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. 2022 Mar 28;20:164. doi: 10.1186/s12951-022-01352-6

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© The Author(s) 2022

Open AccessThis 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/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 10 — Engineered-Exo promoted the survival, proliferation and new cartilage formation of 3D-cultured tissue-engineered cartilage. (A) Cellular uptake assay by confocal microscopy imaging demonstrated rapid uptake of Engineered-Exo (red) at the different time points. (B) Representative images showing the live and dead microtia chondrocytes seeded on Gelma hydrogel, and (C) the comparison for the rate of live cell between Engineered-Exo and PBS control. Scale bar: 500 μm. Green: live microtia chondrocytes. Red: dead microtia chondrocytes. (D) Cell proliferation assessed by the Cell Counting Kit-8 at the specific time points after the exposure to Engineered-Exo or PBS. (E) Quantitative RT-PCR analysis showed regulation of genes associated with anti-apoptosis (PTEN, Survivin, Bcl-2), proliferation (PTEN, PCNA, FGF-2), as well as chondrogenic differentiation and ear cartilage matrix proteins synthesis (PTEN, TGF-β1, COL2A1, ACAN, ELN, SOX9 and COMP). The genes (COL1A1 and MMP 9) unfavorable for the synthesis of hyaline cartilage and elastic cartilage were also assessed. In particular, the tendency of expression of several chondrogenesis-related genes (COL2A1, ACAN, ELN, SOX9 and COL1A1) was described. (F) The protein levels of elastin, collagen II, and SOX9 in microtia chondrocytes were analyzed by western blotting. Data was presented as the mean ±SD of three number of replicates. ∗p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, ∗∗∗∗p < 0.0001