Korean J Orthod.  2009 Aug;39(4):248-256. 10.4041/kjod.2009.39.4.248.

Effects of compressive stress on the expression of M-CSF, IL-1beta, RANKL and OPG mRNA in periodontal ligament cells

• Kim JW
• Lee KS
• Nahm JH
• Kang YG

• KMID: 1762580
• DOI: http://doi.org/10.4041/kjod.2009.39.4.248

Abstract

OBJECTIVE
The aim of this study was to determine if human PDL cells can produce osteoclastogenic mRNA and examine how compressive stress affects the expression of osteoclastogenic mRNA in human PDL cells. METHODS: Human PDL cells were obtained from biscupids extracted for orthodontic treatment. The compressive force was adjusted by increasing the number of cover glasses. PDL cells were subjected to a compressive force of 0.5, 1.0, 2.0, 3.0 or 4.0 g/cm2 for 0.5, 1.5, 6, 24 or 48 hours. Reverse transcription polymerase chain reaction (RT-PCR) analysis was performed to examine levels of M-CSF, IL-1beta, RANKL, OPG mRNA expression. RESULTS: Human PDL cells could produce M-CSF mRNA. Human PDL cells under compressive stress showed increased M-CSF, IL-1beta and RANKL mRNAs expression in a force (up to 2 g/cm2) and time-dependent manner. However, OPG mRNA expression was constant regardless of the level and duration of stress. CONCLUSIONS: Continuous compressive stress induced the mRNA expression of osteoclastogenic cytokines including M-CSF, RANKL, IL-1beta in PDL cells. Together with an unchanged OPG mRNA level, these results suggest that compressive stress-induced osteoclastogenesis in vivo is partly controlled by M-CSF, RANKL and IL-1beta expression in PDL cells.

Keyword

Human PDL cell; Mechanical stress; Osteoclastogenesis

MeSH Terms

Cytokines
Eyeglasses
Glass
Humans
Macrophage Colony-Stimulating Factor
Periodontal Ligament
Polymerase Chain Reaction
Reverse Transcription
RNA, Messenger
Stress, Mechanical
Cytokines
Macrophage Colony-Stimulating Factor
RNA, Messenger

Figure

• 

  Fig 1 Method used to apply a compressive stress. Pre-cultured PDL cells were compressed continuously using a different number of round-shaped cover glasses. Round-shaped cover glasses were placed over a confluent cell layer in each well of a 6-well plate. The amount of compressive force was adjusted by increasing or decreasing the number of cover glasses placed.

• 

  Fig 2 Compressive stress up-regulated M-CSF and IL-1β mRNAs expression in PDL cells. A, RT-PCR analysis of PDL cells. The PDL cells were loaded at different compressive stress (0, 0.5, 1, 2, 3 or 4 g/cm2) for 48 hours; B, Densitometry analysis. The results are expressed as the mean ratio to GAPDH expression of five independent experiments. M-CSF and IL-1β mRNAs expression at 2 g/cm2 was significantly different compared to those of the control.

• 

  Fig 3 Compressive stress up-regulated M-CSF and IL-1β mRNAs expression in a time dependent manner. A, RT-PCR analysis of PDL cells. The PDL cells were loaded at a constant compressive stress (2 g/cm2) for 0, 0.5, 1.5, 6, 24 or 48 hours; B, Densitometry analysis. The results are expressed as the mean ratio to GAPDH expression of five independent experiments. M-CSF and IL-1β mRNAs expression was significantly different from the control after 6 hours and 24 hours, respectively.

• 

  Fig 4 Compressive stress up-regulated RANKL mRNA expression in PDL cells. In contrast, OPG mRNA expression did not change. A, RT-PCR analysis of PDL cells. The PDL cells were loaded at different compressive stresses (0, 0.5, 1, 2, 3 or 4 g/cm2) for 48 hours; B, Densitometry analysis. The results are expressed as the mean ratio to GAPDH expression of five independent experiments. RANKL mRNA expression was significantly different from the control at 2 g/cm2. In contrast, OPG mRNA expression was constant throughout the experiment.

• 

  Fig 5 Compressive stress up-regulated RANKL mRNA expression in a time dependent manner. In contrast, OPG mRNA expression did not change. A, RT-PCR analysis of PDL cells. The PDL cells were loaded at a constant compressive stress (2 g/cm2) for 0, 0.5, 1.5, 6, 24 or 48 hours; B, Densitometry analysis. The results are expressed as the mean ratio to GAPDH expression of five independent experiments. RANKL mRNA expression was significantly different from the control after 6 hours. In contrast, OPG mRNA expression was constant throughout the experiment.

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