I'm testing some hydroxyapatite (HA) based ceramic powders with human MSCs. My study concerns the osteogenic capability of the materials in correspondence to hMSCs. The main goal is to fabricate a scaffold for bone tissue engineering application. The sample contains calcium and phosphate (major elements) and also some carbon, silicone, magnesium and sodium (minor element). I guess Alizarin Red or Von Kossa are not applicable in this case. Does anyone know what are the best bone markers to stain the samples? Thank you in advance.
Hi Hanny,
Perhaps you need to re-think your experiment. As you are using powders, this will make it difficult for you to conduct biological evaluation of the material (HA).
If you are trying to establish the effect of the materials chemical composition, then perhaps you ought to consider conducting an indirect study of the materials.
Briefly, either do an elution study of the powders over various time-points, in culture media only - no serum. Then determine the ion composition of the collected media. Then supplement with Sera etc., and use this to culture the cells. You have to take into consideration the ionic composition of media, powder/media (W/V) ratio.
Or, alternatively, you can use multi-well plates with inserts. The powder loaded (in equal amounts) into the inserts while the cells are grown either directly onto tissue culture plastic wells or on Thermanox discs.
This way you can establish the osteogenic capability of the materials based on chemistry per se, excluding the problem of the materials format (powder) as well as the influence of any topographical features that might be associated with the powder. This will also give you flexibility with the staining protocols you wish to use, and it eliminates the earlier problem you asked about, which was eliminating DNA binding to HA!
This is how I would approach the problem at this stage. Best of luck!
Regards
Basil
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