Fabrication of bone regeneration scaffolds and bone filler material using a perfusion flow system
The present disclosure is directed at a process to form bone grafting material. One may provide a porous collagen scaffold and insert the scaffold into a perfusion chamber of a perfusion flow system. This may then be followed by continuously providing a mineralization perfusion fluid flow through the scaffold at a flow rate to provide dynamic intrafibrillar mineralization of the scaffold and form a collagen/hydroxyapatite composite scaffold. One may optionally provide the scaffold with bone tissue forming cells and then deliver a perfusion fluid including oxygen and one or more nutrients through the collagen/hydroxyapatite composite scaffold and to the bone tissue forming cells at a flow rate such that the bone tissue forming cells remodel the collagen/hydroxyapatite composite scaffold and form a bone tissue extracellular matrix. The bone tissue extracellular matrix may then be decellularized to form an acellular bone repair scaffold.
1. A process to form a bone grafting material, comprising:
providing a porous collagen scaffold;
inserting the collagen scaffold into a perfusion chamber of a perfusion flow system;
continuously providing a mineralization perfusion fluid flow comprising polyaspartic acid through the collagen scaffold at a flow rate to provide dynamic intrafibrillar mineralization of the collagen scaffold;
forming a collagen/hydroxyapatite composite scaffold;
contacting the collagen/hydroxyapatite composite scaffold with mesenchymal stromal cells to form a cell seeded scaffold;
perfusing a fluid including oxygen and one or more nutrients through the cell seeded scaffold;
remodeling the cell seeded scaffold; and
forming a bone tissue extracellular matrix on the cell seeded scaffold comprising fibronectin, type I collagen and biglycan so as to form a remodeled scaffold having 95% porosity comprising pore size of about 250 μm.
2. The process of claim 1 further comprising decellularizing said bone tissue extracellular matrix and forming an acellular bone repair scaffold.
3. The process of claim 2 wherein the acellular bone repair scaffold is placed in a treatment site of a host to be treated.
4. The process of claim 1 wherein said mineralization perfusion fluid comprises polyaspartic acid at a concentration of 15 μM.
5. The process of claim 1 wherein said mineralization perfusion fluid is provided at a continuous flow rate of 20 μm/s to 500 μm/s.
6. The process of claim 1 wherein said mineralization perfusion fluid is provided at a continuous flow for a period of 12 hours to 7 days.
7. The process of claim 1 wherein said perfusion fluid including oxygen and one or more nutrients comprises an osteogenic induction medium.
8. A process to form a bone grafting material, comprising:
providing a porous collagen scaffold;
inserting the collagen scaffold into a perfusion chamber of a perfusion flow system; continuously providing a mineralization perfusion fluid flow comprising polyaspartic acid through the collagen scaffold at a flow rate to provide dynamic intrafibrillar mineralization of the collagen scaffold and forming a collagen/hydroxyapatite composite scaffold; providing the collagen/hydroxyapatite composite scaffold with mesenchymal stromal cells;
delivering a perfusion fluid including oxygen and one or more nutrients through the collagen/hydroxyapatite composite scaffold and to mesenchymal stromal cells at a flow rate;
remodeling the collagen/hydroxyapatite cell seeded composite scaffold;
forming a bone tissue extracellular matrix comprising fibronectin, type I collagen and biglycan so as to form a remodeled scaffold having 95% porosity comprising a pore size of about 250 μm;
removing said mesenchymal stromal cells so as to form an acellular bone repair scaffold;
seeding said acellular bone repair scaffold with autologous bone forming cells of a host to be treated, wherein said host has a treatment site; and
implanting said acellular bone repair scaffold with said autologous bone forming cells into said treatment site of said host.
9. The process of claim 8 wherein said mineralization perfusion fluid is provided at a continuous flow rate of 1 μm/s to 10 mm/s.
10. The process of claim 8 wherein said mineralization perfusion fluid is provided at a continuous flow rate of 20 μm/s to 500 μm/s.
11. The process of claim 8 wherein said mineralization perfusion fluid is provided at a continuous flow for a period of 12 hours to 7 days.
12. The process of claim 8 wherein said perfusion fluid including oxygen and one or more nutrients comprises an osteogenic induction medium.
13. The process of claim 8 wherein the bone grafting material has trabecular bone-like structure.