Compositions using iron excipients and their uses including treatment of cancer

Abstract

To provide a bone graft composition comprising a calcium phosphate putty, and repair a bone defect in a patient by applying the bone graft composition.SOLUTION: A bone graft composition comprising a calcium phosphate putty is provided. A method of repairing a bone defect in a patient by applying the bone graft composition is also provided.SELECTED DRAWING: Figure 1

Description

[Technical Field]

[0001] This application claims the benefit of U.S. Provisional Application No. 63 / 053,277, filed July 17, 2020, the entire contents of which are incorporated herein by reference. [Background technology]

[0002] Bone grafting is a surgical procedure performed to repair fractures that pose significant health risks to the patient or that do not heal properly. While certain small or acute fractures can be healed, larger fractures, such as compound fractures, pose a higher risk. Bone generally has the ability to regenerate completely, but this requires a very small fracture space or some type of scaffolding. Successful bone grafts can result in osteoconduction (the ability of the bone graft material to passively allow bone growth), osteoinduction (the bone graft induces undifferentiated cells to become active osteoblasts, which form new bone tissue), and / or osteogenesis (living bone cells in the graft material contribute to bone remodeling).

[0003] Bone grafts are often autologous bone, i.e., the bone graft material is harvested from the patient's own body, often from the iliac crest. However, the use of autologous bone subjects the patient to increased pain and discomfort and increased risk of infection because the patient must undergo additional surgery to retrieve the autologous bone for use in the grafting procedure.

[0004] The bone graft may be an allograft, i.e., the bone graft material is derived from cadaver bone, usually obtained from a bone bank. Allograft bone exposes the patient to the risk of disease and graft rejection.

[0005] Bone graft materials can be synthetic, and synthetic bone graft materials are often made from hydroxyapatite or other naturally occurring biocompatible materials with mechanical properties similar to those of bone. Most bone grafts are expected to be resorbed and replaced over the course of several months as the natural bone heals. However, synthetic bone graft materials may have lower osteoconductivity and osteoinductivity than autograft or allograft materials. Synthetic materials also carry the risk of donor microbial infection. Therefore, there is a significant medical need for synthetic bone graft materials with improved properties for healing bone injuries. DETAILED DESCRIPTION OF THE INVENTION

[0006] In one aspect, a bone graft composition is provided that includes a calcium phosphate putty and at least one of a hardening agent, a hardening rate control agent, an acidifying agent, an iron excipient, collagen, and a diluent solution.

[0007] In some embodiments, the calcium phosphate putty comprises biphasic calcium phosphate particles. In some embodiments, the biphasic calcium phosphate particles comprise hydroxyapatite and tricalcium phosphate. In some embodiments, the biphasic calcium phosphate particles comprise about 20-60% hydroxyapatite and about 40-80% tricalcium phosphate. In some embodiments, the biphasic calcium phosphate particles have interconnected macropores and micropores. In some embodiments, the biphasic calcium phosphate particles are in the form of spherical particles, fibers, or irregular granules. In some embodiments, the calcium phosphate putty comprises biphasic calcium phosphate particles, the biphasic calcium phosphate particles comprise hydroxyapatite and tricalcium phosphate, and the bone graft composition further comprises collagen and naloxone.

[0008] In some embodiments, the bone graft composition further comprises a bioabsorbable polymer.

[0009] In some embodiments, the bone graft composition has a density of 1-6 g / mL after mixing. In some embodiments, the force required to extrude the bone graft composition through a cannula, tube, or syringe is less than 80 N / m.

[0010] In some embodiments, the bone graft composition does not have an active agent. In some embodiments, the bone graft composition further comprises an active agent. In some embodiments, the bone graft composition is constructed and arranged to deliver an active agent or other agent to a desired site in a patient.

[0011] In some embodiments, the bone graft composition comprises a network of reservoirs and microchannels for storing and delivering an active agent. In some embodiments, the bone graft composition comprises a plurality of reservoirs, microtubes, and / or nanotubes for storing and delivering an active agent. In some embodiments, the active agent is delivered at a controlled rate.

[0012] In some embodiments, the active agent is dissolved in a pharmaceutically suitable carrier. In some embodiments, the active agent dissolved in a pharmaceutically suitable carrier is sprayed or coated onto the synthetic bone graft.

[0013] In some embodiments, the active agent is an opioid growth factor receptor (OGFR) antagonist. In some embodiments, the OGFR antagonist is selected from the group consisting of naloxone, naltrexone, and salts thereof. In some embodiments, the OGFR antagonist is administered with a diluent.

[0014] In some embodiments, the bone graft composition comprises a hardening agent, the hardening agent comprising sodium carbonate (NaCO3) or ferrous sulfate (FeSO4). In some embodiments, the bone graft composition comprises a hardening agent, the hardening agent comprising one or more compounds from the group consisting of calcium sulfate hemihydrate, calcium sulfate dihydrate, calcium sulfate anhydrous, monocalcium phosphate, dicalcium phosphate, tricalcium phosphate, tetracalcium phosphate, octacalcium phosphate, hydroxyapatite, calcium carbonate, magnesium carbonate, strontium carbonate, and sodium hydrogen phosphate.

[0015] In some embodiments, the weight ratio of the hardening agent to the calcium phosphate salt compound is between 1:2 (hardening agent:calcium salt) and 3:4 (hardening agent:calcium salt).

[0016] In some embodiments, the bone graft composition comprises an acidifying agent, the acidifying agent comprising one or more from the group of ascorbic acid, magnesium citrate, potassium citrate, sodium citrate, citric acid monohydrate, and acetic acid.

[0017] In some embodiments, the acidifying agent improves the bioavailability of the active agent in the bone graft composition. In some embodiments, the acidifying agent promotes bone formation. In some embodiments, the acidifying agent increases the adhesiveness of the bone graft material.

[0018] In some embodiments, the bone graft composition comprises a color forming agent. In some embodiments, the color forming agent comprises an activatable color forming agent. In some embodiments, the color forming agent colors the bone graft composition red, blue, orange, green, neon green, purple, brown, black, gray, or battleship gray.

[0019] In some embodiments, the bone graft composition comprises an antimicrobial agent. In some embodiments, the antimicrobial agent comprises one or more from the group of antibiotic agents or antifungal agents.

[0020] In some embodiments, antibiotics include vancomycin, gentamicin, tobramycin, kanamycin, neomycin, ampicillin, methicillin, nafcillin, oxacillin, penicillin, ticarcillin, ciprofloxacin, vancomycin, cefazolin, cefepime, ceftriaxone, clindamycin, aztreonam, imipenem, quinupristin / dalfopristin, chloramphenicol, doxycycline, metronidazole, nitrofurantoin, polymyoxin B, tetracycline, viomycin, chloromycetin, streptomycin, azactam, and pharmaceutically acceptable salts thereof, and combinations thereof.

[0021] In some embodiments, the antifungal agent comprises one or more of the group consisting of polyene antifungals, imidazoles, triazoles, allylamines, and echinocandins. In some embodiments, the bone graft composition exhibits antibacterial and / or antifungal efficacy. In some embodiments, the bone graft composition is not a dental resin.

[0022] In another aspect, there is provided a method of repairing a bone defect in a patient in need thereof, the method comprising applying a bone graft composition disclosed herein to the bone defect.

[0023] In some embodiments, the bone defect is caused by a human disease or condition, and wherein the bone graft composition includes an active agent for treating the human disease or condition. In some embodiments, the bone defect is caused by a human disease or condition, and wherein the bone graft composition does not contain an active agent for treating the human disease or condition. In some embodiments, the human disease or condition is one of several disorders of the vertebrae of the spine, requiring surgical procedures to fuse the vertebrae together. In some embodiments, the human disease or condition is one of several disorders of the bones of the limbs, requiring surgical procedures to fuse the bones together or to repair the defect. In some embodiments, the human disease or condition is a bone void or defect that is not essential to the stability of the bone structure, the bone void or defect is caused by surgery or traumatic injury, and the bone graft composition fills the bone void or defect. In some embodiments, the bone defect comprises a void or gap in the subject's skeletal system, the void or gap is not essential to the stability of the skeletal structure, and the bone graft composition is to fill the void or gap.

[0024] In some embodiments, the surgical procedure comprises a lumbar interbody fusion (LIF) procedure. In some embodiments, the LIF procedure comprises anterior LIF, lateral LIF, transforaminal LIF, and posterior LIF. In some embodiments, the LIF procedure alleviates associated symptoms, such as degenerative disc disease in the subject as determined via radiography, resulting from the formation of bone defects through the surgical procedure. In some embodiments, the LIF procedure alleviates associated neuromuscular or physical deficits in the subject as determined by the subject's Oswestry Disability Index (ODI), neurological assessment of the subject, or radiography of the subject.

[0025] In some embodiments, the human disease or condition is cancer. In some embodiments, the bone defect is caused by cancer. Specific types of cancer include, but are not limited to, skin cancer (e.g., melanoma), connective tissue cancer (e.g., sarcoma, osteosarcoma, Ewing's osteosarcoma, giant cell carcinoma), breast cancer, head and neck cancer, lung cancer (e.g., non-small cell lung cancer), gastric cancer, pancreatic cancer, ovarian cancer, cervical cancer, uterine cancer, anogenital cancer (e.g., testicular cancer), kidney cancer, bladder cancer, colon cancer, prostate cancer, central nervous system (CNS) cancer (e.g., neuroblastoma, etc.), retinal cancer, blood cancer (e.g., multiple myeloma, etc.), and cancer of the lymphatic system (e.g., Hodgkin's lymphoma and non-Hodgkin's lymphoma).

[0026] In some embodiments, the bone defect is caused by a malignant tumor or surgical removal of a malignant tumor via resection. In some embodiments, the malignant tumor comprises a breast cancer metastasis. In some embodiments, the presently disclosed methods utilize the disclosed bone graft compositions for reconstruction of the metaphysis-diaphysis of the humerus in a subject following resection of a malignant tumor, such as a breast cancer metastasis, using an intercalary allograft and plate fixation.

[0027] In some embodiments, the bone graft composition is osteoconductive. In some embodiments, the bone graft composition is osteoinductive. In some embodiments, the bone graft composition is osteopromotive. [Brief explanation of the drawings]

[0028] [Figure 1]MC3T3-E1 subclone 4 mouse calvarial-derived bone-forming cells were cultured as spheroids (micromasses) to induce osteoblast differentiation. Cultures were untreated (negative control), chemically treated to induce osteoblast differentiation (positive control), or cultured with bone graft material containing ferrous sulfate excipient (ferrous sulfate bone graft). Cultures were grown for 7 days before adding bone graft material or chemical induction reagent. After an additional 12 days, cultures were stained for mineral deposition with alizarin red stain (dark stain) (Figure 1A). The ferrous sulfate cultures had significantly more staining than either control group. Cultures were then evaluated for the amount of mineral contained in the spheroids to determine whether mineralization had increased (Figure 1B). The ferrous sulfate bone graft cultures had significantly more mineral in the spheroids compared to the negative control cultures (* = p<0.0001). The minerals in the positive control culture medium were not significantly different from those in the negative control culture medium (ns = not significant). Summary of the Invention

[0029] Described herein are compositions, materials, methods, and kits for bone grafting and repairing and / or filling voids or gaps in bone. The present disclosure provides bone graft compositions with improved material properties and improved bioavailability of drugs contained therein. The present disclosure also provides bone graft compositions containing coloring agents that allow for easy and convenient detection of the implanted material. Additionally, bone graft compositions with improved antibacterial functionality are included herein. Finally, the present disclosure provides bone graft compositions containing active agents with therapeutic properties for treating diseases and conditions in which promoting bone growth, either by stimulating bone formation or preventing bone destruction, is desirable or necessary.

[0030] I. Definition To facilitate understanding of certain terms used herein, the following definitions are provided.

[0031] Unless otherwise defined, technical and scientific terms used herein have the meanings commonly understood by those of ordinary skill in the art. Any suitable materials and / or methodology known to those of ordinary skill in the art may be utilized in carrying out the methods described herein.

[0032] As used in this description and the appended claims, the singular forms "a," "an," and "the" are used interchangeably and are intended to include the plural forms as well and be consistent with their respective meanings, unless the context clearly dictates otherwise. Also, as used herein, "and / or" means and includes all possible combinations of one or more of the listed items, as well as the lack of combination when interpreted in the alternative ("or").

[0033] All numerical designations, e.g., pH, temperature, time, concentration, amount, and molecular weight, including ranges, are approximations that vary by ±10%, 1%, or 0.1%, as appropriate. It is understood, although not always explicitly stated, that all numerical designations may be preceded by the term "about." It is also understood, although not always explicitly stated, that the reagents described herein are merely exemplary and that equivalents of such are known in the art.

[0034] The terms "comprising" or "comprising" are intended to mean that compositions and methods include the recited elements, but do not exclude other elements. "Consisting essentially of," when used to define compositions and methods, is intended to mean excluding other elements that are essential to the combination. For example, a composition consisting essentially of the elements defined herein would not exclude other elements that do not materially affect the basic and novel characteristic(s) of the claimed invention. "Consisting of" is intended to mean excluding more than trace amounts of other ingredients and substantial method steps as described. Embodiments defined by each of these transitions are within the scope of the present invention.

[0035] As used herein, the term "bone graft composition" means any malleable composition suitable for repairing bone defects. The bone graft composition may be a "calcium phosphate putty." In some embodiments, the calcium phosphate putty is a bone void filler.

[0036] As used herein, the term "active agent" means any entity, including chemical or biological entities, approved by a regulatory body, e.g., the USFDA, for the treatment of a particular indication.

[0037] The term "active agent-free" in the context of a composition or formulation means a composition or formulation that does not contain an active agent, as defined herein.

[0038] As used herein, the term "biocompatibility" refers to the ability (e.g., of a composition or material) to perform in a particular application with an appropriate host response, or at least to perform without toxic or other deleterious effects on the host's biological systems, either locally or systemically.

[0039] As used herein, the term "osteoconductive" refers to the ability (e.g., of a composition or material) to passively allow bone growth (e.g., onto and / or into the material). As such, osteoconduction can be characterized as a passive process. Osteoconductive materials or compositions will only contribute to new bone growth in areas where significant bone already exists.

[0040] A material (e.g., a graft or implant) can be osteoconductive because it is configured, for example, to passively allow bone growth on the surface of the material. In another example, a material can be osteoconductive because it is configured to passively allow bone growth into openings (e.g., pores) in the material.

[0041] As used herein, the term "osteoinductive" refers to the ability (e.g., of a composition or material) to actively stimulate a biological response that induces bone formation. As such, osteoinduction can be characterized as an active process. Osteoinductive materials or compositions can induce new bone growth and contribute to new bone growth in areas lacking significant bone.

[0042] Osteoinductivity may include the formation and / or stimulation of osteoprogenitor cells, for example, osteoprogenitor cells in body tissues surrounding or adjacent to the graft or implant.

[0043] As used herein, the term "osteopromoting" refers to the ability (e.g., of a composition or material) to promote new bone formation by enhancing the osteoinductivity of an osteoinductive material. An osteopromoting composition or material enhances osteoinductivity, but is not inherently osteoinductive.

[0044] As used herein, the term "bioactive" refers to the ability (e.g., of a composition or material) to form a hydroxyapatite (HA) surface layer when submerged in simulated body fluid (SBF).

[0045] As used herein, the term "osteostimulatory" refers to the ability (e.g., of a composition, material, or extract thereof) to enhance or positively stimulate osteoblast proliferation and mesenchymal stem cell differentiation.

[0046] As used herein, the terms "antibacterial" or "antimicrobial" refer to the <51> "Inhibition of microbial growth" refers to the ability (e.g., of a composition, material, or extract thereof) to inhibit the growth of microorganisms according to the methods described above.

[0047] As used herein, the term "biodegradable" refers to the ability of a material to be broken down, degraded, and / or digested over time by the action of a biological environment (including the action of an organism, e.g., a patient's body) and / or in response to changes in physiological pH or temperature. In the context of the human body environment, biodegradable means that the material will break down, degrade, and / or be digested under normal physiological conditions.

[0048] As used herein, the terms "resorbable" and "bioresorbable" refer to the ability of a material to be broken down over a period of time and assimilated into the biological environment. Resorbable and bioresorbable in the context of the human body environment means that the material will be broken down over a period of time and assimilated into the body under normal physiological conditions.

[0049] As used herein, the term "moldable" means the property of being flexible and capable of being manually compressed, molded, and manipulated while maintaining integrity, compositional homogeneity, physical properties, and performance characteristics.

[0050] As used herein, a reference to the weight of a component of a bone graft composition or material described herein, such as the phrase "by weight," means the weight of the component before it is added to or mixed with another component of the bone graft composition. For example, the weight can refer to the initial weight of the component measured before further processing of the component into a bone graft composition.

[0051] As used herein, the expression "non-weight-bearing application" refers to an application for the repair of a void or gap in a bone or another bony structure, where the void or gap being repaired is not essential to the stability of the bone or bony structure.

[0052] As used herein, the term "antagonist" is used interchangeably with "inhibitor" and refers to a substance that blocks or suppresses the activity, function, effect, or expression of a target. In some embodiments, the target is a compound, protein, gene, cell, or drug. As used herein, the term "expression" refers to the amount of a target produced by a living cell. In some embodiments, an inhibitor suppresses the expression of a target gene or protein. In some embodiments, an inhibitor includes a compound that blocks the binding of another molecule to an enzyme or molecular pump. In some embodiments, an inhibitor is a compound that causes downregulation of an enzyme. In some embodiments, an inhibitor can be a competitive inhibitor or a non-competitive inhibitor.

[0053] As used herein, the term "administering" includes formulating for administration as well as actually administering, including physically administering by the subject being treated or another.

[0054] As used herein, "subject," "patient," or "individual" refers to any subject, patient, or individual, and the terms are used interchangeably herein. In this regard, the terms "subject," "patient," and "individual" include mammals, particularly humans, dogs, and cats. When used in connection with "in need thereof," the term "subject," "patient," or "individual" contemplates any subject, patient, or individual having or at risk for a particular condition or disorder.

[0055] As used herein, the phrase "therapeutically effective" or "effective" in the context of "dosage" or "amount" means a dosage or amount that provides the particular pharmacological effect for which the compound or compounds are administered. It is emphasized that a therapeutically effective amount may not always be effective in achieving the intended effect in a given subject, even if such a dosage is considered to be a therapeutically effective amount by those skilled in the art. For convenience only, exemplary dosage amounts are provided herein. Those skilled in the art can adjust such amounts according to the methods disclosed herein to treat a particular subject suffering from a particular condition or disorder. A therapeutically effective amount may vary based on the route of administration and dosage form.

[0056] The term "treating" or "treatment" refers to the treatment of conditions that cause or are associated with bone defects, such as cancer. Such treatment includes (i) inhibiting cancer, i.e., preventing its development; (ii) palliating cancer or disorders, i.e., causing cancer regression; (iii) slowing the progression of cancer; and / or (iv) inhibiting, alleviating, or slowing the progression of one or more symptoms of cancer in a subject, such as a human or animal patient. For example, treating cancer includes, but is not limited to, eliminating cancer or a condition caused by cancer, causing tumor remission, inhibiting cancer, or reducing or eliminating at least one symptom of a tumor. Other examples of conditions that cause or are associated with bone defects include, for example, diabetes, osteoporosis, lupus, rheumatoid arthritis, hyperthyroidism, celiac disease, asthma, and multiple sclerosis. Active agents for treating any of the above conditions are well known to those skilled in the art and are within the scope of the present disclosure.

[0057] The terms "treat" or "treatment" can also refer to surgical treatment of conditions causing or associated with skeletal biomechanical instability, age-related degenerative conditions, or bone defects due to trauma. The patient may be a human or veterinary patient. For example, spinal fusion due to degenerative disc disease. In a human subject, such treatment involves removing a degenerated disc, placing an intervertebral spacer or "cage" between two vertebral segments in the treatment segment, placing bone graft material into the spacer / cage, and then mechanically stabilizing the spine using metal rods and screws. After several months, the bones of the vertebral segments fuse, resulting in a mechanically stable vertebral segment. Other conditions include fractures, osteotomies, arthroplasty, reconstructive procedures, oral augmentation (i.e., any dental restoration, including ridge augmentation, sinus lift, or socket repair), and defect (e.g., void filler) repair. In some embodiments, the bone defect comprises a void or gap in the subject's bony skeletal system, where the void or gap is not essential to the stability of the bony skeletal structure. The void or gap in the subject's bony skeletal system may be treated by gently packing the bone graft composition into the void or gap.

[0058] The condition may also be a disorder of a bone in an extremity, in which the bone in the extremity requires surgical treatment to fuse the bone or repair the defect. In some embodiments, the human disease or condition is a bone void or defect that is not essential to the stability of the bone structure, where the bone void or defect is created by surgery or traumatic injury, and the bone graft composition fills the bone void or defect. In some embodiments, the human disease or condition is a bone void or defect that is not essential to the stability of the bone structure, where the bone void or defect is created by surgery or traumatic injury. In some embodiments, the surgical treatment comprises a lumbar interbody fusion (LIF) procedure. In some embodiments, LIF treatment comprises anterior LIF, lateral LIF, transforaminal LIF, and posterior LIF. In some embodiments, the LIF treatment alleviates associated symptoms, such as intervertebral disc degeneration in a subject, as determined via radiography, due to the creation of a bone defect via surgical treatment. In some embodiments, LIF treatment alleviates the subject's associated neuromuscular or physical deficits as determined by the subject's Oswestry Disability Index (ODI), a neurological assessment of the subject, or an x-ray of the subject.

[0059] The term "analog" refers to a compound in which one or more individual atoms or functional groups have been replaced with either different atoms or different functional groups, generally resulting in a compound with similar properties. In some embodiments, an analog refers to a structure that is similar to another but differs in one or two components.

[0060] The term "derivative" refers to a compound formed from a similar starting compound by attaching another molecule or atom to the starting compound. Additionally, a derivative according to the present invention encompasses one or more compounds formed from a precursor compound by the addition of one or more atoms or molecules, or by the combination of two or more precursor compounds.

[0061] II. Calcium Phosphate Bone Graft Compositions Bone graft compositions or materials according to embodiments promote bone repair or regeneration at a target repair site. For example, in some embodiments, the bone graft composition can be osteoconductive, osteoinductive, bioactive, osteostimulating, antibacterial, or any combination thereof. The target repair site can be, for example, a void, gap, or other defect in a bone or other bony structure within a patient. For example, as described in more detail below, the bone graft composition promotes bone growth at a target repair site in the spine, pelvis, limbs, skull, or other bone or bony structure within a patient. The bone graft composition can be implanted, extruded, molded, or otherwise placed at the target repair site. For example, in some embodiments, the bone graft composition can be implanted, extruded, molded, or placed at the target repair site in a non-weight-bearing application. In other embodiments, the bone graft composition can be implanted, extruded, molded, or placed at the target repair site with appropriate orthopedic hardware (i.e., screws, plates, rods, cages / spacers, prostheses, hip implants, knee implants, acetabular implants, etc.) in a load-bearing application.

[0062] In some embodiments, the bone graft composition is a calcium phosphate composition.

[0063] In some embodiments, the calcium phosphate is selected from hydroxyapatite (Ca5(OH)(PO4)3), β-tricalcium phosphate (β-Ca3(PO4)2), calcium phosphate dibasic (CaHPO4), or calcium phosphate tribasic (Ca5(OH)(PO4)3), monocalcium phosphate monohydrate (Ca(H2PO4)2-H2O), dicalcium phosphate dihydrate (CaHPO4·2H2O), octacalcium phosphate (Ca8H2(PO4)6·5H2O), monocalcium phosphate (Ca(H2PO4)2), α-tricalcium phosphate (α-Ca3(PO4)2), sintered hydroxyapatite (Ca 10 (PO4)6(OH)2), oxyapatite (Ca 10 (PO4)6O) or tetracalcium phosphate (Ca4(PO4)2O).

[0064] In some embodiments, hydroxyapatite has the chemical formula Ca 10-x (HPO4) x (PO4) 6-x (OH) 2-x where "x" can vary between 0 and 2.

[0065] In some embodiments, tricalcium phosphate is a precipitated amorphous calcium phosphate having the chemical formula Ca3(PO4)2·nH2O, where "n" = 3-4.5 and the H2O content is 15%-20%.

[0066] In some embodiments, the size of the individual grains (or particles) of the hydroxyapatite material ranges from 1 nm to 5 mm.

[0067] In some preferred embodiments, the particle size for the hydroxyapatite will be selected from the following sizes: 10 μm (microns), 75 μm (microns), 86.4 μm (microns), 125 μm (microns), 147 μm (microns), 212 μm (microns), 368 μm (microns), or 740 μm (microns). In these embodiments, the particle size is understood to represent a maximum size, and the range will include potentially smaller particles of material.

[0068] In some embodiments, the size of the individual grains (or particles) of the β-tricalcium phosphate material ranges from 1 nm to 5 mm.

[0069] In some preferred embodiments, the particle size of the β-tricalcium phosphate will be selected from the following sizes: 100 μm (microns), 125 μm (microns), 212 μm (microns), 304 μm (microns), 645 μm (microns), 500 μm (microns), or 1000 μm (microns). In these embodiments, the particle size is understood to represent a maximum size, and the range will include potentially smaller particles of material.

[0070] In some embodiments, the tricalcium phosphate is a mixture of β-tricalcium phosphate and α-tricalcium phosphate.

[0071] In some embodiments, the calcium phosphate salt can be anhydrous, monohydrate, dihydrate, or any xH2O hydrate isoform.

[0072] In a preferred embodiment, the calcium phosphate salt included in the preferred formulation of the bone graft material is sintered hydroxyapatite (Ca 10 (PO4)6(OH)2), β-tricalcium phosphate (β-Ca3(PO4)2), dicalcium phosphate (CaHPO4), and tricalcium phosphate (Ca5(OH)(PO4)3).

[0073] III. Hardener In some embodiments, the bone graft composition includes a hardening agent, which enhances the degree of hardening of the bone graft composition.

[0074] In some embodiments, the hardening agent may be sodium carbonate (Na2CO3).

[0075] In some embodiments, the hardening agent may be calcium sulfate anhydrous (CaSO), calcium sulfate hemihydrate (CaSO-0.5H0), calcium sulfate dihydrate (CaSO-2H0), calcium carbonate (CaCO), magnesium carbonate (MgCO), strontium carbonate (SrCO), and bioglass.

[0076] In some embodiments, the hardener salt can be anhydrous, monohydrate, dihydrate, or any xH2O hydrate isoform.

[0077] In a preferred embodiment, the hardening agent included in the preferred formulation of bone graft material is sodium carbonate (Na2CO3).

[0078] IV. Agents that control the hardening rate In some embodiments, the bone graft composition includes a hardening agent, which increases the rate of hardening of the bone graft composition.

[0079] In some embodiments, the hardening agent may include calcium oxide (CaO), magnesium oxide (MgO), sodium phosphate dibasic (NaHPO), sodium pyrophosphate tetrabasic (NaPO), sodium orthophosphate (NaPO), or sodium phosphate monobasic (NaHPO).

[0080] In some embodiments, the sodium phosphate salt may include anhydrous, monohydrate, dihydrate, or any xH2O hydrate isoform.

[0081] In a preferred embodiment, the setting rate controlling agents include calcium oxide (CaO), magnesium oxide (MgO), and sodium phosphate dibasic (Na2HPO4).

[0082] V. Acidifying Agents It has been discovered by the inventors that the addition of acidifying agents to bone graft compositions results in improved material properties, dissolution, small molecule bioavailability, enhanced bone formation, and improved handling characteristics. Acidifying agents are defined as chemicals, molecules, or ions that can donate a proton (hydrogen ion, H+; known as a Bronsted-Lowry acid) or, alternatively, can form a covalent bond with an electron pair (known as a Lewis acid).

[0083] In some embodiments, the bone graft composition comprises an acidifying agent, and the acidifying agent is selected from the group consisting of L-ascorbic acid (C6H8O6), ascorbic acid 2-phosphate sesquimagnesium salt hydrate (C 12 H 12 O 10(PO4)2Mg3-xH2O), sodium citrate dihydrate (Na3C6H5O7·2H2O), magnesium citrate (MgC6H6O7), potassium citrate monohydrate (K3C6H5O7·H2O), citric acid monohydrate (C6H8O7·H2O), and acetic acid (C2H4O2).

[0084] In some embodiments, the acidifying agent can be anhydrous, monohydrate, dihydrate, or any xH2O hydrate isoform.

[0085] In some embodiments, the acidifying agent is a component of the bone graft material and is a salt or ester species that produces a polyatomic anion that forms in solution and that otherwise fits the rules defining a chemical species as an acid.

[0086] In a preferred embodiment, the acidifying agent comprises one or more of L-ascorbic acid (C6H8O6), citric acid monohydrate (C6H8O7-H2O), and acetic acid (C2H4O2).

[0087] V. Iron Excipients It has been discovered by the inventors that the addition of iron excipients to bone graft compositions in the form of iron salts, iron oxides, iron compounds, iron alloys, and elemental iron allows for the following: 1) As a coloring agent, the iron excipient allows for easy detection of the putty's position relative to the adjacent bone, and allows visualization of proper mixing by assessing color uniformity during mixing of the liquid and solid precursors of the bone graft composition. 2) Iron excipients modulate the physical properties of bone graft materials by acting as agents that control the rate of setting and as agents that contribute to setting. 3) In radiography (radiography, computed tomography), iron excipients act as contrast agents. 4) Iron excipients contribute to bone formation.

[0088] In some embodiments, the color-forming agent comprises a chromogen that is also an iron excipient. As used herein, the term "chromogen" refers to any chemical species that is an iron excipient that also changes color when dissolved in a solvent.

[0089] In some embodiments, the color former comprises an activatable color former.

[0090] In some embodiments, the coloring agent colors the bone graft composition red, orange, brown, black, or gray. In some preferred embodiments, the coloring agent colors the bone graft composition orange, red, brown, or gray.

[0091] In some embodiments, the iron excipient is selected from the group consisting of iron(III) sulfate hydrate (Fe2(SO4)3), iron(III) chloride (FeCl3), iron(III) citrate (FeC6H5O7), iron(III) oxide (Fe2O3), iron(III) oxide hydroxide (Fe(OH)O), iron(III) phosphate (FePO4), ammonium iron(III) citrate (C6H8O7-xFe3+ -yNH3), elemental iron particles (Fe), iron(III) fluoride (FeF3), iron(II) sulfate heptahydrate (FeSO4·7H2O), ammonium iron(II) sulfate hexahydrate (Fe(NH4)2(SO4)2·6H2O), iron chloride (FeC l2 ), iron(II) disulfide (FeS2), iron(II) sulfate heptahydrate (FeSO4·7H2O), and iron(II) lactate hydrate (Fe(CH3CH(OH)COO)2-xH2O), iron(II) L-ascorbate (FeC 12 H 14 O 12 ) and iron(II) fluoride (FeF2).

[0092] In a preferred embodiment, the iron excipient is ferrous sulfate hydrate (Fe2(SO4)3).

[0093] VII. Collagen The present inventors have discovered that collagen powder and liquid collagen preparations can be used as bone graft materials. Type I collagen is a white, hygroscopic material that is commonly used as a component of bone grafts.

[0094] Collagen powder is a preparation of collagen material in which collagen polymers are chemically or enzymatically cleaved or by any conventional means to produce collagen fragments of a specific size or size range. In this preparation, the collagen can be 1 nm to 100 μm (microns) produced through reverse dialysis. In another preparation, the collagen can be enzymatically cleaved to produce fragments of 100 μm (microns) to 300 μm (microns). In yet another preparation, the collagen can be mechanically crushed or milled and then passed through a filter to produce fragments between 200 μm (microns) and 5 mm. In another embodiment, the collagen has a particle size of 25 μm (microns) to 750 μm (microns). In another embodiment, the collagen has a particle size of 1 μm (microns) to 1.5 mm (millimeters). In yet another preparation, the collagen is crushed or milled at a low temperature (cryogrinding) to produce fragments of 1 μm (microns) to 5 mm.

[0095] Liquid collagen is a collagen preparation that contains a soluble fraction of collagen dissolved in solution. In this preparation, collagen is soaked in an acid solution, and / or heated between 100°C and 300°C, and / or heated in a pressurized environment, and / or filtered to produce fragments of a specific size. In yet another preparation, liquid collagen is produced by grinding or milling collagen suspended in solution using a rotor-stator to produce collagen fragments between 1 μm (micron) and 1 mm.

[0096] In a preferred preparation, collagen is ground to produce a powder having fragments of one of the following sizes: Fragments of 10 μm (microns), 15 μm (microns), 20 μm (microns), 25 μm (microns), 30 μm (microns), 35 μm (microns), 40 μm (microns), 50 μm (microns), 55 μm (microns), 60 μm (microns), 65 μm (microns), 70 μm (microns), 75 μm (microns), 80 μm (microns), 85 μm (microns), 90 μm (microns), 95 μm (microns), 100 μm (microns), 150 μm (microns), 200 μm (microns), 250 μm (microns), 300 μm (microns), 350 μm (microns), 400 μm (microns), 450 μm (microns), 500 μm (microns), 750 μm (microns), or 1000 μm (microns). In another preparation, the collagen powder is composed of fragment sizes in the above ranges.

[0097] VIII. Antibacterial Agents As used herein, the term "antimicrobial agent" refers to an agent that kills or stops the growth of microorganisms. Antibiotics are antibacterial agents that kill bacteria, and antifungals are agents that kill fungi.

[0098] Thus, in some embodiments, the bone graft composition comprises an antimicrobial agent, hi some embodiments, the antimicrobial agent comprises an antibiotic agent or an antifungal agent.

[0099] In some embodiments, the antibiotic comprises vancomycin, gentamicin, tobramycin, kanamycin, neomycin, ampicillin, methicillin, nafcillin, oxacillin, penicillin, ticarcillin, ciprofloxacin, vancomycin, cefazolin, cefepime, ceftriaxone, clindamycin, aztreonam, imipenem, quinupristin / dalfopristin, chloramphenicol, doxycycline, metronidazole, nitrofurantoin, polymyoxin B, tetracycline, viomycin, chloromycetin, streptomycin, azactam, pharmaceutically acceptable salts thereof, and combinations thereof.

[0100] In some embodiments, the antifungal agent comprises a polyene antifungal agent, an imidazole, a triazole, an allylamine, or an echinocandin. In some embodiments, the polyene antifungal agent comprises amphotericin B, candicidin, filipin, hamycin, natamycin, nystatin, rimocidin, an imidazole, a triazole, and a thiazole. In some embodiments, the imidazole comprises bifonazole, butoconazole, clotrimazole, econazole, fenticonazole, isoconazole, ketoconazole, luliconazole, miconazole, omoconazole, oxiconazole, sertaconazole, sulconazole, tioconazole, and a triazole. In some embodiments, triazoles include albaconazole, efinaconazole, epoxiconazole, fluconazole, isavuconazole, itraconazole, posaconazole, propiconazole, ravuconazole, terconazole, and voriconazole. In some embodiments, allylamines include amorolfine, butenafine, naftifine, and terbinafine. In some embodiments, echinocandins include anidulafungin, caspofungin, and micafungin.

[0101] In some embodiments, the bone graft composition exhibits antibacterial or antifungal efficacy.

[0102] IX. Diluted Solution The diluent solution is an aqueous medium consisting of water and other excipients that is capable of solubilizing the powder or dry slurry components of the bone graft composite material.

[0103] In some embodiments, the bone graft composition comprises a diluent or solution that is added to the other components of the bone graft material to create a malleable, moldable putty.

[0104] In some embodiments, the diluent is saline or water.

[0105] In a preferred embodiment, the diluent is 0.9% saline, as commonly used in some medical and scientific applications.

[0106] In another preferred embodiment, the diluent is 0.9% saline containing an acidifying agent.

[0107] In yet another preferred embodiment, the diluent is 0.9% saline containing acetic acid.

[0108] In yet another preferred embodiment, the diluent is 0.9% saline containing acetic acid and any acidifying agent including a citric acid molecule, e.g., citric acid monohydrate or magnesium citrate, potassium citrate, or sodium citrate dihydrate.

[0109] In another preferred embodiment, the diluent is 0.9% saline containing an acidifying agent and a liquid preparation of collagen.

[0110] X. Preparation of Bone Graft Composition As used herein, the term "blending" refers to the physical process of combining or combining non-aqueous materials to form one substance or mass that is a powder or dry slurry, referred to herein as a "powder-dry slurry." Part of the preparation of a bone graft composite requires blending the following components into a single powder-dry slurry composite: calcium phosphate with a setting agent, iron excipient, an agent that controls the setting rate, an acidifying agent, and powdered collagen.

[0111] As used herein, the term "mixing" refers to the physical process of combining a powder dry slurry with a dilute solution to form a paste, putty, or other amorphous solid.

[0112] In some embodiments, 0.1 mL to 15 mL of diluent will be added to 0.1 g to 20 g of powder dry slurry. In some embodiments, 0.5 mL of diluent will be added to 0.5 g of powder dry slurry. In some embodiments, 0.765 mL of diluent will be added to 1 g of powder dry slurry. In some embodiments, 1 mL of diluent will be added to 1 g of powder dry slurry. In some embodiments, 1.5 mL of diluent will be added to 1.5 g of powder dry slurry. In some embodiments, 2 mL of diluent will be added to 2 g of powder dry slurry. In some embodiments, 2.5 mL of diluent will be added to 2.5 g of powder dry slurry. In some embodiments, 3 mL of diluent will be added to 3 g of powder dry slurry. In some embodiments, 3.5 mL of diluent will be added to 3.5 g of powder dry slurry. In some embodiments, 4 mL of diluent would be added to 4 g of powder dry slurry. In some embodiments, 4.5 mL of diluent would be added to 4.5 g of powder dry slurry. In some embodiments, 5 mL of diluent would be added to 5 g of powder dry slurry. In some embodiments, 5.5 mL of diluent would be added to 5.5 g of powder dry slurry. In some embodiments, 6 mL of diluent would be added to 6 g of powder dry slurry. In some embodiments, 6.5 mL of diluent would be added to 6.5 g of powder dry slurry. In some embodiments, 7 mL of diluent would be added to 7 g of powder dry slurry. In some embodiments, 7.5 mL of diluent would be added to 7.5 g of powder dry slurry. In some embodiments, 8 mL of diluent would be added to 8 g of powder dry slurry. In some embodiments, 8.5 mL of diluent would be added to 8.5 g of powder dry slurry. In some embodiments, 9 mL of diluent will be added to 9 g of powder dry slurry. In some embodiments, 9.5 mL of diluent will be added to 9.5 g of powder dry slurry.In some embodiments, 10 mL of diluent will be added to 10 g of powder dry slurry. In some embodiments, 10.5 mL of diluent will be added to 10.5 g of powder dry slurry.

[0113] In some embodiments, the bone graft composition has a density of 1.6 g / mL, 1.65 g / mL, 1.7 g / mL, 1.75 g / mL, 1.76 g / mL, 1.77 g / mL, 1.78 g / mL, 1.79 g / mL, 1.8 g / mL, 1.85 g / mL, 1.9 g / mL, or in the range of 1-6 g / mL after mixing. In some embodiments, the bone graft composition has a density of 2-6 g / mL after mixing. In some embodiments, the bone graft composition has a density of 2-5 g / mL after mixing. In some embodiments, the bone graft composition has a density of 3.33-4.2 g / mL after mixing.

[0114] In some embodiments, the force required to extrude the bone graft composition through a cannula, tube, or syringe is less than 100 N / m. In some embodiments, the force required to extrude the bone graft composition through a cannula, tube, or syringe is less than 90 N / m. In some embodiments, the force required to extrude the bone graft composition through a cannula, tube, or syringe is less than 80 N / m. In some embodiments, the force required to extrude the bone graft composition through a cannula, tube, or syringe is less than 70 N / m. In some embodiments, the force required to extrude the bone graft composition through a cannula, tube, or syringe is less than 60 N / m.

[0115] XI. Active Agent Delivery A. Small molecule surfactants The inventors have discovered that bone graft materials can be designed to more efficiently solubilize, sequester, and deliver small molecule active agents.

[0116] In some embodiments, the bone graft composition includes a small molecule active agent, which is a chemical species that affects some cellular function and is contained or sequestered in the bone graft composite material.

[0117] The small molecule active agent is a chemical species derived from a protein, a synthetic chemical species, or a chemical species derived from a plant or animal source. The chemical species can be chemically altered before incorporation into the bone graft composite material, or the chemical species can be in its native state before incorporation into the bone graft composite material.

[0118] The chemical properties, such as pH and osmolality, of the diluted solution components of the bone graft can be designed to increase the solubility of the small molecule active agent in the diluted solution before mixing with the powder or dry slurry components of the bone graft composition.

[0119] The chemical properties of the powdered dry slurry components of bone grafts can be designed so that when the powdered dry slurry components are mixed with a diluent, the small molecule active agent is less likely to dissolve and is sequestered within the material. This property can then be used to control the elution and bioavailability of the small molecule active agent.

[0120] In some embodiments, the small molecule active agent may be a component of the diluent.

[0121] In some embodiments, the small molecule active agent can be a component of the powder dry slurry component.

[0122] In some embodiments, the small molecule active agent can be a separate preparation, either in liquid form or as a lyophilized solid, that is added to the diluent.

[0123] In some embodiments, the small molecule active agent can be a separate preparation, either a liquid or a lyophilized solid, that is added to the powder dry slurry component.

[0124] In some embodiments, the small molecule is an opioid growth factor antagonist.

[0125] B. Other Biologically Derived Surfactants The present inventors have discovered that bone graft materials can be designed to more efficiently solubilize, sequester, and deliver biologically derived active agents.

[0126] In some embodiments, the bone graft composition includes a bioactive agent, which is a biological species that effects some cellular function and is contained or sequestered in the bone graft composite material.

[0127] Biologically derived active agents are chemical species derived from proteins, synthetically derived chemicals, or plant or animal sources. The biologically derived species can be chemically altered before incorporation into the bone graft composite, or the biologically derived species can be in its native state before incorporation into the bone graft composite.

[0128] The chemical properties, such as pH and osmolality, of the bone graft dilute solution components can be designed to increase the solubility of the bioactive agent in the dilute solution prior to mixing with the powder or dry slurry components of the bone graft composition.

[0129] The chemical properties of the powdered dry slurry components of bone grafts can be engineered so that when the powdered dry slurry components are mixed with a diluent, the bioactive agent is less likely to dissolve and is sequestered within the material, which can be used to control the amount of leaching and bioavailability of the bioactive agent.

[0130] In some embodiments, the bioactive agent may be a component of a dilute solution.

[0131] In some embodiments, the bioactive agent can be a component of the powder dry slurry component.

[0132] In some embodiments, the biologically derived active agent can be a separate preparation, either in liquid form or as a lyophilized solid, that is added to the diluent.

[0133] In some embodiments, the biologically derived active agent can be a separate preparation, either a liquid or a lyophilized solid, that is added to the powdered dry slurry component.

[0134] The additional biologically active agent may be a bone morphogenetic protein (i.e., BMP2, BMP4, BMP7), a platelet-derived growth factor (PDGF) protein (i.e., PDGF-α or PDGF-β), a transforming growth factor (TGF) β protein, a vascular endothelial growth factor (VEGF) protein, cyclopamine, purmorphamine, resolufam, a netrin protein (i.e., netrin-1 or netrin-4), a slit protein (i.e., slit-1, slit-2, slit-3), a repulsive guidance molecule (RGM) protein, or a WNT protein (i.e., WNT1, WNT5, WNT10, WNT11).

[0135] C. Opioid Growth Factor Receptor (OGFR) Antagonists By "OGFR antagonist" is meant any small molecule active agent that inhibits, suppresses, or abolishes at least one OGFR-associated biological activity.

[0136] In some embodiments, the OGFR antagonist is an OGFR binding antagonist, ie, a molecule that interferes with, blocks, or otherwise prevents the interaction or binding of met5-ligand (OGF) to OGFR.

[0137] OGFR binding antagonists can function in two ways. First, they can compete with met5-ligands for binding to OGFR on the nuclear membrane surface, thereby interfering with, blocking, or otherwise preventing the binding of met5-ligands to OGFR, and can bind to OGFR without inducing downstream signaling induced by the binding of met5-ligands to OGFR. Alternatively, OGFR binding antagonists can bind to or sequester PENK or met5-ligands with sufficient affinity and specificity to substantially interfering with, blocking, or otherwise preventing the binding of met5-ligands to OGFR, thereby inhibiting, suppressing, or abolishing at least one OGFR-related biological activity. In general, OGFR binding antagonists can be large molecules (e.g., antibodies) or small molecules (e.g., compounds with a molecular weight of less than 15 kD, 12 kD, 10 kD, or even 8 kD), and can be polypeptides, nucleic acids, or synthetic small molecule compounds. OGFR binding antagonist can be identified by any in vitro assay that can be easily selected by those skilled in the art.For example, OGFR antagonist can be identified by the method described in US Patent No. 5,882,944, US Patent No. 6,007,986, US Patent No. 6,270,979.

[0138] In one embodiment, the OGFR binding antagonist is naloxone or a functional derivative thereof, naltrexone or a functional derivative thereof, or a combination thereof.

[0139] As used herein, the term "functional derivative" refers to a derivative or analog that is structurally and functionally similar to the starting molecule (e.g., that maintains the function of naltrexone or naloxone as an OGFR antagonist). Naloxone and naltrexone analogs can be synthesized using standard synthetic procedures, such as those described in March J., Advanced Organic Chemistry, 3rd Ed. (1985). Examples of naltrexone and naloxone functional derivatives include salt forms, such as naloxone hydrochloride dihydrate or naltrexone hydrochloride. Additional examples of naltrexone and naloxone functional derivatives suitable for use in the methods of the present invention include naltrexone and naloxone analogs described, for example, in U.S. Patent Application Publication No. 2007 / 0197573 A1 and U.S. Patent No. 6,713,488.

[0140] In another embodiment, the OGFR binding antagonist is derived from oxymorphone and binds to OGFR, including naloxone, naltrexone, nalorphine, naloxonazine, levallorphan, nalmefene, cyprodim, cilorphan, cyclazocine, oxilorphan, LY113878, MR2266, diprenorphine, WIN 44,441-3, naltoindole, or norbinaltorphimine.

[0141] In yet another embodiment, the OGFR binding antagonist is derived from trans-3,4-dimethyl-4-phenylpiperidine and binds to OGFR, including LY99335, LY25506, LY117413, or LY255582.

[0142] In another embodiment, the OGFR binding antagonist is derived from a met5-enkephalin or Leu-enkephalin peptide and minimally contains the following amino acid sequence as a means of binding to and targeting the OGFR: Tyr-Gly-Gly-Phe-Met (SEQ ID NO: 1) for met5-enkephalin derived, or Tyr-Gly-Gly-Phe-Leu (SEQ ID NO: 2) for Leu-enkephalin derived.

[0143] In yet another embodiment, the OGFR binding antagonist is derived from the peptide antagonist 101174864 (N,N-Diallyl-Tyr-Aib-Aib-Phe-Leu-OH, SEQ ID NO: 3; Aib = aminoisobutyric acid) or the somatostatin analog CTP (D-Phe-Cys-Tyr-D-Trp-Lys-Thr-Pen-Thr-NH, SEQ ID NO: 4).

[0144] In other embodiments, the OGFR antagonist is not an OGFR binding antagonist, but rather a molecule that inhibits the nuclear localization sequence found within OGFR: 251QSALDYFMFAVRCRHQRRQLVHFAWEHFRPRCKFVWGPQDKLRRFKPSSL (SEQ ID NO: 5).

[0145] In yet another embodiment, the OGFR antagonist employed in the present methods is a small hairpin (sh)-RNA or a small interfering (si)-RNA directed against the OGFR gene and effective to inhibit OGFR gene expression.

[0146] The OGFR antagonists described herein can be administered individually or in combination. Suitable combinations include, for example, naloxone and naltrexone; naloxone and / or naltrexone with another OGFR binding antagonist or another OGFR antagonist.

[0147] D. Delivery Systems and Carriers for Topical Administration In one embodiment of the present disclosure, the bone graft composite material is manufactured by mixing a suitable diluent containing the active agent with a powder dry slurry that is administered directly and locally to the site of the bone injury or surgical procedure where the bone graft material is typically placed to stimulate bone formation.

[0148] The inventors have discovered that specific formulations of diluent and powder dry slurry components can be achieved to optimize the dissolution, sequestration, and bioavailability of OGFR antagonists that share chemical properties with naloxone, naltrexone, a combination of naloxone and naltrexone, or any of their salts.

[0149] In some embodiments, the OGFR antagonist is incorporated into the bone graft composite material so that the concentration in the material is between 1 nanomolar (nM) per cubic centimeter (1 nM / cc) and about 100 mM per cubic centimeter (100 mm / cc). In some embodiments, the concentrations are 1 mM / cc, 1.25 mM / cc, 1.5 mM / cc, 1.75 mM / cc, 2.0 mM / cc, 3.0 mM / cc, 4.0 mM / cc, 5.0 mM / cc, 6.0 mM / cc, 7.0 mM / cc, 8.0 mM / cc, 9.0 mM / cc, and / or 10.0 mM / cc, 20.0 mM / cc, 30.0 mM / cc, 40.0 mM / cc, 50.0 mM / cc, 60.0 mM / cc, 70.0 mM / cc, 80.0 mM / cc, 90.0 mM / cc, 100 mM / cc.

[0150] In some embodiments, the concentration of the OGFR antagonist in the bone graft material is such that the elution rate from the material is between 0.001 pM per second and 10 mM per day.

[0151] In some embodiments, the cumulative dose resulting from elution of the OGFR agonist during the period in which the small molecule is released from the bone graft material by diffusion or other physical effects resulting from the normal degradation of the bone graft material once implanted is any of 0.8 mM, 1 mM, 1.1 mM, 1.15 mM, 1.2 mM, 1.25 mM, 1.3 mM, 1.35 mM, 1.4 mM, 1.45 mM, 1.5 mM, 1.55 mM, 1.6 mM, 1.65 mM, 1.7 mM, 1.75 mM, 1.8 mM, 1.9 mM, 2 mM, 2.7 mM, 3 mM, 3.6 mM, 4 mM, 4.5 mM, 5 mM, 5.4 mM, 6 mM, 6.3 mM, 7 mM, 7.2 mM, 8 mM, 8.1 mM, 9 mM or 10 mM.

[0152] In a preferred embodiment, the OGFR antagonist is naloxone hydrochloride in a concentration of 0.2 mg / cc (0.5 mM / cc), 0.25 mg / cc (0.625 mM / cc), 0.3 mg / cc (0.75 mM / cc), 0.35 mg / cc (0.875 mM / cc), 0.4 mg / cc (1 mM / cc), 0.45 mg / cc (1.125 mM / cc), 0.5 mg / cc (1.25 mM / cc), 0.55 mg / cc (1.37 mM / cc), 0.65 mg / cc (1.65 mM / cc), 0.75 mg / cc (1.875 mM / cc), 0.875 mg / cc (1.95 mM / cc), 0.95 mg / cc (1.95 mM / cc), 1.15 mg / cc (1.15 mM / cc), 1.25 mg / cc (1.375 mM / cc), 1.45 mg / cc (1.45 mM / cc), 1.55 mg / cc (1.55 mM / cc), 1.65 mg / cc (1.65 mM / cc), 1.75 mg / cc (1.75 mM / cc), 1.85 mg / cc (1.85 mM / cc), 1.95 mg / cc (1.9 ... 5mM / cc), 0.6mg / cc(1.5mM / cc), 0.65mg / cc(1.625mM / cc), 0.7mg / cc(1.75mM / cc), 0.75mg / cc(1.875mM / cc), 0.8mg / cc (2mM / cc), 0.85mg / cc (2.125mM / cc), 0.9mg / cc (2.25mM / cc), 0.95mg / cc (2.375mM / cc) or 1mg / cc (2.5mM / cc).

[0153] E. Composition and Administration The methods and compositions herein may be provided in the form of a kit. A "kit" is defined herein as a package containing several individual components that exhibit complementary effects when applied together. In this embodiment, the effects achieved by the kit and the pharmaceutical composition are similar. The kit may optionally include instructions for using the pharmaceutical composition.

[0154] The present invention is further illustrated by, but not limited to, the following examples.

[0155] Example 1: Bone graft material

[0156] In a first embodiment, a preferred embodiment of the bone graft material dry slurry components comprises the following in weight percent: 1) Iron(III) chloride between 1% and 7% 2) Calcium phosphate dibasic between 8.5% and 11% 3) 9.5% to 12.5% ​​calcium phosphate tribasic 4) 15% to 17% hydroxyapatite 5) 3% to 6% β-tricalcium phosphate 6) 5% to 7.5% sodium phosphate dibasic 7) 12% to 16% sodium carbonate 8) 2% to 6% calcium oxide 9) 4% to 6% magnesium oxide 10) 0.5% to 2.5% L-ascorbic acid and 11) 19% to 23% powdered type I collagen. A preferred embodiment of the diluent component is a solution having a volume of 0.5 mL to 12.5 mL consisting of: 1) 0.9% saline 2) 2% to 6% acetic acid 3) 0.1% to 2% citric acid monohydrate and 4) 0.01% to 2% naloxone hydrochloride. Upon mixing the dry slurry components and diluent, the bone graft material becomes moldable after 3 minutes, turns light brown as it solidifies, and reaches a peak temperature of 29.7°C 2 minutes after mixing.

[0157] In a second embodiment, a preferred embodiment of the bone graft material dry slurry components includes the following in weight percent: 1) 5% to 10% iron(III) citrate 2) 8% to 11% calcium phosphate dibasic 3) 9% to 12.5% ​​calcium phosphate tribasic 4) 13% to 17% hydroxyapatite 5) 3% to 6% β-tricalcium phosphate 6) 4% to 7.5% sodium phosphate dibasic 7) 11% to 15% sodium carbonate 8) 2% to 6% calcium oxide 9) 4% to 6% magnesium oxide 10) 0.5% to 2.5% L-ascorbic acid and 11) 19% to 23% powdered type I collagen. A preferred embodiment of the diluent component is a solution having a volume of 0.5 mL to 12.5 mL, consisting of: 1) 0.9% saline 2) 2% to 6% acetic acid 3) 0.1% to 2% citric acid monohydrate and 4) 0.01% to 2% naloxone hydrochloride. Upon mixing the dry slurry components and diluent, the bone graft material becomes moldable after 5 minutes, solidifies to a dark chocolate brown color, and reaches a peak temperature of 28.6°C 30 seconds after mixing.

[0158] In a third embodiment, a preferred embodiment of the bone graft material dry slurry components includes the following in weight percent: 1) 1% to 5% iron(III) oxide 2) 8% to 11% calcium phosphate dibasic 3) 9% to 13% calcium phosphate tribasic 4) 14% to 18% hydroxyapatite 5) 3% to 6.5% β-tricalcium phosphate 6) 4% to 8% sodium phosphate dibasic 7) 12% to 16% sodium carbonate 8) 2% to 6% calcium oxide 9) 4% to 6% magnesium oxide 10) 0.5% to 2.5% L-ascorbic acid and 11) 20% to 24% powdered type I collagen. A preferred embodiment of the diluent component is a solution having a volume of 0.5 mL to 12.5 mL, consisting of: 1) 0.9% saline 2) 2% to 6% acetic acid 3) 0.1% to 2% citric acid monohydrate and 4) 0.01% to 2% naloxone hydrochloride Upon mixing the dry slurry components and diluent, the bone graft material becomes moldable after 3.5 minutes, solidifies to a white with an iron-gray tint, and reaches a peak temperature of 29.8°C 30 seconds after mixing.

[0159] In a fourth embodiment, a preferred embodiment of the bone graft material dry slurry components includes, in weight percent: 1) 5% to 10% iron(III) phosphate 2) 8% to 11% calcium phosphate dibasic 3) 9% to 12.5% ​​calcium phosphate tribasic 4) 13% to 17% hydroxyapatite 5) 3% to 6% β-tricalcium phosphate 6) 4% to 7.5% sodium phosphate dibasic 7) 12% to 16% sodium carbonate 8) 2% to 6% calcium oxide 9) 4% to 6% magnesium oxide 10) 0.5% to 2.5% L-ascorbic acid and 11) 19% to 23% powdered type I collagen. A preferred embodiment of the diluent component is a solution having a volume of 0.5 mL to 12.5 mL, consisting of: 1) 0.9% saline 2) 2% to 6% acetic acid 3) 0.1% to 2% citric acid monohydrate and 4) 0.01% to 2% naloxone hydrochloride. Upon mixing the dry slurry components and diluent, the bone graft material becomes moldable after 3.5 minutes and turns light brown upon solidification, reaching a peak temperature of 33.5°C 1.5 minutes after mixing.

[0160] In a fifth embodiment, a preferred embodiment of the dry slurry components of the bone graft material includes the following in weight percent: 1) 15% to 10% ammonium iron(III) citrate 2) 8% to 11% calcium phosphate dibasic 3) 9% to 12.5% ​​calcium phosphate tribasic 4) 13% to 17% hydroxyapatite 5) 3% to 6% β-tricalcium phosphate 6) 4% to 7.5% sodium phosphate dibasic 7) 11% to 15% sodium carbonate 8) 2% to 6% calcium oxide 9) 4% to 6% magnesium oxide 10) 0.5% to 2.5% L-ascorbic acid and 11) 19% to 23% powdered type I collagen. A preferred embodiment of the diluent component is a solution having a volume of 0.5 mL to 12.5 mL, consisting of: 1) 0.9% saline 2) 2% to 6% acetic acid 3) 0.1% to 2% citric acid monohydrate and 4) 0.01% to 2% naloxane hydrochloride. Upon mixing the dry slurry components and diluent, the bone graft material becomes moldable after 3 minutes and turns brown as it solidifies, reaching a peak temperature of 30.3°C 1.5 minutes after mixing.

[0161] In a sixth embodiment, a preferred embodiment of the bone graft material dry slurry components includes the following in weight percent: 1) Iron(III) sulfate between 5% and 12% 2) 8% to 11% calcium phosphate dibasic 3) 9% to 12.5% ​​calcium phosphate tribasic 4) 13% to 17% hydroxyapatite 5) 3% to 6% β-tricalcium phosphate 6) 4% to 7.5% sodium phosphate dibasic 7) 11% to 15% sodium carbonate 8) 2% to 6% calcium oxide 9) 4% to 6% magnesium oxide 10) 0.5% to 2.5% L-ascorbic acid and 11) powdered type I collagen containing 18% or more and 23% or less. A preferred embodiment of the diluent component is a solution having a volume of 0.5 mL to 12.5 mL, consisting of: 1) 0.9% saline 2) 2% to 6% acetic acid and 3) 0.1% to 2% citric acid monohydrate. In this embodiment, no active agent is incorporated. Upon mixing the dry slurry components and diluent, the bone graft material becomes moldable after approximately 2.5 minutes and turns dark brown as it solidifies, reaching a peak temperature of 38°C 1.5 minutes after mixing.

[0162] In a seventh embodiment, a preferred embodiment of the bone graft material dry slurry components comprises, in weight percent: 1) Iron(III) sulfate between 5% and 12% 2) 8% to 11% calcium phosphate dibasic 3) 9% to 12.5% ​​calcium phosphate tribasic 4) 13% to 17% hydroxyapatite 5) 3% to 6% β-tricalcium phosphate 6) 4% to 7.5% sodium phosphate dibasic 7) 11% to 15% sodium carbonate 8) 2% to 6% calcium oxide 9) 4% to 6% magnesium oxide 10) 0.5% to 2.5% L-ascorbic acid and 11) 18% to 23% powdered type I collagen. A preferred embodiment of the diluent component is a solution having a volume of 0.5 mL to 12.5 mL, consisting of: 1) 0.9% saline 2) 2% to 6% acetic acid 3) 0.1% to 2% citric acid monohydrate and 4) 0.01% to 2% naloxone hydrochloride. The dry slurry ingredients and diluent are mixed together and become moldable after approximately 2.5 minutes, solidifying to a dark brown color and reaching a peak temperature of 38°C 1.5 minutes after mixing.

[0163] Additionally, in a seventh preferred embodiment containing ferrous sulfate, bone graft material was mixed as described and placed in culture to assess osteoblast differentiation according to ASTM F3106-14. MC3T3-E1 (MC3) cells, a murine osteoblast (bone cell) progenitor cell line, were seeded into culture as micromasses (2.5 x 10 cells per micromass suspended in 20 μL of Matrigel + DMEM solution), grown for 120 hours, and maintained in growth medium (DMEM with 20% fetal bovine serum). Control cultures (Control) were maintained in growth medium. Induced control cultures (Induced) were stimulated to become osteocytes in medium supplemented with ascorbic acid, β-glycerophosphate, and dexamethasone. In this example, MC3 micromass spheroids were grown for an additional 7 days, after which experimental groups were stained with Alizarin Red and analyzed for mineral content using one-way ANOVA with Bonferroni correction to assess differences.

[0164] The results demonstrate that cultures containing ferrous sulfate material (with or without activator) significantly increased bone formation compared to induced and control cultures based on alizarin red staining (Figure 1A). The size of the micromass spheroids and the mineral content of each micromass spheroid were quantified to identify the mineral area fraction. The mineral area fraction of micromass spheroids containing ferrous sulfate bone graft material had significantly more mineral than control cultures by 4-fold (* = p < 0.0046) (Figure 1B). From these data, it can be concluded that the ferrous sulfate bone graft material is osteoconductive and osteopromoting. Between-group analysis demonstrated that bone formation in bone grafts without naloxone was a component of the bone graft material, increasing mineral formation by 4-fold (* = p < 0.0046) in cultures compared to control and chemically induced wells. The addition of 0.4 mg of naloxone to the bone graft material increased mineral formation by an additional 25% (x = p<0.0001) relative to the control bone graft material group without naloxone.

[0165] Example 2: Use of bone graft implants in spinal fusion surgery

[0166] Example 2 demonstrates lateral lumbar interbody fusion (LLIF) using the claimed bone graft. Specifically, this study performed LIF using an osteoconductive synthetic bone graft material containing low-molecular-weight naloxone hydrochloride. The synthetic bone graft is composed of bovine-derived type I collagen, hydroxyapatite (HA), beta-tricalcium phosphate, and other calcium salts and some excipients. Hereinafter, the synthetic bone graft composition containing naloxone hydrochloride is referred to as Composition ZF.

[0167] The present disclosure provides methods for repairing a bone defect in a patient in need thereof, comprising applying a bone graft composition, such as Composition ZF, to a surgical procedure, such as a lumbar interbody fusion (LIF). In some embodiments, the LIF procedure includes anterior LIF (ALIF), lateral LIF (LLIF), transforaminal LIF (TLIF), and posterior LIF (PLIF). In some embodiments, the LIF procedure alleviates an associated pathology, such as degenerative disc disease in a subject, as determined through radiography, by creating a bone defect via a surgical procedure. In some embodiments, the LIF treatment alleviates an associated neuromuscular disorder or physical deficit in a subject, as determined through the subject's Oswestry Disability Index (ODI), a neurological evaluation of the subject, or a radiography of the subject.

[0168] Background of LIF treatment

[0169] LIF procedures are differentiated based on the anatomical approach and implanted hardware, including anterior LIF (ALIF), lateral LIF (LLIF), transforaminal LIF (TLIF), and posterior LIF (PLIF). LIF procedures involve removing the disc believed to be causing low back or leg pain and replacing it with an interbody cage or spacer to promote disc-vertebral fusion. LIF procedures have increased significantly over the past 20 years due to advances in surgical techniques, implants, imaging modalities, and demands.

[0170] As a result of the potential challenges of local autografts, allografts and orthobiologics are routinely used in combination with or as a substitute for local bone to meet the bone graft needs in LIF procedures. Although significant progress has been made in the development of bone graft materials, there remains a need for bone graft materials that provide biomechanically strong, rigid fixation and have a favorable safety profile.

[0171] Testing of bone graft compositions

[0172] Indications for synthetic ZF bone grafts include spinal fusion in skeletally mature patients with degenerative disc disease (DDD) at a single level (L2-L5). DDD is defined as discogenic back pain accompanied by disc degeneration and / or grade 2 spondylolisthesis at the involved level, confirmed by patient history and radiographic evaluation. Patients receiving synthetic ZF bone grafts must have failed non-operative treatment for at least 3 months prior to treatment with the synthetic ZF bone graft device. Lateral lumbar interbody fusion (LLIF) involves the implantation of synthetic ZF bone grafts using commercially available polyetheretherketone (PEEK), porous PEEK, or hydroxyapatite (HA)-PEEK spacers and auxiliary fixation systems (lateral plates or bilateral pedicle screws).

[0173] Study design

[0174] Subjects will undergo LLIF with an auxiliary fixation system or lateral plate. In the experimental group, the cavity of the intervertebral body spacer will be filled with synthetic ZF bone graft. In the control group, the cavity of the intervertebral body spacer will be filled with allogeneic bone graft (i.e., DBX putty, demineralized bone matrix) + / - bone marrow fluid. The eligibility criteria for surgery are as follows: 1) Patients aged 22 to 75 years. 2) A diagnosis of degenerative disc disease (DDD) associated with back pain, with or without leg pain, at a single level of the spine between the second lumbar vertebra (L2) and the fifth lumbar vertebra (L5), confirmed by medical history and radiographic evaluation (e.g., plain radiographs, computed tomography (CT), magnetic resonance imaging (MRI)). 3) Spinal instability confirmed by an X-ray taken during flexion and extension, with an angle of 5 degrees or more and / or a translation of 4 mm or more of the affected vertebrae. 4) Imaging findings showing pathology such as osteophyte formation, loss of disc height, ligamentous thickening, disc degeneration / herniation, facet joint degeneration, dark disc appearance on MRI, foraminal narrowing, or loss of foraminal height. 5) DDD is defined as discogenic low back pain with evidence of disc degeneration confirmed by medical history and radiological examination. Patients with DDD may also have spondylolisthesis up to grade 1 at the involved spinal level. 6) Patients who have failed non-surgical treatment for at least 6 months before undergoing surgery. 7) Lateral lumbar interbody fusion (LIF) uses a PEEK or titanium spacer and an adjunctive fixation system (lateral plate or bilateral pedicle screws).

[0175] Example 3: Use of bone void filler for bone voids or gaps that are not essential to the stability of the bone structure.

[0176] Uses of the bone graft compositions disclosed herein include using the bone graft compositions as bone void fillers for bone voids or gaps that are not essential to the stability of bone structures. In particular, synthetic ZB bone graft (a composition identical to synthetic ZF except lacking naloxone) is gently filled into bone voids or gaps in the skeletal system (e.g., the posterolateral spine, pelvis, ilium, and / or extremities).

[0177] Accordingly, the present disclosure provides a method for repairing a bone defect in a patient in need thereof, comprising applying a bone graft composition disclosed herein to the bone defect, wherein the bone defect comprises a void or gap in the bony skeletal system, wherein the void or gap is not essential to the stability of the bony skeletal structure, and the bone graft composition fills the void or gap. In some embodiments, the defects are surgically created bony defects or bony defects resulting from traumatic injury to the bone. The synthetic ZB bone graft resorbs and replaces bone during healing of these defects.

[0178] Example 4: Safety and Efficacy of Synthetic ZF Bone Grafts in Subjects Undergoing Reconstructive Procedures to Stabilize the Humerus After Resection of Metastatic Breast Cancer Tumors

[0179] This study evaluates the safety and efficacy of synthetic ZF bone grafts using intercalary allografts and plate fixation in humeral diaphyseal-epiphyseal reconstruction after resection of malignant tumors, such as breast cancer metastases.

[0180] Study design

[0181] Subjects in the experimental group received a synthetic ZF bone graft containing osteoconductive synthetic bone graft material and the low-molecular-weight naloxone hydrochloride. The synthetic bone graft is composed of bovine-derived type I collagen, hydroxyapatite (HA), beta-tricalcium phosphate, and other calcium salts and several excipients. Subjects in the control group received an allograft fixed with plates and screws, which was filled with polymethyl methacrylate (PMMA) bone cement. Establishing non-inferiority of the experimental group to the control group confirmed the validity of this study.

[0182] Example 5: Safety and Efficacy of Synthetic ZF Bone Grafts in Subjects Undergoing Reconstructive Procedures to Stabilize the Femur, Tibia, or Pelvis After Resection of Metastatic Breast Cancer Tumors

[0183] This study evaluated the safety and effectiveness of synthetic ZF bone grafts for reconstruction of the proximal femur, distal femur, tibia, and pelvis using intramedullary nails, intramedullary locking nails, cephalomedullary nails, plate fixation, or a combination of these orthopedic fixation devices after resection of malignant tumors, such as metastatic breast cancer. After tumor resection, polymethyl methacrylate (PMMA) bone cement was placed into the defect to provide mechanical stability, and the synthetic ZF was placed around the implant / PMMA bone cement construct adjacent to the bone tissue.

[0184] Study design

[0185] After tumor removal, subjects in the experimental group will receive a suitable fixation device using PMMA bone cement and a synthetic ZF bone graft containing osteoconductive synthetic bone graft material and low-molecular-weight naloxone hydrochloride. The synthetic bone graft is composed of bovine-derived type I collagen, hydroxyapatite (HA), beta-tricalcium phosphate, other calcium salts, and some excipients. Subjects in the control group will receive a suitable fixation device and PMMA bone cement after tumor removal. Establishing non-inferiority of the experimental group to the control group confirms the validity of this study.

Claims

1. A bone graft composition comprising a calcium phosphate putty and at least one of the following: a hardening agent, a hardening rate control agent, an acidifying agent, an iron agent excipient, collagen, and a diluent solution.

2. The bone graft composition of claim 1 , wherein the calcium phosphate putty comprises biphasic calcium phosphate particles.

3. The bone graft composition of claim 2 , wherein the biphasic calcium phosphate particles comprise hydroxyapatite and tricalcium phosphate.

4. 4. The bone graft composition of claim 3, wherein the biphasic calcium phosphate particles comprise about 20-60% hydroxyapatite and about 40-80% tricalcium phosphate.

5. The bone graft composition of any one of claims 2 to 4, wherein the biphasic calcium phosphate particles have interconnected macropores and micropores.

6. The bone graft composition of any one of claims 2 to 4, wherein the biphasic calcium phosphate particles are in the form of spherical particles, fibers, or irregular granules.

7. The bone graft composition of any of claims 1 to 6, further comprising a bioabsorbable polymer.

8. The bone graft composition of any of claims 1 to 7, wherein the bone graft composition has a density between 1.68 g / mL and 1.85 g / mL.

9. The bone graft composition of any of claims 1 to 8, wherein the bone graft composition has a compressive resistance of less than about 80 N / m.

10. The bone graft composition of any of claims 1 to 9, wherein the force required to eject the bone graft composition is less than 80 N / m.

11. The bone graft composition of any of claims 1 to 10, further comprising an active agent.

12. The bone graft composition of claim 11 , wherein the bone graft composition is constructed and arranged to deliver an active agent or other drug to a desired site in a patient.

13. The bone graft composition of claim 12 , wherein the bone graft composition comprises a network of reservoirs and microchannels for storing and delivering an active agent.

14. 14. The bone graft composition of claim 13, wherein the bone graft composition comprises a plurality of reservoirs, microtubes and / or nanotubes for storing and delivering active agents.

15. The bone graft composition of claim 13 , wherein the active agent is delivered at a controlled rate.

16. The bone graft composition of claim 13 , wherein the active agent is dissolved in a pharmaceutically suitable carrier.

17. 14. The bone graft composition of claim 13, wherein the active agent dissolved in a pharmaceutically suitable carrier is sprayed or coated onto the bone graft.

18. The bone graft composition of any of claims 11 to 17, wherein the active agent is an opioid growth factor receptor (OGFR) antagonist.

19. 19. The bone graft composition of claim 18, wherein the OGFR antagonist is selected from the group consisting of naloxone, naltrexone, and salts thereof.

20. 20. The bone graft composition of claim 18, wherein the OGFR antagonist is administered with a diluent.

21. 21. The bone graft composition of any of claims 1 to 20, comprising a hardening agent, wherein the hardening agent comprises sodium carbonate (NaCO3).

22. 22. The bone graft composition of any of claims 1 to 21, wherein the bone graft composition comprises the hardening agent, the hardening agent comprising one or more compounds selected from the group consisting of calcium sulfate hemihydrate, calcium sulfate dihydrate, calcium sulfate anhydrous, monocalcium phosphate, dicalcium phosphate, tricalcium phosphate, tetracalcium phosphate, octacalcium phosphate, hydroxyapatite, calcium carbonate, magnesium carbonate, strontium carbonate, and sodium hydrogen phosphate.

23. 23. The bone graft composition of claim 21 or 22, wherein the weight ratio of the hardening agent to the calcium phosphate salt compound is 3:

4.

24. 24. The bone graft composition of any of claims 1 to 23, wherein the bone graft composition comprises an acidifying agent, the acidifying agent comprising one or more from the group of ascorbic acid, magnesium citrate, potassium citrate, sodium citrate, citric acid monohydrate, and acetic acid.

25. 25. The bone graft composition of claim 24, wherein the acidifying agent improves the bioavailability of the bone graft composition.

26. 26. The bone graft composition of claim 24 or 25, wherein the acidifying agent promotes bone formation.

27. The bone graft composition of any of claims 24 to 26, wherein the acidifying agent increases adhesion.

28. The bone graft composition of any one of claims 1 to 27, wherein the bone graft composition comprises a color developing agent.

29. 29. The bone graft composition of any of claims 1-28, comprising iron (III) sulfate hydrate, iron (III) chloride, iron (III) citrate, iron (III) oxide, iron (III) hydroxide oxide, iron (III) phosphate, ammonium iron (III) citrate, elemental iron granules, iron fluoride, iron (II) sulfate heptahydrate, ammonium iron (II) sulfate hexahydrate, iron (II) chloride, iron disulfide, iron (II) sulfate heptahydrate, iron (II) lactate hydrate, iron (II) L-ascorbate, or iron (II) fluoride.

30. 30. The bone graft composition of claims 28 and 29, wherein the color former comprises an activatable color former.

31. 30. The bone graft composition of claim 28 or 29, wherein the coloring agent colors the bone graft composition red, blue, orange, green, neon green, purple, black, brown, gray, or battleship gray.

32. The bone graft composition of claims 1-31, wherein the bone graft composition comprises an antimicrobial agent.

33. 33. The bone graft composition of claim 32, wherein the antimicrobial agent comprises one or more from the group of antibiotic agents or antifungal agents.

34. 34. The bone graft composition of claim 33, wherein the antibiotic comprises vancomycin, gentamicin, tobramycin, kanamycin, neomycin, ampicillin, methicillin, nafcillin, oxacillin, penicillin, ticarcillin, ciprofloxacin, vancomycin, cefazolin, cefepime, ceftriaxone, clindamycin, aztreonam, imipenem, quinupristin / dalfopristin, chloramphenicol, doxycycline, metronidazole, nitrofurantoin, polymyoxin B, tetracycline, viomycin, chloromycetin, streptomycin, azactam, pharmaceutically acceptable salts thereof, and combinations thereof.

35. 34. The bone graft composition of claim 33, wherein the antifungal agent comprises one or more from the group of polyene antifungals, imidazoles, triazoles, allylamines, and echinocandins.

36. The bone graft composition of any of claims 32 to 35, wherein the bone graft composition exhibits antibacterial and / or antifungal efficacy.

37. The bone graft composition of any of claims 1 to 26, wherein the bone graft composition comprises powdered collagen or liquid collagen.

38. The bone graft composition of any of claims 1 to 37, wherein the bone graft composition is not a dental resin.

39. A method of repairing a bone defect in a patient in need thereof, comprising applying the bone graft composition of any of claims 1 to 37 to the bone defect.

40. 40. The method of claim 39, wherein the bone defect is caused by a human disease or condition, and the bone graft composition comprises an active agent for treating the human disease or condition.

41. 40. The method of claim 39, wherein the bone defect is caused by a human disease or condition and the bone graft composition does not contain an active agent for treating the human disease or condition.

42. 42. The method of claim 40 or 41, wherein the human disease or condition is cancer.

43. 42. The method of claim 40 or 41, wherein the human disease or condition is one of several disorders of the vertebrae of the spinal column that require surgical procedures to fuse the vertebrae.

44. 42. The method of claim 40 or 41, wherein the human disease or condition is one of several disorders of the bones of the limbs, which require surgical intervention to fuse the bones or repair the defect.

45. 42. The method of claim 40 or 41, wherein the human disease or condition is a bone void or defect that is not essential to the stability of bone structure, the bone void or defect being caused by surgery or trauma, and the bone graft composition fills the bone void or defect.

46. 42. The method of claim 40 or 41, wherein the bone graft composition is osteoconductive.

47. 42. The method of claim 40 or 41, wherein the bone graft composition is osteoinductive.

48. 42. The method of claim 40 or 41, wherein the bone graft composition is bone promoting.

49. 44. The method of claim 43, wherein the surgical procedure comprises a lumbar interbody fusion procedure (LIF).

50. 50. The method of claim 49, wherein the LIF procedure includes anterior LIF, lateral LIF, transforaminal LIF, and posterior LIF.

51. 50. The method of claim 49, wherein the LIF treatment alleviates associated symptoms, such as degenerative disc disease in a subject determined via radiography, resulting from the formation of bone defects via surgical procedures.

52. 50. The method of claim 49, wherein the LIF treatment alleviates the subject's associated neuromuscular or physical deficits as determined by the subject's Oswestry Disability Index (ODI), the subject's neurological assessment, or the subject's radiography.

53. 40. The method of claim 39, wherein the bone defect comprises a void or gap in the subject's bony skeletal system, the void or gap not being essential to the stability of the bony skeletal structure, and the bone graft composition is filled into the void or gap.

54. 40. The method of claim 39, wherein the bone defect is caused by a malignant tumor or resection of a malignant tumor.

55. 54. The method of claim 53, wherein the malignant tumor is a breast cancer metastasis tumor.

56. 40. The bone graft composition of any of claims 1-39, wherein the calcium phosphate putty comprises biphasic calcium phosphate particles, the biphasic calcium phosphate particles comprising hydroxyapatite and tricalcium phosphate, and the bone graft composition comprises collagen and naloxone.

57. The bone graft composition of claim 1 , wherein the collagen has a particle size of 25 μm to 750 μm.

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