and 2 other(s)
Demineralized bone matrix is a widely used allograft from where not only the inorganic mineral but also embedded growth factors are removed by hydrochloric acid. The cellular response to the growth factors released during the preparation of demineralized bone matrix has not been studied. Here we investigated the in vitro impact of acid bone lysate (ABL) prepared from porcine cortical bone chips on oral fibroblasts. Whole genome microarray together with the pharmacologic blocking of TGF-β receptor type I kinase with SB431542 revealed that acid bony lysates activate TGF-β signaling in oral fibroblasts. These findings were further confirmed with RT-PCR, by the expression of interleukin 11, proteoglycan4 and NADPH oxidase 4 on human oral fibroblasts and by the expression of interleukin 11 on periosteal pig cells. The effect was independent of ABL source; mandible, calvaria or tibia. Gene expression was, however, dependent on the concentration of hydrochloric acid. ABL prepared with 1M hydrochloric acid reduced the TGF-β activity in ABL. In addition, interleukin 11 expression was confirmed at the protein level by ELISA, supporting further the TGF-β activity. Furthermore, ABL inhibited osteogenic differentiation of calvaria cells, as shown by histological staining and gene expression. In summary, these results demonstrate that TGF-β is a major growth factor of bone grafts being released upon preparation of demineralized bone matrix.
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