Dear all,
I want to set up experiments with hFOB1.19 cells. I would like to test the attachment of these cells to implants (eg. titanium disks or ceramic implants) to analyze if the implant would osseointegrate or not. Is there a common scense wether the tests should be performed at the permissive temperatur of 34°C oder at the restrictive temperatur of 39°C? Do you have any suggestions?
Many thanks in advance,
Jara
Dear Jara, I would say that if you want to evaluate proliferation 33-34ºC should be selected, in order to allow the cells to expand along the implant. On the other hand, if you want to evaluate differentiation processes the restrictive temperature (39ºC) or with some differentation agents (osteogenic medium with ascorbic acid (AA) and β-glycerophosphate (β-GP)) should be selected. A mineralization study (Alizarin red, Von Kossa), ALP or collagen secretion can be used to verify their osseointegration potential.
Dear Ana,
many thanks for your helpful answer. I think I will go first for the attachment and proliferation at 34°C. I could shift the cells to 39°C after that and measure differentiation. Do you have any suggestions for a read out of the differentiation status?
Many thanks, Jara
Dear Jara,
It would depend on the amount of time you would evaluate them, usually at different time points (1, 3, 7, 14, 21, and 28 days). Mineralization (Alizarin red, Von Kossa) studies can be very useful at long-term differentiation state. Alkaline phosphatase levels at early differentiation time points. You can also evaluate the gene expression of collagen, osteopontin or osteocalcin, among many others. Nevertheless, you have to be aware of the limitations of your own materials as If you have a ceramic coating with Ca, the evaluation of mineralization by Ca deposits can be a challenge.
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