Sulfated hyaluronic acid and dexamethasone possess a synergistic potential in the differentiation of osteoblasts from human bone marrow stromal cells

Abstract

The development of novel bioactive biomaterials is urgently needed to meet the needs of an aging population. Both sulfated hyaluronic acid and dexamethasone are candidates for the functionalization of bone grafts, as they have been shown to enhance the differentiation of osteoblasts from bone marrow stromal cells in vitro and in vivo. However, the underlying mechanisms are not fully understood. Furthermore, studies combining different approaches to assess synergistic potentials are rare. In this study, we aim to gain insights into the mode of action of both sulfated hyaluronic acid and dexamethasone by a comprehensive analysis of the cellular fraction, released matrix vesicles, and the extracellular matrix, combining classical biochemical assays with mass spectrometry–based proteomics, supported by novel bioinformatical computations. We found elevated differentiation levels for both treatments, which were further enhanced by a combination of sulfated hyaluronic acid and dexamethasone. Single treatments revealed specific effects on osteogenic differentiation. Dexamethasone activates signalling pathways involved in the differentiation of osteoblasts, for example, CXC‐motif chemokine receptor type 4 and mitogen‐activated protein kinases. The effects of sulfated hyaluronic acid were predominantly linked to an alteration in the composition of the extracellular matrix, affecting the synthesis, secretion, and/or activity of fibrillary (fibronectin and thrombospondin‐2) and nonfibrillary (transglutaminase‐2, periostin, and lysyloxidase) extracellular matrix components, including proteases and their inhibitors (matrix metalloproteinase‐2, tissue inhibitor of metalloproteinase‐3). The effects were treatment specific, and less additive or contrary effects were found. Thus, we anticipate that the synergistic action of the treatment‐specific effects is the key driver in elevated osteogenesis.

Glucocorticoid Treatment Leads to Aberrant Ion and Macromolecular Transport in Regenerating Zebrafish Fins

“…Transcriptome analysis of osteoblasts isolated from fin regenerates undergoing prednisolone treatment for 4 days shows the same trend with more genes being upregulated than downregulated (data not shown). Notably, a similar phenomenon has been observed in MC3T3-E1 and hBMSC cells undergoing dexamethasone treatment and subsequent LC-MS SILAC analysis (41, 42). This may illustrate the strong transactivational capacities of glucocorticoids in selected tissues and demonstrate increased gene expression during the course of regeneration.…”

“…Our protein abundance-focused approach yielded interesting results. Differential expression analysis, functional clustering and WGCNA, which recently has been implemented in the proteome analysis of dexamethasone treated human bone marrow stromal cells (hBMSC) in vitro (41), point to profound changes in ECM composition and (calcium) ion binding capacity during regeneration, as well as altered cation transmembrane transporter activity and macromolecule localization upon prednisolone exposure. Interestingly, both injury and glucocorticoid treatment predominantly increase the abundance of sensitive proteins rather than decreasing it (197 vs. 106 proteins and 67 vs. 36 proteins, respectively).…”

Prospects and challenges of multi-omics data integration in toxicology

“…Untargeted proteomic measurements currently mainly rely on liquid chromatography (LC), coupled to mass spectrometry (MS). Most frequently, proteins are extracted, optionally separated, digested and the proteolytic peptides analyzed by LC-MS/MS, which typically allows for the detection of 3000-5000 proteins per experiment, e.g., Schmidt et al (2018). Quantitative detection can be improved in terms of reliability using labeling approaches such as stable isotope labeling (Ong et al 2002).…”

Osteogenic and Chondrogenic Potential of the Supramolecular Aggregate T-LysYal®

“…Stem cell differentiation is affected by the micro-environment: the ECM governs maintenance and propagation of MSCs (24) and it contains macromolecules such as collagen, adhesive glycoprotein, and GAG. HA, which is a ubiquitarian GAG, has been demonstrated to stimulate MSCs differentiation by affecting the ECM composition (25).…”