Fig. 5. Representation of the death of primary subcutaneous adipocytes after incubation with bile salt particles.
Cell viability of the primary subcutaneous adipocytes after being incubated with (A) different concentrations of the sodium deoxycholate and sodium cholate salts in RPMI media and (B) different concentrations of cholate-based and deoxycholate-based composite microparticles for different time points. Shown on top of the bars in each number concentration group are the correspondent mass concentrations for each condition calculated by counting the number of particles in a known mass. (C) Viability values for cells treated with supernatant from 106 particles incubated in media for 3 hours (the degradation products of the particles). (D) Inhibitory effect of bovine serum albumin (BSA) on the lysis effect of deoxycholate-based microparticles, where 105 particles suspended either in RPMI media or 5% BSA solution in RPMI media were added to each well. n = 3 for all assays. Cell viabilities were measured using MTS assay and calculated by dividing the net cell titer absorbance of the test wells by the untreated control wells. Two-way ANOVA with Tukey’s posttest and confidence interval of 95% were used to analyze (A) and (B), and unpaired t test was used to analyze (D). “*” represents the significance compared to 0.01% salt or 105 particles per well of treatments, “#” compares the significance between 1-hour and 3-hour treatments, and “$” represents the difference between the treatment in 5% BSA versus RPMI media. No significant difference was observed between 1-hour and 3-hour treatments in (A). **P < 0.01, ***P < 0.001, and ****P < 0.0001.