In vitro investigation of novel calcium phosphates using osteogenic cultures.

A rat bone marrow stromal cell (RBM) culture was used to evaluate novel bioactive calcium phosphate ceramics. Three rapidly resorbable, glassy crystalline materials with the main crystalline phase Ca2KNa(PO4)2 were investigated (sample code GB 1a, GB 14, GB 9). These materials were designed to exhibit a higher degree of biodegradability than tricalcium phosphate. Additionally, a bioactive glass ceramic of low biodegradability was examined (sample code AP 40). RBM cells were cultured on the disc-shaped test substrata for 14 d. The culture medium was changed and calcium and phosphate concentrations of the medium were determined daily. Specimens were evaluated using light microscopy and morphometry of the cell-covered substrate surface, scanning electron microscopy and energy dispersive X-ray analysis. Except for GB 1a, the rat bone marrow cells attached and grew on all substrate surfaces. Of the different calcium phosphate ceramics tested, AP 40 facilitated osteoblast growth and the elaboration of the extracellular matrix to the highest degree followed by GB 9 and GB 14. The inhibition of cell growth encountered with GB 1a seemed to be related to its high phosphate ion release.

Morphological evaluation of osteoblasts cultured on different calcium phosphate ceramics.

The objective of these investigations was to develop an in vitro test system for evaluating novel rapidly resorbable calcium phosphate ceramics of varying composition. Rat bone marrow cells were cultured on the disc-shaped test substrates for 14 days. Five calcium phosphates were examined: R1 CaNaPO4; R1/M2, composed of CaNaPO4 and MgNaPO4; R1/2, composed of CaNaPO4 and Mg2SiO4; R1 + 9% SiO2 consisting of CaNaPO4 and 9% SiO2 (wt%) and R17, Ca2KNa(PO4)2. Two studies were performed. In study I cultures were re-fed every two to three days. In study II the medium was changed daily, and calcium and phosphate concentrations of the medium were determined daily. Specimens were prepared for light microscopy and morphometric evaluation of the cell-covered substrate area, scanning electron microscopy and energy-dispersive X-ray analysis. With all materials tested except for R1/2, an increase of cellular growth was observed after changing the medium daily. Of the different calcium phosphate ceramics tested, R17 and R1/M2 facilitated osteoblast growth and elaboration of extracellular matrix to the highest degree. The inhibition of cell growth encountered with R1 in study I and R1/2 in both studies seemed to be related to a high phosphate-ion release from these materials.