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Platelet lysate induces chondrogenic differentiation of umbilical cord-derived mesenchymal stem cells by regulating the lncRNA H19/miR-29b-3p/SOX9 axis.

ABSTRACT: Platelet lysate (PL) has been shown to induce chondrogenic differentiation of human umbilical cord-derived mesenchymal stem cells (hUCMSCs). However, the underlying mechanism is still not clear. The aim of this study was to investigate whether long noncoding RNA H19 is involved in this process. hUCMSCs were isolated, identified and cultured in 5% PL-supplemented chondrogenic differentiation medium. Chondrogenic differentiation was assessed by Alcian blue staining. The expressions of H19, miR-29b-3p, SRY-related high-mobility-group box 9 (SOX9), collagen II and aggrecan were determined by quantitative real-time PCR and western blot. The interaction between miR-29b-3p and H19 or SOX9 was analyzed by luciferase reporter assay. During PL-induced chondrogenic differentiation of hUCMSCs, expressions of H19 and SOX9 were increased, whereas miR-29b-3p expression was decreased. H19 overexpression promoted, whereas H19 silencing attenuated the PL-induced chondrogenic differentiation of hUCMSCs. SOX9 was identified as a direct target of miR-29b-3p, and H19 was observed to act as a sponge of miR-29b-3p to up-regulate SOX9 expression. The chondrogenic differentiation-promoting effect of H19 overexpression was negated when miR-29b-3p expression was up-regulated by Lenti-miR-29b-3p infection. In conclusion, PL induced chondrogenic differentiation of hUCMSCs by regulating the H19/miR-29b-3p/SOX9 axis.

SUBMITTER: Cao B

PROVIDER: S-EPMC7714074 | biostudies-literature | 2020 Dec

REPOSITORIES: biostudies-literature

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Platelet lysate induces chondrogenic differentiation of umbilical cord-derived mesenchymal stem cells by regulating the lncRNA H19/miR-29b-3p/SOX9 axis.

Cao Boran B Dai Xin X

FEBS open bio 20201106 12

Platelet lysate (PL) has been shown to induce chondrogenic differentiation of human umbilical cord-derived mesenchymal stem cells (hUCMSCs). However, the underlying mechanism is still not clear. The aim of this study was to investigate whether long noncoding RNA H19 is involved in this process. hUCMSCs were isolated, identified and cultured in 5% PL-supplemented chondrogenic differentiation medium. Chondrogenic differentiation was assessed by Alcian blue staining. The expressions of H19, miR-29b ...[more]

PMID: 33058414

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Project description:Background Exosomes derived from stem cells have been demonstrated to be good candidates for the treatment of osteochondral injury. Our previous studies have demonstrated that mechanical stimulation could be crucial for the secretion of exosomes derived from umbilical cord mesenchymal stem cells (U-MSCs). Therefore, we explore whether mechanical stimulation caused by a rotary cell culture system (RCCS) has a beneficial effect on exosome yield and biological function. Methods U-MSCs were subjected to an RCCS at different rotational speeds and exosomes were characterised by transmission electron microscopy, nanoparticle tracking analysis and western blotting. small-interfering RNAs of Rab27a (siRNA-Rab27a) was used to reduce exosome production. Quantitative real-time PCR (qRT-PCR) was used to detect the expression of mechanically sensitive long non-coding RNA H19 (LncRNA H19). The effects of exosomes on chondrocyte proliferation were examined using cell counting kit-8 (CCK-8), toluidine blue staining and a series of related genes. Annexin V-FITC and PI (V-FITC/PI) flow cytometry was used to detect the effect of exosomes on the inhibition of chondrocyte apoptosis. Macroscopic evaluation, MRI quantification and immunohistochemical staining were conducted to investigate the in vivo effects of exosomal LncRNA H19 through SD rat cartilage defect models. Results RCCS significantly promoted exosome production at 36 rpm/min within 196 h. Mechanical stimulation was able to increase the expression level of exosomes. The exosomal LncRNA H19 was found to promote chondrocyte proliferation and matrix synthesis and inhibit apoptosis in vitro. Chondral regeneration activity was lost in LncRNA H19-defective exosomes. The injection of exosomal LncRNA H19 in vivo resulted in improved macroscopic assessment, MRI quantification and histological analysis. Moreover, exosomal LncRNA H19 was able to relieve pain levels during the early stages of cartilage repair in an animal experiment. Conclusion Our findings confirmed that mechanical stimulation can enhance exosome yield as well as biological function for the repair of cartilage defects. The underlying mechanism may be related to the high expression of LncRNA H19 in exosomes. The translational potential of this article: This study provides a theoretical support of optimizing exosome production. It advances the yield of mesenchymal stem cell exosome and facilitate the clinical application to repair of osteochondral damage.

Dataset Information

Platelet lysate induces chondrogenic differentiation of umbilical cord-derived mesenchymal stem cells by regulating the lncRNA H19/miR-29b-3p/SOX9 axis.

Publications

Platelet lysate induces chondrogenic differentiation of umbilical cord-derived mesenchymal stem cells by regulating the lncRNA H19/miR-29b-3p/SOX9 axis.

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