Hydrostatic Pressure Regulates MicroRNA Expression Levels in Osteoarthritic Chondrocyte Cultures via the Wnt/β-Catenin Pathway

Abstract

Mechanical loading and hydrostatic pressure (HP) regulate chondrocytes’ metabolism; however, how mechanical stimulation acts remain unclear. MicroRNAs (miRNAs) play an important role in cartilage homeostasis, mechanotransduction, and in the pathogenesis of osteoarthritis (OA). This study investigated the effects of a cyclic HP (1–5 MPa), in both normal and OA human chondrocytes, on the expression of miR-27a/b, miR-140, miR-146a/b, and miR-365, and of their target genes (MMP-13, ADAMTS-5, IGFBP-5, and HDAC-4). Furthermore, we assessed the possible involvement of Wnt/β-catenin pathway in response to HP. Chondrocytes were exposed to HP for 3h and the evaluations were performed immediately after pressurization, and following 12, 24, and 48 h. Total RNA was extracted and used for real-time PCR. β-catenin was detected by Western blotting analysis and immunofluorescence. In OA chondrocytes, HP induced a significant increase (p < 0.01) of the expression levels of miR-27a/b, miR-140, and miR-146a, and a significant reduction (p < 0.01) of miR-365 at all analyzed time points. MMP-13, ADAMTS-5, and HDAC-4 were significantly downregulated following HP, while no significant modification was found for IGFBP-5. β-catenin levels were significantly increased (p < 0.001) in OA chondrocytes at basal conditions and significantly reduced (p < 0.01) by HP. Pressurization did not cause any significant modification in normal cells. In conclusion, in OA chondrocytes, HP restores the expression levels of some miRNAs, downregulates MMP-13, ADAMTS-5, and HDAC-4, and modulates the Wnt/β-catenin pathway activation.

Elevated hydrostatic pressure promotes ameloblastoma cell invasion through upregulation of MMP‐2 and MMP‐9 expression via Wnt/β‐catenin signalling

“…Hens et al reported that Wnt/β‐catenin signalling was stimulated in calvarial cells in vitro after being subjected to mechanical loading such as stretching. In contrast, Cheleschi et al suggested that β‐catenin levels were significantly reduced by increased HP in osteoarthritis chondrocytes. In human osteoblastic cells subjected to 15 minutes of stretching, Wnt/β‐catenin signalling was downregulated, despite an initial early increase in β‐catenin levels .…”

The effects of different hydrostaticpressures on gene expression in vertebral cartilage endplate cells

“…After previus studies focused on IVD cells, we noticed the cartilage endplate is the vital organ for IVD [1,2,8,9]. There are two nutritional pathways for the IVD: the annulus fibrous pathway and the cartilage endplate pathway which is much more important for nutrition supply [12,13].…”

“…We thought the cartilage endplate was disturbed by hydrostatic pressurization in human IVD. There is a little apparent study on the gene expression of cartilage endplate cells with the application of hydrostatic pressurization [8,9]. Therefore, we aimed at anabolic/catabolic metabolic alterations of cartilage endplate cells in response to various magnitudes and durations of compression.…”

Effect of centrifugal force on the development of articular neocartilage with bovine primary chondrocytes

“…However, even though dynamic compressive loading of chondrocytes has been shown to have a positive effect on the expression of chondrogenic genes in the majority of studies, some controversial and negative effects have been shown as well (Anderson and Johnstone 2017). Various models, such as hydrostatic pressure (Cheleschi et al 2017;De Palma et al 2018;Parkkinen et al 1993;Pascarelli et al 2015) and centrifugation (Kim et al 2017), have been used to investigate the influence of mechanical forces at the cellular level in normal and/or osteoarthritic human chondrocytes. Hydrostatic pressure (1-5 MPa, frequency 0.25 Hz for a period of 3 h) upregulated some microRNAs but downregulated MMP-13, ADAMTS-5 and HDAC-4 via Wnt/β-catenin or NF-kB pathway (Cheleschi et al 2017;De Palma et al 2018).…”

“…Various models, such as hydrostatic pressure (Cheleschi et al 2017;De Palma et al 2018;Parkkinen et al 1993;Pascarelli et al 2015) and centrifugation (Kim et al 2017), have been used to investigate the influence of mechanical forces at the cellular level in normal and/or osteoarthritic human chondrocytes. Hydrostatic pressure (1-5 MPa, frequency 0.25 Hz for a period of 3 h) upregulated some microRNAs but downregulated MMP-13, ADAMTS-5 and HDAC-4 via Wnt/β-catenin or NF-kB pathway (Cheleschi et al 2017;De Palma et al 2018). Centrifugation is the most useful and simplest method, which does not need complicated devices and is not affected by the surrounding environment.…”