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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.

No MeSH data available.


Related in: MedlinePlus

MTs expression in hMSCs. (A) hMSCs isolated from 12 patients with AIS (P1-P12). Protein expression of MT1 and MT2 was irregular in hMSCs from patients with AIS. hMSCs from four subjects (P2, P3, P6, and P10) demonstrated low MT1 expression, and those from six subjects (P2-P5, P10, and P11) exhibited low MT2 expression. Expression was extremely low in two subjects (P2 and P10). (B) hMSCs isolated from 12 control subjects (N1-N12). All hMSCs isolated from control subjects exhibited uniform bands for MT1 and MT2. (C) Overall, MT2 demonstrated reduced expression in the AIS group compared with control group. n=12. *P<0.05 vs. control group. hMSCs, human mesenchymal stem cells; AIS, adolescent idiopathic scoliosis; MT, melatonin receptor.
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f1-mmr-14-02-1201: MTs expression in hMSCs. (A) hMSCs isolated from 12 patients with AIS (P1-P12). Protein expression of MT1 and MT2 was irregular in hMSCs from patients with AIS. hMSCs from four subjects (P2, P3, P6, and P10) demonstrated low MT1 expression, and those from six subjects (P2-P5, P10, and P11) exhibited low MT2 expression. Expression was extremely low in two subjects (P2 and P10). (B) hMSCs isolated from 12 control subjects (N1-N12). All hMSCs isolated from control subjects exhibited uniform bands for MT1 and MT2. (C) Overall, MT2 demonstrated reduced expression in the AIS group compared with control group. n=12. *P<0.05 vs. control group. hMSCs, human mesenchymal stem cells; AIS, adolescent idiopathic scoliosis; MT, melatonin receptor.

Mentions: Study participants were 12 patients with severe AIS (8 females, 4 males; age 14–19 years) and 12 control subjects (8 females, 4 males; age 15–24 years; Tables II and III). hMSCs isolated from bone marrow were defined morphologically by a fibroblast-like appearance. Overall, expression levels of MT1 and MT2 were irregular in hMSCs from patients with AIS. MT1 expression was low in MSCs from 4 subjects with AIS (P2, P3, P6 and P10), and MT2 expression was low in MSCs from 6 subjects with AIS (P2-P5, P10 and P11; extremely low in P2 and P10; Fig. 1A). By contrast, all MSC samples isolated from control subjects exhibited uniform bands for MT1 and MT2 (Fig. 1B). When the levels from all samples were combined, MT2 demonstrated significantly reduced expression in the AIS group compared with the control group (P=0.00261; Fig. 1C).


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)

MTs expression in hMSCs. (A) hMSCs isolated from 12 patients with AIS (P1-P12). Protein expression of MT1 and MT2 was irregular in hMSCs from patients with AIS. hMSCs from four subjects (P2, P3, P6, and P10) demonstrated low MT1 expression, and those from six subjects (P2-P5, P10, and P11) exhibited low MT2 expression. Expression was extremely low in two subjects (P2 and P10). (B) hMSCs isolated from 12 control subjects (N1-N12). All hMSCs isolated from control subjects exhibited uniform bands for MT1 and MT2. (C) Overall, MT2 demonstrated reduced expression in the AIS group compared with control group. n=12. *P<0.05 vs. control group. hMSCs, human mesenchymal stem cells; AIS, adolescent idiopathic scoliosis; MT, melatonin receptor.
© Copyright Policy - open-access
Related In: Results  -  Collection

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

f1-mmr-14-02-1201: MTs expression in hMSCs. (A) hMSCs isolated from 12 patients with AIS (P1-P12). Protein expression of MT1 and MT2 was irregular in hMSCs from patients with AIS. hMSCs from four subjects (P2, P3, P6, and P10) demonstrated low MT1 expression, and those from six subjects (P2-P5, P10, and P11) exhibited low MT2 expression. Expression was extremely low in two subjects (P2 and P10). (B) hMSCs isolated from 12 control subjects (N1-N12). All hMSCs isolated from control subjects exhibited uniform bands for MT1 and MT2. (C) Overall, MT2 demonstrated reduced expression in the AIS group compared with control group. n=12. *P<0.05 vs. control group. hMSCs, human mesenchymal stem cells; AIS, adolescent idiopathic scoliosis; MT, melatonin receptor.
Mentions: Study participants were 12 patients with severe AIS (8 females, 4 males; age 14–19 years) and 12 control subjects (8 females, 4 males; age 15–24 years; Tables II and III). hMSCs isolated from bone marrow were defined morphologically by a fibroblast-like appearance. Overall, expression levels of MT1 and MT2 were irregular in hMSCs from patients with AIS. MT1 expression was low in MSCs from 4 subjects with AIS (P2, P3, P6 and P10), and MT2 expression was low in MSCs from 6 subjects with AIS (P2-P5, P10 and P11; extremely low in P2 and P10; Fig. 1A). By contrast, all MSC samples isolated from control subjects exhibited uniform bands for MT1 and MT2 (Fig. 1B). When the levels from all samples were combined, MT2 demonstrated significantly reduced expression in the AIS group compared with the control group (P=0.00261; Fig. 1C).

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