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. 2018 Jul 19;9:1639. doi: 10.3389/fimmu.2018.01639

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Copyright © 2018 Kholia, Herrera Sanchez, Cedrino, Papadimitriou, Tapparo, Deregibus, Brizzi, Tetta and Camussi.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Histological analyses of aristolochic acid nephropathy (AAN) in vivo model. (A) Representative micrographs of H&E-stained renal tissue from healthy mice injected with vehicle alone (control), or mice injected with aristolochic acid (AA), or AA mice treated with human liver stem cell-derived extracellular vesicles (HLSC-EVs). (B) Representative micrographs of Masson’s trichrome stained renal sections from control, AA, or HLSC-EVs treated AA mice. The blue stain represents collagen fibres considered to be a marker for interstitial fibrosis. Original magnification at 400×. (C) Representative micrographs of proliferating cell nuclear antigen (PCNA) stained renal tissue of control, AA, or AA mice treated with HLSC-EVs original magnification at 400×. (D) Histological score of tubular necrosis in AAN mice experimental groups. Mice treated with AA had significantly elevated levels of tubular necrosis, which was alleviated on treatment with HLSC-EVs. No significant reduction in tubular necrosis was observed in mice treated with Fibro-EVs. Data represent mean ± SD of tubular necrosis observed under high power field (original magnification: 400×). A one-way analyses of variance (ANOVA) with Bonferroni’s multi comparison test was performed. ***p < 0.001 AA vs control or HLSC-EVs vs AA, ^#p < 0.01 Fibro-EVs vs HLSC-EVs. (E) Histological quantification of interstitial fibrosis in AAN mice experimental groups by multiphase image analysis of 10 fields per section. Data represent mean ± SD; a one-way ANOVA with Bonferroni’s multi comparison test was performed. ***p < 0.001 AA vs control, **p < 0.01 HLSC-EVs vs AA, ^#p < 0.01 Fibro-EVs vs HLSC-EVs. (F) Histological score of PCNA positive cells in AAN mice experimental groups observed under high power with an original magnification of 400×. An increase in PCNA positive cells (marker for proliferation) was observed in HLSC-EVs treated mice renal tissue. No significant increase in PCNA was observed in AA mice treated with Fibro-EVs compared to control. Data represent mean ± SD. A one-way ANOVA with Bonferroni’s multi comparison test was performed. **p < 0.01 AA vs control, or Fibro-EVs vs HLSC-EVs, *p < 0.05 HLSC-EVs vs AA, ^#p < 0.01 Fibro-EVs vs HLSC-EVs (n = 9 mice per group).