Assessment of the proliferation of human mesenchymal stromal cells in the presence of human demineralised bone matrix

Tissue engineering integrates discoveries from biochemistry, cell and molecular biology, material science and biomedical engineering to produce innovative three-dimensional composites that can be used to replace or correct damaged tissues and organs. Precise classification of osteoinductive properties of human demineralised bone is often the problem, because it varies from batch to batch. An in vitro assay using bone marrow derived human mesenchymal stromal cells (hMSCs) was developed to improve the classification of the osteoinductive quality of demineralised bone matrix. In this study, three-dimensional, partially demineralised bone scaffolds were investigated for their ability to induce osteogenic differentiation of hMSCs in vitro. Proliferation of the hMSCs was measured by the CelTiter 96A (R) AQueous One Solution Cell assay. Chemical structure was evaluated using quantitative and qualitative X-ray analysis. Scanning electron microscopy revealed primary proliferation of the cells cultivated 14 days and showed elevations in the content of Ca2+. These results demonstrate that partially demineralised human bone material supports osteogenic differentiation of hMSCs in vitro. This study documents that in vitro test using hMSCs can be used for classification of the osteoinductive quality of human demineralised bone matrix.