Osteogenically-induced exosomes stimulate osteogenesis of human adipose-derived stem cells

Abstract

Exosomes exhibit great therapeutic potential in bone tissue engineering. The study aimed to investigate whether the exosomes derived from human adipose-derived stem cells (hADSCs-Exos) during different time-span of osteogenic differentiation could promote osteogenesis. The appropriate concentrations of hADSCs-Exos to enhance the proliferation, migration and osteogenesis of hADSCs-Exos were also examined. PKH67 labelled hADSCs-Exos was used to detect the internalization ability of hADSCs. The osteogenic differentiation abilities of hADSCs after treatment with hADSCs-Exos was evaluated by Alizarin red staining (ARS). The proliferation and migration of hADSCs was examined by cell counting kit-8 and wound healing assay, respectively. The expression of exosomal surface markers and osteoblast-related protein of hADSCs was assessed by Western blot. PKH67-labelled exosomes were internalized by hADSCs after 4 h incubation. ARS showed that the amount of mineralized nodules in Exo1−14d group was significantly higher than that in Exo15−28d group. hADSCs-Exos could promote the proliferation and migration capacity of hADSCs. Western blot analysis showed that after hADSCs-Exos treatment, ALP and RUNX2 were significantly enhanced. Specially, the Exo1−14d group of 15 μg/mL significantly upregulated the expression of RUNX2 than the other exosomes treated groups. Our findings suggest that exosomes secreted by hADSCs during osteogenic induction for 1–14 days could be efficiently internalized by hADSCs and could induce osteogenic differentiation of hADSCs. Moreover, administration of Exo1−14d at 15 μg/mL promoted the proliferation and migration of hADSCs. In conclusion, our research confirmed that comprised of hADSCs-Exos and hADSCs may provide a new therapeutic paradigm for bone tissue engineering.

Evidence for a Novel Antiviral Mechanism of Teleost Fish: Serum-Derived Exosomes Inhibit Virus Replication through Incorporating Mx1 Protein

“…Furthermore, whether exosomes could internalize into mandarin fish fry (MFF-1) cells was investigated. Exosomes were labeled with green-fluorescent dye PKH67 [ 14 ]. PKH67-labeled exosomes were incubated with cells for 2 h, and green fluorescence was detected around the nucleus in MFF-1 cells ( Figure 1 D).…”

Adipose-Derived Stem Cells Secretome and Its Potential Application in “Stem Cell-Free Therapy”

“…This may be due to the presence of soluble proteins and exosomal immunomodulatory miR-21 [165]. On the other hand, ASC-exos has been shown to induce osteogenic differentiation [166] and also suppress inflammation by upregulating miR-145 and miR-221, and therefore can be helpful in the treatment of osteoarthritis [167]. It has already been shown in the case of chondrocytes isolated from osteoarthritic patients that treatment with ASCs extracellular vesicles reduces MMP-1, MMP-3, MMP-13, and ADAMTS-5 and increases collagen II expression [168,169].…”

Adipose Stem Cells in Regenerative Medicine: Looking Forward

“…In addition to ASCs therapeutic potential, studies have shown that ASCs-Exo can stimulate bone regeneration through a paracrine mechanism ( Zhu et al, 2021 ). Furthermore, in mouse models, decellularized adipose tissue hydrogels have shown promise, especially when combined with ASCs.…”