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Up-regulated osteogenic transcription factors during early response of human periodontal ligament stem cells to cyclic tensile strain.

Tang N, Zhao Z, Zhang L, Yu Q, Li J, Xu Z, Li X - Arch Med Sci (2012)

Bottom Line: Besides, the direction of MSCs differentiation has been verified regulated by mechanical signals.Then osteogenic related genes and proteins were analyzed by RT-PCR and western blot.The significant increase of Runx2, Osx and Satb2 expressions may suggest an early response toward osteogenic orientation of hPDLSCs.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Oral Biomedical Engineering, Sichuan University, China.

ABSTRACT

Introduction: As one group of periodontal ligament (PDL) cells, human periodontal ligament stem cells (hPDLSCs) have been isolated and identified as mesenchymal adult stem cells (MSCs) since 2004. It has been well accepted that PDL sensitively mediates the transmission of stress stimuli to the alveolar bone for periodontal tissue remolding. Besides, the direction of MSCs differentiation has been verified regulated by mechanical signals. Therefore, we hypothesized that tensile strain might act on hPDLSCs differentiation, and the early response to mechanical stress should be investigated.

Material and methods: The hPDLSCs were cultured in vitro and isolated via a magnetic activated CD146 cell sorting system. After investigation of surface markers and other experiments for identification, hPDLSCs were subjected to cyclic tensile strain at 3,000 µstrain for 3 h, 6 h, 12 h, and 24 h, without addition of osteogenic supplements. In the control groups, the cells were cultured in similar conditions without mechanical stimulation. Then osteogenic related genes and proteins were analyzed by RT-PCR and western blot.

Results: Cyclic tensile strain at 3,000 µstrain of 6 h, 12 h, and 24 h durations significantly increased mRNA and protein expressions of Satb2, Runx2, and Osx, which were not affected in unloaded hPDLSCs.

Conclusions: We indicate that hPDLSCs might be sensitive to cyclic tensile strain. The significant increase of Runx2, Osx and Satb2 expressions may suggest an early response toward osteogenic orientation of hPDLSCs.

No MeSH data available.


Related in: MedlinePlus

Cloning formation experiments of CD146+ PDL cells (A, B) and CD146- PDL cells (C, D)
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Figure 0003: Cloning formation experiments of CD146+ PDL cells (A, B) and CD146- PDL cells (C, D)

Mentions: The hPDLSCs were isolated from the 3rd passage PDL cells via a magnetic activated cell sorting system (Miltenyi Biotec, Germany). CD146 cell sorting columns (Miltenyi Biotec, Germany) were used to collect the CD146 (+) cells, the percentage of which in the total cell volume was about 1.5%. CD146 (–) cells were collected as a control for cell identification. After centrifugation at 1000 rpm, the sorted cells were re-suspended in culture medium composed of α-MEM (Gibco, USA) supplemented with 15% (v/v) FBS (Hyclone, USA), and 0.3% (v/v) bovine pituitary extract (BPE; M&C Gene Technology, China), followed by cultivation in humidified air with 5% CO2 at 37°C. The medium was changed three times a week. On reaching confluence, the cells were passaged or seeded into dishes or plates for subsequent experiments. The CD146 (+) cells were identified as hPDLSCs (Figures 1–3).


Up-regulated osteogenic transcription factors during early response of human periodontal ligament stem cells to cyclic tensile strain.

Tang N, Zhao Z, Zhang L, Yu Q, Li J, Xu Z, Li X - Arch Med Sci (2012)

Cloning formation experiments of CD146+ PDL cells (A, B) and CD146- PDL cells (C, D)
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3400899&req=5

Figure 0003: Cloning formation experiments of CD146+ PDL cells (A, B) and CD146- PDL cells (C, D)
Mentions: The hPDLSCs were isolated from the 3rd passage PDL cells via a magnetic activated cell sorting system (Miltenyi Biotec, Germany). CD146 cell sorting columns (Miltenyi Biotec, Germany) were used to collect the CD146 (+) cells, the percentage of which in the total cell volume was about 1.5%. CD146 (–) cells were collected as a control for cell identification. After centrifugation at 1000 rpm, the sorted cells were re-suspended in culture medium composed of α-MEM (Gibco, USA) supplemented with 15% (v/v) FBS (Hyclone, USA), and 0.3% (v/v) bovine pituitary extract (BPE; M&C Gene Technology, China), followed by cultivation in humidified air with 5% CO2 at 37°C. The medium was changed three times a week. On reaching confluence, the cells were passaged or seeded into dishes or plates for subsequent experiments. The CD146 (+) cells were identified as hPDLSCs (Figures 1–3).

Bottom Line: Besides, the direction of MSCs differentiation has been verified regulated by mechanical signals.Then osteogenic related genes and proteins were analyzed by RT-PCR and western blot.The significant increase of Runx2, Osx and Satb2 expressions may suggest an early response toward osteogenic orientation of hPDLSCs.

View Article: PubMed Central - PubMed

Affiliation: State Key Laboratory of Oral Biomedical Engineering, Sichuan University, China.

ABSTRACT

Introduction: As one group of periodontal ligament (PDL) cells, human periodontal ligament stem cells (hPDLSCs) have been isolated and identified as mesenchymal adult stem cells (MSCs) since 2004. It has been well accepted that PDL sensitively mediates the transmission of stress stimuli to the alveolar bone for periodontal tissue remolding. Besides, the direction of MSCs differentiation has been verified regulated by mechanical signals. Therefore, we hypothesized that tensile strain might act on hPDLSCs differentiation, and the early response to mechanical stress should be investigated.

Material and methods: The hPDLSCs were cultured in vitro and isolated via a magnetic activated CD146 cell sorting system. After investigation of surface markers and other experiments for identification, hPDLSCs were subjected to cyclic tensile strain at 3,000 µstrain for 3 h, 6 h, 12 h, and 24 h, without addition of osteogenic supplements. In the control groups, the cells were cultured in similar conditions without mechanical stimulation. Then osteogenic related genes and proteins were analyzed by RT-PCR and western blot.

Results: Cyclic tensile strain at 3,000 µstrain of 6 h, 12 h, and 24 h durations significantly increased mRNA and protein expressions of Satb2, Runx2, and Osx, which were not affected in unloaded hPDLSCs.

Conclusions: We indicate that hPDLSCs might be sensitive to cyclic tensile strain. The significant increase of Runx2, Osx and Satb2 expressions may suggest an early response toward osteogenic orientation of hPDLSCs.

No MeSH data available.


Related in: MedlinePlus