Exosomes derived from bone marrow mesenchymal stem cells with chondromodulin-1 overexpression affect the proliferation of chondrocytes in osteoarthritis

doi:10.12307/2023.754

Abstract

Abstract: BACKGROUND: Chondromodulin-1 (Chm-1), which is highly expressed in normal cartilage, can inhibit angiogenesis, prevent chondrocyte hypertrophy and alleviate the development of osteoarthritis. Chm-1 plays an important role in the occurrence and development of osteoarthritis.
OBJECTIVE: To investigate the effect of exosomes (Exos) derived from bone marrow mesenchymal stem cells with high expression of Chm-1 on the proliferation of chondrocytes in osteoarthritis.
METHODS: Rat bone marrow mesenchymal stem cells were isolated, cultured and identified. Bone marrow mesenchymal stem cells were transfected with Chm-1 lentivirus and no-load lentivirus. NC-Exos and Chm-1-Exos in the supernatant of cell culture were extracted by ultra-high speed centrifugation. The model of osteoarthritis in vitro was established by treating chondrocytes with interleukin-1 β for 24 hours. Then osteoarthritis chondrocytes were treated with NC-Exos and Chm-1-Exos for 7 days, respectively, and PBS-treating osteoarthritis chondrocytes were used as a control group. The Exos uptake of osteoarthritis chondrocytes was observed by a confocal microscope. The proliferation of osteoarthritis chondrocytes was evaluated by CCK-8 assay. Expression levels of Chm-1 in NC-Exos and Chm-1-Exos were detected by western blot assay. The protein and mRNA expression levels of Col II, SOX9 and AGG in osteoarthritis chondrocytes were detected by western blot assay and RT-qPCR, respectively.
RESULTS AND CONCLUSION: (1) Bone marrow mesenchymal stem cells showed a typical fusiform shape and had the ability to differentiate into three lines. (2) Lentivirus with green fluorescence was successfully transfected into bone marrow mesenchymal stem cells. (3) Exos, whose size was between 40-150 nm, could be absorbed by osteoarthritis chondrocytes and expressed their specific marker proteins CD63, CD81 and TSG101. (4) Compared with NC-Exos, Chm-1-Exos expressed higher proteins of Chm-1 (P < 0.05). (5) At 24 hours after treatment, compared with the NC-Exos group, the Chm-1-Exos group presented higher cell viability. (6) At 7 days after treatment, compared with the NC-Exos group, the Chm-1-Exos group showed higher protein and mRNA expression levels of Col II, SOX9 and AGG (P < 0.05). (7) The above experimental results have shown that Chm-1-Exos can promote the proliferation of osteoarthritis chondrocytes.

Key words: osteoarthritis, bone marrow mesenchymal stem cell, exosome, chondrocyte, Chm-1

CLC Number:

Wu Zhiwen, Shen Enpu, Li Beibei, Liu Danping, Qi Hui. Exosomes derived from bone marrow mesenchymal stem cells with chondromodulin-1 overexpression affect the proliferation of chondrocytes in osteoarthritis[J]. Chinese Journal of Tissue Engineering Research, 2023, 27(33): 5277-5282.

Figures/Tables 7

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