Combination metabolic therapy decreases VM-M3 cell proliferation <i>in vitro</i>.

<p>VM-M3 cells were grown in conditions mimicking metabolic therapy, including combinations of varied glucose concentration (25mM, 15mM, or 3mM) media, ketone supplementation (5mM βHB), and daily HBOT sessions (100% O₂, 2.5 ATA, 90 min). Cell density was measured with trypan blue hemocytometry to produce a growth curve over 96 hours. <b>(A)</b> Proliferation of VM-M3 cells was inhibited with decreasing glucose concentration and the addition of HBOT. At 24 hours, cell density of 3mM glu and 3mM glu + HBOT cells was significantly less than control (25mM) glucose treated cells. At 48, 72, and 96 hours, the cell density of all treatment groups was significantly reduced compared to control (*p<0.05; Two-way ANOVA). <b>(B)</b> Combining low glucose, ketone supplementation, and HBOT resulted in a marked decrease in cell proliferation which was significant from control at all time points tested. (p<0.05; one-way ANOVA). Cells receiving combination therapy had a significant decrease in proliferation compared to cells treated with LG only at 24, 48, and 72 hours, and compared to βHB only at 48, 72, and 96 hours, and exhibited a non-significant trend of decreased proliferation compared to LG + HBOT treated cells for the duration of the growth curve. Results were considered significant when p<0.05. Error bars represent <b>±</b>SEM.