Copper Does Not Induce Tenogenic Differentiation but Promotes Migration and Increases Lysyl Oxidase Activity in Adipose-Derived Mesenchymal Stromal Cells

Abstract

Background. Copper belongs to the essential trace metals that play a key role in the course of cellular processes maintaining the whole body’s homeostasis. As there is a growing interest in transplanting mesenchymal stromal cells (MSCs) into the site of injury to improve the regeneration of damaged tendons, the purpose of the study was to verify whether copper supplementation may have a positive effect on the properties of human adipose tissue-derived MSCs (hASCs) which potentially can contribute to improvement of tendon healing. Results. Cellular respiration of hASCs decreased with increasing cupric sulfate concentrations after 5 days of incubation. The treatment with CuSO4 did not positively affect the expression of genes associated with tenogenesis (COL1α1, COL3α1, MKX, and SCX). However, the level of COL1α1 protein, whose transcript was decreased in comparison to a control, was elevated after a 5-day exposition to 25 μM CuSO4. The content of the MKX and SCX protein in hASCs exposed to cupric sulfate was reduced compared to that of untreated control cells, and the level of the COL3α1 protein remained unchanged. The addition of cupric sulfate to hASCs’ medium increased the activity of lysyl oxidase which was positively correlated with concentration of CuSO4. Moreover, a high level of CuSO4 stimulated the action of intracellular superoxide dysmutase. The hASC secretion profile after a 5-day exposure to 50 μM cupric sulfate differed from that of untreated cells and was similar to the secretion profile of human tenocytes. Additionally, cupric sulfate increased secretion of CXCL12 in hASCs. Furthermore, the exposition to the CuSO4 significantly increased directed migration of human ASCs in a dose-dependent manner. Conclusion. Copper sulfate supplementation can have a beneficial effect on tendon regeneration not by inducing tenogenic differentiation, but by improving the recruitment of MSCs to the site of injury, where they can secrete growth factors, cytokines and chemokines, and prevent the effects of oxidative stress at the site of inflammation, as well as improve the stabilization of collagen fibers, thereby accelerating the process of tendon healing.

Pro-angiogenic and antibacterial copper containing nanoparticles in PLGA/amorphous calcium phosphate bone nanocomposites

“…We conclude that ASCs are highly sensitive towards the CuO NPs incorporated in the nanocomposite. It has been reported that after a 5-day exposition to 25 μm CuSO 4 , cellular respiration of human ASCs was decreased as well as gene expression of collagen I [ 62 ], supporting our determination of lower ECM formation in the 5 w/w % CuO containing material compared to the control.…”

ATP7A-dependent copper sequestration contributes to termination of β-CATENIN signaling during early adipogenesis

“…We propose that Atp7a-mediated transient Cu deficit is beneficial for initiation of lipogenesis and lipid accumulation, and serves as an integral component of the entire differentiation program. The effects of Cu on cytoskeleton and cell morphology were previously observed in 3T3-L1 cells [ 23 ], as well as in other cell types [ 32 , 33 ]. Our studies suggest that β-catenin, which coordinates gene transcription and cell adhesion, could be an important factor in the observed morphological changes; however, the role of Cu in adipocyte morphology need to be studied further.…”

Integrative single-cell RNA and ATAC sequencing reveals that the FOXO1-PRDX2-TNF axis regulates tendinopathy

“…However, the cuproptosis level of the TDSC-7 cluster cells was abnormally decreased, indicating that these cells were in a precarious state. Copper is an essential trace metal, and proper copper supplementation has a beneficial effect on tendon regeneration and self-renewal ( 39 ). Herchenhan et al.…”

“…However, the cuproptosis level of the TDSC-7 cluster cells was abnormally decreased, indicating that these cells were in a precarious state. Copper is an essential trace metal, and proper copper supplementation has a beneficial effect on tendon regeneration and self-renewal (39). Herchenhan et al found that an appropriate copper concentration was crucial for maintaining lysyl oxidase activity, which was required for orderly collagen fibril formation in tenocytes (40).…”