Melatonin enhances chondrogenic differentiation of human mesenchymal stem cells

Abstract

Intramembranous ossification and endochondral ossification are two ways through which bone formation and fracture healing occur. Accumulating amounts of evidence suggests that melatonin affects osteoblast differentiation, but little is known about the effects of melatonin on the process of chondrogenic differentiation. In this study, the effects of melatonin on human mesenchymal stem cells (MSCs) undergoing chondrogenic differentiation were investigated. Cells were induced along chondrogenic differentiation via high-density micromass culture in chondrogenic medium containing vehicle or 50 nm melatonin. Histological study and quantitative analysis of glycosaminoglycan (GAG) showed induced cartilage tissues to be larger and richer in GAG, collagen type II and collagen type X in the melatonin group than in the untreated controls. Real-time RT-PCR analysis demonstrated that melatonin treatment significantly up-regulated the expression of the genes involved in chondrogenic differentiation, including aggrecan (ACAN), collagen type II (COL2A1), collagen type X (COL10A1), SRY (sex-determining region Y)-box 9 (SOX9), runt-related transcription factor 2 (RUNX2) and the potent inducer of chondrogenic differentiation, bone morphogenetic protein 2 (BMP2). And the expression of melatonin membrane receptors (MT) MT1 and MT2 were detected in the chondrogenic-induced-MSCs by immunofluorescence staining. Luzindole, a melatonin receptor antagonist, was found to partially block the ability of melatonin to increase the size and GAG synthesis of the induced cartilage tissues, as well as to completely reverse the effect of melatonin on the gene expression of ACAN, COL2A1, COL10A1, SOX9 and BMP2 after 7 days of differentiation. These findings demonstrate that melatonin enhances chondrogenic differentiation of human MSCs at least partially through melatonin receptors.

Melatonin promotes osteoblast differentiation by regulating Osterix protein stability and expression

“…Melatonin has confirmed to increase glycosaminoglycan synthesis and cartilage tissue, as well as to enhance the expression of chondrogenic marker genes including aggrecan, collagen type II and X, SOX9, and BMP2 during the chondrogenic differentiation at least partially through melatonin receptors (MT) including MT1 and MT2 55 . Meanwhile, it has been reported that Osterix regulates chondrocyte differentiation and bone growth in growth plate chondrocytes during endochondral ossification 56 .…”

Melatonin reversed tumor necrosis factor‐alpha‐inhibited osteogenesis of human mesenchymal stem cells by stabilizing SMAD1 protein

“…Melatonin is a highly evolutionarily conserved molecule with multiple biological functions, such as regulation of circadian rhythms, inhibition of tumor growth, immunomodulation, and antioxidation . In addition, melatonin plays a vital role in regulating skeleton formation and development and has been well established to promote both osteogenic and chondrogenic differentiations of MSCs . During in vitro osteogenic differentiation of MSCs, melatonin treatment dramatically stimulated osteogenic differentiation and increased expressions of osteogenic markers .…”

Melatonin promotes goat spermatogonia stem cells (SSCs) proliferation by stimulating glial cell line-derived neurotrophic factor (GDNF) production in Sertoli cells

“…Studies have shown evidences that melatonin can enhance the production of iPSCs due to its antioxidant activity [8, 9]. In human adult MSCs, for instance, melatonin combined with the extracellular matrix can enhance cellular alkaline phosphatase (AP) activity, induce osteogenesis via the melatonin receptor 2 (MT2), and also reverse stress-induced MSC injuries [10–13]. Meanwhile, melatonin can elevate the viability of rat neural stem cells (NSCs) and induce NSCs differentiate into dopaminergic neurons and decrease astrocyte production [14, 15].…”