In vitro biomineralization by osteoblast-like cells. II. Characterization of cellular culture supernatants.
The quantification of total calcium, phosphorus, iron, chromium and nickel in cell culture medium by electrochemical or spectroscopic means may require digestion of samples. Nevertheless, when pH adjustment is performed for values higher than about 6.5, the formation of two phases occurs: a white precipitate and a clear solution. Analysing both phases using microelectrodes, atomic absorption spectrometry (AAS), diffuse reflectance infrared Fourier transform (DRIFT) spectroscopy, X-ray dispersive (XRD) analysis, scanning electron microscopy (SEM) and energy dispersive spectroscopic (EDS) analysis, it was observed that iron, chromium and nickel are not co-precipitating with the white solid phase. If quantification of calcium, phosphorus and magnesium is intended, a ten-fold dilution at least, must be performed to avoid most of these elements going into the precipitate. This knowledge is crucial if a mineralization study is going to be made.
INEB/FEUP, Instituto de Engenharia Biomédica, Departamento de Engenharia Química/Faculdade de Engenharia da Universidade do Porto, Portugal.
S
S
Morais
G S
GS
Carvalho
J L
JL
Faria
H T
HT
Gomes
J P
JP
Sousa
D000818:Animals; D002113:Calcification, Physiologic; D002118:Calcium / Q000032:analysis; D002478:Cells, Cultured; D002857:Chromium / Q000032:analysis; D003470:Culture Media; D004563:Electrochemistry / Q000379:methods; D006863:Hydrogen-Ion Concentration; D007501:Iron / Q000032:analysis; D008670:Metals / Q000032:analysis*; D009532:Nickel / Q000032:analysis; D010006:Osteoblasts / Q000737:chemistry* / Q000187:drug effects / Q000378:metabolism; D010758:Phosphorus / Q000032:analysis; D011817:Rabbits; D013054:Spectrophotometry, Atomic; D013193:Stainless Steel
D003470:Culture Media; D008670:Metals; D002857:Chromium; D013193:Stainless Steel; D010758:Phosphorus; D009532:Nickel; D007501:Iron; D002118:Calcium