I prepare macrophages from mouse bone marrow and grow them in a sponge-like porous hydrogel that we make.

I am accumulating data that indicates different tightly controlled pore sizes affects resting phenotype and inflammatory response to toll-like receptor stimulation (LPS in my case).

I want to look at cellular metabolism in the materials with different pore sizes with and without stimulation.

I specifically want to compare the balance of glycolysis to oxidative phosphorylation.

As a rough start on the glycolysis side I can collect conditioned media from the cultures over time and measure lactate production before and after LPS.

What has me stumped is the oxidative phosphorylation. Oxygen consumption rate seems hard to get at here. The implant material is opaque. Most of the assays I see need a clear light path through a well in a 96 well plate.

I frequently grind up our cell colonized porous hydrogel to obtain cell lysates for protein and/or RNA studies.

Is there anything that can be done with conditioned media samples as far as oxidative phosphorylation? Is there any substrate that could be fed in to these cultures generate a stable soluble end-point product?