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. 2021 Aug 19;11:16839. doi: 10.1038/s41598-021-96107-y

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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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b-ITON and “pre-blast” injections had no effect on retinal thickness, but there were differential effects on vitreal inflammation. (A) There were no significant differences in GCC between any groups (P = 0.1016, ANOVA). (B) There were also no differences in total retinal thickness across any groups (P = 0.7667, ANOVA). (C) Representative OCT retinal scans used to quantify vitreal haze (arrows show examples of vitreous haze). (D) Vitreous intensity normalized to RPE intensity increased after b-ITON and siEGFP compared to b-ITON alone (P = 0.002), but did not increase with siCASP2 injections and remained similar levels to b-ITON alone (P > 0.05). There was a significant decrease in vitreous intensity between siEGFP and siCASP2 injected eyes (P = 0.033, post-hoc Tukey). (E) Vitreous intensity normalised to RPE intensity was also compared between grouped siCASP2 and siEGFP injections administered “pre-blast” and “post-blast”. There was greater vitreal intensity after b-ITON with “post-blast” intravitreal injections at 28 dpi compared to b-ITON with “pre-blast” intravitreal injections at 14 dpi (P = 0.001, GEE). (F) Representative retinal IHC images stained with DAPI (blue), ED1 (red) and GFAP (green) showing retinal ED1⁺ cell infiltration after b-ITON and greater after b-ITON with siCASP2 and siEGFP intravitreal injections. (G) Representative ON IHC images for DAPI (blue) and ED1 (red). (H) There was weak evidence for an increase in ED1⁺ cells in the ON in b-ITON groups (P = 0.075, ANOVA). Scale bars in (F,G) = 100 μm. Error bars represent mean ± SEM.