The effect of inhibiting exosomes derived from adipose-derived stem cells via the TGF-β1/Smad pathway on the fibrosis of keloid fibroblasts

Abstract

Background: The main mechanism of keloid formation is that keloid fibroblasts (KFs) apoptosis is inhibited, leading to excessive proliferation. Transforming growth factor-β1 (TGF-β1) is a key signal molecule in the process of regulating cell fibrosis. This paper discusses the effect of adipose-derived stem cell exosomes (ADSCs-EXO) on the proliferation and apoptosis of KFS and its possible mechanism, in order to provide reference for the clinical intervention of hypertrophic scar.Methods: ADSCs were isolated and cultured from human adipose tissue, the supernatant was collected, and the exosomes secreted by ADSCs-EXO were extracted by ultracentrifugation. At the same time, KFs were cultured from human keloid tissue to P3 generation, and then divided into four groups: control group, experimental group A, experimental group B and experimental group C. KFs were then cultured with four concentrations of ADSCs-EXO (0, 1, 10, and 100 μg/mL, respectively). After 24 hours, cells in each group were taken to detect the following: proliferation of cells in each group using the cell counting Kit 8 (CCK-8) method, cell migration ability via the Transwell test, cell apoptosis by flow cytometry, collagen synthesis using the hydroxyproline method, messenger ribonucleic acid (mRNA) expression of fibrosis-related genes in each group by real-time fluorescent polymerase chain amplification, and the expression of fibrosis-related proteins in the cells of each group by western blotting.Results: Compared with the control group, the proliferation rate, migration rate, and collagen synthesis levels in the three experimental groups decreased with the increase of ADSCs-EXO concentration, while the apoptosis rate in the three experimental groups increased with the increase of ADSCs-EXO concentration, and the differences were statistically significant (P<0.05). Also, compared with the control group, the relative mRNA and protein expression of alpha-smooth muscle actin (α-SMA), TGF-β1, and Smad3 in the three groups decreased significantly, while the expression of three kinds of mRNA and protein decreased with the increase of ADSCs-EXO concentration, and the differences were statistically significant (P<0.05).Conclusions: ADSCs-EXO may inhibit the proliferation and migration, and promote the apoptosis of KFs by inhibiting the expression of the TGF-β1/Smad pathway.

Adipose stem cells-released extracellular vesicles as a next-generation cargo delivery vehicles: a survey of minimal information implementation, mass production and functional modification

“…Only one study revealed that the intravenous injections of ASCs EVs could effectively slow-down the course of the systemic sclerosis via regulating miR-29a-3p/Dnmt3a/Pdgfrbb/Bcl2/Bcl-xl axis [ 35 ]. Two studies found that ASCs EVs could inhibit the proliferation/migration, and promote the apoptosis of keloid/hypertrophic scar fibroblasts via the regulation of miR-192-5p/IL-17RA/Smad axis [ 23 ] or inhibiting TGF-β1/Smad pathway [ 174 ]. Another two studies reported the essential roles of ASCs EVs in promoting the vascularization of skin flaps [ 163 , 176 ], and one study found that ASCs EVs were comparable to parent ASCs in the inhibition of alloimmune response for vascularized composite allotransplantation [ 13 ].…”

Umbilical cord mesenchymal stem cell-derived exosomal Follistatin inhibits fibrosis and promotes muscle regeneration in mice by influencing Smad2 and AKT signaling

“…The UMSCs-derived Exo may serve as potential tools for diabetes [30], regulate cancer P r e p r i n t [31], and regulate fibroblast fibrosis [32]. Therefore, we collected Exo secreted by UMSCs.…”

Signaling pathways of adipose stem cell-derived exosomes promoting muscle regeneration

“…[23] In addition, ASC-exos can improve ovarian function in premature ovarian failure, [24] accelerate the repair of bone defects, [25] and inhibit the formation of keloids. [26] Several studies have confirmed the function of ASC-exos in promoting adult skeletal muscle regeneration. Zhang and Wang et al [27][28][29] demonstrated that ASC-exos can reduce muscle atrophy, degeneration, and fat infiltration, enhance healing, and improve regeneration and biomechanical characteristics in rotator cuff tears.…”

“…In renal conditions, ASC-exos can protect the kidney from acute ischemia-reperfusion injury, [22] and transfer miR-486 to prevent podocytes injury through the small mothers against decapentaplegic 1 (Smad1)/mechanistic target of rapamycin (mTOR) pathway [23] . In addition, ASC-exos can improve ovarian function in premature ovarian failure, [24] accelerate the repair of bone defects, [25] and inhibit the formation of keloids [26] …”

“…[ 23 ] In addition, ASC-exos can improve ovarian function in premature ovarian failure, [ 24 ] accelerate the repair of bone defects, [ 25 ] and inhibit the formation of keloids. [ 26 ]…”