TIMP3/TGF‑β1 axis regulates mechanical loading‑induced chondrocyte degeneration and angiogenesis

Abstract

chondrocytes in injured cartilage tissue are susceptible to mechanical loading; mechanical overloading can induce cartilage degeneration. The aim of the present study was to investigate whether mechanical loading can regulate chondrocyte degeneration and angiogenesis via the tissue inhibitor of matrix metalloproteinase-3 (TiMP3)/transforming growth factor (TGF)-β1 axis. Primary human chondrocytes were obtained from knee articular cartilage of a healthy donor. Then, normal chondrocytes or TiMP3 lentivirus-transfected (lV-TiMP3) chondrocytes were subjected to mechanical loading (10 MPa compression). Then, chondrocytes were stimulated with 1 µg/ml lipopolysaccharide (lPS) or treated with ldn-193189 (inhibitor of TGF-β1 signaling pathway). in addition, human umbilical vein endothelial cells (HuVecs) were co-cultured with chondrocytes or lV-TiMP3 chondrocytes. The expression levels of collagen-i, proteoglycan, TiMP3, TGF-β1, Smad2 and Smad3 were detected by reverse transcription-quantitative Pcr and western blotting. Moreover, cell apoptosis and viability were determined using flow cytometry and MTT analysis, while cell migration was observed by Transwell assays. in addition, the vascular endothelial growth factor (VeGF)/VeGF receptor (r)2 binding rate in HuVecs was detected by a solid-phase binding assay. it was demonstrated that mechanical loading significantly inhibited the expression levels of collagen-i and proteoglycan in chondrocytes, as well as reducing cell proliferation and promoting cell apoptosis. in addition, the expression levels of TiMP3, TGF-β1, phosphorylated (p)-Smad2 and p-Smad3 were significantly decreased in degenerated chondrocytes that were induced by lPS, as well as in chondrocytes treated with ldn-193189. Furthermore, TiMP3 overexpression suppressed cell migration and reduced the VeGF/VeGFr2 binding rate in HuVecs. Mechanical loading significantly inhibited the expression levels of TiMP3, TGF-β1, p-Smad2 and p-Smad3 in chondrocytes, and also increased cell migration of HuVecs; TGF-β1 treatment or TiMP3 overexpression reversed these effects. Thus, the TiMP3/TGF-β1 axis may be a vital signaling pathway in mechanical loading-induced chondrocyte degeneration and angiogenesis.

Single-cell RNA sequence presents atlas analysis for chondrocytes in the talus and reveals the potential mechanism in coping with mechanical stress

“…One study has elucidated that ENO1 and NFKBIA are important factors in OA progression (Bhosale and Richardson, 2008), which may indicate a double-sided impact of ProCs, MirCs, and RegCs. VEGFA , a hub gene for MirCs and SpCs, and THBS1 , a hub gene for SpCs and RegCs, have been shown to be involved in angiogenesis ( Zhan and Cai, 2019 ; Zhao et al, 2020 ), which can be induced by mechanical loading ( Beckmann et al, 2014 ). Briefly, the 10 hub genes may indicate special functions for identified cell types and the whole talus cartilage, especially in managing mechanical loading.…”

“…However, in FCs, preHTCs and MirCs, it was identified as a hub gene; possibly the three cells manage to cope with hypoxia and mechanical loading, especially MirCs. (Zhan and Cai, 2019;Zhao et al, 2020), which can be induced by mechanical loading (Beckmann et al, 2014). Briefly, the 10 hub genes may indicate special functions for identified cell types and the whole talus cartilage, especially in managing mechanical loading.…”

Extracellular Vesicles Allow Epigenetic Mechanotransduction between Chondrocytes and Osteoblasts

“…Additionally, some reports propose that downstream target TIMP-3 is not only involved in the regulation of the matrix metalloproteinases but also is related to angiogenesis during the invasion and metastasis of cancer cells and the protection of myocardial infarction [58,59]. Recently, Zhao et al reported the TIMP3/TGF-β1 axis may be responsi-ble for the deterioration and angiogenesis of chondrocytes under mechanical loading [60], which may be related to the regulatory role of miR-221-3p as well in this context. contain miR-221-3p may affect osteoblasts via the subchondral bone microchannel network.…”

Senescent skeletal cells cross-talk with synovial cells plays a key role in the pathogenesis of osteoarthritis

“…This cycle was exemplified in a recent study showing that adipokine apelin (APLN) in synovial fibroblasts of patients with OA regulates the activity of cartilage, synovium, bone, and various immune cells, and is related to the pathogenesis of OA [103]. Mechanistically, APLN inhibits the expression of miRNA-144-3p and stimulates TGF β/Smad pathway Promote the hypertrophic differentiation of chondrocyte [93,94] Osteocyte RANKL/OPG; RANK-RANKL-OPG system Induce osteoclast differentiation and enhance bone resorption [95][96][97] VEGF; mTORC1 pathway Indirect regulation of chondrocytes by stimulating angiogenesis [93] Synovium fibroblasts AMPK and p38 pathways; APLN; PI3K and ERK pathway Knockdown of APLN expression could ameliorated changes in OA cartilage severity [103] Musculoskeletal cells Sarcolipin Sarcolipin secreted by senescent muscle cells promote skeletal muscle fibrosis and ultimately lead to sarcopenia, thereby accelerating the development of OA in terms of biomechanical mechanisms [104] Myokine, myostatin; Wnt/β-catenin signaling The muscle released myokine, myostatin inhibits osteogenic differentiation by suppressing Wnt/β-catenin signaling [105] the expression of IL-1β by activating the PI3K and ERK pathways [103]. Thus, downregulating the expression of APLN may ameliorate changes in OA cartilage severity.…”

“…The lack of SOST aggravates OA by modulating the turnover and cell apoptosis of the subchondral bone, thus destructing the bone homeostasis [100,101]. In addition, researchers have shown that osteoblasts can induce subchondral angiogenesis via the TGF-β1/Smad axis to indirectly affect the chondrocytes [93,94]. Thus, inhibiting the damage to subchondral bone at the early stage via these pathways may be considered a method to treat OA.…”