Observation of osteogenic differentiation cascade of living mesenchymal stem cells on transparent hydroxyapatite ceramics

The use of bioceramics and cultured cells for tissue engineering is a novel approach, which is available in a wide variety of clinical situations. The approach requires apparent verification of the cellular functions occurring on the ceramic surface, and these functions could be monitored by microscopic observation of the cultured living cells on the ceramic material. However, such observation is difficult due to the opaque nature of ordinary ceramics. To overcome this drawback, we used transparent hydroxyapatite (tHA) ceramics as a culture substrate and a transgenic rat having an enhanced green fluorescent protein (EGFP)-expressing gene as the cell source. Marrow mesenchymal stem cells (MSC) were obtained from the rat and cultured on both tHA ceramics and a tissue culture polystyrene (TCPS) dish. One hour after the cell seeding, many MSC had attached and showed initial cell spreading. The attachment and spreading were more obvious 5h after the seeding. Following the culture in the osteogenic condition, the cells differentiated into osteoblasts, which fabricated bone matrix on the culture substrate. The phenomena were similarly observed on both the tHA ceramics and TCPS substrata. These results confirm the excellent properties of tHA ceramics, which support cell attachment, proliferation, and differentiation. Transparent materials make us know the biological usefulness of ceramics in tissue-engineering field.