Preparation of EGCG decorated, injectable extracellular vesicles for cartilage repair in rat arthritis

Abstract Arthritis is a kind of chronic inflammatory autoimmune disease, which can destroy joint cartilage and bone, leading to joint pain, joint swelling, and limited mobility. Traditional therapies have many side effects or focus too much on anti-inflammation while neglecting joint repair. In this experiment, we combined EGCG (Epigallocatechin gallate) with extracellular vesicles derived from macrophages to treat rheumatoid arthritis. Sustained-release resulted in a significant decrease in chondrocyte expression of HIF-1α, a decrease in apoptosis-related proteins Cytochrome C, Caspase-3, Caspase-9, and Bax. Molecular biological analysis showed that extracellular vesicles-encapsulated EGCG (EVs-EGCG) more significantly upregulated type II collagen expression by about 1.8-fold than EGCG alone, which was more beneficial for arthritis repair. Animal experiments revealed that these EGCG-coated extracellular vesicles significantly reduced swelling, decreased synovial hyperplasia, repaired cartilage, and attenuated arthritis-related pathology scores in arthritic rats. Measurement data showed that EVs-EGCG treatment reduced joint swelling by approximately 39.5% in rheumatoid rats. In vitro studies have shown that this EVs-EGCG can increase the expression of cartilage type II collagen and reduce apoptosis of chondrocytes. Moreover, it was demonstrated in vivo experiments to reduce cartilage destruction in rheumatoid arthritis rats, providing a solution for the treatment of rheumatoid arthritis.

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Comparison of the Therapeutic Effects of Gold Nanoclusters and Gold Nanoparticles on Rheumatoid Arthritis

Journal of Biomedical Nanotechnology ◽

10.1166/jbn.2019.2848 ◽

2019 ◽

Vol 15 (11) ◽

pp. 2281-2290 ◽

Cited By ~ 3

Author(s):

Yao Zhao ◽

Zhesheng He ◽

Ruoping Wang ◽

Pengju Cai ◽

Xiangchun Zhang ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

Au Nanoparticles ◽

Type Ii Collagen ◽

Therapeutic Effects ◽

Type Ii ◽

Au Clusters ◽

Anti Inflammatory ◽

Inflammatory Effect

Rheumatoid arthritis (RA) is a chronic autoimmune disease characterized by synovial inflammation and progressive cartilage and bone damage. In our previous studies, we found that Au clusters using glutathione as a template (GACs) produced profound anti-inflammatory effects in vitro on lipopolysaccharide (LPS)-induced inflammation in mouse macrophage RAW 264.7 cells and type II collagen-induced rat RA in vivo. In this study, we examined whether the template for Au clusters synthesis has an effect on its anti-inflammatory effect and whether Au nanoparticles with larger particle diameter produce the same anti-inflammatory effect. We synthesized Au clusters with bovine serum albumin (BSA) as a template (BACs), Au clusters with glutathione (GSH) as a template (GACs), and Au nanoparticles with glutathione as a template (GANs) and compared their anti-inflammatory effects in vitro and in vivo. These three Au nanomaterials can inhibit the production of lipopolysaccharide (LPS)-induced proinflammatory mediators and ameliorate type II collagen-induced rat RA. However, although the three Au nanomaterials produced similar anti-inflammatory effects, the GANs with larger particle sizes were less stable in vivo and accumulated in the peritoneum after intraperitoneal injection, resulting in poor absorption in vivo. The BACs showed relatively high liver accumulation due to the larger molecular weight of the outer shell. Therefore, we believe that the GACs are potential reliable nanodrugs for the treatment of RA.

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Discovery and development of a type II collagen neoepitope (TIINE) biomarker for matrix metalloproteinase activity: From in vitro to in vivo

Analytical Biochemistry ◽

10.1016/j.ab.2006.10.034 ◽

2007 ◽

Vol 361 (1) ◽

pp. 93-101 ◽

Cited By ~ 59

Author(s):

O.V. Nemirovskiy ◽

D.R. Dufield ◽

T. Sunyer ◽

P. Aggarwal ◽

D.J. Welsch ◽

...

Keyword(s):

Matrix Metalloproteinase ◽

Type Ii Collagen ◽

Type Ii ◽

Metalloproteinase Activity

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Inhibitory effects of polyphenol punicalagin on type-II collagen degradation in vitro and inflammation in vivo

Chemico-Biological Interactions ◽

10.1016/j.cbi.2013.06.018 ◽

2013 ◽

Vol 205 (2) ◽

pp. 90-99 ◽

Cited By ~ 31

Author(s):

Dinorah Jean-Gilles ◽

Liya Li ◽

V.G. Vaidyanathan ◽

Roberta King ◽

Bongsup Cho ◽

...

Keyword(s):

Type Ii Collagen ◽

Collagen Degradation ◽

Type Ii ◽

Inhibitory Effects

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MicroRNA-21 from bone marrow mesenchymal stem cell-derived extracellular vesicles targets TET1 to suppress KLF4 and alleviate rheumatoid arthritis

Therapeutic Advances in Chronic Disease ◽

10.1177/20406223211007369 ◽

2021 ◽

Vol 12 ◽

pp. 204062232110073

Author(s):

Guo-Qing Li ◽

Yu-Xuan Fang ◽

Ying Liu ◽

Fan-Ru Meng ◽

Xia Wu ◽

...

Keyword(s):

Rheumatoid Arthritis ◽

Bone Marrow ◽

Extracellular Vesicles ◽

Mouse Fibroblast ◽

Luciferase Assay ◽

Loss Of Function ◽

Regulatory Relationship ◽

Dual Luciferase Assay

Background: Accumulating evidence has demonstrated that bone marrow mesenchymal stem cells (BMSCs)-derived extracellular vesicles (EVs) can be used effectively to transfer drugs and biomolecules to target lesions. Meanwhile, BMSCs have been reported to be beneficial in the treatment of rheumatoid arthritis (RA). In this study, we employ gain- and loss-of-function experiments to determine how BMSCs-derived EVs alleviate RA in vitro and in vivo. Methods: We isolated EVs from BMSCs and characterized them by transmission electron microscopy and western blot analysis. The regulatory relationship between miR-21 and TET1 was predicted by bioinformatics analysis and validated by dual luciferase assay. Next, we utilized bisulfite sequencing PCR to decipher how TET1 promoted KLF4 transcription. Then, we established an RA mouse model and determined the role of miR-21 in RA progression. Functional assays were used to validate the role the miR-21-TET1-KLF4 regulatory axis in controlling mouse fibroblast-like synoviocytes (mFLS) cell proliferation and inflammatory cytokines secretion in vitro. Results: RT-qPCR results revealed that miR-21 was highly expressed in BMSCs-derived EVs, and confirmed that BMSCs-derived EVs transferred miR-21 into mFLS cells. Bioinformatic analysis predicted that TET1 was the directly downstream target of miR-21, which was further validated by dual luciferase assay. TET1 promoted KLF4 promoter methylation to increase its expression. Collectively, BMSCs-derived EVs relieved RA by delivering miR-21, while the exosomal miR-21 alleviated RA through targeting the TET1/KLF4 regulatory axis. Conclusion: miR-21 from BMSCs-derived EVs suppresses KLF4 to relive RA by targeting TET1.

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A Novel in Vitro Model to Investigate Behavior of Articular Chondrocytes in Osteoarthritis

The Journal of Rheumatology ◽

10.3899/jrheum.080080 ◽

2009 ◽

Vol 37 (2) ◽

pp. 426-431 ◽

Cited By ~ 2

Author(s):

KENNETH S. RANKIN ◽

RACHEL L. LAKEY ◽

CRAIG H. GERRAND ◽

ANDREW P. SPROWSON ◽

ANDREW W. McCASKIE ◽

...

Keyword(s):

Molecular Mechanisms ◽

Articular Chondrocytes ◽

Type Ii Collagen ◽

Type Ii ◽

Transcriptional Expression ◽

Matrix Metalloproteinase 1 ◽

Standard Culture ◽

Cells Cultured

Objective. To investigate in vivo simulation of the microenvironment in which osteoarthritis (OA) chondrocytes are cultured in vitro.Methods. Human articular chondrocytes were cultured under normoxic and hypoxic conditions. Cells were cultured on standard culture plastic or a porous polyHEMA surface that closely resembles the in vivo cartilage microarchitecture. Morphological changes to the cells were demonstrated by fluorescent staining with DAPI and vinculin. Proteoglycan and type II collagen protein levels were assessed using established techniques. Matrix metalloproteinase-1 (MMP-1) production was assessed by ELISA. The gene expression of type II collagen and SOX9 was measured using real-time polymerase chain reaction.Results. Cells grown on culture plastic were seen to be flat and hexagonal. Cells cultured on the porous polyHEMA surface exhibited morphology in keeping with the in vivo microenvironment. Glycosaminoglycan release in hypoxia was high from cells cultured on standard culture plastic. Transcriptional expression of type II collagen was upregulated in hypoxia and by culture on the polyHEMA surface. Transcriptional expression of SOX9 in hypoxia was upregulated compared to normoxia; no significant effect was seen by varying the culture surface. Translational expression of type II collagen was upregulated at 20% oxygen on the polyHEMA surface compared to culture plastic and this was related to MMP-1 expression.Conclusion. Culture of chondrocytes in hypoxia and on a porous surface simulates the in vivo microenvironment and illustrates the molecular mechanisms of OA.

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Erratum to “Discovery and development of a type II collagen neoepitope (TIINE) biomarker for matrix metalloproteinase activity: From in vitro to in vivo” [Anal. Biochem. 361 (2007) 93–113]

Analytical Biochemistry ◽

10.1016/j.ab.2007.04.026 ◽

2007 ◽

Vol 370 (2) ◽

pp. 258 ◽

Cited By ~ 1

Author(s):

O.V. Nemirovskiy ◽

D.R. Dufield ◽

T. Sunyer ◽

P. Aggarwal ◽

D.J. Welsch ◽

...

Keyword(s):

Matrix Metalloproteinase ◽

Type Ii Collagen ◽

Type Ii ◽

Metalloproteinase Activity

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Method for in vitro production of cartilage microtissues from scaffold-free spheroids composed of human adipose-derived stem cells

Biomedical Research and Therapy ◽

10.15419/bmrat.v7i4.597 ◽

2020 ◽

Vol 7 (4) ◽

pp. 3697-3708

Author(s):

Vy Thi-Kieu Tu ◽

Ha Thi-Ngan Le ◽

Xuan Hoang-Viet To ◽

Phuc Dang-Ngoc Nguyen ◽

Phat Duc Huynh ◽

...

Keyword(s):

Stem Cells ◽

Cartilage Damage ◽

Cartilage Regeneration ◽

Type Ii Collagen ◽

Adipose Derived Stem Cells ◽

Type Ii ◽

In Vitro Production ◽

Engineered Cartilage

Introduction: Cartilage damage is one of the injuries that is difficult for the human body to self-repair due to the avascular and completely mature tissue with only few stem or progenitor cells present. Recently, some studies showed that engineered cartilage tissues could be used to treat or improve this injury. This study aimed to produce the cartilage microtissues from the differentiation of scaffold-free spheroids composed of human adipose-derived stem cells. Methods: Human adipose-derived stem cells (ADSCs) were isolated and expanded following the previously published study. They were then cultured in the non-adherent condition to produce spheroids. The spheroids of the ADSCs were collected and induced into cartilage microtissues in the inducible medium for 21 days. The cartilage microtissue was characterized by some cartilage phenotype markers, including the accumulation of extracellular matrix proteins (aggrecan, glycosaminoglycan, and type II collagen), and the expression of certain genes specific to chondrocytes (Sox9, Col2, Col1, and Acan). Results: The results showed that the expression of chondrocyte-specific genes gradually increased during the 21 days of culture for differentiation. On day 21, the microtissues expressed aggrecan, glycosaminoglycan, and type II collagen proteins. Conclusion: This study demonstrated that cartilage microtissues could easily be produced from scaffold-free spheroids from ADSCs. Thus, cartilage microtissues can be produced in this manner for in vivo transplantation to promote cartilage regeneration, or to produce cartilage tissues for in vitro studies.

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