I want to quantify the rate of mineralization of preosteoblasts (
MC3T3-E1) on a 3D scaffold. However, the scaffold is mainly composed of Hydroxyappatite (HA). Thus, I cannot use Alizarin Red Staining because HA is partially composed of Calcium.
Hi Carlos
I hope you are well
This is good question which should be considered and this is true, scaffolds containing hydroxyapatite is composed of Ca and O that have the ability to mineralize. The ratio of Ca/p in HA is 1.67 so, in every scaffolds we try to obtain this ratio. Also, this ratio for mineralized scaffolds is so important. I think the Alizarin Red is not good choice but article at below can be helpful.
Micro-CT based quantification of non-mineralized tissue on cultured hydroxyapatite scaffolds.
Hi Carlos,
Enzymatic studies such as Alkaline phosphatase activity, osteocalcine or osteopontine level. apart from these, you can also perform PCR analyses in order to reveal the osteogenic gene expression levels. Thus, you would have measure the mineralization by cell spesific methods.
Best regards,
Looking to quantitative evaluation of calcification, direct chemical determination of total calcium after culturing the scaffold in presence and absence of preosteoblasts would appear as the master way. Provided the actual variation is detectable by the adopted analytical system. AA based analytical procedures are likely to be explored.
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