STEM CELL IMPREGNATED CORTICAL FIBERS

1 . A method for making a bone augmentation composition, comprising:

hydrating an allograft comprising cortical fibers with a cell culture media;

seeding the hydrated allograft with human mesenchymal stem cell solution derived from a source other than the patient being treated.

culturing the stem-cell seeded allograft for a predetermined time;

freezing the stem-cell seeded allograft containing the cell culture media with a cryopreservation solution; and

after cryopreservation. thawing the stem-cell seeded allograft containing the cell culture media to provide the bone augmentation composition for application to a bone treatment site,

wherein a surviving stem cell population in the bone augmentation composition after thawing is between 50% and 90% of an initial stem cell population in the bone augmentation composition prior to freezing.

2 . The method of claim 1 , wherein the surviving stem cell population in the bone augmentation composition is between 1 and 5×10 5 cells/ml.

3 . The method of claim 1 , wherein the culturing the stem-cell seeded allograft for a limited time occurs for a period of about 0.5 to 5 hrs at a temperature of about 20 to 50° C.

4 . The method of claim 1 , wherein the seeding comprises shaking the hydrated allograft with the stem solution.

5 . The method of claim 1 , wherein the stem cell solution applied during the seeding has a stem cell density of about 1×10 5 to 1×10 6 cells/ml as counted by an Alamar Blue Assay.

6 . The method of claim 1 , wherein the freezing comprises storing the stem-cell seeded allograft containing the cell culture media at about −80° C.

7 . The method of claim 1 , wherein the cell culture media comprises a stem cell nourishment solution.

8 . The method of claim 1 , wherein the seeding comprises shaking or rotating the hydrated allograft while applying the stem cell solution.

9 . The method of claim 8 , wherein the shaking agitates the hydrated allograft at about 2 to 500 rpm.

10 . The method of claim 1 , wherein the hydrated allograft containing the stem cell solution is frozen with dimethyl sulfoxide DMSO having a concentration of less than about 10 weight percent.

11 . The method of claim 1 , wherein the cortical fibers comprise a loose, intertangled mixture of the cortical fibers.

12 . A bone augmentation composition, comprising:

i) human mesenchymal stem cells derived from a source other than the patient being treated,

ii) a cell culture media. and

ii) an allograft comprising cortical fibers

wherein the allograft comprising the stem cells and the cell culture media, after freezing and thawing, has a surviving stem cell population in the bone augmentation composition between 1 and 5×10 5 cells/ml, as counted by an Alamar Blue Assay.

13 . The composition of claim 12 , wherein the allograft comprising the stem cells and the cell culture media comprises a frozen composition stored in a container while awaiting application of the bone augmentation composition to a bone treatment site.

14 . The composition of claim 13 , wherein the frozen composition comprises dimethyl sulfoxide DMSO as a cryo-preservative added to the composition prior to freezing.

15 . The composition of claim 12 , wherein the allograft comprises demineralized cortical fibers.

16 . The composition of claim 12 , wherein the allograft comprises a loose intertangled collection of the cortical fibers.

17 . The composition of claim 12 , wherein the allograft comprising the stem cells and the cell culture media comprises a moldable product.

18 . The composition of claim 12 , wherein the allograft comprising the stem cells and the cell culture media is prepared using any of the methods of claims 1 - 11 .

19 . The composition of claim 12 , wherein a surviving stem cell population in the bone augmentation composition after thawing is between 50% and 90% of an initial stem cell population in the bone augmentation composition prior to freezing.

20 . A bone augmentation composition, comprising:

i) stem cells,

ii) a cell culture media. and

ii) an allograft comprising bone fibers

wherein the allograft comprising the stem cells and the cell culture media, after freezing and thawing, has a surviving stem cell population in the bone augmentation composition between 1 and 5×10 5 cells/ml, as counted by an Alamar Blue Assay.