Mineralized nodule formation in rat bone marrow stromal cell culture without beta-glycerophosphate.
Rat bone marrow stromal cells were cultured in the presence of 10 nM dexamethasone and various concentrations of beta-glycerophosphate. At day 12-15, some nodules consisting of polygonal cells were formed in all culture conditions, and these nodules were mineralized 2-3 days later. beta-Glycerophosphate significantly enhanced nodule formation at concentrations of not less than 5 mM. The mineralized nodules formed in the absence of beta-glycerophosphate were examined using phase-contrast microscopy, undemineralized and demineralized tissue histology, histochemistry for alkaline phosphatase, immunohistochemistry for type I, II, and III collagen, energy dispersive X-ray microanalysis, electron diffraction, and Fourier transform infrared spectroscopy (FT-IR). Mineralized nodules had histological characteristic similar to bone. Cells associated with nodules exhibited high alkaline phosphatase activity, and extracellular matrix of the nodules predominantly consisted of type I collagen. X-Ray microanalysis showed the presence of Ca and P in the mineralized area, and electron diffraction pattern showed the mineral to have apatite crystal structure. Moreover FT-IR indicated that the mineral was a mixture of hydroxyapatite and carbonateapatite. From these observations, it is concluded that the mineralized nodules formed in our culture system are truly bone-like.
Satomura
K
K
First Department of Oral and Maxillofacial Surgery, School of Dentistry, Tokushima University, Japan.
Hiraiwa
K
K
Nagayama
M
M
D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D001853:Bone Marrow / Q000187:drug effects / Q000378:metabolism; D001854:Bone Marrow Cells; D001842:Bone and Bones / Q000166:cytology*; D002113:Calcification, Physiologic / Q000187:drug effects; D002118:Calcium / Q000032:analysis; D002478:Cells, Cultured; D003094:Collagen / Q000032:analysis; D003907:Dexamethasone / Q000494:pharmacology; D004577:Electron Probe Microanalysis; D005994:Glycerophosphates / Q000494:pharmacology*; D006651:Histocytochemistry; D008297:Male; D008858:Microscopy, Phase-Contrast; D010758:Phosphorus / Q000032:analysis; D051381:Rats; D013055:Spectrophotometry, Infrared
D005994:Glycerophosphates; D010758:Phosphorus; D003907:Dexamethasone; D003094:Collagen; D000469:Alkaline Phosphatase; D002118:Calcium; C031463:beta-glycerophosphoric acid