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J. Biol. Chem., Vol. 280, Issue 3, 1733-1739, January 21, 2005

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Apoptosis and Survival of Osteoblast-like Cells Are Regulated by Surface Attachment^*

Vavara Grigoriou, Irving M. Shapiro, Elisabeta A. Cavalcanti-Adam¶, Russell J. Composto||**, Paul Ducheyne||, and Christopher S. Adams

From the Department of Orthodontics, School of Dental Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, Department of Orthopaedic Surgery, Thomas Jefferson Medical College, Philadelphia, Pennsylvania 19107-5099, ^¶Institut für Physikalische Chemie, Universität Heidelberg, D-69120 Heidelberg, Germany, and ^||Center for Bioactive Materials and Tissue Engineering and Departments of ^**Material Science and Engineering and Bioengineering, University of Pennsylvania, Philadelphia, Pennsylvania 19104

We tested the hypothesis that RGDS peptides regulate osteoblast survival in culture. Osteoblast-like MC3T3-E1 cells were allowed to attach to RGDS peptides that had been tethered to a silicone surface utilizing a previously described grafting technique. The RGDS-modified surface caused up-regulation of _v3 integrin. We noted that there was an increase in expression of activated focal adhesion kinase and activated Akt. There was no change in the expression level of the anti-apoptotic protein Bcl-2, the pro-apoptotic protein Bad, or the inactivated form of Bad, pBad. Attachment to the RGDS-treated membrane completely abolished apoptosis induced by staurosporine, the Ca²⁺·P_i ion pair, and sodium nitroprusside. However, the surface modification did not interfere with apoptosis mediated by the free RGDS peptide or serum-free medium. When the activity of the phosphatidylinositol 3-kinase pathway was inhibited, RGDS-dependent resistance to apoptosis was eliminated. These results indicated that the binding of cells to RGDS abrogated apoptosis via the mitochondrial pathway and that the suppression of apoptosis was dependent on the activity of phosphatidylinositol 3-kinase.

Received for publication, March 5, 2004 , and in revised form, August 16, 2004.

^* This work was supported by National Institutes of Health Grants DE-13319, DE-10875, DE-05748, HL-60426, DAMD17-03-1-0713, and NASA Grant NRA 00-MEDS. The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

To whom correspondence should be addressed: Dept. of Orthopaedic Surgery, Thomas Jefferson University, 501 Curtis Bldg., 1015 Walnut St., Philadelphia, PA 19107. Tel.: 215-955-8754; Fax: 215-955-9159; E-mail: Christopher.Adams{at}jefferson.edu.

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