There is significant interest in the development of injectable carriers for cell transplantation to engineer bony tissues. In this study, we hypothesized that adhesion ligands covalently coupled to hydrogel carriers would allow one to control pre-osteoblast cell attachment, proliferation, and differentiation. Modification of alginate with an RGDcontaining peptide promoted osteoblast adhesion and spreading, whereas minimal cell adhesion was observed on unmodified hydrogels. Raising the adhesion ligand density increased osteoblast proliferation, and a minimum ligand density (1.5-15 femtomoles/cm2) was needed to elicit this effect. MC3T3-E1 cells demonstrated increased osteoblast differentiation with the peptide-modified hydrogels, as confirmed by the up-regulation of bone-specific differentiation markers. Further, transplantation of primary rat calvarial osteoblasts revealed statistically significant increases of in vivo bone formation at 16 and 24 weeks with G4RGDY-modified alginate compared with unmodified alginate. These findings demonstrate that biomaterials may be designed to control bone development from transplanted cells.
Alsberg E., Hill EE, Mooney DJ (2001). Craniofacial tissue engineering. Crit Rev Oral Biol Med12:64-75.
2.
Anselme K. (2000). Osteoblast adhesion on biomaterials. Biomaterials21:667-681.
3.
Dee KC, Rueger DC, Andersen TT, Bizios R. (1996). Conditions which promote mineralization at the bone-implant interface: a model in vitro study. Biomaterials17:209-215.
4.
Franceschi RT, Iyer BS ( 1992). Relationship between collagen synthesis and expression of the osteoblast phenotype in MC3T3-E cells. J Bone Miner Res7:235-246.
5.
Gronowicz GA, Derome ME (1994). Synthetic peptide containing Arg-Gly-Asp inhibits bone formation and resorption in a mineralizing organ culture system of fetal rat parietal bones. JBone Miner Res9:193-201.
6.
Gronowicz GA, McCarthy MB (1995). Glucocorticoids inhibit the attachment of osteoblasts to bone extracellular matrix proteins and decrease beta 1-integrin levels . Endocrinology136:598-608.
7.
Grzesik WJ, Robey PG (1994). Bone matrix RGD glycoproteins: immunolocalization and interaction with human primary osteoblastic bone cells in vitro. J Bone Miner Res9:487-496.
8.
Guo JF, Jourdian GW, MacCallum DK (1989). Culture and growth characteristics of chondrocytes encapsulated in alginate beads. Connect Tissue Res19:277-297.
9.
Hem DL, Hubbell JA (1998). Incorporation of adhesion peptides into nonadhesive hydrogels useful for tissue resurfacing. J Biomed Mater Res39:266-276.
10.
Hubbell JA, Massia SP, Drumheller PD (1992). Surface-grafted cell-binding peptides in tissue engineering of the vascular graft. Ann NY Acad Sci665:253-258.
11.
Jikko A., Harris SE, Chen D., Mendrick DL, Damsky CH (1999). Collagen integrin receptors regulate early osteoblast differentiation induced by BMP-2. J Bone Miner Res14:1075-1083.
12.
Kim WS, Vacanti CA, Upton J., Vacanti JP (1994). Bone defect repair with tissue-engineered cartilage . Plast Reconstr Surg94:580-584.
13.
Krebsbach PH, Mankani MH, Satomura K., Kuznetsov SA, Robey PG (1998). Repair of craniotomy defects using bone marrow stromal cells. Transplantation66:1272-1278.
14.
Maheshwari G., Brown G., Lauffenburger DA, Wells A., Griffith LG (2000). Cell adhesion and motility depend on nanoscale RGD clustering. J Cell Sci 113(Pt10): 1677-1686.
15.
Mann BK, Tsai AT, Scott-Burden T., West JL ( 1999). Modification of surfaces with cell adhesion peptides alters extracellular matrix deposition. Biomaterials20:2281-2286.
16.
Manolagas SC, Burton DW, Deftos LJ (1981). 1,25-Dihydroxyvitamin D3 stimulates the alkaline phosphatase activity of osteoblast-like cells. J Biol Chem256:7115-7117.
17.
Martins-Green M. (2000). Dynamics of cell-ECM interaction. In: Principles of tissue engineering. Lanza RP, Langer R, Vacanti J, editors. San Diego, CA: Academic Press, pp. 33-55.
18.
Massia SP, Hubbell JA (1991). An RGD spacing of 440 nm is sufficient for integrin alpha V beta 3-mediated fibroblast spreading and 140 nm for focal contact and stress fiber formation. J Cell Biol114:1089-1100.
19.
Ohgushi H., Goldberg VM, Caplan AI (1989). Heterotopic osteogenesis in porous ceramics induced by marrow cells. J Orthop Res7:568-578.
20.
Pockwinse SM, Wilming LG, Conlon DM, Stein GS, Lian JB ( 1992). Expression of cell growth and bone specific genes at single cell resolution during development of bone tissue-like organization in primary osteoblast cultures. J Cell Biochem49:310-323.
21.
Rezania A., Johnson R., Lefkow AR, Healy KE (1999). Bioactivation of metal oxide surfaces. 1. Surface characterization and cell response. Langmuir15:6931-6939.
22.
Rowley JA, Madlambayan G., Mooney DJ (1999). Alginate hydrogels as synthetic extracellular matrix materials. Biomaterials20:45-53.
23.
Schneider WC (1957). Determination of nucleic acids in tissues by pentose analysis. Meth Enzymol3:680-684.
24.
Smetana K. Jr (1993). Cell biology of hydrogels. Biomaterials14 :1046-1050.
25.
Stile RA, Healy KE (2001). Thermo-responsive peptide-modified hydrogels for tissue regeneration. Biomacromolecules2:185-194.
26.
Wang D., Christensen K., Chawla K., Xiao G., Krebsbach PH, Franceschi RT (1999). Isolation and characterization of MC3T3-El preosteoblast subclones with distinct in vitro and in vivo differentiation/mineralization potential. J Bone Miner Res14:893-903.
27.
Xiao G., Wang D., Benson MD, Karsenty G., Franceschi RT (1998). Role of the alpha2-integrin in osteoblast-specific gene expression and activation of the Osf2 transcription factor. J Biol Chem273:32988-32994.
28.
Yaszemski MJ, Yasko AW (1998). Musculoskeletal tissue engineering for orthopedic surgical applications. In: Frontiers in tissue engineering . Patrick CWJ, Mikos AG, McIntire LV, editors. New York, NY: Elsevier Science, Ltd., pp. 197-212.
