Crosslinked compositions comprising collagen and demineralized bone matrix, methods of making and methods of use
A composition comprising a collagen protein and demineralized bone matrix is described wherein the composition is chemically cross-linked with a carbodiimide such as N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC). The crosslinking reaction can be conducted in the presence of N-hydroxysuccinimide (NHS). The collagen can be in a porous matrix or scaffolding. The DBM can be in the form of particles dispersed in the collagen. A method of making the composition is also described wherein a collagen slurry is cast into the desired shape, freeze dried to form a porous scaffolding and infitrated with a solution comprising the cross-linking agent. The composition can be used as an implant for tissue (e.g., soft tissue or bone) engineering.
1 . A composition comprising:
demineralized bone matrix (DBM); and
a collagen protein;
wherein the composition is cross-linked.
2 . The composition of claim 1 , wherein the composition is chemically crosslinked with a carbodiimide crosslinking agent.
3 . The composition of claim 2 , wherein the carbodiimide crosslinking agent is N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC).
4 . The composition of claim 2 , wherein the composition is chemically cross-linked in the presence of N-hydroxysuccinimide (NHS).
5 . The composition of claim 1 , further comprising one or more growth factors.
6 . The composition of claim 1 , wherein the composition comprises from 2 to 95 wt/% DBM.
7 . The composition of claim 1 , wherein the composition comprises from 55 to 85 wt/% DBM.
8 . The composition of claim 1 , wherein the DBM comprises particles of the DBM dispersed in the collagen.
9 . The composition of claim 1 , wherein the collagen protein is in a porous scaffolding.
10 . The composition of claim 9 , wherein the DBM comprises particles of DBM dispersed in the porous scaffolding.
11 . The composition of claim 8 , wherein the DBM particles have an average particle size of up to 5 mm.
12 . The composition of claim 8 , wherein the DBM particles have an average particle size ranging from 53 to 850 μm.
13 . The composition of claim 1 , wherein the composition is chemically crosslinked with a compound selected from the group consisting of gluteraldehyde, formaldehyde, 1,4-butanediol diglycidyl ether, hydroxypyridinium, hydroxylysylpyridinium, and formalin.
14 . The composition of claim 1 , wherein the composition is crosslinked by irradiation.
15 . The composition of claim 1 , wherein the composition is crosslinked by photooxidation.
16 . The composition of claim 1 , wherein the composition is crosslinked via an enzymatic process.
17 . The composition of claim 16 , wherein the collagen protein is crosslinked via the action of tissue transglutaminase.
18 . The composition of claim 16 , wherein the composition is crosslinked with lysyl oxidase.
19 . The composition of claim 1 , wherein the composition is crosslinked by a dehydrothermal treatment.
20 . The composition of claim 1 , wherein the composition is crosslinked under acidic conditions.
21 . The composition of claim 1 , wherein the collagen protein is crosslinked using e-beam irradiation, gamma irradiation, or light.
22 . The composition of claim 21 , wherein the collagen protein is crosslinked using pulsed light.
23 . The composition of claim 1 , further comprising a spacer.
24 . The composition of claim 23 , wherein the spacer is a polyoxyalkyleneamine spacer or a polyethylene glycol spacer.
25 . The composition of claim 1 , wherein the composition further comprises vinyl pyrrolidinone or methyl methacrylate.
26 . The composition of claim 1 , further comprising an additive selected from the group consisting of collagenase inhibitors, growth factors, antibodies, metalloproteinases, cell attachment fragment(s), and combinations thereof.
27 . The composition of claim 26 , wherein the additive is bound to the collagen or DBM.
28 . The composition of claim 26 , wherein the additive is not bound to the collagen or DBM.
29 . The composition of claim 1 , wherein the composition is crosslinked by glycation or glycosylation.
30 . The composition of claim 1 , wherein the crosslinks are pentosidine crosslinks.
31 . The composition of claim 1 , wherein the crosslinks are epsilon(gamma-glutamyl)lysine crosslinks.
32 . A method of making a composition comprising a collagen protein and demineralized bone matrix, the method comprising:
crosslinking the composition.
33 . The method of claim 32 , wherein the composition is chemically crosslinked with a carbodiimide crosslinking agent.
34 . The method of claim 33 , wherein the carbodiimide is N-(3-dimethylaminopropyl)-N-ethylcarbodiimide hydrochloride (EDC).
35 . The method of claim 33 , wherein the composition is chemically cross-linked in the presence of N-hydroxysuccinimide (NHS).
36 . The method of claim 35 , wherein the NHS is present at an EDC/NHS ratio of 1:2 to 2:5.
37 . The method of claim 35 , wherein the NHS is present at an EDC/NHS ratio of 1:2, 2:3 or 2:5.
38 . The method of claim 32 , further comprising dispersing particles of the demineralized bone matrix in a collagen slurry, casting the slurry into the cavity of a mold and freeze drying the cast slurry to form a porous scaffolding comprising the collagen protein and particles of the demineralized bone matrix.
39 . The method of claim 38 , wherein the slurry is an aqueous slurry.
40 . The method of claim 38 , wherein crosslinking comprises:
infiltrating a carbodiimide crosslinking agent into pores of the porous scaffolding; and
allowing the carbodiimide cross-linking agent to react with the collagen protein and/or the DBM to form cross-links.
41 . The composition of claim 32 , wherein the crosslinking results from culturing a non-crosslinked matrix in vivo to allow collagen crosslinking by cellular mechanisms.
42 . A method of treatment comprising implanting a composition comprising demineralized bone matrix (DBM) and a collagen protein into a mammal, wherein the composition is crosslinked.
43 . The method of claim 42 , wherein the composition is chemically crosslinked with a carbodiimide crosslinking agent.
44 . The method of claim 42 , wherein the composition is implanted into the spine of the mammal.
45 . The method of claim 42 , wherein the composition is implanted into an intervertebral space of the mammal.
46 . The method of claim 42 , wherein the composition is implanted into the site of a trauma injury.
47 . The method of claim 42 , wherein the composition is implanted into a craniomaxillofacial cavity.
48 . The method of claim 42 , wherein the mammal is a human.
49 . A composition comprising:
demineralized bone matrix (DBM); and
a collagen protein;
wherein the composition is cross-linked via an amide linkage.
50 . The composition of claim 49 , further comprising one or more growth factors.
51 . The composition of claim 49 , wherein the composition comprises from 2 to 95 wt/% DBM.
52 . The composition of claim 49 , wherein the composition comprises from 55 to 85 wt/% DBM.
53 . The composition of claim 49 , wherein the composition comprises particles of the DBM dispersed in the collagen protein.
54 . The composition of claim 49 , wherein the collagen protein is in a porous scaffolding.
55 . The composition of claim 54 , wherein the composition comprises particles of the DBM dispersed in the porous scaffolding.
56 . The composition of claim 55 , wherein the DBM particles have a particle size of up to 5 mm.
57 . The composition of claim 55 , wherein the DBM particles have a particle size of from 53 to 850 μm.