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Unformatted text preview: ferentiation factor; OPG, osteoprotegerin; MMP, matrix
metalloproteinase Organ culture
Granulation tissue obtained from a patient with massive
osteolysis was cut into small pieces and cultured in
Dulbecco modiﬁed Eagle’s medium (DMEM) without
serum for 48 h according to the method described by
Goldring et al.5 The cultures were incubated at 37°C in
an atmosphere of 5% CO2 and 95% air. The conditioned medium was collected and used for the following
Human osteoblasts (Cambrex Bio Science Walkersville,
Walkersville, MD, USA) were cultured in DMEM
containing 10% fetal calf serum. At conﬂuence, the
medium was replaced with serum-free medium and cultured for an additional 24 h. The conditioned medium of
granulation tissue previously collected was then added
to the medium at various dilutions and cultured for an
additional 24 h.
Enzyme-linked immunosorbent assay
The concentration of IL-6 and TNF-α in conditioned
medium was measured by enzyme-linked immunosorbent assay (ELISA, BioSource Internati...
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- Spring '08