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. 2022 Aug 3;13:4496. doi: 10.1038/s41467-022-32147-w

Fig. 4. Multiphasic temporal effects of intermittent actuation (IA).

Fig. 4

a Timeline of multiphasic cellular and fibrous capsule (FC) changes induced by IA. b Representative fluorescent images of the FC stained with Ly-6G + (green) and DAPI (blue). Scale bars are 20 µm. c Quantification of neutrophils present within FC+/− IA at day 3 and 5. d Representative fluorescent images of the FC stained with α-SMA (green) and CD31 (red). Scale bars are 50 µm. e Quantification of myofibroblasts present within FC+/− IA at 2 weeks. f Representative histologic images of the FC stained with haematoxylin and eosin. Scale bars are 20 µm. g Quantification of total cells/capsular area +/− IA at day 3, day 5, and 2 weeks. h Representative topographical reconstructions of µCT images showing the differences in FC thickness+/− IA at 2 weeks. i Average FC thickness of the control and actuated groups at day 3, day 5, 2 weeks, and 8 weeks with two measurements taken per animal. j Representative polarised light microscopy images of the FC obtained after picrosirius red staining at 8 weeks. Scale bars are 100 µm. k, Quantification of the FC collagen fibre orientation by optical coherency with 60 ROIs per animal. l Representative SEM images demonstrating reduced cellular invasion with actuation at the 8-week timepoint. Scale bars are 500 µm. n = 2–6 animals per group; data are means ± standard error of mean; *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001. See Supplementary Note 1 for detailed statistical analyses.