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. 2020 Mar 24;10:5313. doi: 10.1038/s41598-020-62140-6

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© The Author(s) 2020

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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Animal model of fat grafting (n = 12). (A) The fat grafts harvested at post-grafted week 12. (B) Line charts were plotted to compare the volume retention rate of fat grafts at post-grafted week 12. (C,D); The results of CD31(green)/human mitochondria immunofluorescence(red) staining indicated that, at week 4 post-transplantation, increased exogenous angiogenesis was observed in fat grafts co-transplanted with hADSC-EVs and hypoxic hADSC-EVs, compared with those co-transplanted with PBS. Moreover, co-transplanting with hypoxic hADSC-EVs can further enhance the exogenous angiogenesis in the early stage, compared with co-transplanting with normal hADSC-EVs. Scale bar: 200 μm. (E,F) The results of immunofluorescence staining (CD31/human mitochondria) indicated that host-derived CD31-positive vessels (human mitochondria-) were observed in the peripheral region of fat grafts. Semi-quantitative analysis for the calculation of host-derived neovascularisation indicated that co-transplantation with hADSC-EVs or hypoxic hADSC-EVs exerted no significant effect on host-derived angiogenesis at post-grafted week 4, compared with those co-transplanted with PBS. Scale bar: 200 μm. (G,H) The results of immunofluorescence staining (human mitochondria) indicated that, at week 12 post-transplantation, significant increases in retention rate of exogenous adipose tissue were observed in grafts co-transplanted with hADSC-EVs and hypoxic hADSC-EVs, compared with that co-transplanted with PBS. The retention rate of exogenous adipose tissue can be further improved by co-transplanted hypoxic hADSC-EVs compared with normal hADSC-EVs. Scale bar: 500 μm. Data are presented as the mean ± SD (n = 12). *P < 0.05, **P < 0.01.