Adhesion, growth and differentiation of human bone marrow stromal cells on non-porous calcium carbonate and plastic substrata: Effects of dexamethasone and 1,25dihydroxyvitamin D3

Bone marrow contains stromal fibroblastic stem cells which have the potential to differentiate into bone-forming cells. Therefore addition of bone marrow to porous bone substitutes such as coral or coralline hydroxyapatite may be expected to enhance bone ingrowth into these implants. This study was designed to evaluate the possibility of growing human bone marrow stromal fibroblastic cells (HBMC) on a calcium carbonate substrate. For this purpose, HBMC were cultured for 20 days on plastic or calcium carbonate and cell adhesion, growth, and differentiation were studied. It was concluded that calcium carbonate is a highly compatible material for the growth of HBMC. Cells were capable of adhesion within 30 min and were spread within 24 h on this material. However, plating efficiency was decreased in comparison to plastic. Population doubling times (PDT) showed that they were similar when the cells were grown on plastic or calcium carbonate as substratum (PDT=4, 5.5 days). Early protein synthesis included collagen I, collagen III, osteopontin and bone sialoprotein. To induce differentiation of HBMC on plastic and calcium carbonate the influence of dexamethasone (Dex) and 1,25dihydroxyvitamin D3 (1,25(OH)2D3) on alkaline phosphatase (ALP) expression was studied. Basic ALP activity was similar when cells were grown on plastic or calcium carbonate. However, Dex and 1,25(OH)2D3 increased ALP activity of HBMC which could be driven best towards osteogenesis in the presence of Dex and 1,25(OH)2D3.