Abnormal osteogenic and chondrogenic differentiation of human mesenchymal stem cells from patients with adolescent idiopathic scoliosis in response to melatonin

Abstract

Abnormalities of membranous and endochondral ossification in patients with adolescent idiopathic scoliosis (AIS) remain incompletely understood. To investigate abnormalities in the melatonin signaling pathway and cellular response to melatonin in AIS, a case-control study of osteogenic and chondrogenic differentiation was performed using human mesenchymal stem cells (hMSCs). AIS was diagnosed by physical and radiographic examination. hMSCs were isolated from the bone marrow of patients with AIS and control subjects (n=12 each), and purified by density gradient centrifugation. The expression levels of melatonin receptors (MTs) 1 and 2 were detected by western blotting. Osteogenic and chondrogenic differentiation was induced by culturing hMSCs in osteogenic and chondrogenic media containing vehicle or 50 nM melatonin. Alkaline phosphatase (ALP) activity assays, quantitative glycosaminoglycan (GAG) analysis, and reverse transcription-quantitative polymerase chain reaction analysis were performed. Compared with controls, MT2 demonstrated low expression in the AIS group. Melatonin increased ALP activity, GAG synthesis and upregulated the expression of genes involved in osteogenic and chondrogenic differentiation including, ALP, osteopontin, osteocalcin, runt-related transcription factor 2, collagen type II, collagen type X, aggrecan and sex-determining region Y-box 9 in the normal control hMSCs, but did not affect the AIS groups. Thus, AIS hMSCs exhibit abnormal cellular responses to melatonin during osteogenic and chondrogenic differentiation, which may be associated with abnormal membranous and endochondral ossification, and skeletal growth. These results indicate a potential modulating role of melatonin via the MT2 receptor on abnormal osteogenic and chondrogenic differentiaation in patients with AIS.

Melatonin: Another avenue for treating osteoporosis?

“…Chen et al found that treatment with 50 nmol/L melatonin enhanced the mRNA expression levels of osteogenic marker genes, including ALP, osteopontin, osteocalcin, and RUNX2, in the normal control hMSC group compared with the untreated normal control. During chondrogenic differentiation, melatonin treatment increases quantitative glycosaminoglycan synthesis in normal control hMSC, which is significantly elevated on day 21 compared with untreated normal control cells . Multiple sclerosis (MS) is a common autoimmune and neurodegenerative disease of the central nervous system.…”

“…During chondrogenic differentiation, melatonin treatment increases quantitative glycosaminoglycan synthesis in normal control hMSC, which is significantly elevated on day 21 compared with untreated normal control cells. 144 Multiple sclerosis (MS) is a common autoimmune and neurodegenerative disease of the central nervous system. Serum levels of melatonin usually decrease in MS patients who are also at risk of osteoporosis.…”

Melatonin plays critical role in mesenchymal stem cell-based regenerative medicine in vitro and in vivo

“…For example, MT stimulates MT1 in cultured placenta-derived MSCs, but not MT2, to increase the cell proliferation and survival rate and further enhance the degree of neuronal differentiation [44]. MSCs isolated from adolescent idiopathic scoliosis patients express lower levels of MT2 and demonstrate lower expression levels of osteogenic and chondrogenic markers than MSCs derived from control healthy individuals after MT treatment [45]. Thus, the MT receptor-dependent pathway also plays a critical role in enhancing MSC stemness and differentiation in regenerative medicine.…”

Long noncoding RNA lncAIS downregulation in mesenchymal stem cells is implicated in the pathogenesis of adolescent idiopathic scoliosis

“…AIS is a complex, three dimensional deformity of the spine that commonly occurs in pubescent girls. We and others showed that MSCs in AIS patients display decreased osteogenic differentiation ability [17,18,29,30]. Thus abnormal osteogenic differentiation of MSCs is implicated in the pathogenesis of AIS.…”

“…It has been considered that osteopenia may be a primary contributing factor to the spinal deformity of AIS. We and others showed that MSCs from AIS patients undergo abnormal differentiation during development of osteoblasts, chondrocytes and adipocytes [17,18,29,30], suggesting that abnormal osteogenic differentiation of MSCs is implicated in the pathogenesis of AIS. However, how MSCs is abnormally regulated in the patients with AIS remains elusive.…”