Mh. Fernandes et al., MINERALIZATION IN SERIALLY PASSAGED HUMAN ALVEOLAR BONE-CELLS, Journal of materials science. Materials in medicine, 8(2), 1997, pp. 61-65
Well-characterized human bone cell cultures have been regarded as a us
eful tool to study bone control mechanisms and also to analyse bone/bi
omaterials interactions. In the present study, human alveolar bone cel
ls were cultured in alpha-minimal essential medium (alpha-MEM) contain
ing 10% foetal bovine serum (FBS), 50 mu g/ml ascorbic acid, 10 mM sod
ium beta-glycerophosphate and either in the presence or in the absence
of 10 nM dexamethasone (Dexa). Cultures were characterized concerning
cell viability/proliferation, alkaline phosphatase (ALP), acid phosph
atase (ACP) and tartraric acid resistant phosphatase (TRAP) activities
, and formation of mineralized areas. Cell proliferation increased gra
dually for approximately 20 days. In the presence of Dexa, cells forme
d isolated or interconnected multilayered clusters that increased with
culture time. Histochemical assays revealed strong positive reactions
for ALP and calcium and phosphates deposits, mainly in relation to ce
lls associated with the clusters. High levels of ALP activity (biochem
ical determination) were observed. Cells cultured in the absence of De
xa showed significantly lower ALP activity and no calcium and phosphat
es deposits were present. Serially passaged cells kept the proliferati
on rate constant but a decrease in ALP activity was observed either in
the presence or in the absence of Dexa. The ability to form mineraliz
ed areas (cultures fed with Dexa) also decreased on serial subculture.