Enzyme-activated collagen and tissue matrices
Devices and methods for treating defects in connective tissue are provided along with methods for making such devices. The devices can include enzyme-activated acellular tissue matrices that facilitate regrowth of the damaged tissue.
1. A method of treatment comprising:
selecting an anatomical site for treatment;
selecting a collagen-based acellular tissue matrix, wherein the matrix has been contacted with a composition containing at least one protease, wherein the protease consists essentially of at least one of MMP-1, MMP-3, or MMP-9, wherein the protease can cleave one or more matrikines from the tissue matrix, and wherein the composition further contains a deactivating agent comprising one or more chelators for reducing the activity of the at least one protease by binding metal ions needed for activity of the at least one protease; and
implanting the collagen-based acellular tissue matrix into the anatomic site.
2. The method of claim 1 , wherein the acellular tissue matrix comprises an acellular dermal matrix.
3. The method of claim 2 , wherein the acellular tissue matrix comprises an acellular porcine dermal matrix.
4. The method of claim 1 , wherein the protease is a matrix metalloproteinase (MMP).
5. The method of claim 1 , wherein the at least one protease is in a solution.
6. The method of claim 1 , wherein the composition further contains at least one of an elastase or a dispase.
7. The method of claim 1 , wherein the chelating agent comprises ethylenediaminetetraacetic acid (EDTA), ethylene glycol tetraacetic acid (EGTA), or orthophenanthroline.
8. The method of claim 1 , wherein the chelating agent regulates the degradation of the tissue matrix by modulating protease activity.
9. The method of claim 1 , further comprising exposing the acellular tissue matrix to a disinfectant.
10. The method of claim 9 , wherein the disinfectant is isopropyl alcohol.
11. The method of claim 1 , further comprising sterilizing the acellular tissue matrix.
12. The method of claim 11 , wherein sterilizing the acellular tissue matrix comprises application of ethylene oxide, propylene, gamma irradiation, or e-beam irradiation.
13. The method of claim 1 , further comprising lyophilizing the acellular tissue matrix.
14. The method of claim 1 , further comprising storing the acellular tissue matrix in a hydrated state prior to implanting the acellular tissue matrix into the anatomical site.
15. The method of claim 1 , wherein the composition further contains a bioactive substance.
16. The method of claim 15 , wherein the bioactive substance comprises an antibacterial agent, a cytokine, a growth factor, non-collagenous tissue, or cells.
17. The method of claim 1 , wherein the activity of the protease is restored automatically upon implantation into a subject.
18. The method of claim 17 , wherein the restoration of protease activity results in the release of matrikines from the tissue matrix.
19. The method of claim 1 , wherein the matrikines comprise GHK (SEQ ID NO: 1), CNYYSNS (SEQ ID NO: 2), VGPVG (SEQ ID NO: 3), VGVAPG (SEQ ID NO: 4), KKGHK (SEQ ID NO: 5), or DGGRYY (SEQ ID NO: 6).
20. The method of claim 1 , wherein the chelator is present in the composition at a concentration between 4 mM and 20 mM.