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KMID : 1377020230200050753
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Tissue Engineering and Regenerative Medicine
2023 Volume.20 No. 5 p.753 ~ p.766
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Thrombospondin-2 Couples Pressure-Promoted Chondrogenesis through NF-κB Signaling
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Jing Niu
Fan Feng
Songbai Zhang
Yue Zhu
Runfang Song
Junrong Li
Liang Zhao
Hui Wang
Ying Zhao
Min Zhang
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Abstract
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BACKGROUND : Our previous studies found that the mechanical stimulation promote chondrogenic differentiation of bone marrow mesenchymal stem cells (BMSCs), along with up-regulation of thrombospondin-2 (TSP-2). The aim of this study was to explore the effect of thrombospondin-2 (TSP-2) on the mechanical pressure-stimulated chondrogenic differentiation of BMSCs and the possible role of NF-κB signaling in the mechano-chemical coupling regulation toward chondrogenesis.
METHODS : Rat BMSCs were isolated, cultured and identified. The time-dependent expressions of TSP-2 and Sox9 in BMSCs under a dynamic mechanical pressure of 0?120 kPa at 0.1 Hz for 1 h were tested by qPCR and Western blotting. The role of TSP-2 in chondrogenic differentiation of BMSCs under mechanical pressure was validated by using small interfering RNA. The impact of TSP-2 and mechanical pressure on chondrogenesis were detected and the downstream signaling molecules were explored using Western blotting.
RESULTS : Mechanical pressure stimulation of 0?120 kPa for 1 h significantly upregulated the expression of TSP-2 in BMSCs. The expression of the chondrogenesis markers Sox9, Aggrecan, and Col-II were all upregulated under dynamic mechanical pressure or TSP-2 stimulation. Additional exogenous TSP-2 may potentiate the chondrogenic effect of mechanical stimulation. After knock down TSP-2, the upregulation of Sox9, Aggrecan and Col-II under mechanical pressure was inhibited. The NF-κB signaling pathway responded to both dynamic pressure and TSP-2 stimulation, and the cartilage-promoting effect was blocked by an NF-κB signaling inhibitor.
CONCLUSION : TSP-2 plays an essential role in the chondrogenic differentiation of BMSCs under mechanical pressure. NF-κB signaling is involved in the mechano-chemical coupling of TSP-2 and mechanical pressure for the chondrogenic differentiation of BMSCs.
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KEYWORD
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Thrombospondin-2 (TSP-2), Mechanical pressure, Bone marrow mesenchymal stem cells (BMSCs), Chondrogenesis, Nuclear factor kappa-B (NF-jB) signaling
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