 10mol% polyglycolide), and about 5-15% by weight 8000 MW PEG, about 5-15% by weight 35,000 MW PEG.
  • the polymer of the formulation is a copolymer, such as a poly(D-L-lactide) and glycolide copolymer.
  • the copolymer comprises about 30-50, 31-49, 32-48, 33-47, 34-46, 35-45, 36-44, 37-43, 38-42, 39-41, 80-99, 80-98, 80-97, 80-96, 80- 95, 80-94, 80-93, 80-92, 80-91, 80-90, 80-89, 80-88, 80-87, 80-86, 80-85, 85-99, 85-98, 85-97, 85-96, 85-95, 85-94, 85-93, 85-92, 85-91, 85-90, 90-99, 90-98, 90-97, 90-96, 90-95, 90-94, 90- 93, 90-92, 90-91, 95-99, 95-98, 95-97, or 95-96 percent molar poly(D-L-lactide), and about 30- 50, 31-49, 32-48, 33-47, 34-46, 35-45, 36-44, 37-43, 38-42, 39-41,
  • the copolymer comprises about 50 percent by mole poly(D-L-lactide), and about 50 percent by mole glycolide.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight poly(D-L-lactide) and glycolide copolymer (e.g., 50mol% poly(D-L- lactide), 50mol% glycolide), and about 10-30% by weight PEG.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight poly(D-L-lactide) and glycolide copolymer (e.g., 50mol% poly(D-L-lactide, 50mol% polyglycolide), and about 5-15% by weight 8000 MW PEG, about 5-15% by weight 35,000 MW PEG.
  • the polymer of the formulation is a copolymer, such as a PDS and lactide copolymer.
  • the copolymer comprises about 80-99, 80-98, 80-97, 80-96, 80-95, 80-94, 80-93, 80-92, 80-91, 80-90, 80-89, 80-88, 80-87, 80-86, 80-85, 85-99, 85- 98, 85-97, 85-96, 85-95, 85-94, 85-93, 85-92, 85-91, 85-90, 90-99, 90-98, 90-97, 90-96, 90-95, 90-94, 90-93, 90-92, 90-91, 95-99, 95-98, 95-97, or 95-96 percent molar PDS, and about 1-20, 1- 18, 1-16, 1-14, 1-12, 1-10, 1-8, 1-6, 1-4, 1-2, 2-20, 2-18, 2-16, 2-14, 2-12, 2-10, 2-8, 2-6, 2-4, 3- 20, 3-18, 3-16, 3-14, 3-12, 3-10, 3-8, 3-6, 3-4, 4
  • the copolymer comprises about 90-95 percent by mole PDS and about 5-10 percent by mole lactide.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight PDS and lactide copolymer (e.g., 90mol% PDS, 10mol% lactide), and about 10-30% by weight PEG.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight PDS and lactide copolymer (e.g., 90mol% PDS, 10mol% lactide), and about 5-15% by weight 8000 MW PEG, about 5-15% by weight 35,000 MW PEG.
  • the polymer of the formulation is a copolymer, such as a glycolide and lactide copolymer.
  • the copolymer comprises about 80-99, 80-98, 80- 97, 80-96, 80-95, 80-94, 80-93, 80-92, 80-91, 80-90, 80-89, 80-88, 80-87, 80-86, 80-85, 85-99, 85-98, 85-97, 85-96, 85-95, 85-94, 85-93, 85-92, 85-91, 85-90, 90-99, 90-98, 90-97, 90-96, 90- 95, 90-94, 90-93, 90-92, 90-91, 95-99, 95-98, 95-97, or 95-96 percent molar glycolide, and about 1-20, 1-18, 1-16, 1-14, 1-12, 1-10, 1-8, 1-6, 1-4, 1-2, 2-20, 2
  • the copolymer comprises caprolactone/glycolide (e.g., 90:10, 95:5) copolymer, poly(D,L-lactide-co-glycolide) (e.g., 50:50) copolymer, dioxanone/L-lactide (e.g., 90:10) copolymer, or glycolide/L-lactide (e.g., 95:5) copolymer.
  • the formulation comprises a second polymer, e.g., about 10-30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20-30, 20-25, or 25-30 percent by weight second polymer.
  • the second polymer may be polyethylene glycol (PEG).
  • the formulation comprises about 30-70% by weight Attorney Docket No: 50222-711.601 ceramic, about 10-30% by weight a first polymer, and about 10-30% by weight a second polymer.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight copolymer, and about 10-30% by weight PEG.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight copolymer, and about 5-15% by weight 8000 MW PEG, about 5-15% by weight 35,000 MW PEG.
  • a formulation comprises a ceramic material and one or more polymers.
  • the formulation comprises about 30% to about 70% a ceramic material (e.g., ⁇ -TCP).
  • a ceramic material e.g., ⁇ -TCP
  • the formulation comprises about 30-70, 30-65, 30-60, 30-55, 30-50, 30-45, 30-40, 30-35, 35-70, 35-65, 35-60, 35-55, 35-50, 35-45, 35-40, 40-70, 40-65, 40- 60, 40-55, 40-50, 40-45, 45-70, 45-65, 45-60, 45-55, 45-50, 50-70, 50-65, 50-60, 50-55, 60-70, 60-65, 65-70, 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, or 70
  • the formulation comprises a first polymer, e.g., about 10-30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20-30, 20-25, or 25-30 percent by weight first polymer.
  • the first polymer may be a copolymer, such as a dioxanone and lactide (e.g., L-lactide) copolymer.
  • the copolymer comprises about 80-99, 80-98, 80-97, 80-96, 80-95, 80-94, 80-93, 80-92, 80-91, 80-90, 80-89, 80-88, 80-87, 80-86, 80-85, 85-99, 85-98, 85- 97, 85-96, 85-95, 85-94, 85-93, 85-92, 85-91, 85-90, 90-99, 90-98, 90-97, 90-96, 90-95, 90-94, 90-93, 90-92, 90-91, 95-99, 95-98, 95-97, or 95-96 percent molar dioxanone, and about 1-20, 1- 18, 1-16, 1-14, 1-12, 1-10, 1-8, 1-6, 1-4, 1-2, 2-20, 2-18, 2-16, 2-14, 2-12, 2-10, 2-8, 2-6, 2-4, 3- 20, 3-18, 3-16, 3-14, 3-12, 3-10, 3-8, 3-6, 3
  • the copolymer comprises about 90-95 percent by mole dioxanone and about 5-10 percent by mole lactide.
  • the formulation comprises a second polymer, e.g., about 10-30, 10-25, 10-20, 10-15, 15-30, 15-25, 15-20, 20-30, 20-25, or 25-30 percent by weight second polymer.
  • the second polymer may be polyethylene glycol (PEG).
  • the formulation comprises about 30-70% by weight ceramic, about 10-30% by weight a first polymer, and about 10-30% by weight a second polymer.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight copolymer, and about 10-30% by weight PEG
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight PDS and lactide copolymer (e.g., 90mol% dioxanone, 10mol% L-lactide), and about 10-30% by weight PEG.
  • the formulation may comprise about 30-70% by weight ⁇ -TCP, about 10-30% by weight PDS and lactide Attorney Docket No: 50222-711.601 copolymer (e.g., 90mol% dioxanone, 10mol% L-lactide), and about 5-15% by weight 8000 MW PEG, about 5-15% by weight 35,000 MW PEG.
  • the formulation has a low viscosity that may be useful during manufacture for extruding through a small diameter nozzle.
  • the nozzle may have a diameter of about 240 ⁇ m to about 500 ⁇ m or about 280 to about 450 ⁇ m, or about 240 to about 850 ⁇ m e.g., about 240, 260, 280, 300, 320, 340, 360, 380, 400, 420, 440, 460, 480, 500, 520, 540, 560, 580, 600, 620, 640, 660, 680, 700, 720, 740, 760, 780, 800, 820, 840, or 850 ⁇ m.
  • the formulation is melt-mixed in a dual asymmetric centrifugal mixer to create a homogenous liquid ink.
  • a mixture of low and high viscosity PEG can be used to tailor the molten ink viscosity such that oozing of molten ink out of the 3D printer nozzle during non-print motions is minimized while still enabling flow through a 100’s of microns diameter nozzle when the screw extruder is engaged for print motions.
  • Higher molecular weight PEG generally is stiffer and stronger than low molecular weight PEG, the incorporation of which also improves the mechanical strength and stiffness of the feedstock material.
  • the formulation has a higher viscosity that may be useful during manufacture by forming the formulation into a filament. The filament may then be used for fused filament fabrication.
  • the formulation is in the form of a filament.
  • the filament formulation has a diameter of about 1.5 mm to about 2 mm, or about 1.5 mm, about 1.75 mm, or about 2 mm.
  • the formulation is in the form of a pellet.
  • ink formulations were prepared into pellets for use in 3D structures.
  • the pellets can be made into filaments.
  • pellets can be made Attorney Docket No: 50222-711.601 into powders.
  • pellets have a length of about 1 to about 6 mm, or about 1 to about 5.5 mm, about 1 to about 5 mm, about 1 to about 4.5 mm, about 1 to about 4 mm, about 1 to about 3.5 mm, about 1 to about 3 mm, about 1 to about 2.5 mm, about 1 to about 2 mm, about 1 to about 1.5 mm, about 1.5 to about 6 mm, about 1.5 to about 5.5 mm, about 1.5 to about 5 mm, about 1.5 to about 4.5 mm, about 1.5 to about 4 mm, about 1.5 to about 3.5 mm, about 1.5 to about 3 mm, about 1.5 to about 2.5 mm, about 2 to about 6 mm, about 2 to about 5.5 mm, about 2 to about 5 mm, about 2 to about 4.5 mm, about 2 to about 4 mm, about 2 to about 3.5 mm, about 2 to about 3 mm, about 2 to about 2.5 mm, about 2 to about 6 mm, about 2 to about 5.5 mm, about 2 to about 5 mm, about 2 to about
  • the pellets have a length of about 2.5 mm to about 4.5 mm, or about 2.5 mm, about 3 mm, about 3.5 mm, about 4 mm, or about 4.5 mm.
  • a pellet encompasses a variety of different shapes including spears, rods, granules, blocks, particles, and particles of any suitable shape.
  • the formulation is in the form of a powder.
  • the powder could be produced from a pellet.
  • components of the formulation are melt-mixed into homogenous ink, and cryomilled to form powder.
  • components of the formulation are dissolved in a solvent-based slurry and spray dried to form a powder.
  • powders are used in selective laser sintering.
  • 3D Printed Structures In another aspect, provided herein are 3D printed structures.
  • the structures may be prepared using a formulation and/or method of manufacture described herein.
  • structures include scaffolds, and vice versa.
  • a three-dimensional structure has micropores. The micropores may be formed after removal of a particulate or pore former. The micropores may be formed by a use of a blowing agent during formulation.
  • the micropores provide additional surface area to the structure for contact with a therapeutic agent as compared to a structure lacking Attorney Docket No: 50222-711.601 micropores.
  • the therapeutic agent comprises a targeting moiety that is non-covalently bound to a ceramic material of the structure.
  • the micropores have an average diameter of about 1 micron to about 500 microns.
  • the micropores have an average diameter of about 50 microns to about 250 microns, about 60 microns to about 240 microns, about 70 microns to about 230 microns, about 80 microns to about 220 microns, or about 90 microns to about 210 microns.
  • the micropores have an average diameter of about 1 micron to about 50 microns. For instance, about 1 micron to about 45 microns, about 1 micron to about 40 microns, about 1 micron to about 35 microns, about 1 micron to about 30 microns, about 1 micron to about 25 microns, about 1 micron to about 20 microns, about 1 micron to about 15 microns, about 1 micron to about 10 microns, about 10 microns to about 50 microns, about 10 microns to about 45 microns, about 10 microns to about 40 microns, about 10 microns to about 35 microns, about 10 microns to about 30 microns, about 10 microns to about 25 microns, about 10 microns to about 20 microns, about 10 microns to about 15 microns, about 20 microns to about 50 microns, about 20 microns to about 45 microns, about 20 microns to about 40 microns, about 20 microns Attorney Dock
  • the microporosity of the scaffold results in a hydrophilic scaffold, i.e. liquid readily wicks throughout the scaffold via capillary forces from the interconnected microporosity.
  • a three-dimensional structure has a density of about 1 g/cm 3 to about 3 g/cm 3 . In some embodiments a three-dimensional structure has a density of about 1 g/cm 3 to about 2 g/cm 3 .
  • the structure has about 10-50% copolymer such as polycaprolactone/polyglycolide copolymer (PCL/PGA, e.g., 90:10, 95:5), poly(D,L-lactide-co- glycolide) copolymer (PLGA, e.g., 50:50), PDS-glycolide copolymer (PDS/PGA, e.g., 90:10), PDS-L-lactide copolymer (PDS/PLA, e.g., 90:10), or Dioxanone/L-lactide copolymer (e.g., 90:10).
  • PCL/PGA polycaprolactone/polyglycolide copolymer
  • PLGA poly(D,L-lactide-co- glycolide) copolymer
  • PDS-glycolide copolymer PDS-glycolide copolymer
  • PDS/PLA PDS-L-lactide copoly
  • the structure has about 10, 15, 20, 25, 30, 35, 40, 45, or 50% polymer such as PCL/PGA, PDS/PGA, PDS/PLA, PLGA, Dioxanone/L-lactide, Caprolactone/Glycolide, Glycolide/L-lactide, or Poly(D,L-lactide-co-glycolide).
  • polymer such as PCL/PGA, PDS/PGA, PDS/PLA, PLGA, Dioxanone/L-lactide, Caprolactone/Glycolide, Glycolide/L-lactide, or Poly(D,L-lactide-co-glycolide).
  • Example structures include those having: about 85-90% ceramic (e.g., ⁇ -TCP) and about 10-15% polymer (e.g., PCL/PGA, PDS/PGA, PDS/PLA, PLGA, or Dioxanone/L-lactide) by weight, about 80-85% ceramic (e.g., ⁇ -TCP) and about 15-20% polymer (e.g., PCL/PGA, PDS/PGA, PDS/PLA, PLGA, Dioxanone/L-lactide, Caprolactone/Glycolide, Glycolide/L-lactide, or Poly(D,L-lactide-co-glycolide)) by weight, about 75-80% ceramic (e.g., ⁇ -TCP) and about 20-25% polymer (e.g., PCL/PGA, PDS/PGA, PDS/PLA, Attorney Docket No: 50222-711.601 PLGA, Dioxanone/L-lactide, Caprolactone/
  • compositions of ink formulations herein are varied to optimize specific surface area.
  • the surface area may be optimized for combination with a certain therapeutic agent.
  • the structure has a surface area of about 0.2-2 m 2 /g for combination with a BMP protein (e.g., tBMP-2).
  • the surface area is calculated by Brunauer-Emmett-Teller (BET) by gas physisorption.
  • BET Brunauer-Emmett-Teller
  • the compositions of ink formulations herein are varied to optimize resorption rate of one or more materials of the scaffold.
  • the polymers are selected based on resorption rate. The resorption rates vary from slowest to fastest as: polycaprolactone, polycaprolactone/polyglycolide copolymer (95:5), polycaprolactone/glycolide copolymer (90:10), polydioxanone/L-lactide copolymer (90:10), poly(D,L-lactide-co-glycolide) copolymer (50:50).
  • the method comprises syringe-based melt extrusion bioprinting.
  • Example inks for this method may be low in viscosity for extrusion of the ink through a narrow nozzle.
  • Non-limiting example methods of manufacturing using this method are described in Example 2, for instance, with regard to printing ink formulations #1, #2, #3, #4.
  • Attorney Docket No: 50222-711.601 the method is an extrusion based method comprising a 3D printing method that extrudes the material out of a nozzle.
  • the extrusion based method encompasses bioprinting (syringe- based pneumatic printing) or Fused Granular Fabrication (FGF) where pellets of feedstock are fed from a hopper into a mini-screw extrusion head which melts and pushes the material out of a fine nozzle.
  • the inks are formed into small (e.g., 2-5 mm) pellets or granules.
  • the ink comprises a plurality of pellets or granules having an average diameter of X, wherein at least 90% of the plurality of pellets or granules have an individual diameter of X +/- 0.5 mm.
  • the plurality of pellets or granules have an average diameter of 2 mm, where at least 90% of the plurality of pellets or granules have an individual diameter of 1.5-2.5 mm.
  • the plurality of pellets or granules have an average diameter of 5 mm, where at least 90% of the plurality of pellets or granules have an individual diameter of 4.5-5.5 mm.
  • the method comprises fused filament fabrication (FFF).
  • Example inks for this method may be formed into filaments for printing on FFF 3D printers. Non-limiting example methods of manufacturing using this method are described in Example 2, for instance, with regard to printing ink formulations #5 and #6.
  • the method comprises pelletized fused deposition modeling.
  • Fused deposition modeling is an additive manufacturing process. Three dimensional objects are formed through extrusion and deposition of individual layers of thermoplastic materials. FDM involves the melt extrusion of filament materials through a heated nozzle and deposition as thin solid layers on a platform. A thermoplastic polymer material is fed into a temperature-controlled FDM extrusion head and it is heated to a semi-liquid state. Afterward, the FDM extrusion head extrudes and deposits the material in ultra-thin layers onto a base with precision. The material solidifies, laminating to the preceding layer.
  • each layer is built by extruding a small bead of material, called a road, in a particular pattern, such that the layer is covered with the adjacent roads.
  • extrusion head height is increased and subsequent layers are built to construct the part.
  • FDM is used to fabricate solid models.
  • raster fill gaps have a positive value which is applied to impart a channel within a build layer. Arranged in a regular manner, the channels are interconnected even in three dimensions.
  • Layer by layer fabrication allows design of a pore morphology which varies across a scaffold structure.
  • the method comprises selective laser sintering (SLS).
  • Selective laser sintering is a process wherein a dispenser deposits layers of powdered material into a Attorney Docket No: 50222-711.601 target area.
  • a laser control mechanism typically includes a computer with the article design stored on it. The laser control mechanism modulates and moves a laser beam to selectively irradiate the powder layer within defined boundaries of the design, melting the powder on which the laser beam falls. This is done to selectively sinter sequential powder layers.
  • the method produces a completed article comprised of a plurality of layers sintered together.
  • the resulting subject is soaked in water to dissolve certain components of the ink, e.g., PEG, particulate (e.g., pore forming agent, sucrose), blowing agent (e.g., sodium bicarbonate), or a combination thereof.
  • the structure may then be dried, sterilized, treated with a therapeutic as described elsewhere herein, or a combination thereof.
  • Any of the 3D-printed structures described herein can be coated with a tetherable protein (for example, tBMP2).
  • the structures can be washed in an acidic sodium acetate buffer. This can be one, two, or more washes.
  • the washing can then be followed by a two-hour incubation of the structures in sodium acetate buffer that contains a 1 mg/mL concentration of tBMP2 protein.
  • the tetherable tBMP2 binds to the ⁇ -TCP surface of the implantable structures in a monolayer.
  • the ink formulations discussed herein can include a light-sensitive resin that is mixed with the ceramic powder for digital light processing (DLP), an additive manufacturing technique that is faster than robocasting or melt extrusion.
  • DLP digital light processing
  • Components in a photosensitive, ceramic-filled resin for DLP 3D printing of bone implants typically include ceramic powder (e.g., ⁇ -TCP, hydroxyapatite, bioglass, typically ⁇ 10 ⁇ m particle size), one or more crosslinking acrylates or methacrylates (e.g., polyethylene glycol diacrylate, polycaprolactone methacrylate), a plasticizer to reduce resin viscosity (e.g., water), a dispersant to promote breakdown of powder agglomerates (e.g., Darvan® 821-A), photoinitiator to initiate the photocrosslinking reaction (e.g., Lithium phenyl-2,4,6-trimethylbenzoylphosphinate), and a photoabsorber to retain high x-y resolution (e.g., tartrazine).
  • ceramic powder e.g., ⁇ -TCP, hydroxyapatite, bioglass, typically ⁇ 10 ⁇ m particle size
  • the ink is exposed layer by layer to a DLP image, causing the lighted pixels to selectively solidify when the resin encounters the light.
  • the implantable structure can be thermally processed to burn out the included polymer and densify the ceramic (e.g., a polyethylene glycol diacrylate- containing resin), or left as-is, resulting in a flexible ceramic/polymer composite implant (e.g., a polycaprolactone methacrylate-containing resin).
  • a 3D printed structure e.g., scaffold
  • a device comprises the Attorney Docket No: 50222-711.601 therapeutic agent connected to, dispersed within, or otherwise combined with the 3D printed structure.
  • a therapeutic agent is inclusive of a plurality of therapeutic agents, such as 2, 3, 4, or 5 therapeutic agents.
  • Therapeutic agents In some embodiments, the therapeutic agent comprises a mammalian growth factor or a functional portion thereof.
  • Mammalian growth factors can be osteoinductive molecules that are capable of initiating and enhancing the bone repair process.
  • a functional portion of the mammalian growth factor is a region that has a therapeutic effect. For instance, a functional portion of a mammalian growth factor is osteoinductive.
  • a functional portion of a mammalian growth factor is capable of initiating and/or enhancing bone repair.
  • a functional portion of a mammalian growth factor may have osteogenic activity.
  • Non-limiting examples of mammalian growth factors are described herein.
  • the mammalian growth factor comprises: epidermal growth factor (EGF), platelet derived growth factor (PDGF), insulin like growth factor (IGF-1), fibroblast growth factor (FGF), fibroblast growth factor 2 (FGF2), fibroblast growth factor 18 (FGF18), transforming growth factor alpha (TGF- ⁇ ), transforming growth factor beta (TGF- ⁇ ), transforming growth factor beta 1 (TGF- ⁇ 1), transforming growth factor beta 3 (TGF- ⁇ 3), osteogenic protein 1 (OP-1), osteogenic protein 2 (OP-2), osteogenic protein 3 (OP-3), bone morphogenetic protein 2 (BMP-2), bone morphogenetic protein 3 (BMP-3), bone morphogenetic protein 4 (BMP-4), bone morphogenetic protein 5 (BMP-5), bone morphogenetic protein 6 (BMP-6), bone morphogenetic protein 7 (BMP- 7), bone morphogenetic protein (BMP-9), bone morphogenetic protein 10 (BMP-10), bone morphogenetic protein 11 (BMP-11), bone
  • a targeting peptide comprises one or more sequences of Table 3.
  • the targeting peptide comprises a sequence at least about 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to a sequence of Table 3.
  • Additional targeting peptides useful in the present disclosure include any one of SEQ ID NO: 1 to SEQ ID NO: 558 of US 7,572,766.
  • the targeting peptide comprises a sequence at least about 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to any one of SEQ ID NO: 1 to SEQ ID NO: 558 of US 7,572,766.
  • the cells can be added to the completed implantable structures.
  • a structure e.g., scaffold
  • a therapeutic agent e.g., a chimeric polypeptide comprising the therapeutic agent and a targeting moiety
  • a second solution such as phosphate buffered saline (PBS).
  • PBS phosphate buffered saline
  • the method further comprises drying the 3D structure of step (c) or step (d).
  • the mass of the therapeutic agent e.g., a therapeutic agent alone or a therapeutic agent connected to a targeting moiety
  • the mass of the therapeutic agent per cubic centimeter of the structure in a device is between about 0.05 and 50 (mg/cc), e.g., about 0.05, 0.06, 0.07, 0.08, 0.09, 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, 10, 15, 20, 25, 30, 35, 40, 45, or 50 mg/cc or any number therebetween.
  • the therapeutic agent is about 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, or 2 mg per cubic centimeter device.
  • One method of measuring the amount of therapeutic peptide bound to the structure includes: (1) measuring the mass of therapeutic peptide input in the first solution, (2) measuring the mass of the therapeutic agent remaining in the first solution after combination with and removal from the structure, (3) measuring the mass of the therapeutic agent in the second solution if a wash step is included, (4) summing (2) and (3); and subtracting the sum of (4) from (1).
  • the subject has a bone fracture or a bone defect. In some instances, the subject requires a vertebral fusion of the spine. In some instances, the subject has a cartilage tear or cartilage defect. In some instances, the subject has cartilage loss. In some embodiments, the subject is suffering from a defect in bone, cartilage, soft tissue, tendon, fascia, ligament, organ, osteotendinous tissue, dermal, or osteochondral, or a combination of one or more of the aforementioned defects. In some embodiments, a defect is a lack of bone, cartilage, soft tissue, tendon, fascia, ligament, organ, osteotendinous tissue, dermal, or osteochondral, or a combination of one or more of the aforementioned defects.
  • a defect in the subject arises from trauma. In some embodiments, a defect in the subject arises due to a congenital condition. In some embodiments, a defect in the subject arises due to an acquired condition. In some embodiments, a defect refers to the absence, loss, and/or break in a tissue and/or organ of the body. In some embodiments, a “bone defect” refers to the Attorney Docket No: 50222-711.601 absence or loss (e.g., partial loss) of bone at an anatomical location in a subject where it would otherwise be present in a control healthy subject. A bone defect may be the result of an infection (e.g., osteomyelitis), a tumor, a trauma, or an adverse event of a treatment.
  • an infection e.g., osteomyelitis
  • a bone defect may also affect the muscles, soft tissue, tendons, or joints surrounding the bone defect and cause injury.
  • a bone defect includes damage to a soft tissue.
  • a “cartilage defect” refers to the absence or loss (e.g., partial loss) of cartilage at an anatomical location in a subject where it would otherwise be present in a control healthy subject.
  • a cartilage defect may be the result of disease, osteochondritis, osteonecrosis, or trauma. For example, a cartilage defect may affect the knee joint.
  • Non-limiting examples of conditions suitable for treatment with a structure or device described herein include osteoarthritis, disc degeneration, congenital defect, spinal stenosis, spondylolisthesis, spondylosis, bone fracture, scoliosis, kyphosis, spinal fusion (PLF, and interbody fusions), trauma repair of bone, dental repair, craniomaxillofacial repair, ankle fusion, kyphoplasty, balloon osteoplasty, scaphoid facture repair, tendon-osseous repair, osteoporosis, avascular necrosis, congenital skeletal malformations, costal reconstruction, subchondral bone repair, cartilage repair (e.g., at low doses), or trauma, or a combination thereof.
  • the methods may comprise treatment to hair follicles.
  • the trauma may be to the bone, cartilage, soft tissue, tendon, fascia, ligament, organ, osteotendinous tissue, or dermal tissue, or osteochondral tissue.
  • the method is to treat an osteochondral injury.
  • the methods of treatment may comprise spinal fusion.
  • spinal fusion is a surgical technique to join two or more vertebrae.
  • the spinal fusion comprises PLF.
  • the spinal fusion comprises interbody fusions.
  • Some embodiments of these methods can further include first selecting a subject in need of bone or cartilage formation.
  • the structure or device is administered to the subject proximal to the desired site of bone or cartilage formation in the subject.
  • methods of replacing and/or repairing bone or cartilage in a subject in need thereof that include administering to the subject a therapeutically effective amount of any of the structure or devices described herein.
  • Some embodiments of these methods can further include first selecting a subject in need of bone replacement, bone repair, cartilage Attorney Docket No: 50222-711.601 replacement, or cartilage repair.
  • the structure or device is administered to the subject proximal to the desired site of bone or cartilage replacement or repair in the subject.
  • methods of treating a bone fracture or bone loss in a subject in need thereof comprising administering to the subject a therapeutically effective amount of any of the structure or devices described herein. Some embodiments of these methods can further include first selecting a subject having a bone fracture or bone loss.
  • the structure or device is administered to the subject proximal to the bone fracture or the site of bone loss in the subject.
  • methods of repairing soft tissue in a subject in need thereof that include administering to the subject a therapeutically effective amount of any of the structure or devices described herein.
  • Some embodiments of these methods can further include first selecting a subject having a bone fracture or bone loss.
  • the composition is administered to the subject proximal to the bone fracture or the site of bone loss in the subject.
  • methods of localized delivery of a therapeutic to a subject in need thereof that include administering to the subject a therapeutically effective amount of any of the structure or devices described herein.
  • Some embodiments of these methods can further include first selecting a subject having a bone fracture or bone loss.
  • the structure or device is administered to the subject proximal to the bone fracture or the site of bone loss in the subject.
  • Methods of determining the efficacy of treatment of a bone fracture or bone loss in a subject include, e.g., imaging techniques (e.g., magnetic resonance imaging, X-ray, or computed tomography).
  • imaging techniques e.g., magnetic resonance imaging, X-ray, or computed tomography
  • Methods of detecting bone or cartilage formation, or replacement or repair of bone or cartilage in a subject are also known in the art and include, e.g., imaging techniques (e.g., magnetic resonance imaging, X-ray, or computed tomography).
  • imaging techniques e.g., magnetic resonance imaging, X-ray, or computed tomography
  • Suitable animal models for treatment of a bone fraction or bone loss, bone or cartilage formation, or bone or cartilage replacement or repair are known in the art.
  • a method of treatment comprises administering to the subject a structure or device herein.
  • administration comprises implanting a polypeptide or composition herein.
  • Attorney Docket No: 50222-711.601 In some embodiments, a polypeptide and/or composition herein comprising BMP-2 is administered to the subject.
  • the BMP2 comprises a sequence at least about 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to SEQ ID NO: 454.
  • the BMP-2 is administered to induce formation of bone in the subject.
  • the BMP-2 is administered to induce formation of cartilage.
  • the BMP-2 is administered in a spinal fusion.
  • the term “about” or “approximately” means within an acceptable error range for the particular value as determined by one of ordinary skill in the art, which will depend in part on how the value is measured or determined, e.g., the limitations of the measurement system. For example, “about” can mean within 1 or more than 1 standard deviation, per the practice in the given value.
  • subject refers to any mammal.
  • a subject therefore refers to, for example, mice, rats, dogs, cats, horses, cows, pigs, guinea pigs, rats, humans, monkeys, and the like.
  • the subject When the subject is a human, the subject may be referred to herein as a patient.
  • the subject or “subject in need of treatment” may be a canine (e.g., a dog), feline (e.g., a cat), equine (e.g., a horse), ovine, bovine, porcine, caprine, primate, e.g., a simian (e.g., a monkey (e.g., marmoset, baboon), or an ape (e.g., a gorilla, chimpanzee, orangutan, or gibbon), a human, or a rodent (e.g., a mouse, a guinea pig, a hamster, or a rat).
  • a canine e.g., a dog
  • feline e.g., a cat
  • equine e.g., a horse
  • ovine, bovine, porcine caprine
  • primate e.g., a simian (e.g.
  • the subject or “subject in need of treatment” may be a non-human mammal, especially mammals that are conventionally used as models for demonstrating therapeutic efficacy in humans (e.g., murine, lapine, porcine, canine, or primate animals) may be employed.
  • the term “therapeutically effective amount” refers to an amount of a polypeptide or composition effective to “treat” a disease, condition or disorder in a subject. In some cases, therapeutically effective amount of the polypeptide or composition reduces the severity of symptoms of the disease, condition or disorder. In some instances, the disease, condition or disorder comprises a defect in an organ or tissue.
  • affinity refers to the strength of the sum total of non-covalent interactions between a ⁇ -TCP binding sequence (or a chimeric polypeptide or polypeptide Attorney Docket No: 50222-711.601 comprising a ⁇ -TCP binding sequence) and its binding partner (e.g., ⁇ -TCP).
  • Affinity can be measured by common methods known in the art, including those described herein. Affinity can be determined, for example, using surface plasmon resonance (SPR) technology (e.g., BIACORE®) or biolayer interferometry (e.g., FORTEBIO®).
  • SPR surface plasmon resonance
  • BIACORE® BIACORE®
  • biolayer interferometry e.g., FORTEBIO®
  • Percent (%) sequence identity with respect to a reference polypeptide sequence is the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the reference polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are known for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Appropriate parameters for aligning sequences are able to be determined, including algorithms needed to achieve maximal alignment over the full length of the sequences being compared.
  • the ink formulation of embodiment 31, comprising about 60% by weight ⁇ TCP, about 20% by weight caprolactone/glycolide copolymer, about 10% by weight PEG having a molecular weight from about 500 g/mol to about 15,000 g/mol, and about 10% PEG having a molecular weight from about 25,000 g/mol to about 50,000 g/mol. 33.
  • the ink formulation of embodiment 32 comprising about 60% by weight ⁇ TCP, about 20% by weight caprolactone/glycolide copolymer, about 10% by weight PEG having a molecular weight of about 8,000 g/mol, and about 10% PEG having a molecular weight of about 35,000 g/mol. 34.
  • An ink formulation comprising about 55% to about 65% by weight ⁇ TCP, about 15% to about 25% by weight PDS, about 5% to about 15% PEG having a molecular weight from about 500 g/mol to about 15,000 g/mol, and about 5% to about 15% PEG having a molecular weight from about 25,000 g/mol to about 50,000 g/mol. 41.
  • the ink formulation of embodiment 40 comprising about 60% by weight ⁇ TCP, about 20% by weight PDS, about 10% by weight PEG having a molecular weight from about 500 g/mol to about 15,000 g/mol, and about 10% PEG having a molecular weight from about 25,000 g/mol to about 50,000 g/mol. 42.
  • the ink formulation of embodiment 41 comprising about 60% by weight ⁇ TCP, about 20% by weight PDS, about 10% by weight PEG having a molecular weight of about 8,000 g/mol, and about 10% PEG having a molecular weight of about 35,000 g/mol. 43.
  • An ink formulation comprising about 55% to about 65% by weight ⁇ TCP, about 15% to about 25% by weight dioxanone/L-lactide copolymer, about 5% to about 15% PEG having a molecular weight from about 500 g/mol to about 15,000 g/mol, and about 5% to about 15% PEG having a molecular weight from about 25,000 g/mol to about 50,000 g/mol. 44.
  • the ink formulation of embodiment 43 comprising about 60% by weight ⁇ TCP, about 20% by weight dioxanone/L-lactide copolymer, about 10% by weight PEG having a molecular weight from about 500 g/mol to about 15,000 g/mol, and about 10% PEG having a molecular weight from about 25,000 g/mol to about 50,000 g/mol. 45.
  • the ink formulation of embodiment 44 comprising about 60% by weight ⁇ TCP, about 20% by weight dioxanone/L-lactide copolymer, about 10% by weight PEG having a molecular weight of about 8,000 g/mol, and about 10% PEG having a molecular weight of about 35,000 g/mol. 46.
  • An ink formulation comprising about 55% to about 65% by weight ⁇ TCP, about 15% to about 25% by weight glycolide/L-lactide copolymer, about 5% to about 15% PEG having a molecular weight from about 500 g/mol to about 15,000 g/mol, and about 5% to about 15% PEG having a molecular weight from about 25,000 g/mol to about 50,000 g/mol. 48.
  • the ink formulation of embodiment 47 comprising about 60% by weight ⁇ TCP, about 20% by weight glycolide/L-lactide copolymer, about 10% by weight PEG having a molecular weight from about 500 g/mol to about 15,000 g/mol, and about 10% PEG having a molecular weight from about 25,000 g/mol to about 50,000 g/mol.
  • Attorney Docket No: 50222-711.601 49 The ink formulation of embodiment 48, comprising about 60% by weight ⁇ TCP, about 20% by weight glycolide/L-lactide copolymer, about 10% by weight PEG having a molecular weight of about 8,000 g/mol, and about 10% PEG having a molecular weight of about 35,000 g/mol. 50.
  • An ink formulation comprising about 55% to about 65% by weight ⁇ TCP, about 15% to about 25% by weight poly-l-lactide, about 5% to about 15% PEG having a molecular weight from about 500 g/mol to about 15,000 g/mol, and about 5% to about 15% PEG having a molecular weight from about 25,000 g/mol to about 50,000 g/mol. 52.
  • the ink formulation of embodiment 51 comprising about 60% by weight ⁇ TCP, about 20% by weight poly-l-lactide, about 10% by weight PEG having a molecular weight from about 500 g/mol to about 15,000 g/mol, and about 10% PEG having a molecular weight from about 25,000 g/mol to about 50,000 g/mol. 53.
  • the ink formulation of embodiment 52 comprising about 60% by weight ⁇ TCP, about 20% by weight poly-l-lactide, about 10% by weight PEG having a molecular weight of about 8,000 g/mol, and about 10% PEG having a molecular weight of about 35,000 g/mol. 54.
  • a method of preparing a three-dimensional structure comprises performing additive manufacturing with the ink formulation of any one of embodiments 26-53. 55. The method of embodiment 54, wherein the ink formulation is in the form of a pellet. 56. The method of embodiment 55, wherein the additive manufacturing comprises fused granular fabrication (FGF). 57. The method of embodiment 54, wherein the ink formulation is in the form of a filament. 58. The method of embodiment 57, wherein the additive manufacturing comprises fused filament fabrication (FFF). 59.
  • the growth factor is selected from Table 1.
  • the therapeutic agent comprises a bone morphogenetic protein (BMP).
  • BMP bone morphogenetic protein
  • the therapeutic agent comprises a targeting moiety, and the targeting moiety is non-covalently bound to the structure.
  • the targeting moiety comprises a polypeptide at least about 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to any one of the sequences of Tables 2-3.
  • the therapeutic agent comprises a chimeric polypeptide comprising a sequence at least about 70%, 75%, 80%, 85%, 90%, 95%, or 100% identical to any one of SEQ ID NOS: 433-441.
  • a method of treating a bone defect in a subject in need thereof the method comprising application of the structure of any one of embodiments 1-25 or embodiments 54-58 to the defect of the subject.
  • 68. A method of treating a bone defect in a subject in need thereof, the method comprising application of the device of any one of embodiments 59-66 to the defect of the subject.
  • 69 The method of embodiment 67 or embodiment 68, wherein the defect is present in the spine.
  • Example 1 Ink Formulations and 3D Printed Scaffolds
  • Ink formulation and scaffold #1 This 3D printing ink material is a flexible, polymer-ceramic composite material containing ⁇ -TCP that may be tethered to a therapeutic agent, such as tBMP2.
  • This formulation contains two sacrificial pore formers (water soluble polyethylene glycol and water soluble sucrose) to expose more ⁇ -TCP surface area for tBMP2 binding.
  • This ink is a low viscosity formulation that was extruded through a 400 ⁇ m diameter nozzle on an Allevi 3 pneumatic bioprinter.
  • the resulting scaffold was soaked in water to dissolve the sacrificial polyethylene glycol and sucrose pore formers.
  • the resulting scaffold #1 contains 75% by weight ⁇ -TCP powder, and 25% by weight polycaprolactone.
  • This ink is also converted into a filament form and utilized to prepare a 3D printed scaffold using a fused filament fabrication (FFF or FDM) 3D printer.
  • FFF or FDM fused filament fabrication
  • the resulting scaffold is soaked in water to dissolve the sacrificial polyethylene glycol and sucrose pore formers.
  • the resulting scaffold contains 75% by weight ⁇ -TCP powder, and 25% by weight polycaprolactone.
  • This ink is also converted into pellets (e.g., a filament is formed into small pellets) and fed from a hopper into a mini-screw extrusion head which melts and pushes the material out of a fine nozzle.
  • This ink is also cryomilled into a fine powder.
  • a laser heats the powder into the desired configuration using the SLS method.
  • Table 4 Example ink formulation #1 Ink formulation #2
  • This 3D printing ink material is a flexible, polymer-ceramic composite material containing ⁇ -TCP that may be tethered to a therapeutic agent, such as tBMP2.
  • This formulation contains a sacrificial pore former (water soluble polyethylene glycol) to expose more ⁇ -TCP surface area for agent binding.
  • This formulation employs a 95mol% caprolactone 5mol% glycolide copolymer for faster bioresorption characteristics compared to polycaprolactone.
  • This ink is a low viscosity formulation that was extruded through a 320 ⁇ m diameter nozzle on an Allevi 3 pneumatic bioprinter. After 3D printing is complete, the resulting scaffold was soaked in water to dissolve the sacrificial polyethylene glycol pore former. The resulting scaffold #2 contains 75% by weight ⁇ -TCP powder, and 25% by weight caprolactone/glycolide copolymer (95:5).
  • This ink is also converted into a filament form and utilized to prepare a 3D printed scaffold using a fused filament fabrication (FFF or FDM) 3D printer. After 3D printing is complete, the resulting scaffold is soaked in water to dissolve the sacrificial polyethylene glycol pore former. The resulting scaffold contains 75% by weight ⁇ -TCP powder, and 25% by weight caprolactone/glycolide copolymer (95:5).
  • This ink is also converted into pellets (e.g., a filament is formed into small pellets) and fed from a hopper into a mini-screw extrusion head which melts and pushes the material out of a fine nozzle.
  • This ink is also cryomilled into a fine powder.
  • a laser heats the powder into the desired configuration using the SLS method.
  • Table 5 Example ink formulation #2 Ink formulation #3
  • This 3D printing ink material is a flexible, polymer-ceramic composite material containing ⁇ -TCP that may be tethered to a therapeutic agent, such as tBMP2.
  • This formulation contains a sacrificial pore former (water soluble polyethylene glycol) to expose more ⁇ -TCP surface area for agent binding.
  • This formulation employs a 90mol% caprolactone 10mol% glycolide copolymer for fast bioresorption characteristics compared to polycaprolactone.
  • This ink is a low viscosity formulation that was extruded through a 320 ⁇ m diameter nozzle on an Allevi 3 pneumatic bioprinter. After 3D printing was complete, the resulting scaffold was soaked in water to dissolve the sacrificial polyethylene glycol pore former. The resulting scaffold #3 contains 75% by weight ⁇ -TCP powder, and 25% by weight caprolactone/glycolide copolymer (90:10). This ink is also converted into a filament form and utilized to prepare a 3D printed scaffold using a fused filament fabrication (FFF or FDM) 3D printer. After 3D printing is complete, the resulting scaffold is soaked in water to dissolve the sacrificial polyethylene glycol pore former.
  • FFF or FDM fused filament fabrication
  • the resulting scaffold contains 75% by weight ⁇ -TCP powder, and 25% by weight caprolactone/glycolide copolymer (90:10).
  • This ink is also converted into pellets (e.g., a filament is formed into small pellets) and fed from a hopper into a mini-screw extrusion head which melts and pushes the material out of a fine nozzle.
  • This ink is also cryomilled into a fine powder.
  • a laser heats the powder into the desired configuration using the SLS method. Table 6.
  • Example ink formulation #3 Ink formulation #4 This 3D printing ink material is a flexible, polymer-ceramic composite material containing ⁇ -TCP that may be tethered to a therapeutic agent, such as tBMP2.
  • This formulation contains a sacrificial pore former (water soluble polyethylene glycol) to expose more ⁇ -TCP surface area for agent binding.
  • This formulation employs a 50mol%:50mol% poly(D,L-lactide-co-glycolide) copolymer for fast bioresorption characteristics compared to polycaprolactone.
  • This ink is a low viscosity formulation that was extruded through a 400 ⁇ m diameter nozzle on an Allevi 3 pneumatic bioprinter.
  • the resulting scaffold #4 contains 75% by weight ⁇ -TCP powder, and 25% by weight poly(D,L-lactide-co-glycolide) copolymer (50:50). This ink is also converted into a filament form and utilized to prepare a 3D printed scaffold using a fused filament fabrication (FFF or FDM) 3D printer. After 3D printing is complete, the resulting scaffold is soaked in water to dissolve the sacrificial polyethylene glycol pore former.
  • FFF or FDM fused filament fabrication
  • the resulting scaffold contains 75% by weight ⁇ -TCP powder, and 25% by weight poly(D,L- lactide-co-glycolide) copolymer (50:50).
  • Attorney Docket No: 50222-711.601 This ink is also converted into pellets (e.g., a filament is formed into small pellets) and fed from a hopper into a mini-screw extrusion head which melts and pushes the material out of a fine nozzle. This ink is also cryomilled into a fine powder. A laser heats the powder into the desired configuration using the SLS method. Table 7.
  • Example ink formulation #4 Ink formulation #5 This 3D printing ink material is a flexible, polymer-ceramic composite material containing ⁇ -TCP that may be tethered to a therapeutic agent, such as tBMP2.
  • This formulation contains a sacrificial pore former (water soluble polyethylene glycol) to expose more ⁇ -TCP surface area for agent binding.
  • This ink is a moderate viscosity formulation that was formed into a 1.75 mm filament for 3D printing on a RepRap style FFF 3D printers (e.g. Prusa i3 MK3S 3D printer) with a 400 ⁇ m diameter nozzle.
  • the higher molecular weight polyethylene glycol (8000 MW for FFF 3D printing vs.
  • This ink is a low viscosity formulation that can be extruded through a 400 ⁇ m diameter nozzle on an Allevi 3 pneumatic bioprinter. After 3D printing is complete, the resulting scaffold is soaked in water to dissolve the sacrificial polyethylene glycol pore former. The resulting scaffold contains 75% by weight ⁇ -TCP powder, and 25% by weight dioxanone/L-lactide copolymer (90:10). This ink is also converted into a filament form and utilized to prepare a 3D printed scaffold using a fused filament fabrication (FFF or FDM) 3D printer. After 3D printing is complete, the resulting scaffold is soaked in water to dissolve the sacrificial polyethylene glycol pore former.
  • FFF or FDM fused filament fabrication
  • This ink is a moderate viscosity formulation that can be formed into a 1.75 mm filament for 3D printing on a RepRap style FFF 3D printers (e.g. Prusa i3 MK3S 3D printer) with a 400 ⁇ m diameter nozzle.
  • the higher molecular weight blend of polyethylene glycol (8000 MW and 20000 MW for FFF 3D printing vs.1500 MW for syringe- based bioprinting) results in a higher viscosity material. which aids in extrusion of 1.75 mm diameter filaments.
  • the resulting scaffold is soaked in water to dissolve the sacrificial polyethylene glycol and sucrose pore formers.
  • Example ink formulation #10 Ink formulation #11 This 3D printing ink material is a flexible, polymer-ceramic composite material containing ⁇ -TCP that may be tether to a therapeutic agent, such as tBMP2.
  • This formulation contains a sacrificial pore former (water soluble polyethylene glycol) to expose more ⁇ -TCP surface area for agent binding.
  • This formulation employs a 90mol% caprolactone 10mol% glycolide copolymer for fast bioresorption characteristics compared to polycaprolactone.
  • Example ink formulation #21 Attorney Docket No: 50222-711.601 Ink formulation #22
  • This 3D printing ink material is a flexible, polymer-ceramic composite material containing ⁇ -TCP that may be tethered to a therapeutic agent, such as tBMP2.
  • This formulation contains sacrificial pore formers (water soluble polyethylene glycol components) to expose more ⁇ -TCP surface area for agent binding.
  • This ink is a moderate viscosity formulation that can be formed into ⁇ 3-4mm diameter pellets to be used as feedstock in a fused granular fabrication (FGF) 3D printer (e.g. Piocreat G5) with a 300 – 1,000 ⁇ m diameter nozzle.
  • FGF fused granular fabrication
  • This ink is cryomilled into a fine powder, e.g., for SLS printing.
  • Example ink formulation #22 Ink formulation #23 This 3D printing ink material is a flexible, polymer-ceramic composite material containing ⁇ -TCP that may be tethered to a therapeutic agent, such as tBMP2.
  • This formulation contains sacrificial pore formers (water soluble polyethylene glycol components) to expose more ⁇ -TCP surface area for agent binding.
  • This ink is a moderate viscosity formulation that can be formed into ⁇ 3-4mm diameter pellets to be used as feedstock in a fused granular fabrication (FGF) 3D printer (e.g.
  • FGF fused granular fabrication
  • Piocreat G5 with a 300 – 1,000 ⁇ m diameter nozzle.
  • the resulting scaffold is soaked in water to dissolve the sacrificial polyethylene glycol pore former.
  • the higher molecular weight blend of polyethylene glycol (8000 MW and 35000 MW for FGF 3D printing vs. 1500 MW for syringe-based bioprinting) results in a higher viscosity material.
  • the resulting scaffold is soaked in water to dissolve the sacrificial PEG.
  • the resulting scaffold contains 55-88% by weight ⁇ -TCP powder, and 13-50% by weight Glycolide/L-lactide copolymer (95:5).
  • This ink is prepared in pellet or filament form for FGF or FFF printing, respectively. This ink is cryomilled into a fine powder, e.g., for SLS printing. Table 32.
  • Example ink formulation #23 Ink formulation #24 This 3D printing ink material is a flexible, polymer-ceramic composite material containing ⁇ -TCP that may be tethered to a therapeutic agent, such as tBMP2. This formulation contains sacrificial pore formers (water soluble polyethylene glycol components) to expose more ⁇ -TCP surface area for agent binding.
  • This ink is a moderate viscosity formulation that can be formed into ⁇ 3-4mm diameter pellets to be used as feedstock in a fused granular fabrication (FGF) 3D printer (e.g. Piocreat G5) with a 300 – 1,000 ⁇ m diameter nozzle.
  • FGF fused granular fabrication
  • the resulting scaffold is soaked in water to dissolve the sacrificial polyethylene glycol pore former.
  • the higher molecular weight blend of polyethylene glycol (8000 MW and 35000 MW for FGF 3D printing vs. 1500 MW for syringe-based bioprinting) results in a higher viscosity material.
  • the resulting scaffold is soaked in water to dissolve the sacrificial PEG.
  • the glass jar was placed in a dual asymmetric centrifugal mixer (FlackTek Speedmixer) and mixed at low intensity (300 rpm) for 2 min to homogenize the powder blend Attorney Docket No: 50222-711.601 before high rpm mixing.
  • the mixer was mixed for 5 min at high intensity (3500 rpm).
  • the internal friction causes the polycaprolactone and polyethylene glycol to melt, changing the ink to a viscous molten liquid.
  • This liquid phase mixing facilitates intimate dispersion of the ⁇ -TCP powder and sucrose powder into the molten polymer blend.
  • the blended ink was allowed to cool for 10-15 min, then mixed for 5 more minutes at 3500 rpm.
  • the scaffold was 3D printed on painter’s tape applied to a smooth glass or polymer surface, such as a glass microscope slide, larger glass plate, or 96 well plate lid.
  • Post Processing 3D printed structures were soaked overnight in distilled water to dissolve the polyethylene glycol and sucrose from the printed material, thus creating a porous and flexible ⁇ -TCP/polycaprolactone composite. Scaffolds were dried for at least twelve hours to ensure residual water has evaporated from the porous scaffold before binding with a therapeutic agent such as tBMP2 protein.
  • FIGS.1A-1C Images of the scaffold are shown in FIGS.1A-1C.
  • 3D printing is also performed using a FFF 3D printer.
  • the ink #1 is extruded into filaments and the filaments are loaded in a FFF 3D printer to generate a 3D printed scaffold.
  • the scaffold is processed using the post processing method outlined above.
  • Ink formulation and scaffold #2 Method To make a 5 cc batch of ink, 5.6 g of ⁇ -TCP powder, 1.87 g of 95:5 caprolactone/glycolide copolymer pellets, and 1.87 g of polyethylene glycol flake were added to a glass mixing container. The glass jar was placed in dual asymmetric centrifugal mixer (FlackTek Speedmixer) and mixed at low intensity (300 rpm) for 2 min to homogenize the powder blend before high rpm mixing. The mixture was mixed for 5 min at high intensity (3500 rpm).
  • the internal friction causes the 95:5 caprolactone/glycolide copolymer and polyethylene glycol to melt, changing the ink to a viscous molten liquid.
  • This liquid phase mixing facilitates intimate dispersion of the ⁇ -TCP powder into the molten polymer blend.
  • the blended ink was Attorney Docket No: 50222-711.601 allowed to cool for 10-15 min, and then mixed for 5 more minutes at 3500 rpm. The mixing/cooling process was repeated for a total of four 5 min mixes at 3500 rpm. After the fourth mix at 3500 rpm, the ink charge was poured out on a glass plate and two spatulas were used to form into a roughly 1 cm diameter x 6 cm long cylinder.
  • 3D Printing The solid polymer/ ⁇ -TCP pieces were transferred to a 5 cc stainless steel syringe (for use in Allevi 3 Bioprinter). Extruder CORE printing head was heated to 130°C and allowed to dwell for approximately 30 min to ensure melting of the ink. Ink was printed with 320 micron I.D. conical metallic Luer lock tip using 80 psi pressure and 6 mm/s nozzle velocity. Scaffolds were 3D printed on painter’s tape applied to a smooth glass or polymer surface, such as a glass microscope slide, larger glass plate, or 96 well plate lid.
  • 3D printed structures were soaked overnight in distilled water to dissolve the polyethylene glycol from the printed material, thus creating a porous and flexible ⁇ -TCP/95:5 caprolactone/glycolide copolymer composite.
  • the scaffolds were dried for at least twelve hours to ensure residual water evaporated from the porous scaffold before binding with a therapeutic agent like tBMP2.
  • Scaffolds were sterilized by soaking for 2-4 hours in a 70% ethanol solution and allowed to dry in biosafety cabinet for approximately 12 hours. Images of the scaffold are shown in FIGS.2A-2C. 3D printing is also performed using a FFF 3D printer.
  • the ink #2 is extruded into filaments and the filaments are loaded in a FFF 3D printer to generate a 3D printed scaffold.
  • the scaffold is processed using the post processing method outlined above.
  • Ink formulation and scaffold #3 Method To make a 5 cc batch of ink, 5.6 g of ⁇ -TCP powder, 1.87 g of 90:10 caprolactone/glycolide copolymer chips, and 1.87 g of polyethylene glycol flake were added to a glass mixing container. The mixture was placed in a glass jar in dual asymmetric centrifugal mixer (FlackTek Speedmixer) and mixed at low intensity (300 rpm) for 2 min to homogenize the powder blend before high rpm mixing.
  • the ink charge was poured out on a glass plate and two spatulas were used to Attorney Docket No: 50222-711.601 form into a roughly 1 cm diameter x 6 cm long cylinder. While the ink was still semi-molten, it was cut into several ⁇ 1-2 cm long pieces with straight razor.
  • 3D Printing the solid polymer/ ⁇ -TCP pieces were transferred to a 5 cc stainless steel syringe (for use in Allevi 3 Bioprinter). Extruder CORE printing head was heated to 130°C and allowed to dwell for approximately 30 min to ensure melting of the ink. Ink was printed with 320 micron I.D.
  • FIGS.3A-3C Images of the scaffold are shown in FIGS.3A-3C.
  • 3D printing is also performed using a FFF 3D printer.
  • the ink #3 is extruded into filaments and the filaments are loaded in a FFF 3D printer to generate a 3D printed scaffold.
  • the scaffold is processed using the post processing method outlined above.
  • Ink formulation and scaffold #4 Method To make a 2.5 cc batch of ink, 2.8 g of ⁇ -TCP powder, 0.94 g of 50:50 poly(D,L- lactide-co-glycolide) copolymer chunks, and 0.94 g of polyethylene glycol flake were added to a glass mixing container. The glass jar was placed in dual asymmetric centrifugal mixer (FlackTek Speedmixer) and mixed at low intensity (300 rpm) for 2 min to homogenize the powder blend before high rpm mixing. It was mixed for 5 min at high intensity (3500 rpm).
  • the internal friction causes the 50:50 poly(D,L-lactide-co-glycolide) copolymer and polyethylene glycol to flow, changing the ink to a viscous molten liquid.
  • This liquid phase mixing facilitates intimate dispersion of the ⁇ -TCP powder into the molten polymer blend.
  • the blended ink was allowed to cool for 10-15 min and then mixed for 5 more minutes at 3500 rpm. The mixing/cooling process was repeated for a total of four 5 min mixes at 3500 rpm. After the fourth mix at 3500 rpm, the ink charge was poured out on a glass plate and two spatulas were used to form into a roughly 1 cm diameter x 3 cm long cylinder.
  • 3D printed structures are soaked overnight in distilled water to dissolve the polyethylene glycol from the printed material, thus creating a porous and flexible ⁇ -TCP/50:50 poly(D,L-lactide-co-glycolide) composite. Scaffolds were dried for at least twelve hours to ensure residual water has evaporated from the porous scaffold before binding with a therapeutic agent like tBMP2. Scaffolds were sterilized by soaking for 2-4 hours in a 70% ethanol solution and allowed to dry in biosafety cabinet for approximately 12 hours. Images of the scaffold are shown in FIGS.4A-4C. 3D printing is also performed using a FFF 3D printer.
  • the ink #4 is extruded into filaments and the filaments are loaded in a FFF 3D printer to generate a 3D printed scaffold.
  • the scaffold is processed using the post processing method outlined above.
  • Ink formulation and scaffold #5 Method To make a 5 cc batch of ink, 5.6 g of ⁇ -TCP powder, 1.87 g of polycaprolactone powder, and 1.87 g of polyethylene glycol flake were added to a glass mixing container. The glass jar was placed in dual asymmetric centrifugal mixer (FlackTek Speedmixer) and mix at low intensity (300 rpm) for 2 min to homogenize the powder blend before high rpm mixing. It was mixed for 5 min at high intensity (3500 rpm).
  • a 3x length extended melt filter nozzle (with filter screens removed) with 1.75 mm diameter hole size was used.
  • a cooling fan was placed near the extrusion nozzle to speed up solidification of the extruded filament.
  • the 1.75 mm diameter filament was extruded at extrusion temperature to 62°C and speed at 1 ⁇ 2 on the analog dial (approximately 1 cm/second extrusion speed).
  • Attorney Docket No: 50222-711.601 3D Printing The 1.75 mm filament was loaded in a Prusa i3 MK3S 3D printer. This filament material was printed using a 400 micron brass nozzle, 105°C extruder temperature, and 15 mm/s print speed.
  • Ink formulation and scaffold #6 Method To make a 5.5 cc batch of ink, 5.6 g of ⁇ -TCP powder, 1.87 g of polycaprolactone powder, 1.87 g of polyethylene glycol flake, and 1.04 g of sodium bicarbonate were added to a glass mixing container. The glass jar was placed in dual asymmetric centrifugal mixer (FlackTek Speedmixer) and mixed at low intensity (300 rpm) for 2 min to homogenize the powder blend before high rpm mixing. It was mixed for 2 min at high intensity (3500 rpm). During mixing, the internal friction causes the polycaprolactone and polyethylene glycol to melt, changing the ink to a viscous molten liquid.
  • FlackTek Speedmixer Fraction Mixer
  • This liquid phase mixing facilitates intimate dispersion of the ⁇ -TCP powder and sodium bicarbonate powder into the molten polymer blend.
  • the molten ink was poured onto a glass plate and flattened with a spatula to approximately 3 mm thick layer for cooling. After cooling, shears were used to cut into approximately 3-4 mm pellets. The was repeated for two additional 5.5 cc batches to create a total 16.5 cc of pellets for filament extrusion.
  • Filament Fabrication 16.5 cc of 3-4 mm pellets were loaded into the hopper of a filament extruder (Filabot EX2 Filament Extruder). A 3x length extended melt filter nozzle (with filter screens removed) with 1.75 mm diameter hole size was used.
  • a cooling fan was placed near the extrusion nozzle to speed up solidification of the extruded filament.
  • the 1.75 mm diameter filament was extruded at extrusion temperature to 62°C and speed at 1 ⁇ 2 on the analog dial (approximately 1 cm/second extrusion speed).
  • 3D Printing The 1.75 mm filament was loaded in a Prusa i3 MK3S 3D printer. This filament material was printed using a 400 micron brass nozzle, 155°C extruder temperature, and 15 mm/s print speed.
  • Ink formulation and scaffold #7 Method To make a 10 cc batch of ink, 11.2 g of ⁇ -TCP powder, 3.74 g of Dioxanone/L- lactide (90:10) copolymer chips, and 3.74 g of polyethylene glycol flake were added to a glass mixing container. The glass jar was placed in a dual asymmetric centrifugal mixer (FlackTek Speedmixer) and mixed at low intensity (300 rpm) for 2 min to homogenize the powder blend before high rpm mixing. The glass jar was transferred to a hot plate and heated until it reached 185°C (measured with IR thermometer).
  • the glass jar was immediately transferred back to the dual asymmetric centrifugal mixer and mixed for 5 min at high intensity (3500 rpm). Liquid phase mixing facilitates intimate dispersion of the ⁇ -TCP powder into the molten polymer blend.
  • the glass jar was transferred back to the hotplate and the temperature was increased to 185°C. Upon reaching the temperature, the glass jar was immediately transferred back to the dual asymmetric centrifugal mixer and mixed for 5 more minutes at 3500 rpm. The mixing/heating process was repeated for a total of four 5 min mixes at 3500 rpm.
  • the ink charge was poured out on a glass plate and two spatulas were used to form into a roughly 1 cm diameter x 6 cm long cylinder. While ink was still semi-molten, it was cut into several ⁇ 1-2 cm long pieces with straight razor.
  • 3D Printing solid polymer/ ⁇ -TCP pieces were transferred to a 5 cc stainless steel syringe (for use in Allevi 3 Bioprinter). Extruder CORE printing head was heated to 110°C and allowed to dwell for approximately 30 min to ensure melting of the ink. The ink was printed with 400 micron I.D. conical metallic Luer lock tip using 15 psi pressure and 10 mm/s nozzle velocity.
  • the scaffold was 3D printed on painter’s tape applied to a smooth glass or polymer surface, such as a glass microscope slide, larger glass plate, or 96 well plate lid.
  • Post Processing 3D printed structures were soaked overnight in distilled water to dissolve the polyethylene glycol from the printed material. The process created a porous and flexible ⁇ - TCP/90:10 dioxanone-L-lactide copolymer composite. Scaffolds were dried for at least twelve hours to ensure residual water has evaporated from the porous scaffold before binding with a therapeutic agent such as tBMP2 protein. Scaffolds were sterilized by soaking for 2-4 hours in a 70% ethanol solution and allowed to dry in biosafety cabinet for approximately 12 hours.
  • FIGS.7A-7C Images of the scaffold are shown in FIGS.7A-7C.
  • Attorney Docket No: 50222-711.601 3D printing is also performed using a FFF 3D printer.
  • the ink #7 is extruded into filaments and the filaments are loaded in a FFF 3D printer to generate a 3D printed scaffold.
  • the scaffold is processed using the post processing method outlined above.
  • Ink formulation and scaffold #8 Method To make a 16 cc batch of ink, 18 g of ⁇ -TCP powder, 6 g of polycaprolactone powder, 3 g of polyethylene glycol (8,000 MW) flake and 3 g of polyethylene glycol (20,000 MW) flake were added to a teflon mixing container.
  • the teflon container was placed in dual asymmetric centrifugal mixer (FlackTek Speedmixer) and mixed at low intensity (300 rpm) for 2 min to homogenize the powder blend before high rpm mixing.
  • the teflon container was mixed for 2.5 min at high intensity (3500 rpm).
  • the internal friction causes the polycaprolactone and polyethylene glycol to melt, changing the ink to a viscous molten liquid.
  • This liquid phase mixing facilitates intimate dispersion of the ⁇ -TCP powder into the molten polymer blend.
  • the molten ink was poured onto a glass plate and flatten with a spatula to approximately 3 mm thick layer for cooling. It was cooled for 10-15 min.
  • the 1.75 mm diameter filament was extruded at extrusion temperature to 62°C and speed at 1 ⁇ 2 on the analog dial (approximately 1 cm/second extrusion speed).
  • 3D Printing The 1.75 mm filament was loaded in a Prusa i3 MK3S 3D printer. This filament material was printed using a 400 micron brass nozzle, 105°C extruder temperature, and 15 mm/s print speed.
  • Post Processing 3D printed structures were soaked overnight in distilled water to dissolve the polyethylene glycol from the printed material, thus creating a porous and flexible ⁇ - TCP/polycaprolactone composite.
  • Scaffolds were dried for at least twelve hours to ensure residual water has evaporated from the porous scaffold before binding with a therapeutic agent like tBMP2. Scaffolds were sterilized by soaking for 2-4 hours in a 70% ethanol solution and allowed to dry in biosafety cabinet for approximately 12 hours. Images of the scaffold are shown in FIGS. 8A- 8C.
  • the teflon container was mixed for 2 min at high intensity (3500 rpm).
  • high intensity 3500 rpm
  • the internal friction causes the polycaprolactone and polyethylene glycol to melt, changing the ink to a viscous molten liquid.
  • This liquid phase mixing facilitates intimate dispersion of the ⁇ -TCP powder into the molten polymer blend.
  • the molten ink was poured onto a glass plate and flatten with a spatula to approximately 3 mm thick layer for cooling. It was cooled for 10-15 min.
  • the mixture was returned to the teflon container and mixed for 2 more minutes at a high intensity (3500 rpm).
  • the mixing/cooling process was repeated for a total of four 2 min mixes at 3500 rpm.
  • 3D Printing The 1.75 mm filament was loaded in a Prusa i3 MK3S 3D printer. This filament material was printed using a 400 micron brass nozzle, 140°C extruder temperature, and 10 mm/s print speed.
  • Post Processing 3D printed structures were soaked overnight in distilled water to dissolve the polyethylene glycol and sucrose from the printed material, thus creating a porous and flexible ⁇ -TCP/polycaprolactone composite. Scaffolds were dried for at least twelve hours to ensure residual water has evaporated from the porous scaffold before binding with a therapeutic agent like tBMP2. Scaffolds were sterilized by soaking for 2-4 hours in a 70% ethanol solution and allowed to dry in biosafety cabinet for approximately 12 hours.
  • FIGS.9A-9C Images of the scaffold are shown in FIGS.9A-9C.
  • Ink formulation and scaffold #10 Method To make a 16 cc batch of ink, 18 g of ⁇ -TCP powder, 6 g of caprolactone/glycolide copolymer (95:5) pellets, 3 g of polyethylene glycol (8,000 MW) flake and Attorney Docket No: 50222-711.601 3 g of polyethylene glycol (20,000 MW) flake were added to a teflon mixing container. The teflon container was placed in dual asymmetric centrifugal mixer (FlackTek Speedmixer) and mixed at low intensity (300 rpm) for 2 min to homogenize the powder blend before high rpm mixing.
  • FlackTek Speedmixer Fraction Mixer
  • the teflon container was mixed for 2.5 min at high intensity (3500 rpm).
  • the internal friction causes the caprolactone/glycolide copolymer (95:5) and polyethylene glycol to melt, changing the ink to a viscous molten liquid.
  • This liquid phase mixing facilitates intimate dispersion of the ⁇ -TCP powder into the molten polymer blend.
  • the molten ink was poured onto a glass plate and flatten with a spatula to approximately 3 mm thick layer for cooling. It was cooled for 10-15 min. The mixture was returned to the teflon container and mixed for 4 more minutes at a high intensity (3500 rpm).
  • the 1.75 mm diameter filament was extruded at extrusion temperature to 62°C and speed at 1 ⁇ 2 on the analog dial (approximately 1 cm/second extrusion speed).
  • 3D Printing The 1.75 mm filament was loaded in a Prusa i3 MK3S 3D printer. This filament material was printed using a 400 micron brass nozzle, 150°C extruder temperature, and 10 mm/s print speed.
  • Post Processing 3D printed structures were soaked overnight in distilled water to dissolve the polyethylene glycol from the printed material, thus creating a porous and flexible ⁇ - TCP/ caprolactone/glycolide copolymer (95:5) composite.
  • the teflon container was placed in dual asymmetric centrifugal mixer (FlackTek Speedmixer) and mixed at low intensity (300 rpm) for 2 min to homogenize the powder blend before high rpm mixing.
  • Attorney Docket No: 50222-711.601 the teflon container was mixed for 2 min at high intensity (3500 rpm).
  • the internal friction causes the caprolactone/glycolide copolymer (90:10) and polyethylene glycol to melt, changing the ink to a viscous molten liquid. This liquid phase mixing facilitates intimate dispersion of the ⁇ -TCP powder into the molten polymer blend.
  • the 1.75 mm diameter filament was extruded at extrusion temperature to 62°C and speed at 1 ⁇ 2 on the analog dial (approximately 1 cm/second extrusion speed).
  • 3D Printing The 1.75 mm filament was loaded in a Prusa i3 MK3S 3D printer. This filament material was printed using a 400 micron brass nozzle, 140°C extruder temperature, and 10 mm/s print speed.
  • Post Processing 3D printed structures were soaked overnight in distilled water to dissolve the polyethylene glycol from the printed material, thus creating a porous and flexible ⁇ - TCP/ Poly(D,L-lactide-co-glycolide) copolymer (50:50) composite.
  • Ink formulation and scaffold #14 is printed using a syringe-based melt extrusion printing method, for example, using methods as described for printing inks #1-#4.
  • Ink #14 is printed using fused filament fabrication 3D printing, for example, using methods as described for printing ink#5 and ink#6.
  • Ink formulation and scaffold #15 Ink #9 is printed using a syringe-based melt extrusion printing method, for example, using methods as described for printing inks #1-#4.
  • Ink #15 is printed using fused filament fabrication 3D printing, for example, using methods as described for printing ink#5 and ink#6.
  • the internal friction caused the caprolactone/glycolide copolymer and polyethylene glycol to melt, changing the ink to a viscous molten liquid.
  • This liquid phase mixing facilitated intimate dispersion of the ⁇ -TCP powder into the molten polymer blend.
  • the molten ink was poured onto a glass plate and flattened with a spatula to an approximately 3 mm thick layer for cooling. The ink was cooled for 10-15 min, and mixed for 1.5 more minutes at 3500 rpm. This mixing/cooling process was repeated for a total of four 1.5-minute mixes at 3500 rpm.
  • FIGS 15A-15B Images of example scaffolds are shown in FIGS 15A-15B. After soaking and drying, the scaffold contained 75 Wt% ⁇ -TCP powder and 25 Wt% Caprolactone/Glycolide copolymer (95:5).
  • Ink formulation and scaffold #19 Method To make a 16cc batch of ink, 18 g of ⁇ -TCP powder, 6 g of Caprolactone/Glycolide copolymer (90:10) chips, 3 g of polyethylene glycol (8,000 MW) flake and 3 g of polyethylene glycol (35,000 MW) flake was added to a teflon mixing container.
  • the teflon container was placed in dual asymmetric centrifugal mixer (FlackTek Speedmixer) and mixed at low intensity (300 rpm) for 2 min to homogenize the powder blend before high rpm mixing.
  • the composition was mixed for 1.5 min at high intensity (3500 rpm).
  • the internal friction caused the caprolactone/glycolide copolymer and polyethylene glycol to melt, changing the ink to a viscous molten liquid.
  • This liquid phase mixing facilitated intimate dispersion of the ⁇ -TCP powder into the molten polymer blend.
  • the molten ink was poured onto a glass plate and flattened with a spatula to an approximately 3 mm thick layer for cooling.
  • Pellet Fabrication An 850 ⁇ m diameter nozzle was attached to the metal barrel of a heated, pneumatic extrusion device (e.g. an Allevi 3 bioprinter with a 5 mL stainless steel syringe barrel) and 3-4 cc of 4-6 mm ink feedstock squares were loaded into the barrel. The barrel was heated to 80-120°C temperature, for example 100°C, and left for 15 minutes.
  • a heated, pneumatic extrusion device e.g. an Allevi 3 bioprinter with a 5 mL stainless steel syringe barrel
  • the pneumatic pressure of the pneumatic extrusion device was then set to about 50-100 psi, for example 55 psi.
  • a platform was installed below the extrusion tip as a substrate on Attorney Docket No: 50222-711.601 which to extrude pellets.
  • the platform can be smooth silicone sheet material.
  • a manually generated Marlin G-code was used to extrude up to 120 pellets in a rectangular array, resulting in pellets having about 2 mm to about 3 mm diameter and being roughly equiaxed.
  • 3D Printing The pellets were then loaded into the hopper of a Piocreat G5 FGF 3D printer. The material was printed using a nozzle of about 300 ⁇ m to about 1000 ⁇ m at about 105 to about 145°C nozzle temperature using an about 5mm/s to about 30mm/s printing speed.
  • Post Processing The 3D printed structures were soaked for at least 16 hours in distilled water to dissolve the polyethylene glycol from the printed material, thus creating a porous and flexible ⁇ -TCP and caprolactone/glycolide copolymer composite. The scaffolds were dried for at least twelve hours to ensure residual water has evaporated from the porous scaffold before binding with TBMP2 protein.
  • Scaffolds were sterilized by soaking for 2-4 hours in a 70% ethanol solution and allowed to dry in a biosafety cabinet for about 12 hours. Images of example scaffolds are shown in FIGS 16A-16B. After soaking and drying, the scaffold contained 75 Wt% ⁇ -TCP powder and 25 Wt% Caprolactone/Glycolide copolymer (90:10). Structure properties Physical properties of example scaffolds made using inks #1-#12 and 18-19 were determined and are outlined in Table 34. Physical properties of example scaffolds made using inks #16, 18, and 19 were determined and outline in Table 35. Table 34. Properties of Examples Scaffolds Attorney Docket No: 50222-711.601 Table 35.
  • Example Scaffolds The structures of this example are tested using Brunauer-Emmett-Teller (BET) surface area analysis by gas physisorption. A compression test is also performed on the structures.
  • BET Brunauer-Emmett-Teller
  • a compression test is also performed on the structures.
  • Example 3 Therapeutic Agent A chimeric polypeptide comprising the BMP therapeutic peptide connected to five beta- tricalcium phosphate binding peptides was expressed and purified using standard expression and purification methods.
  • the chimeric polypeptide is referred to as tBMP-2 and has the following sequence: MPIGSLLADTTHHRPWTVIGESTHHRPWSIIGESSHHKPFTGLGDTTHHRPWGILAESTH HKPWTASGAGGSEGGGSEGGTSGATGAGTSTSGGGASTGGGTGQAKHKQRKRLKSSC KRHPLYVDFSDVGWNDWIVAPPGYHAFYCHGECPFPLADHLNSTNHAIVQTLVNSVNS KIPKACCVPTELSAISMLYLDENEKVVLKNYQDMVVEGCGCR (SEQ ID NO: 434).
  • Example 4 Device Manufacture The 3D printed structure “Flexible 3-layer membrane 3D printed with a 400 um nozzle” of Example 2 was combined with the tBMP-2 therapeutic agent of Example 3 to create a device. The scaffold was combined with tBMP-2 in a binding solution and unbound tBMP-2 was washed off the scaffold. The resulting device comprised the scaffold with bound tBMP-2. The device was lyophilized. Similar devices are prepared using 3D printed structures and growth factors herein. Attorney Docket No: 50222-711.601
  • Example 5 Animal Models One or more devices of Example 4 are tested in an animal model to demonstrate bone regeneration through ⁇ CT imaging and histological analysis.
  • Indications can include lumbar spinal fusion (a 3D printed insert for spinal fusion cage), posterolateral (PLF) spine fusion (3D printed scaffold that spans transverse processes), tibial segmental defects (a 3D printed scaffold based on patient CT data), and/or alveolar ridge augmentation (a 3D printed thin barrier membrane).
  • the first study is a lapine posterolateral fusion model. The objective of this study is to evaluate the in vivo performance of test devices at varying concentrations of growth factor and masses of scaffold. The test groups are evaluated for spine fusion rate, new bone formation, and residual graft using radiographic plain films, microCT, biomechanical and histological endpoints at 8 weeks following implantation.
  • the TPs are then decorticated with a high-speed burr.
  • the test devices are placed over the transverse processes and fascia and skin are closed and stapled.
  • analgesics are administered according to IACUC approval. Animals are fed ad libitum and allowed to move about their cages without restriction. No postoperative immobilization devices are used. During the following weeks, the animals are observed closely and given additional pain medication based on their mobility, diet, disposition, and general activity that would signify increased pain.
  • the rabbits are radiographed postoperatively and at 8 weeks.
  • MicroCT morphometry analysis is performed using a region of interest (ROI) placed across the fusion site and areas of bone are calculated. Fusion sites of each animal are processed for histology at 8 weeks.
  • the second study is a sheep interbody fusion model to evaluate test devices in the interbody space of sheep lumbar spine. Sheep undergo interbody fusion through a lateral approach. Implanted motion segments are stabilized with pedicle screw and rod fixation. PEEK interbody spacers are placed in the interbody space following discectomy and endplate prep. Spacers are filled with test device. Sheep are euthanized at 6 months post-op after review of in- life MDCTs at 4, 8, and 12 weeks post operation.
  • L929 mouse fibroblast cells were seeded in 24-well plates at seeding density of 1x10 5 cells/per well and placed in a humidified incubator at 37°C and 5% CO2 overnight.
  • Sterile scaffold extracts (scaffolds shown in table 36) were prepared by first submerging scaffolds in media for 24 hours then adding these extracts to the 24-well plates seeded with the L929 mouse fibroblast cell line. After 24 hours, visual examination of cells exposed to the extracts was used to determine if there was a cytotoxic cell response after cells were exposed to scaffolds. Cytotoxicity was considered present if cells were detached, lysed, or had changes in morphology.
  • FIG. 17A Microscopy images of cells exposed to the scaffolds of table 36 are shown in FIG. 17A.
  • High Density Polyethylene (negative control) scored an average of less than 1 which is not cytotoxic to slightly cytotoxic.0.1% Zincdiethyldithiocarbamate (ZDEC) (positive control) scored an average of 4 which is severely cytotoxic, and had widespread cell detachment.95:5 and 90:10 scored 0 indicating that they are non cytotoxic.
  • the OT sample scored less than 1, indicating that it is non cytotoxic to slightly cytotoxic.

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  • Optics & Photonics (AREA)
  • Inorganic Chemistry (AREA)
  • Composite Materials (AREA)
  • Medicinal Preparation (AREA)
  • Materials For Medical Uses (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

La présente invention concerne des dispositifs comprenant un agent thérapeutique lié à une structure tridimensionnelle imprimée. L'invention concerne également des formulations d'encre pour une impression tridimensionnelle. De plus, l'invention concerne des procédés de fabrication de ces dispositifs et leurs utilisations, par exemple dans le traitement d'un état pathologique chez un sujet en ayant besoin. <i />
EP23858242.3A 2022-08-23 2023-08-22 Compositions céramiques imprimées en 3d et procédés d'utilisation Pending EP4577255A1 (fr)

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US202263373278P 2022-08-23 2022-08-23
PCT/US2023/072672 WO2024044600A1 (fr) 2022-08-23 2023-08-22 Compositions céramiques imprimées en 3d et procédés d'utilisation

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EP (1) EP4577255A1 (fr)
JP (1) JP2025528402A (fr)
CN (1) CN120112315A (fr)
AU (1) AU2023330160A1 (fr)
WO (1) WO2024044600A1 (fr)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5641502A (en) * 1995-06-07 1997-06-24 United States Surgical Corporation Biodegradable moldable surgical material
US20070053954A1 (en) * 2003-10-24 2007-03-08 Rowe Stephen C Macromer-melt formulations
US20220323641A1 (en) * 2019-08-20 2022-10-13 Theradaptive, Inc. Materials for delivery of tetherable proteins in bone implants

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WO2024044600A1 (fr) 2024-02-29
JP2025528402A (ja) 2025-08-28
AU2023330160A1 (en) 2025-03-20
CN120112315A (zh) 2025-06-06

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