O3124 ADDITION OF HUMAN MESENCHYMAL STEM CELLS & OSTEOGENIC PROTEIN-1 TO DEMINERALISED BONE MATRIX AND INSOLUBLE COLLAGENOUS MATRIX RESULTS IN DE NOVO BONE FORMATION

Abstract

Aim:. The objective of this study was to improve the osseoinductive capacity of human demineralised bone matrix (DBM) and human insoluble collagenous matrix (ICM), following incorporation of recombinant human osteogenic protein 1 (rhOP-1) and human mesenchymal stem cells (MSCs). Ethical Committee approval has been obtained by our Institution. Methods: Recombinant human osteogenic protein Ð 1 (400ng/0.25g of bone) was seeded onto DBM and ICM together with human MSCs (1 x 105). Cellular proliferation was quantitatively evaluated in vitrousing Alamar Blue and 3H-TdR assays. Quantitative cellular differentiation was assessed using the alkaline phosphatase assay. Von Kossa staining, X-ray analysis, and PCR were used for qualitative evaluation of cellular differentiation. Qualitative analysis of proliferation and differentiation was assessed using scanning electron microscopy (SEM). Results: MSC proliferation and differentiation down the osteogenic lineage was observed on DBM and ICM in the presence of OP-1, and also on DBM alone. Alamar blue and 3H-TdR assays conþrmed that MSC proliferation occurred on both DBM and ICM, with the values being signiþcantly greater with addition of OP-1. The ALP activity showed that MSCs differentiated into osteo-blasts on DBM alone, and on DBM and ICM with OP-1. In all cases, OP-1 had a signiþcant effect on MSCs. Discussion: DBM and ICM when seeded with MSCs and OP-1 provide an improved osteoconductive and osteoinductive graft material resulting in de novo-bone formation.