Chondrogenic differentiation of human bone marrow-derived mesenchymal stem cells treated by GSK-3 inhibitors

Abstract

A study of the cartilage differentiation of mesenchymal stem cells (MSCs) would be of particular interest since one strategy for cell-based treatment of cartilage defects emphasizes the use of cells that are in a differentiated state. The present study has attempted to evaluate the effects of two well-known glycogen synthase kinase-3 inhibitors, including lithium chloride (LiCl) and SB216763 on a human marrow-derived MSC (hMSC) chondrogenic culture. Passaged-3 MSCs were condensed into small pellets and cultivated in the following groups based on the supplementation of chondrogenic medium: transforming growth factor (TGF)-β1, TGF-β1 + LiCl, TGF-β1 + SB216763, TGF-β3, TGF-β3 + LiCl, and TGF-β3 + SB216763. The cultures were maintained for 21 days and then analyzed for expression of Sox9, aggrecan, collagen II, β-catenin, and axin genes. Deposition of glycosaminoglycan (GAG) in the cartilage matrix was also measured for certain cultures. The presence of both LiCl and SB216763 along with TGF-β in the MSC chondrogenic culture led to the up-regulation of cartilage-specific genes. TGF-β3 appeared much better than TGF-β1. Based on our findings, SB216763 was more effective in up-regulation of cartilage-specific genes. These chondrogenic effects appeared to be mediated through the Wnt signaling pathway since β-catenin and axin tended to be up-regulated and down-regulated, respectively. In the culture with SB216763 + TGF-β3, significantly more GAG was deposited (P < 0.05). In conclusion, addition of either SB216763 or LiCl to hMSC chondrogenic culture up-regulates cartilage-specific gene expression and enhances GAG deposition in the culture.

Lithium-silicate sol–gel bioactive glass and the effect of lithium precursor on structure–property relationships

“…Arioka et al showed that lithium chloride inhibits GSK-3, one of the main regulators of the Wnt/b-catenin pathway, which plays a crucial role in the differentiation of osteoblasts. In addition, Hui et al [2] demonstrated that lithium can reduce the degradation of collagen via a cytokine-induced pro-inflammatory response in cartilage and Eslaminejad et al [8] showed that lithium enhances the formation of proteoglycan-rich extracellular matrix in chondrogenic culture of mesenchymal stem cells. These findings are of particular interest since cartilage is one of the most challenging tissues to regenerate, since it is avascular.…”

“…2 Results and discussion 2.1 Effect of the lithium salt on the sol-gel process Eslaminejad et al [5,8] studied the effect of Li þ concentration on mesenchymal stem cells, from 1 to 10 mM. They found 5 mM to be optimal in terms of inducing chondrogenic differentiation of MSCs in a pellet culture system.…”

Sol–gel derived lithium-releasing glass for cartilage regeneration

“…In other studies, the stimulatory effects of GSK-3 antagonist on chondrogenic differntiation of human bone marrow derived stromal cells in 3-D micromass/pellet culture have been reported 9,10 . These studies, however, suggested that the interaction between pathways initiated by both TGF-β3 and Li was a prerequisite for the enhanced .…”

“…SiO2 -Li2O glass used in the present study was designed to deliver 5 mM of Li. The reason this dose was chosen was that previous studies demonstrated that the effects of lithium on cellular behaviour is dose dependent with optimal cell viability when culture medium contained 5 mM lithium and higher concentration of Li (7 mM) reduced cell viability 9,10 .…”

“…There is increasing evidence that activation of Wnt-signalling by means of GSK-3 antagonist lithium (e.g. in the form of lithium chloride) in cell lines and mesenchymal stem cells has the ability to stimulate the expression of chondrogenic markers [9][10][11] . Further, lithium has been shown capable of reducing catabolic events in the pathogenesis of OA [12][13][14] .…”