Single step polymerization of covalently bound multilayer matrices
A method is provided for forming complex tissue using density gradient multilayer polymerization (DGMP) to form strong hydrogels with smooth transitions between layers. The multicompartment hydrogel is formed by co-dissolving a polymer precurser with a constituent in multiple solvent fractions to a create prepolymer solutions with different densities, layering the prepolymer solutions on top of each other from high to low solvent density, and irradiating the prepolymer solutions to form a polymer. The hydrogels may be used as biomimetic matrices.
1. A method for preparing a multicompartment hydrogel comprising:
a. co-dissolving a polymer precursor with an inert density modifier in multiple solvent fractions to a create prepolymer solutions with different densities;
b. layering the prepolymer solutions on top of each other in an order proceeding from high to low solvent density; and
c. irradiating the prepolymer solutions to form a polymer.
2. The method of claim 1 , wherein the polymer precursor comprises acrylamide and bisacrylamide.
3. The method of claim 1 , wherein the polymer precursor comprises polyethylene glycol diacrylate (PEGda).
4. The method of claim 1 , wherein co-dissolving further comprises adding an acrylated fluorophore or a conjugated RGD-containing adhesion peptide to the prepolymer solutions.
5. The method of claim 1 , wherein one or more of the layers of the prepolymer solutions are irradiated at different times.
6. The method of claim 1 , wherein the inert density modifier comprises sucrose, iodixanol, or a mixture thereof.
7. The method of claim 1 , wherein the inert density modifier is present in an amount of between about 0% to about 15% or between about 10% to about 50%.
8. The method of claim 1 , further comprising washing out the inert density modifier after the polymer is formed.
9. The method of claim 1 , wherein the multicompartment hydrogel is configured for culturing tissue in a three-dimensional arrangement.
10. The method of claim 1 , wherein co-dissolving comprises serially diluting the polymer precursor with the inert density modifier to progressively create the prepolymer solutions in order of decreasing densities.
11. A method for creating a multicompartment polymer matrix, comprising:
creating a plurality of prepolymer solutions of decreasing densities by serially co-dissolving an inert density modifier with a prepolymer;
progressively layering the plurality of prepolymer solutions in a mold in order of decreasing density, wherein density differences between adjacent layers induce phase separations between layers; and
exposing the layered prepolymer solutions to a polymerizing radiation source to form a polymer.
12. The method of claim 11 , wherein the step of creating further comprises co-dissolving one or more of cells, ligands, proteins and peptides.
13. The method of claim 11 , wherein the prepolymer comprises acrylamide and bisacrylamide.
14. The method of claim 11 , wherein the prepolymer comprises polyethylene glycol diacrylate (PEGda).
15. The method of claim 11 , wherein co-dissolving further comprises adding an acrylated fluorophore or a conjugated RGD-containing adhesion peptide.
16. The method of claim 11 , further comprising washing the polymer to remove the inert density modifier.
17. The method of claim 11 , wherein the inert density modifier comprises sucrose, iodixanol, or a mixture thereof.
18. A method for creating a multicompartment polymer matrix, comprising:
creating a plurality of prepolymer solutions of decreasing densities by serially co-dissolving an inert density modifier with a prepolymer;
layering the plurality of prepolymer solutions in a mold in layers progressing from higher to lower density solutions, wherein density differences between adjacent layers induce phase separations between layers; and
exposing the layered prepolymer solutions to a polymerizing radiation source to form a polymer.
19. The method of claim 18 , wherein the layering is repeated to produce layered combinations of higher to lower density layers.
20. The method of claim 18 , wherein the step of creating further comprises co-dissolving one or more of cells, ligands, proteins and peptides with the prepolymer and the inert density modifier.