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. 2021 Jul 15;11:14506. doi: 10.1038/s41598-021-93881-7

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© The Author(s) 2021

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

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Comparison of histopathologic changes in patient liver biopsies before and after DAA treatment. Liver biopsies from the study patients (n = 10) were evaluated for inflammation, steatosis, and fibrosis by a board-certified pathologist. We also compared differences in the CPA and percent of steatosis (using Visiopharm) before and after treatment. (A) The MHAI was used to grade the amount of hepatitis activity in each patient’s liver biopsy²⁵. Liver biopsies post-DAA treatment had significantly decreased MHAI scores (P < 0.001). (B) We then compared changes in each patient’s individual MHAI scores pre- and post-DAA treatment. Even though all the patients showed decreased MHAI scores, 60% of the patients still had mild, persistent lymphocytic inflammation. (C) Next, we compared the stages of hepatic fibrosis using the Ishak criteria (scale of 0 to 6) as determined by the pathologist. No significant difference was observed between pre- and post-DAA treatment (P < 0.063). (D) The changes in hepatic fibrosis are shown for each individual patient. (E) There were no significant differences in the CPAs between the patients before and after treatment. However, when the patients were evaluated individually (F), patient 4 showed a marked increase in fibrosis in her post-treatment liver biopsy when compared to her initial biopsy (yellow line). (G) Next, we compared differences in the amount of steatosis in the patients’ pre- and post-DAA liver biopsies using a custom algorithm from Visiopharm, which calculated the amount of steatosis per total biopsy area (see Fig. 2). The majority of patients had decreased or similar amounts of steatosis in their post-treatment liver biopsy when compared to the one collected prior to treatment. However, since Patient 4 had such a marked increase in hepatic steatosis in her post-treatment biopsy, no significant difference was observed. This was best highlighted when the patients were compared on an individual basis (H). *Significant differences between values pre- and post- DAA treatment (P value < 0.05). The colored lines include patients 1 through 8 and 10. ns: not significant. MHAI: Modified hepatitis activity index; Pre: pre-DAA treatment; Post: post-DAA treatment; CPA: Collagen proportionate area, “Eq. (1)”.