Peptides for supporting endothelial progenitor cell rolling and capture and endothelialization of biomaterials
The present invention relates to the production of endothelialized matrices and materials from immature endothelial cells using substrates to which particular peptides have been grafted. The resultant substrates can be used to capture and support immature endothelial cells. Further, the methods and compositions of the present invention provide viable cell delivery platforms that allow for production and provision of endothelialized medical devices and implants, including vascular grafts, stents, shunts, and valves, endothelialized surfaces and channels for in vitro testing devices, including microfluidic chips, and materials that support vascularization such as for use in engineered tissues. The present invention includes novel methods required for the successful production of cellularized substrates, systems and components used for the same, and methods of using the resultant cell and tissue compositions.
1. A composition for inducing endothelialization or vascularization, comprising:
a substrate;
at least one first peptide consisting of SEQ ID NO: 1 operatively linked to the surface of said substrate; and
at least one second peptide consisting of SEQ ID NO: 2 operatively linked to the surface of said substrate.
2. The composition of claim 1 , further comprising at least one additional peptide selected from the group consisting of SEQ ID NOs: 8, 11 and 18 and combinations thereof.
3. The composition of claim 1 , wherein said substrate is selected from the group consisting of biomimetic material, a polymer, hydrogel, material suitable for use in a medical device or implant and decellularized extracellular matrix (dECM).
4. The composition of claim 1 , wherein said substrate comprises a hydrogel.
5. The composition of claim 1 , wherein said substrate comprises biomimetic material.
6. The composition of claim 5 , wherein the substrate comprises decellularized extracellular matrix (dECM).
7. The composition of claim 1 , wherein the molar ratio of the second peptide to the first peptide is between 3:1 and 1:3.
8. A method for capturing endothelial cells or supporting immature endothelial cells under physiological shear, the method comprising:
providing a substrate that is operably linked to at least one first peptide consisting of SEQ ID NO: 1 and at least one second peptide consisting of SEQ ID NO: 2; and
exposing said substrate to a source of endothelial cells.
9. The method of claim 8 , wherein said endothelial cells are immature endothelial cells, endothelial progenitor cells, or pluripotent stem cell-derived endothelial cells.
10. The method of claim 8 , wherein said substrate is further operably linked to at least one additional peptide selected from the group consisting of SEQ ID NOs: 8, 11 and 18 and combinations thereof.
11. The method of claim 8 , further comprising subjecting the endothelial cells to physiologic shear in the presence of said substrate.
12. A method for producing an endothelial cell capture and support surface, the method comprising:
providing a substrate;
operably linking at least one first adhesion ligand peptide and at least one second adhesion ligand peptide to the surface of said substrate to produce a prepared surface, wherein said at least one first adhesion ligand peptide consists of SEQ ID NO: 1 and said at least one second adhesion ligand peptide consists of SEQ ID NO: 2.
13. The method of claim 12 , further comprising operably linking said surface of said substrate to at least one additional adhesion ligand selected from the group consisting of SEQ ID NOs: 8, 11 and 18 and combinations thereof.
14. The method of claim 12 , wherein said substrate comprises a material suitable for use in a medical device or implant.
15. The method of claim 12 , wherein the endothelial cell capture and support surface comprises or is incorporated into an in vitro testing device.
16. The method of claim 15 , wherein said in vitro testing device is a microfluidic chip.
17. The method of claim 12 , wherein said substrate is selected from the group consisting of biomimetic material, a polymer, hydrogel, material suitable for use in a medical device or implant and decellularized extracellular matrix (dECM).
18. An endothelial cell capture and support surface produced by the method of claim 12 .
19. A medical device or implant comprising the composition of claim 1 .
20. The medical device or implant of claim 19 , wherein said medical device or implant is a vascular graft, stent, shunt, or valve.