Incorporation of plant virus particles and polymers as 2D and 3D scaffolds to manipulate cellular behaviors
Methods are generally disclosed for attaching a cell binding motif to a carboxy end of a coat protein of a Tobacco Mosaic Virus particle to form a modified-TMV particle; and attaching a cell to the cell binding motif of the modified-TMV particle. Methods are also disclosed for incorporated virus particles, e.g., TMV virus particles into hydrogels.
1. A method of forming a hydrogel, the method comprising:
forming modified Tobacco Mosaic Virus particles, the modification comprising including a cell binding motif on a carboxy end of a coat protein of the Tobacco Mosaic Virus particles;
incorporating a plurality of the modified Tobacco Mosaic Virus particles into a hydrogel precursor formulation; and
polymerizing the hydrogel precursor formulation to form a hydrogel matrix with the modified Tobacco Mosaic Virus particles incorporated therein.
2. The method of claim 1 , wherein the hydrogel precursor formulation comprises a mixture of alginate in bicarbonate solution and a mild acid.
3. The method of claim 1 , further comprising crosslinking the hydrogel matrix.
4. The method of claim 1 , wherein the cell binding motif comprises an RGD tripeptide sequence derived from fibronectin.
5. The method of claim 1 , wherein the cell binding motif comprises an RGD tripeptide sequence derived from vitronectin.
6. The method of claim 1 , wherein the cell binding motif comprises an RGD tripeptide sequence derived from osteocalcin.
7. The method of claim 1 , wherein the cell binding motif comprises a PHSRN (SEQ ID NO: 55) sequence derived from fibronectin.
8. The method of claim 1 , wherein the cell binding motif comprises an DGEA (SEQ ID NO: 4) peptide sequence derived from collagen.
9. The method of claim 1 , wherein the cell binding motif comprises a P15 peptide sequence derived from collagen.
10. The method of claim 1 , wherein the cell binding motif is selected from the group consisting of GRGDSPG (SEQ ID NO: 1), AVTGRGDSPASS (SEQ ID NO: 2), EDRVPHSRNSIT (SEQ ID NO: 3), DGEA (SEQ ID NO: 4), and GTPGPQGIAGQRGVV (SEQ ID NO: 5).
11. The method of claim 1 , further comprising maintaining the hydrogel in a serum-free environment supplemented with growth factors.
12. The method of claim 11 , wherein the growth factors comprise a fibroblast growth factor, a transforming growth factor-beta 1, or an epidermal growth factor.
13. The method of claim 1 , further comprising seeding the hydrogel with cells.
14. The method of claim 13 , wherein upon the seeding, the cells bind to the virus particles.
15. The method of claim 1 , wherein the method is free of covalent modification of the hydrogel.
16. A method of forming a hydrogel, the method comprising:
incorporating a plurality of modified Tobacco Mosaic Virus particles into a hydrogel precursor formulation;
polymerizing the hydrogel precursor formulation to form a hydrogel matrix with the modified Tobacco Mosaic Virus particles incorporated therein; and
maintaining the hydrogel incorporating the modified Tobacco Mosaic Virus particles therein in a serum-free environment supplemented with growth factors.
17. The method of claim 16 , wherein the growth factors comprise a fibroblast growth factor, a transforming growth factor-beta1, or an epidermal growth factor.
18. The method of claim 16 , wherein the modified Tobacco Mosaic Virus particles comprise a cell binding motif on a carboxy end of a coat protein of the modified Tobacco Mosaic Virus particles.
19. A method of forming a hydrogel, the method comprising:
incorporating a plurality of modified Tobacco Mosaic Virus particles into a hydrogel precursor formulation;
polymerizing the hydrogel precursor formulation to form a hydrogel matrix with the modified Tobacco Mosaic Virus particles incorporated therein; and
seeding the hydrogel incorporating the modified Tobacco Mosaic Virus particles therein with cells.
20. The method of claim 19 , wherein upon the seeding, the cells bind to the modified Tobacco Mosaic Virus particles.
21. The method of claim 19 , wherein the modified Tobacco Mosaic Virus particles comprise a cell binding motif on a carboxy end of a coat protein of the Tobacco Mosaic Virus particles.