High yield engineered nanovesicles from ADSC with enriched miR-21-5p promote angiogenesis in adipose tissue regeneration

Di Sun; Shan Mou; Lifeng Chen; Jie Yang; Rongrong Wang; Aimei Zhong; Wei Wang; Jing Tong; Zhenxing Wang; Jiaming Sun

doi:10.1186/s40824-022-00325-y

High yield engineered nanovesicles from ADSC with enriched miR-21-5p promote angiogenesis in adipose tissue regeneration

Biomater Res. 2022 Dec 17;26(1):83. doi: 10.1186/s40824-022-00325-y.

Authors

Di Sun^#^{1

2}, Shan Mou^#^{1

2}, Lifeng Chen^{1

2}, Jie Yang^{1

2}, Rongrong Wang^{1

2}, Aimei Zhong^{1

2}, Wei Wang^{1

2}, Jing Tong^{1

2}, Zhenxing Wang^#^{3

4}, Jiaming Sun^#^{5

6}

Affiliations

¹ Department of Plastic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China.
² Wuhan Clinical Research Center for Superficial Organ Reconstruction, Wuhan, 430022, China.
³ Department of Plastic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China. benjamin.wzx@163.com.
⁴ Wuhan Clinical Research Center for Superficial Organ Reconstruction, Wuhan, 430022, China. benjamin.wzx@163.com.
⁵ Department of Plastic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, 430022, China. sunjm1592@sina.com.
⁶ Wuhan Clinical Research Center for Superficial Organ Reconstruction, Wuhan, 430022, China. sunjm1592@sina.com.

^# Contributed equally.

Abstract

Background: Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) have been found to have a great potential for soft tissue repair due to various biological functions, including pro-angiogenesis and low immunogenicity. However, the low yield and heterogeneity of MSC-EVs limited their clinical transformation. This study was designed to develop a novel adipose-derived stem cell engineered nanovesicles (ADSC-NVs) with high production and explore its pro-angiogenetic effect and application in adipose tissue regeneration.

Methods: Adipose-derived stem cell-derived extracellular vesicles (ADSC-EVs) were isolated from an EVs-free culture medium for human ADSCs (hADSCs). ADSC-NVs were prepared by sequentially extruding ADSCs followed by iodixanol density gradient ultracentrifugation and were compared with ADSC-EVs in morphology, size distribution, protein contents and yield. The pro-angiogenetic effect of ADSC-NVs in different doses (0, 5, 20 and 80 μg/mL) in vitro was determined using transwell assay, tube formation assay, western blot and qRT-PCR. In vivo, BALB/c nude mice were administered injection of a mixture of fat granules and different dose of ADSC-NVs and grafts were harvested at 12 weeks post-transplantation for further analysis. By analyzing the weight and volume of grafts and histological evaluation, we investigated the effect of ADSC-NVs in vessel formation and adipose tissue regeneration.

Results: Our results showed yield of purified ADSC-NVs was approximately 20 times more than that of ADSC-EVs secreted by the same number of ADSCs. In vitro, both ADSC-NVs and ADSC-EVs exhibited a dose-dependent pro-angiogenetic effect, despite their distinct miRNA profiles. These effects of ADSC-NVs may be mediated by enriched miR-21-5p via PTEN inhibition and PI3K/p-Akt signaling activation. Furthermore, after a mixed injection of ADSC-NVs, vessel formation and adipose regeneration were observed in vivo in fat implants.

Conclusions: Our study developed a potent alternative of ADSC-EVs. ADSC-NVs have a high pro-angiogenesis potential and can be used as cell-free therapeutic biomaterials in soft tissue regeneration.

Keywords: Adipose tissue regeneration; Angiogenesis; Engineered nanovesicles; MSCs; miRNAs.

Abstract

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