i want to check any synergistic or additive effect but my two indicators are not two drugs rather drug is applied with hyperthermia to enhance the apoptosis. so in this case can i apply isobolograph to check the synergistic effects or not? please guide me in this respect..
Hi Shahbaz,
Could you please give more details about your study? Are you going to use in vitro or in vivo models or clinical study?
Either way, you can use isobologram for apoptosis induction effects of two factors which you described. Because hyperthermia is already known as an apoptosis inducer factor, so you wrote the drug is also used for enhancing the apoptosis. Under these circumstances, I think, you can check their synergistic, additive effects on apoptosis induction.
So you have to decide which parameter you are going to use for 100% apoptosis induction. In this point, I need to know your model to decide.
Best
i am doing in vitro study on cell cultures...so as you said i can apply this to check the synergism then how we will evaluate the effect value of both inducers? beacause i download the calcusyn demo version in which the required drug and effects individually and then mixtures...but the value of effect should be between 0~1. please guide me further Cigdem Ozen.
thanks
First of all, you have to be sure your factors really increase apoptosis. I suggest you do flow cytometer with annexin V with different concentrations or conditions.
For checking apoptosis also means checking cytotoxicity. You have to decide a cytotoxicity assay to find the factors' cytotoxic effect in a concentration range of 0%-100% cytotoxicity. So, you have to conduct a cytotoxicity assay like SRB or MTT with the factors (hyperthermia and the drug). First of all, you have to find the concentrations of the factors to induce 0 to 100% growth inhibition. So when you find this concentrations you have to plan another experiment of cytotoxicity with each factor and combination. For example if a certain concentration of the drug inhibits 40% of growth, and hypothermia inhibits 40% of growth individually. In these certain conditions, if you combine them, they should give 80% if they are additive, if more than 80%, like 100% they are synergitic, if the result is less than 80%, so they are antagonistic. You can just transform this percent numbers to 0-1, like 80% = 0.8
Good luck
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