Biomolecular patterning of three dimensional tissue scaffolds
The invention provides methods and compositions for making and using collagen-glycosaminoglycan three-dimensional scaffolds immobilized with biomolecules that are spatially and temporally patterned. The method comprises adding benzophenone to a collagen-glycosaminoglycan three dimensional scaffold in the dark; adding one or more biomolecules to one or more areas of the collagen-glycosaminoglycan three-dimensional scaffold (which can be done optionally in the dark or in the light); and exposing the collagen-2glycosaminoglycan three-dimensional scaffold to light at a wavelength of about 350 to about 365 nm.
1. A method of immobilizing one or more types of biomolecules onto a collagen-glycosaminoglycan three-dimensional scaffold comprising:
(a) adding benzophenone to a collagen-glycosaminoglycan three-dimensional scaffold in the dark;
(b) adding one or more biomolecules to one or more areas of the collagen-glycosaminoglycan three-dimensional scaffold in the dark; and
(c) exposing the collagen-glycosaminoglycan three-dimensional scaffold to light at a wavelength of about 350 to about 365 nm.
2. The method of claim 1 , wherein one or more portions of the collagen-glycosaminoglycan three-dimensional scaffold are exposed to the light while one or more other portions of the scaffold remain in the dark.
3. The method of claim 2 , wherein the one or more types of biomolecules are immobilized onto the scaffold at two or more different depths into the scaffold.
4. The method of claim 1 , wherein two or more types of biomolecules are immobilized to the scaffold, and wherein the two or more types of biomolecules are immobilized to two or more areas of the scaffold, or to two or more depths in the scaffold, or both.
5. The method of claim 1 , wherein a first mask is placed over the collagen-glycosaminoglycan three-dimensional scaffold prior to exposing the collagen-glycosaminoglycan three-dimensional scaffold to the light.
6. The method of claim 5 , further comprising:
(d) placing the collagen-glycosaminoglycan three-dimensional scaffold in the dark and removing the first mask from the collagen-glycosaminoglycan three-dimensional scaffold;
(e) adding one or more biomolecules to one or more areas the collagen-glycosaminoglycan three-dimensional scaffold in the dark;
(f) placing a second mask over the collagen-glycosaminoglycan three-dimensional scaffold in the dark;
(g) exposing the collagen-glycosaminoglycan three-dimensional scaffold to light at a wavelength of about 350 to about 365 nm; and optionally repeating steps (d) to (g) with additional masks and biomolecules.
7. The method of claim 1 , wherein the one or more of the biomolecules are immobilized to the scaffold via a cleavable linker.
8. The method of claim 7 , wherein the cleavable linker is cleavable by an exogenous cue.
9. The method of claim 1 , wherein the one or more biomolecules are peptides, proteins, carbohydrates, cytokines, hormones, nucleic acid molecules, integrins, antibodies, specific binding fragments of antibodies, or a biological sample.
10. The method of claim 7 , wherein the cleavable linker is cleavable by an endogenous cue.
11. The method of claim 1 , further comprising (d) seeding the collagen-glycosaminoglycan three-dimensional scaffold with cells.