# US 2016/0354518 A1 · Bone pastes comprising biofunctionalized calcium phosphate cements with enhanced cell functions for bone repair > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2016/0354518 A1 - Document ID: US20160354518A1 - Status: Published application - Kind: A1 - Application number: 15/241,950 - Filed: 2016-08-19 - Published: 2016-12-08 - Earliest claimed: 2011-10-21 - Assignees: University of Maryland, Baltimore - Inventors: Huakun XU - Web page: https://patentgrep.com/patent/US20160354518A1 ## CPC - A61L 27/365: {Bones} - A61L 24/0005: {Ingredients of undetermined constitution or reaction products thereof} - A61L 24/0015: {Medicaments; Biocides} - A61L 24/0031: {Hydrogels or hydrocolloids} - A61L 24/0036: {Porous materials, e.g. foams or sponges} - A61L 24/0042: {Materials resorbable by the body} - A61L 24/02: containing inorganic materials - A61L 24/043: {Mixtures of macromolecular materials} - A61L 24/08: Polysaccharides {(A61L24/043 takes precedence)} - A61L 24/106: {Fibrin; Fibrinogen} - A61L 24/108: {Specific proteins or polypeptides not covered by groups A61L24/102 - A61L24/106} - A61L 27/12: Phosphorus-containing materials, e.g. apatite - A61L 27/20: Polysaccharides - A61L 27/225: {Fibrin; Fibrinogen} - A61L 27/227: {Other specific proteins or polypeptides not covered by A61L27/222, A61L27/225 or A61L27/24} - A61L 27/3616: {Blood, e.g. platelet-rich plasma} - A61L 27/3633: {Extracellular matrix [ECM]} - A61L 27/3834: {Cells able to produce different cell types, e.g. hematopoietic stem cells, mesenchymal stem cells, marrow stromal cells, embryonic stem cells} - A61L 27/54: Biologically active materials, e.g. therapeutic substances {(A61L27/227 takes precedence)} - A61L 27/56: Porous materials, {e.g. foams or sponges} - C04B 28/34: containing cold phosphate binders - A61L 2400/06: Flowable or injectable implant compositions - A61L 2430/02: for reconstruction of bones; weight-bearing implants - C04B 2111/00836: {for medical or dental applications} - 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 (preservation of bodies or disinfecting characterised by the agents employed A01N; preserving, e.g. sterilising, food or foodstuffs A23; preparations for medical, dental or toiletry purposes A61K) - C04B: LIME, MAGNESIA; SLAG; CEMENTS; COMPOSITIONS THEREOF, e.g. MORTARS, CONCRETE OR LIKE BUILDING MATERIALS; ARTIFICIAL STONE {(roofing granules E04D7/005)}; CERAMICS (devitrified glass-ceramics C03C10/00); REFRACTORIES; TREATMENT OF NATURAL STONE ## Abstract The invention provides injectable, biofunctional agent-containing calcium phosphate cement bone pastes for bone tissue engineering, and methods of making and using the same. ## Claims ### Claim 1 A bone paste comprising calcium phosphate cement and one or more biofunctional agents, wherein the biofunctional agents are selected from the group consisting of RGD-containing peptides, fibronectin, fibronectin-like engineered polymer protein (FEPP), derived extracellular matrix (dECM), and platelet concentrate. ### Claim 2 (depends on 1) The bone paste of claim 1, wherein the bone paste comprises calcium phosphate cement and an RGD-containing peptide, wherein RGD-containing peptide is selected from the group consisting of RGD, G4RGDSP, RGDS, GRGD, GRGDGY, RGDSGGC, and GRGDS, and wherein RGD-containing peptide is present within a range of about 0.0005% to about 5% by mass. ### Claim 3 (depends on 1) The bone paste of claim 1, wherein the bone paste comprises calcium phosphate cement and fibronectin, wherein fibronectin is present within a range of about 0.0005% to about 5% by mass. ### Claim 4 (depends on 1) The bone paste of claim 1, wherein the bone paste comprises calcium phosphate cement and FEPP, wherein FEPP is present within a range of about 0.0005% to about 5% by mass. ### Claim 5 (depends on 1) The bone paste of claim 1, wherein the bone paste comprises calcium phosphate cement and dECM, wherein dECM is present within a range of about 0.001% to about 10% by mass. ### Claim 6 (depends on 1) The bone paste of claim 1, wherein the bone paste comprises calcium phosphate cement and platelet concentrate, wherein platelet concentrate is present within a range of about 0.001% to about 10% by mass. ### Claim 7 (depends on 1) The bone paste of claim 1, wherein the bone paste comprises calcium phosphate cement and any two of the biofunctional agents. ### Claim 8 (depends on 1) The bone paste of claim 1, wherein the bone paste comprises calcium phosphate cement and any three of the biofunctional agents. ### Claim 9 (depends on 1) The bone paste of claim 1, wherein the bone paste comprises calcium phosphate cement and any four of the biofunctional agents. ### Claim 10 (depends on 1) The bone paste of claim 1, wherein the bone paste comprises calcium phosphate cement and each of the five biofunctional agents. ### Claim 11 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) The bone paste of any one of claims 1-10, wherein the calcium phosphate cement comprises one or more ingredients selected from the group consisting of tetracalcium phosphate (TTCP) (Ca4(PO4)2O), dicalcium phosphate anhydrous (DCPA) (CaHPO4), dicalcium phosphate dihydrate (CaHPO4.2H2O), tricalcium phosphate (Ca3[PO4]2), α-tricalcium phosphate (α-Ca3(PO4)2), β-tricalcium phosphate (β-Ca3(PO4)2), octacalcium phosphate (Ca8H2(PO4)6.5H2O), amorphous calcium phosphate (Ca3(PO4)2), calcium carbonate (CaCO3), calcium hydroxide (Ca[OH]2), and hydroxyapatite (Ca10(PO4)6(OH)2), and mixtures thereof. ### Claim 12 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) The bone paste of any one of claims 1-10, wherein the calcium phosphate cement comprises tetracalcium phosphate and dicalcium phosphate anhydrous. ### Claim 13 (depends on 12) The bone paste of claim 12, wherein the calcium phosphate cement comprises a molar ratio of tetracalcium phosphate to dicalcium phosphate anhydrous of about 1:5 to about 5:1. ### Claim 14 (depends on 12) The bone paste of claim 12, wherein the calcium phosphate cement comprises a molar ratio of tetracalcium phosphate to dicalcium phosphate anhydrous of about 1:3 to about 1:1. ### Claim 15 (depends on 12) The bone paste of claim 12, wherein the calcium phosphate cement comprises an approximately 1:1 molar ratio of tetracalcium phosphate to dicalcium phosphate anhydrous. ### Claim 16 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) The bone paste of any one of claims 1-10, wherein the calcium phosphate cement further comprises chitosan. ### Claim 17 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) The bone paste of any one of claims 1-10, wherein the calcium phosphate cement further comprises fibers. ### Claim 18 (depends on 17) The bone paste of claim 17, wherein the fibers are degradable. ### Claim 19 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) The bone paste of any one of claims 1-10, wherein the calcium phosphate cement further comprises chitosan and degradable fibers. ### Claim 20 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) The bone paste of any one of claims 1-10, wherein the calcium phosphate cement further comprises a porogen. ### Claim 21 (depends on 20) The bone paste of claim 20, wherein the porogen is NaHCO3 and citric acid. ### Claim 22 (depends on 21) The bone paste of claim 21, wherein the mass fraction of NaHCO3 is from about 5% to about 30%, and the mass fraction of citric acid is from about 50% to about 60%. ### Claim 23 (depends on 16) The bone paste of claim 16, wherein the bone paste comprises calcium phosphate cement and a RGD-containing peptide, wherein the RGD-containing peptide is present within a range of about 0.0005% to about 0.05% by mass, and wherein the RGD-containing peptide is covalently linked to said chitosan. ### Claim 24 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) The bone paste of any one of claims 1-10, wherein the bone paste further comprises stem cells. ### Claim 25 (depends on 24) The bone paste of claim 24, wherein the cells are selected from the group consisting of human umbilical cord mesenchymal stem cells, bone marrow stem cells, embryonic stem cells, pluripotent stem cells, induced pluripotent stem cells, multipotent stem cells, progenitor cells, and osteoblasts. ### Claim 26 (depends on 24) The bone paste of claim 24, wherein the cells are attached to a surface of the bone paste, or the cells are interspersed throughout the bone paste, or both. ### Claim 27 (depends on 1, 2, 3, 4, 5, 6, 7, 8, 9, 10) The bone paste of any one of claims 1-10, wherein the bone paste is injectable. ### Claim 28 A bone paste comprising calcium phosphate cement, one or more biofunctional agents and cell-encapsulating microbeads, wherein the biofunctional agents are selected from the group consisting of RGD peptide, fibronectin, fibronectin-like engineered polymer protein (FEPP), derived extracellular matrix (dECM), and platelet concentrate. ### Claim 29 (depends on 28) The bone paste of claim 28, wherein the bone paste comprises calcium phosphate cement and an RGD-containing peptide, wherein RGD-containing peptide is selected from the group consisting of RGD, G4RGDSP, RGDS, GRGD, GRGDGY, RGDSGGC, and GRGDS, and wherein RGD-containing peptide is present within a range of about 0.0005% to about 5% by mass. ### Claim 30 (depends on 28) The bone paste of claim 28, wherein the bone paste comprises calcium phosphate cement and fibronectin, wherein fibronectin is present within a range of about 0.0005% to about 5% by mass. ### Claim 31 (depends on 28) The bone paste of claim 28, wherein the bone paste comprises calcium phosphate cement and FEPP, wherein FEPP is present within a range of about 0.0005% to about 5% by mass. ### Claim 32 (depends on 28) The bone paste of claim 28, wherein the bone paste comprises calcium phosphate cement and dECM, wherein dECM is present within a range of about 0.001% to about 10% by mass. ### Claim 33 (depends on 28) The bone paste of claim 28, wherein the bone paste comprises calcium phosphate cement and platelet concentrate, wherein platelet concentrate is present within a range of about 0.001% to about 10% by mass. ### Claim 34 (depends on 28) The bone paste of claim 28, wherein the bone paste comprises calcium phosphate cement and any two of the biofunctional agents. ### Claim 35 (depends on 28) The bone paste of claim 28, wherein the bone paste comprises calcium phosphate cement and any three of the biofunctional agents. ### Claim 36 (depends on 28) The bone paste of claim 28, wherein the bone paste comprises calcium phosphate cement and any four of the biofunctional agents. ### Claim 37 (depends on 28) The bone paste of claim 28, wherein the bone paste comprises calcium phosphate cement and each of the five biofunctional agents. ### Claim 38 (depends on 28, 29, 30, 31, 32, 33, 34, 35, 36, 37) The bone paste of any one of claims 28-37, wherein the calcium phosphate cement comprises one or more ingredients selected from the group consisting of tetracalcium phosphate (TTCP) (Ca4(PO4)2O), dicalcium phosphate anhydrous (DCPA) (CaHPO4), dicalcium phosphate dihydrate (CaHPO4.2H2O), tricalcium phosphate (Ca3[PO4]2), α-tricalcium phosphate (α-Ca3(PO4)2), β-tricalcium phosphate (β-Ca3(PO4)2), octacalcium phosphate (Ca8H2(PO4)65H2O), amorphous calcium phosphate (Ca3(PO4)2), calcium carbonate (CaCO3), calcium hydroxide (Ca[OH]2), and hydroxyapatite (Ca10(PO4)6(OH)2), and mixtures thereof. ### Claim 39 (depends on 28, 29, 30, 31, 32, 33, 34, 35, 36, 37) The bone paste of any one of claims 28-37, wherein the calcium phosphate cement comprises tetracalcium phosphate and dicalcium phosphate anhydrous. ### Claim 40 (depends on 39) The bone paste of claim 39, wherein the calcium phosphate cement comprises a molar ratio of tetracalcium phosphate to dicalcium phosphate anhydrous of about 1:5 to about 5:1. ### Claim 41 (depends on 39) The bone paste of claim 39, wherein the calcium phosphate cement comprises a molar ratio of tetracalcium phosphate to dicalcium phosphate anhydrous of about 1:3 to about 1:1. ### Claim 42 (depends on 39) The bone paste of claim 39, wherein the calcium phosphate cement comprises an approximately 1:1 molar ratio of tetracalcium phosphate to dicalcium phosphate anhydrous. ### Claim 43 (depends on 28, 29, 30, 31, 32, 33, 34, 35, 36, 37) The bone paste of any one of claims 28-37, wherein the calcium phosphate cement further comprises chitosan. ### Claim 44 (depends on 28, 29, 30, 31, 32, 33, 34, 35, 36, 37) The bone paste of any one of claims 28-37, wherein the calcium phosphate cement further comprises fibers. ### Claim 45 (depends on 40) The bone paste of claim 40, wherein the fibers are degradable. ### Claim 46 (depends on 28, 29, 30, 31, 32, 33, 34, 35, 36, 37) The bone paste of any one of claims 28-37, wherein the calcium phosphate cement further comprises chitosan and degradable fibers. ### Claim 47 (depends on 28, 29, 30, 31, 32, 33, 34, 35, 36, 37) The bone paste of any one of claims 28-37, wherein the calcium phosphate cement further comprises a porogen. ### Claim 48 (depends on 47) The bone paste of claim 47, wherein the porogen is NaHCO3 and citric acid. ### Claim 49 (depends on 48) The bone paste of claim 48, wherein the mass fraction of NaHCO3 is from about 5% to about 30%, and the mass fraction of citric acid is from about 50% to about 60%. ### Claim 50 (depends on 43) The bone paste of claim 43, wherein the bone paste comprises calcium phosphate cement and a RGD-containing peptide, wherein the RGD-containing peptide is present within a range of about 0.0005% to about 0.05% by mass, and wherein the RGD-containing peptide is covalently linked to said chitosan. ### Claim 51 (depends on 28, 29, 30, 31, 32, 33, 34, 35, 36, 37) The bone paste of any one of claims 28-37, wherein the microbeads are hydrogel microbeads. ### Claim 52 (depends on 51) The bone paste of claim 51, wherein the microbeads are comprised of alginate, partially oxidized alginate, oxidized alginate, alginate-fibrin, partially oxidized alginate-fibrin, oxidized alginate-fibrin, poly(ethylene glycol diacrylate), poly(ethylene glycol)-anhydride dimethacrylate, gelatin, chemically cross-linked polymers, ionically cross-linked polymers, heat-polymerized polymers, or photopolymerized polymers. ### Claim 53 (depends on 51) The bone paste of claim 51, wherein the microbeads are alginate-fibrin microbeads. ### Claim 54 (depends on 53) The bone paste of claim 53, wherein the alginate-fibrin microbeads comprise a fibrinogen mass fraction of from about 0.05% to about 1%. ### Claim 55 (depends on 53) The bone paste of claim 53, wherein the alginate-fibrin microbeads comprise a fibrinogen mass fraction of about 0.1%. ### Claim 56 (depends on 53) The bone paste of claim 53, wherein the alginate is at about 7.5% oxidation. ### Claim 57 (depends on 28, 29, 30, 31, 32, 33, 34, 35, 36, 37) The bone paste of any one of claims 28-37, wherein the microbeads are present in a volume of about 40 to 60%. ### Claim 58 (depends on 28, 29, 30, 31, 32, 33, 34, 35, 36, 37) The bone paste of any one of claims 28-37, wherein the microbeads have an average diameter of less than about 2 millimeters. ### Claim 59 (depends on 28, 29, 30, 31, 32, 33, 34, 35, 36, 37) The bone paste of any one of claims 28-37, wherein the cells are stem cells. ### Claim 60 (depends on 28, 29, 30, 31, 32, 33, 34, 35, 36, 37) The bone paste of any one of claims 28-37, wherein the cells are selected from the group consisting of human umbilical cord mesenchymal stem cells, bone marrow stem cells, embryonic stem cells, pluripotent stem cells, induced pluripotent stem cells, multipotent stem cells, progenitor cells, and osteoblasts. ### Claim 61 (depends on 28, 29, 30, 31, 32, 33, 34, 35, 36, 37) The bone paste of any one of claims 28-37, wherein the bone paste is injectable. ### Claim 62 A method for preparing a bone paste, comprising covalently linking an RGD-containing peptide to chitosan to form RGD-grafted chitosan, dissolving the RGD-grafted chitosan in water to form a chitosan liquid, and mixing calcium phosphate cement into the chitosan liquid, thereby preparing a bone paste. ### Claim 63 (depends on 62) The method of claim 62, wherein the RGD-containing peptide is selected from the group consisting of RGD, G4RGDSP, RGDS, GRGD, GRGDGY, RGDSGGC, and GRGDS. ### Claim 64 (depends on 62, 63) The method of claim 62 or 63, wherein the RGD-containing peptide is present within a range of about 0.0005% to about 5% by mass. ### Claim 65 (depends on 62, 63) The method of claim 62 or 63, wherein the chitosan is chitosan lactate. ### Claim 66 (depends on 62, 63) The method of claim 62 or 63, wherein a degradable fiber is added to the chitosan liquid prior to mixing calcium phosphate cement into the chitosan liquid. ### Claim 67 (depends on 62, 63) The method of claim 62 or 63, wherein the calcium phosphate cement comprises an approximately 1:1 molar ratio of tetracalcium phosphate to dicalcium phosphate anhydrous.