Circular RNA CREBBP modulates cartilage degradation by activating the Smad1/5 pathway through the TGFβ2/ALK1 axis

Xu Yiyang,Mao Guping,Long Dianbo,Deng Zengfa,Xin Ruobin,Zhang Ziji,Xue Ting,Liao Weiming,Xu Jie,Kang Yan 생화학분자생물학회 2022 Experimental and molecular medicine Vol.54 No.-

Osteoarthritis, characterized by articular cartilage degradation, is the leading cause of chronic disability in older adults. Studies have indicated that circular RNAs are crucial regulators of chondrocyte development and are involved in the progression of osteoarthritis. In this study, we investigated the function and mechanism of a circular RNA and its potential for osteoarthritis therapy. The expression levels of circCREBBP, screened by circular RNA sequencing during chondrogenic differentiation in adipose tissue-derived stem cells, and TGFβ2 were significantly increased in the cartilage of patients with osteoarthritis and IL-1β-induced chondrocytes. circCREBBP knockdown increased anabolism in the extracellular matrix and inhibited chondrocyte degeneration, whereas circCREBBP overexpression led to the opposite effects. Luciferase reporter assays, rescue experiments, RNA immunoprecipitation, and RNA pulldown assays confirmed that circCREBBP upregulated TGFβ2 expression by sponging miR-1208, resulting in significantly enhanced phosphorylation of Smad1/5 in chondrocytes. Moreover, intra-articular injection of adeno-associated virus-sh-circCrebbp alleviated osteoarthritis in a mouse model of destabilization of the medial meniscus. Our findings reveal a critical role for circCREBBP in the progression of osteoarthritis and provide a potential target for osteoarthritis therapy.

Polymerized Ionic Liquid for the Regulation of Phase Structure of PLA/PCL Blends

Yiyang Zhou,Qiuyue Meng,Ping Wang,Haibing Wei,Pei Xu,Yunsheng Ding 한국고분자학회 2022 Macromolecular Research Vol.30 No.9

Polymerized ionic liquid containing block structure (PIL) has been used as regulator for the phase structure of poly(lactide) (PLA)/poly(ε-caprolactone) (PCL) blends. The phase structure, crystallization, rheology behavior and mechanical properties of PLA/PCL and PLA/PCL/PIL blends were systematically investigated. PIL could be located in PCL phase or at the PLA/PCL interface, enhancing the interaction between polymer components. The crystallization ability of PLA and PCL was simultaneously enhanced with the addition of PIL, since the plasticization effect of ionic moiety and PEO segments in PIL as well as the nucleation effect of PIL-formed ionic cluster. When PIL content was 0.5 wt%, the PLA/PCL/0.5PIL blend system exhibited much better mechanical properties than additivefree PLA/PCL blend. But as PIL contents increased, the significantly change in viscosity ratio between PLA and PCL would lead to obviously change in phase structure of PLA/PCL blend, thus the mechanical properties of the blends would be degraded.

Exosomes Derived from Human Adipose Mesenchymal Stem Cells Inhibits Fibrosis and Treats Oral Submucous Fibrosis via the miR-181a-5p/Smad2 Axis

Shao Zifei,Xu Jinhao,Xu Xiaoyang,Wang Xiang,Zhou Yuxi,Li Yiyang,Li Kun 한국조직공학과 재생의학회 2024 조직공학과 재생의학 Vol.21 No.1

BACKGROUND: Oral submucous fibrosis (OSF) is a chronic disease with carcinogenic tendency that poses a non-negligible threat to human health. Exosomes derived from human adipose mesenchymal stem cells (ADSC-Exo) reduces visceral and cutaneous fibroses, but their role in OSF has received little attention. The aim of this study was to investigate the effects of ADSC-Exo on OSF and elucidate the mechanism. METHODS: In brief, ADSCs were extracted from adipose tissues and subjected to flow cytometry and induction culture. Fibroblasts were isolated from human buccal mucosa and subjected to immunofluorescence. Myofibroblasts were obtained from fibroblasts induced by arecoline and identified. Immunofluorescence assay confirmed that myofibroblasts could take up ADSC-Exo. The effects of ADSC-Exo on the proliferative and migratory capacities of myofibroblasts were examined using the Cell Counting Kit-8 and scratch assay. Real-time quantitative polymerase chain reaction (qPCR) was performed to evaluate mothers against decapentaplegic homolog 2 (Smad2), Smad3, Smad7, collagen type 1 (Col1), Col3, alpha smooth muscle actin (α-SMA), fibronectin, and vimentin. Western blotting was performed to detect phospho (p)-Smad2, Smad2, p-Smad2/3, Smad2/3, Smad7, Col1, Col3, α-SMA, fibronectin, and vimentin. Furthermore, the dual-luciferase reporter assay was performed to prove that miR-181a-5p in ADSC-Exo directly inhibited the expression of Smad2 mRNA to regulate the transforming growth factor beta (TGF-β) pathway. We also performed qPCR and western blotting to verify the results. RESULTS: ADSC-Exo could promote the proliferation and migration of myofibroblasts, reduce the expressions of p-smad2, Smad2, p-smad2/3, Smad2/3, Col1, αSMA, fibronectin, and vimentin and elevated the levels of Smad7 and Col3. In addition, miR-181a-5p was highly expressed in ADSC-Exo and bound to the 3'-untranslated region of Smad2. ADSC-Exo enriched with miR-181a-5p reduced collagen production in myofibroblasts and modulated the TGF-β pathway. CONCLUSIONS: ADSC-Exo promoted the proliferative and migratory capacities of myofibroblasts and inhibited collagen deposition and trans-differentiation of myofibroblasts in vitro. miR-181a-5p in exosomes targets Smad2 to regulate the TGF-β pathway in myofibroblasts. ADSC-Exo perform antifibrotic actions through the miR-181a-5p/Smad2 axis and may be a promising clinical treatment for OSF. BACKGROUND: Oral submucous fibrosis (OSF) is a chronic disease with carcinogenic tendency that poses a non-negligible threat to human health. Exosomes derived from human adipose mesenchymal stem cells (ADSC-Exo) reduces visceral and cutaneous fibroses, but their role in OSF has received little attention. The aim of this study was to investigate the effects of ADSC-Exo on OSF and elucidate the mechanism. METHODS: In brief, ADSCs were extracted from adipose tissues and subjected to flow cytometry and induction culture. Fibroblasts were isolated from human buccal mucosa and subjected to immunofluorescence. Myofibroblasts were obtained from fibroblasts induced by arecoline and identified. Immunofluorescence assay confirmed that myofibroblasts could take up ADSC-Exo. The effects of ADSC-Exo on the proliferative and migratory capacities of myofibroblasts were examined using the Cell Counting Kit-8 and scratch assay. Real-time quantitative polymerase chain reaction (qPCR) was performed to evaluate mothers against decapentaplegic homolog 2 (Smad2), Smad3, Smad7, collagen type 1 (Col1), Col3, alpha smooth muscle actin (α-SMA), fibronectin, and vimentin. Western blotting was performed to detect phospho (p)-Smad2, Smad2, p-Smad2/3, Smad2/3, Smad7, Col1, Col3, α-SMA, fibronectin, and vimentin. Furthermore, the dual-luciferase reporter assay was performed to prove that miR-181a-5p in ADSC-Exo directly inhibited the expression of Smad2 mRNA to regulate the transforming growth factor beta (TGF-β) pathway. We also performed qPCR and western blotting to verify the results. RESULTS: ADSC-Exo could promote the proliferation and migration of myofibroblasts, reduce the expressions of p-smad2, Smad2, p-smad2/3, Smad2/3, Col1, αSMA, fibronectin, and vimentin and elevated the levels of Smad7 and Col3. In addition, miR-181a-5p was highly expressed in ADSC-Exo and bound to the 3'-untranslated region of Smad2. ADSC-Exo enriched with miR-181a-5p reduced collagen production in myofibroblasts and modulated the TGF-β pathway. CONCLUSIONS: ADSC-Exo promoted the proliferative and migratory capacities of myofibroblasts and inhibited collagen deposition and trans-differentiation of myofibroblasts in vitro. miR-181a-5p in exosomes targets Smad2 to regulate the TGF-β pathway in myofibroblasts. ADSC-Exo perform antifibrotic actions through the miR-181a-5p/Smad2 axis and may be a promising clinical treatment for OSF.

Electrospinning Inorganic/Organic Nanohybridization Membranes with Hydrophobic and Oleophobic Performance

Tingting Zhang,Zihui Li,Ya Liu,Kangjia Ding,Yangfeng Guo,Yiyang Xu,Mengfan Sun,Dongfang Wang,Qian Li 한국섬유공학회 2023 Fibers and polymers Vol.24 No.12

Polytetrafluoroethylene (PTFE) nano-porous membrane has been widely used in various fields due to its excellent thermal stability and chemical stability. However, PTFE nanofiber membranes with simultaneous hydrophobic and oleophobic properties are essential to promote the application of PTFE. Here, based on inorganic/organic nanohybridization, we report a strategy for constructing nanostructures on fibers by adding nanoparticles, such as MOF and ZrO2. The results of FTIR and XRD confirmed MOF was synthesized successfully. At the same time, the SEM results showed UiO-66-(COOH)2 is spherical with an average diameter of 152 nm, and there is no agglomeration, which is suitable for electrospinning. Further, MOF and ZrO2 were payload into PTFE nanofibers. The results of SEM and AFM confirmed nanostructures will be more uniform and pronounced with the increase of UiO-66-(COOH)2 content, and nanostructures are most obvious when the content of UiO-66-(COOH)2 is 15%. The introduced nanostructures can increase the oil contact angle of the PTFE nano-porous membrane to 110° without introducing other groups, and further improve the water contact angle from 133° to 145°. Meanwhile, the introduction of a certain amount of hydrophilia groups can increase the oil contact angle to more than 120°. The simple strategy is of great significance to expand the application of PTFE fiber membrane in dealing with waste water treatment fields.