Exosomes from Kartogenin-Pretreated Infrapatellar Fat Pad Mesenchymal Stem Cells Enhance Chondrocyte Anabolism and Articular Cartilage Regeneration

Abstract

Objective. To evaluate the effect of Kartogenin-pretreated exosomes derived from infrapatellar fat pad mesenchymal stem cells on chondrocyte in vitro and articular cartilage regeneration in vivo. Methods. Infrapatellar fat pad mesenchymal stem cells (IPFP-MSCs) were isolated from rabbits to harvest exosomes. After identification of mesenchymal stem cells and exosomes, rabbit chondrocytes were divided into three groups for further treatment: the EXO group (chondrocytes treated with exosomes isolated from infrapatellar fat pad mesenchymal stem cells), KGN-EXO group (chondrocytes treated with exosomes isolated from infrapatellar fat pad mesenchymal stem cells pretreated with KGN), and control group. After processing and proliferation, phenotypic changes of chondrocytes were measured. In the in vivo study, 4 groups of rabbits with articular cartilage injury were treated with KGN-EXO, EXO, IPFP-MSCs, and control. Macroscopic evaluation and histological evaluation were made to figure out the different effects of the 4 groups on cartilage regeneration in vivo. Results. The proliferation rate of chondrocytes in the EXO or KGN-EXO group was significantly higher than that in the control group ( P < 0.05 ). The qRT-PCR results showed that the expression of Sox-9, Aggrecan, and Col II was the highest in the KGN-EXO group compared with the EXO group and the control group ( P < 0.05 ). The results of Western blot were consistent with the results of qRT-PCR. In vivo, the cartilage defects in the KGN-EXO group showed better gross appearance and improved histological score than those in IPFP-MSC groups, EXO groups, and control groups ( P < 0.05 ). At 12 weeks, the defect site in the KGN-EXO group was almost completely repaired with a flat and smooth surface, while a large amount of hyaline cartilage-like structures and no obvious cracks were observed. Conclusion. Our study demonstrates that the exosomes isolated from infrapatellar fat pad mesenchymal stem cells pretreated with KGN have potent ability to induce chondrogenic differentiation of stem cells, effectively promoting the proliferation and the expression of chondrogenic proteins and genes of chondrocytes. The KGN-EXO can also promote the repair of articular cartilage defects more effectively, which can be used as a potential therapeutic method in the future.

Scrapie-Responsive Gene 1 Promotes Chondrogenic Differentiation of Umbilical Cord Mesenchymal Stem Cells via Wnt5a

“…At 12 weeks after surgery, the rabbits were sacrificed under deep anesthesia, and femoral heads were isolated for imaging and macroscopic evaluation according to the guidelines of the International Cartilage Repair Society (ICRS) scoring system [ 20 ]. Subsequently, each femoral head was fixed in 4% PFA for 48 h and decalcified for 3 weeks.…”

The Role of Extracellular Vesicles in the Pathogenesis, Diagnosis, and Treatment of Osteoarthritis

“…Qiu et al found that miR-143 and miR-124 were upregulated in the mouse OA model after treatment of EVs derived from curcumin-treated MSC, and the normal expression of NF-kB and Rho-associated coiled-coil containing protein kinase 1 (ROCK1) was significantly restored, resulting in weakened progression of OA [ 55 ]. Studies have shown that EVs isolated from rabbit subpatellar fat pad mesenchymal stem cells pretreated with Kartogenin (KGN) have the effective ability to induce chondrogenic differentiation of stem cells, effectively promote the proliferation and expression of chondrogenic proteins and genes of chondrocytes, and are more effective to promote the repair of articular cartilage defects [ 56 ]. Similarly, EVs derived from KGN pretreated SD rat BMSC are more effective than EVs derived from BMSC in cartilage repair [ 57 ].…”

Extracellular Vesicles: a Promising cell-free Therapy for Cartilage Repair

“…reported that EVs derived from kartogenin-preconditioned BMSCs enhanced chondral matrix synthesis and reduced degradation; thus this approach appears more effective for cartilage repair than the use of EVs from BMSCs without pretreatment with kartogenin [ 103 ]. A study using infrapatellar fat pad MSCs showed a similar result: EVs pretreated with kartogenin more effectively promoted articular cartilage defect repair [ 56 ]. Thus by altering the cellular environment through the addition of chemical compounds to the cell culture medium, the efficiency of the resultant EVs is improved.…”