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

Chen C, Xu C, Zhou T, Gao B, Zhou H, Chen C, Zhang C, Huang D, Su P - Mol Med Rep (2016)

Bottom Line: Alkaline phosphatase (ALP) activity assays, quantitative glycosaminoglycan (GAG) analysis, and reverse transcription‑quantitative polymerase chain reaction analysis were performed.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.These results indicate a potential modulating role of melatonin via the MT2 receptor on abnormal osteogenic and chondrogenic differentiaation in patients with AIS.

View Article: PubMed Central - PubMed

Affiliation: Department of Spine Surgery, The First Affiliated Hospital of Sun Yat‑sen University, Guangzhou, Guangdong 510080, P.R. China.

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.

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Related in: MedlinePlus

Effect of melatonin on osteogenic differentiation and the expression of marker genes in human mesenchymal stem cells. mRNA expression was measured at day 12, and relative expression levels were calculated using the 2−ΔΔCt method. Melatonin treatment enhanced the expression of osteogenic marker genes, including (A) ALP, (B) osteopontin, (C) osteocalcin, and (D) RUNX2, in the control groups, but not in the AIS groups. *P<0.05 vs. control-Os group. ALP, alkaline phosphatase; Os, osteogenesis; AIS, adolescent idiopathic scoliosis; RUNX2, runt-related transcription factor 2.
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f3-mmr-14-02-1201: Effect of melatonin on osteogenic differentiation and the expression of marker genes in human mesenchymal stem cells. mRNA expression was measured at day 12, and relative expression levels were calculated using the 2−ΔΔCt method. Melatonin treatment enhanced the expression of osteogenic marker genes, including (A) ALP, (B) osteopontin, (C) osteocalcin, and (D) RUNX2, in the control groups, but not in the AIS groups. *P<0.05 vs. control-Os group. ALP, alkaline phosphatase; Os, osteogenesis; AIS, adolescent idiopathic scoliosis; RUNX2, runt-related transcription factor 2.

Mentions: Following osteogenic differentiation for 12 days, treatment with 50 nM melatonin significantly enhanced ALP activity in normal control MSCs compared with untreated normal controls (P=0.01685), indicating that these MSCs were sensitive to melatonin during osteogenesis. By contrast, melatonin did not promote ALP activity in the AIS groups (Fig. 2). Similarly, melatonin treatment enhanced the mRNA expression levels of osteogenic marker genes, including ALP, osteopontin, osteocalcin and RUNX2, in the normal control group compared with the untreated normal control (P= 0.00472, P= 0.0177, P= 0.0285 and P=0.00726, respectively), however melatonin exhibited no effect on the AIS groups (Fig. 3). Thus, MSCs from patients with AIS appeared to be unresponsive to melatonin treatment during osteogenesis.


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

Chen C, Xu C, Zhou T, Gao B, Zhou H, Chen C, Zhang C, Huang D, Su P - Mol Med Rep (2016)

Effect of melatonin on osteogenic differentiation and the expression of marker genes in human mesenchymal stem cells. mRNA expression was measured at day 12, and relative expression levels were calculated using the 2−ΔΔCt method. Melatonin treatment enhanced the expression of osteogenic marker genes, including (A) ALP, (B) osteopontin, (C) osteocalcin, and (D) RUNX2, in the control groups, but not in the AIS groups. *P<0.05 vs. control-Os group. ALP, alkaline phosphatase; Os, osteogenesis; AIS, adolescent idiopathic scoliosis; RUNX2, runt-related transcription factor 2.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4940077&req=5

f3-mmr-14-02-1201: Effect of melatonin on osteogenic differentiation and the expression of marker genes in human mesenchymal stem cells. mRNA expression was measured at day 12, and relative expression levels were calculated using the 2−ΔΔCt method. Melatonin treatment enhanced the expression of osteogenic marker genes, including (A) ALP, (B) osteopontin, (C) osteocalcin, and (D) RUNX2, in the control groups, but not in the AIS groups. *P<0.05 vs. control-Os group. ALP, alkaline phosphatase; Os, osteogenesis; AIS, adolescent idiopathic scoliosis; RUNX2, runt-related transcription factor 2.
Mentions: Following osteogenic differentiation for 12 days, treatment with 50 nM melatonin significantly enhanced ALP activity in normal control MSCs compared with untreated normal controls (P=0.01685), indicating that these MSCs were sensitive to melatonin during osteogenesis. By contrast, melatonin did not promote ALP activity in the AIS groups (Fig. 2). Similarly, melatonin treatment enhanced the mRNA expression levels of osteogenic marker genes, including ALP, osteopontin, osteocalcin and RUNX2, in the normal control group compared with the untreated normal control (P= 0.00472, P= 0.0177, P= 0.0285 and P=0.00726, respectively), however melatonin exhibited no effect on the AIS groups (Fig. 3). Thus, MSCs from patients with AIS appeared to be unresponsive to melatonin treatment during osteogenesis.

Bottom Line: Alkaline phosphatase (ALP) activity assays, quantitative glycosaminoglycan (GAG) analysis, and reverse transcription‑quantitative polymerase chain reaction analysis were performed.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.These results indicate a potential modulating role of melatonin via the MT2 receptor on abnormal osteogenic and chondrogenic differentiaation in patients with AIS.

View Article: PubMed Central - PubMed

Affiliation: Department of Spine Surgery, The First Affiliated Hospital of Sun Yat‑sen University, Guangzhou, Guangdong 510080, P.R. China.

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.

No MeSH data available.


Related in: MedlinePlus