In vitro production of bone-like material
View Patent ↗Embodiments of the invention is directed to methods of forming a bone-like material, by placing a matrix into a culture container, adding cells to the matrix, placing the tissue-cell construct into the culturing environment and incubating the construct under appropriate conditions for the time required to produce an amount of bone-like material of sufficient volume to be harvested for further processing. Other embodiments of the invention are directed to the use of the bone-like material formed by methods of the invention in implantation procedures.
1. A method of forming a bone-like material, comprising:
placing a matrix comprising calcium phosphate into a culture container;
adding mesenchymal stem cells to the matrix;
placing the matrix-cell construct into a culturing environment and incubating the construct under appropriate conditions for a time required to produce an amount of bone-like material of sufficient volume to be harvested for further processing.
2. The method of claim 1 , further comprising de-cellularizing the bone-like material using freeze thaw cycling or washing with detergents and solvents.
3. The method of claim 2 , further comprising forming the de-cellularized bone-like material into an implant.
4. The method of claim 3 , wherein the implant is a spinal fusion implant.
5. The method of claim 1 , wherein the matrix is tri-calcium phosphate, hydroxyapatite, demineralized bone matrix, collagen or polymers.
6. The method of claim 1 , wherein the calcium phosphate comprises allogenic bone fragments.
7. The method of claim 1 , wherein the calcium phosphate comprises one or more of hydroxyapatite, tricalcium phosphate, and carbonate apatite.
8. The method of claim 1 , wherein the matrix further comprises an orthopedic protein that comprises glycosaminoglycans or aggrecan.
9. A composition comprising:
an implant and bone-like material, wherein the bone-like material is bound to the implant and has the ability to bind to a component of an extracellular matrix of a biological tissue, whereby the implant and the biological tissue become attached upon introduction of the composition to biological tissue; and
wherein the bone-like material is formed by placing a matrix comprising calcium phosphate into a culture container, adding mesenchymal stem cells to the matrix, placing the tissue-cell construct into the culturing environment and incubating the construct under appropriate conditions for a time required to produce an amount of bone-like material.
10. The composition of claim 9 , wherein the bone-like material is formed in vitro.
11. The composition of claim 9 , wherein the implant is porous.
12. The composition of claim 9 , wherein the implant allows for migration of the mesenchymal stem cells into the implant.
13. The composition of claim 9 , wherein the implant comprises hydroxyapatite, tricalcium phosphate, fibrin or fibrinogen.
14. The composition of claim 9 , further comprising a growth factor.
15. A method of using the composition of claim 9 for treatment, repair or replacement of musculoskeletal tissue, by implantation of the composition into a subject in need thereof.
16. The method of claim 15 , wherein a subject receiving treatment is a mammal.
17. The method of claim 16 , wherein the mammal is a human.
18. The composition of claim 9 , wherein the calcium phosphate comprises allogenic bone fragments.
19. The composition of claim 9 , wherein the calcium phosphate comprises one or more of hydroxyapatite, tricalcium phosphate, and carbonate apatite.
20. The composition of claim 9 , wherein the matrix further comprises an orthopedic protein that comprises glycosaminoglycans or aggrecan.