Korean J Orthod.  2020 May;50(3):188-196. 10.4041/kjod.2020.50.3.188.

Periodontal ligament proliferation and expressions of bone biomolecules upon orthodontic preloading: Clinical implications for tooth autotransplantation

• Phutinart S
• Krisanaprakornkit S
• Makeudom A
• Suzuki B
• Suzuki EY

• KMID: 2502628
• DOI: http://doi.org/10.4041/kjod.2020.50.3.188

Abstract

Objective
Preservation of the periodontal ligament (PDL) is vital to the success of tooth autotransplantation (TAT). Increased PDL volumes and facilitated tooth extraction have been observed upon orthodontic preloading. However, it is unclear whether any changes occur in the expressions of bone biomolecules in the increased PDL volumes. This study aimed to determine the expressions of runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), receptor activator of nuclear factor kappa-B ligand (RANKL), and osteoprotegerin (OPG) in PDL upon preloading.
Methods
Seventy-two premolars from 18 patients were randomly assigned to experimental groups that received a leveling force for 1, 2, or 4 weeks or to a control unloaded group. Following extraction, PDL volumes from 32 premolars of eight patients (21.0 ± 3.8 years) were evaluated using toluidine blue staining. The expressions of the biomolecules in the PDL from 40 premolars of ten patients (21.4 ± 4.0 years) were analyzed via immunoblotting.
Results
The median percentage of stained PDL was significantly higher at 2 and 4 weeks after preloading than in the unloaded condition (p < 0.05). The median RUNX2 and ALP expression levels were significantly higher at 2 and 4 weeks after preloading than in the unloaded condition (p < 0.05), whereas the median RANKL/OPG ratios were significantly higher at 1 and 4 weeks after preloading (p < 0.05).
Conclusions
Orthodontic preloading for 4 weeks enhances PDL volumes as well as the expressions of RUNX2, ALP and the RANKL/OPG ratio in the PDL, suggesting this loading period is suitable for successful TAT.

Keyword

Bone biomolecules; Orthodontic preloading; Periodontal ligament; Tooth autotransplantation

Figure

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  Figure 1 Representative image illustrating the stained periodontal ligament in an unloaded tooth (control) and in teeth after orthodontic loading for 1, 2, or 4 weeks.

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  Figure 2 Percentage of stained periodontal ligament (PDL) in unloaded teeth compared to that in loaded teeth for different loading durations. *p < 0.05.

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  Figure 3 Representative immunoblotting results of the expressions of runt-related transcription factor 2 (RUNX2), alkaline phosphatase (ALP), receptor activator of nuclear factor kappa-B ligand (RANKL), osteoprotegerin (OPG), and beta actin in human periodontal ligament tissues after orthodontic loading for 1, 2, or 4 weeks, or unloaded as a control.

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  Figure 4 Box plot diagrams showing significantly enhanced expressions of runt-related transcription factor 2 (RUNX2) (A) and alkaline phosphatase (ALP) (B) normalized by the expression of beta actin, as well as the receptor activator of nuclear factor kappa-B ligand (RANKL)/ osteoprotegerin (OPG) ratio (C), at different loading durations (gray boxes) compared to those of controls that are unloaded (empty boxes). Moderately positive correlations are found between the expressions of RUNX2 (A) and ALP (B) or the RANKL/OPG ratio (C) and loading durations from 1 to 4 weeks. The horizontal line within each box represents the median. *p < 0.05; **p < 0.01.

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