Fine structural observations of calcium storage in human dental pulp cells in primary culture

Primary culture of explants of human dental pulp tissue allows the study of the cytophysiology and differentiation of the cultured cells over a two-week period. The distribution of calcium was found in two different experimental conditions : with and without a calcium loading, by mean of a lead technique checked by microprobe analysis. The existence of two cell populations was revealed. Intra-mitochondrial ring-like granules characterize type 1 cells when overloaded, while a strong calcium storage is detected in the rough endoplasmic reticulum, Golgi apparatus and mitochondrial (without any inner organization of the deposits) of the type 2 cells. Our results also show the presence of calcium on gap-junctions (revealed) by lanthanum method), and on the extracellular matrix (collagen fibres and complex carbohydrates). The ability of some mitochondria to store calcium (ring-like granules) suggests that the type 1 cells are fully differentiated in odontoblast-like cells and perhaps engaged in mineralization processes. The calcium binding sites, localized on the extracellular matrix may therefore be considered as the earliest foci of calcification.