Figure 3

Quantitative imaging analysis highlights differences in hepatic steatosis before and after DAA treatment for viral hepatitis C. H&E-stained slides were digitally scanned on an Aperio ImageScope Digital slide scanner at low-power (×1.2). We used an algorithm developed by Visiopharm (imaging analysis software) to more objectively quantity steatosis droplets in the liver biopsies. Lipid droplets that were considered as macrovesicular steatosis droplets measured > 250 square µm in greatest dimension (turquoise-colored circles) and microvesicular steatosis “Eq. (2)” droplets were those that measured < 250 square µm (magenta circles). All lipid droplets smaller than 25 square microns were removed to diminish background. Vessels and artifacts in the biopsy tissue were marked with black dashed lines (see G and H), allowing them to be removed from analysis. The entire core was outlined by the software, which enabled measurement of the surface area of each biopsy. Patients 2 and 4 are shown as representative examples of patients whose post-treatment liver biopsies showed decreased and increased macrovesicular steatosis when compared to their pre-treatment biopsies, respectively. Patient 2 (A,B) presented with more macrovesicular steatosis pre-treatment (6.99% using Visiopharm analysis E) than post-treatment (1.56% using Visiopharm analysis F). Patient 4 (C,D) had minimal macrovesicular steatosis pre-treatment (0.86% using Visiopharm analysis G) that increased substantially post-treatment (6.32% using Visiopharm analysis H). Table 2 shows all the histopathologic findings in the patient biopsies (n = 10). Magnification: (A) and (E), ×40; (B) and (F), ×100; (C,D) and (G,H) ×400. Pre-: pre-DAA treatment; Post: post-DAA treatment.