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

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.

Clay Sculpture‐Inspired 3D Printed Microcage Module Using Bioadhesion Assembly for Specific‐Shaped Tissue Vascularization and Regeneration

“…[55,56] Sun et al showed that the addition of exosomes and growth factors with angiogenic properties improves the survival rate of grafts by promoting angiogenesis. [57,58] In some studies, graft survival was improved by transplanting vascular loops or vascular-like organs into the graft area. [59] These studies suggest that the in vivo survival rate of grafts can be improved by angiogenesis of grafts in vivo.…”

De novo design of a nanoregulator for the dynamic restoration of ovarian tissue in cryopreservation and transplantation

“…MSC-derived extracellular vesicles (MSC EVs) are nanoscale paracrine mediators secreted by MSCs, which contain a variety of actives that promote angiogenesis and tissue regeneration [ 204 , 205 ]. In 2022, Sun et al [ 206 ] developed adipose-derived stem cell-engineered nanovesicles, which were found to be superior to conventional MSC EVs in promoting adipose tissue angiogenesis, suggesting their potential as cell-free therapeutic biomaterials in soft tissue regeneration. It is hypothesized that if MSC nanovesicles are applied to ovarian tissue transplantation, the survival rate of transplanted tissues may be significantly enhanced.…”

Engineered Extracellular Vesicles in Wound Healing: Design, Paradigms, and Clinical Application

“…[ 118 ] DGC is often utilized in the separation of non‐engineered EVs in wound repair and in eEV studies in areas like tumors, viral infections, soft tissue regeneration, and non‐invasive imaging. [ 118–121 ] This suggests the method doesn't damage EVs' structure and function, and the engineered part of eEVs is preserved during the DGC process. These factors point towards DGC's potential application in wh‐eEVs research. Ultrafiltration (UF), a pressurized membrane separation technique, allows small molecule substances to pass through a membrane when a certain pressure is applied, while large molecule substances cannot, leading to the target substance's purification.…”

“…[118] DGC is often utilized in the separation of non-engineered EVs in wound repair and in eEV studies in areas like tumors, viral infections, soft tissue regeneration, and non-invasive imaging. [118][119][120][121] This suggests the method doesn't damage EVs' structure and function, and the engineered part of eEVs is preserved during the DGC process. These factors point towards DGC's potential application in wh-eEVs research.…”