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. 2022 Sep 16;7:53. doi: 10.1038/s41536-022-00250-7

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© The Author(s) 2022

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 license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license 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 license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 6 — A Rat T3-EV were derived for T3-EV hydrogel formation and delivery. T3-EVs were derived for embedding into composite BMSC hydrogel produced with a mixture of gelatin, fibrinogen, and HA. Cartilage defect injury was created, and hydrogel injection was performed in situ to deliver T3-EV hydrogel for defect repair. B Before transplantation, T3-EV hydrogel was cross-linked by the addition of thrombin to further maintain the shape fidelity of the hydrogel. (a) The cross-linked hydrogel demonstrated good shape fidelity and b–d) good distribution of EVs internalized within BMSCs (b: PKH26-labeled EVs stained red; c: nucleus stained blue; d: merged image with b and c). C Cell viability >95% was demonstrated for 7 days with live/dead assay for the T3-EV hydrogel. D Histological assessment of cartilage repair with T3-EV hydrogel gel at 24 weeks. Neo-cartilage formation in different groups was compared with tissue integrity (HE staining in the 1st row) and GAG deposition with Toluidine blue and Safranin-O staining (second and third row). E–G Histological grading of repaired cartilage in different groups (n = 6 for each) over 24 weeks. F Articular cartilage in both un-EV-gel and T3-EV-gel groups showed declined Mankin score and G higher ICRS histological score compared to the control group in the femoral condyle (FC) and tibial plateau (TP) over the 24 weeks in vivo. H Immunostaining of chondrogenic markers SOX9 (first row), ACAN (second row), and MMP13 (third row) were also conducted and observed (red for the protein, blue for the nucleus) in the generated cartilage in different groups. I Immunostaining of miR-455 (FISH), SOX11, and FOXO1 was also performed and observed (red for the protein, blue for the nucleus) to examine the miR-455/SOX11/FOXO axis in the generated cartilage in different groups. *P < 0.05, **P < 0.01, ***P < 0.001, NS not significant. #p < 0.05 compared to the un-EV gel group.