Impact of dexamethasone concentration on cartilage tissue formation from human synovial derived stem cells in vitro

Abstract

Human synovial mesenchymal stem cells (hSMSCs) are a promising cell source for cartilage regeneration because of their superior chondrogenic potential in vitro. This study aimed to further optimize the conditions for inducing chondrogenesis of hSMSCs, focusing on the dose of dexamethasone in combination with transforming growth factor-β3 (TGFβ3) and/or bone morphogenetic protein-2 (BMP2). When hSMSCs-derived aggregates were cultured with TGFβ3, dexamethasone up to 10 nM promoted chondrogenesis, but attenuated it with heterogeneous tissue formation when used at concentrations over than 100 nM. On the other hands, BMP2-induced chondrogenesis was remarkably disturbed in the presence of more than 10 nM dexamethasone along with unexpected adipogenic differentiation. In the presence of both TGFβ3 and BMP2, dexamethasone dose dependently promoted cartilaginous tissue formation as judged by tissue volume, proteoglycan content, and type 2 collagen expression, whereas few adipocytes were detected in the formed tissue when cultures were supplemented with over 100 nM dexamethasone. Even in chondrogenic conditions, dexamethasone thus affected hSMSCs differentiation not only toward chondrocytes, but also towards adipocytes dependent on the dose and combined growth factor. These findings have important implications regarding the use of glucocorticoids in in vitro tissue engineering for cartilage regeneration using hSMSCs.

Effect of the small compound TD ‐198946 on glycosaminoglycan synthesis and transforming growth factor β3‐associated chondrogenesis of human synovium‐derived stem cells in vitro

“…Due to the lack of a significant effect of TD‐198946 on induction of chondrogenesis from hSSCs, we next investigated the potential synergistic effects of the molecule in TGFβ‐associated chondrogenic induction with hSSCs. TGFβs are some of the most popular cytokines to drive the differentiation of many stem cells towards chondrocytes; however, their ability to achieve this with hSSCs is known to be limited (Chijimatsu et al, ; Shirasawa et al, ). Consistently, TGFβ3 alone induced less chondrogenesis with a lack of staining for proteoglycan based on safranin O and aggrecan, as well as type 2 collagen staining (Figure a–c).…”

“…However, the current cartilaginous particles sometimes contained a heterogenic region at their centre exhibiting an absence of type 2 collagen. We recently reported that inadequate glucocorticoid signalling results in centric impairment in TGFβ3‐associated cartilage tissue formation from hSSCs and the concentration of dexamethasone possibly affected the results (Chijimatsu et al, ). TGFβs are known to upregulate not only type 2 collagen but also type 1 collagen and type 10 collagen (Dexheimer et al, ; Hara et al, ).…”

“…Thus, cotreatment with TD‐198946 and TGFβ3 is not effective regarding production of a genuine hyaline‐like cartilaginous tissue. Therefore, in the future, we need to address the use of TD‐198946 with other chondrogenic growth factors such as using bone morphogenetic proteins, insulin‐like growth factors, and growth differentiation factors (Chijimatsu et al, ; Yu, Han, & Kim, ).…”

“…In this regard, human synovium‐derived stem cells (hSSCs) have been identified as a good cell source based on their superior potential for chondrogenesis and proliferation compared with other adult stem cells in vitro (Sakaguchi, Sekiya, Yagishita, & Muneta, ). We have been developing several strategies for cartilage repair using hSSCs in vivo (Shimomura et al, ; Yasui, Ando, et al, ) and, moreover, have shown successful formation of cartilage‐like tissue from hSSCs in vitro (Chijimatsu et al, ; Koizumi et al, ; Yasui, Chijimatsu, et al, ). Although it will be potential alternatives for ACI, the cost to induce chondrogenesis using growth factors is not a negligible issue for clinical applications.…”

Molecular and Lifestyle Factors Modulating Obesity Disease

“…Several studies have demonstrated that a subtle crosstalk occurs between chondrogenesis and adipogenesis. The addition of dexamethasone to human synovium-derived stem cells during chondrogenic induction promotes also adipogenesis [10]. Adipogenic features, such as signet-ring morphology, have been observed in pericytes cultured in chondrogenic medium [11].…”