[1,25 (OH), dihydroxyvitamin D3 induces the formation of osteoclasts from mononuclear cells of rat marrow].
OBJECTIVE
To investigate the effect of 1,25 (OH)2 dihydroxyvitamin D3 on the generation of osteoclasts from mononuclear cells of adult rats.


METHODS
With density gradient centrifugation, the mononuclear cells were isolated from rat bone marrow and cultured in the alpha-MEM with 10(-8) mol/L 1,25 (OH)2 dihydroxyvitamin D3. The induced cells were fixed and stained with tartrate-resistant acid phosphatase (TRAP). The bone resorption pits were examined by scanning electron microscope, and the morphology of osteoclasts were examined with inverted phase contrast microscope.


RESULTS
The TRAP+ multinucleated cells could be observed on the seventh culturing day of mononuclear cell group. The number of TRAP+ multinucleated cells went up to a peak on the fourteenth culturing day, and then began to decrease on the twenty-first culturing day. The number of osteoclasts induced from mononuclear cell group was more than that from bone marrow group (P < 0.05) when it was during the fourteenth culturing day to the twenty-third culturing day. The number of bone resorption pits in the mononuclear cell group was much more than that in the bone marrow group (P < 0.05) when it was in period from the tenth culturing day to the twenty-third culturing day. The number of osteoclasts induced from mononuclear cell group could keep the peak value of lasting 7 days, but that induced from bone marrow group could only last its peak value for 3 days.


CONCLUSIONS
The method with 1,25(OH)2 dihydroxyvitamin D3 inducing the formation of osteoclasts from the marrow mononuclear cells is better than that from the whole bone marrow.
Ge
Dong-Xia
DX
Institute of Biomedical Engineering, West China School of Preclinical and Forensic Medicine, Sichuan University, Chengdu 610041, China.
Li
Liang
L
Wu
Jiang
J
Chen
Huai-Qing
HQ
D000135:Acid Phosphatase / Q000378:metabolism; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D001862:Bone Resorption / Q000378:metabolism; D002117:Calcitriol / Q000494:pharmacology*; D005260:Female; D007527:Isoenzymes / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000166:cytology* / Q000187:drug effects*; D010010:Osteoclasts / Q000166:cytology* / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D000071681:Tartrate-Resistant Acid Phosphatase; D013997:Time Factors
D007527:Isoenzymes; D000135:Acid Phosphatase; D000071681:Tartrate-Resistant Acid Phosphatase; D002117:Calcitriol