Abstract
Journal of Bone and Mineral Research, Journal of Bone and Mineral Research December 2002:17:2130-2140 (doi: 10.1359/jbmr.2002.17.12.2130)

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RGD Peptides Immobilized on a Mechanically Deformable Surface Promote Osteoblast Differentiation

E. A. CAVALCANTI-ADAM, 1,2   I. M. SHAPIRO, 1,3   R. J. COMPOSTO, 3,4   E. J. MACARAK, 2   C. S. ADAMS1,3  

1Department of Orthopaedic Surgery, Thomas Jefferson Medical College, Philadelphia, Pennsylvania, USA.

2School of Dental Medicine, University of Pennsylvania, Philadelphia, Pennsylvania, USA.

3Center for Bioactive Materials and Tissue Engineering, University of Pennsylvania, Philadelphia, Pennsylvania, USA.

4Department of Material Science and Engineering, University of Pennsylvania, Philadelphia, Pennsylvania, USA.

Address reprint requests to: Elisabetta Ada Cavalcanti-Adam, D.M.D., M.S. Department of Orthopedic Surgery Orthopedic Research Laboratory Room 501, Curtis Building Jefferson Medical College 1015 Walnut Street Philadelphia PA 19107-5099, USA




The major objective of this work was to attach bone cells to a deformable surface for the effective transmission of force. We functionalized a silastic membrane and treated it with 3-aminopropyltriethoxysilane (APTS). A minimal RGD peptide was then covalently linked to the aminated surface. MC3T3-E1 osteoblast-like cells were cultured on the arginine-glycine-aspartic acid (RGD)-treated membrane for 3-15 days and cell attachment and proliferation was evaluated. We observed that cells were immediately bound to the membrane and proliferated. After 8 days on the material surface, osteoblasts exhibited high levels of ALP staining, indicating that the cells were undergoing maturation. Alizarin red staining and Fourier transform infrared (FTIR) analysis showed that the mineral formed by the cells was a biological apatite. The second objective was to apply a mechanical force to cells cultured on the modified silicone membrane. Dynamic equibiaxial strain, 2% magnitude, and a 0.25-Hz frequency were applied to bone cells for 2 h. Osteoblasts elicited increased phalloidin fluorescence, suggesting that there was reorganization of the cytoskeleton. Furthermore, the applied strain elicited increased expression of the αvβ3 integrin receptor. We concluded that the covalent binding of RGD peptides to a silicone membrane provides a compatible surface for the attachment and subsequent differentiation of osteoblasts. Moreover, the engineered surface transduces applied mechanical forces directly to the adherent cells via integrin receptors.

Cited by

Tetsuyuki Hayashibara, Toru Hiraga, Bing Yi, Motoyoshi Nomizu, Yoshinari Kumagai, Riko Nishimura and Toshiyuki Yoneda. (2004) A Synthetic Peptide Fragment of Human MEPE Stimulates New Bone Formation In Vitro and In Vivo. Journal of Bone and Mineral Research 19:3, 455-462
Online publication date: 1-Mar-2004.
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Authors:
E. A. CAVALCANTI-ADAM,
I. M. SHAPIRO,
R. J. COMPOSTO,
E. J. MACARAK,
C. S. ADAMS
Keywords:
arginine-glycine-aspartic acid peptides
osteoblast
silicone membrane
cytoskeleton
mechanical forces