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OCN is the primary noncollagenous protein in the bone cells secreted in the late stage of osteogenesis and known as a specific indicator of osteogenic differentiation.
21, 28, and 35 day cultures were washed twice with DPBS and fixed in methanol-acetone (1 1) at 4°C for 10 min. Mineralization of the extracellular matrix served as an indicator of osteogenic differentiation.
The increased osterix expression in the BM-MSC and AD-MSC cultures subjected to differentiation is a relevant indicator of osteogenic differentiation because this transcription factor is indispensable for differentiation.
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Both parameters were determined as indicators of osteogenic differentiation [ 12]. aECM containing sCS2, sCS3, sHA1Δ4S, sHA2, and sHA3 caused a significant increase of TNAP activity by about threefold in comparison to col-aECM) whereas aECM containing sCS1, hyaluronan, and sHA1Δ6S did not alter TNAP activity.
Expression of ECM proteins is a reliable indicator of early osteogenic activity [19] and was identified in all four studies at days 7 and 14.
These proteins are important indicators of late osteogenic differentiation [ 41, 51- 57].
They are widely considered as a significant indicator of the biocompatibility and osteogenic properties of the material due to their multipotent abilities to differentiate into bone precursor cells [ 5, 8, 9].
As a key component of bone matrix vesicles, ALP is an early indicator of immature osteoblast activity and osteogenic differentiation [ 36, 39, 40].
Such nodules are considered reliable indicators of in vitro osteogenic differentiation [ 18].
Figure 5 Evaluation of osteogenic differentiation.
These results reminded on the differentiation of osteogenic progenitor cells.
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