Korean J Orthod.  2018 Jul;48(4):253-261. 10.4041/kjod.2018.48.4.253.

Effect of caspases and RANKL induced by heavy force in orthodontic root resorption

• Minato Y
• Yamaguchi
• Shimizu M
• Kikuta J
• Hikida T
• Hikida M
• Suemitsu M
• Kuyama K
• Kasai K

• KMID: 2417967
• DOI: http://doi.org/10.4041/kjod.2018.48.4.253

Abstract

OBJECTIVE
Orthodontic root resorption (ORR) due to orthodontic tooth movement is a difficult treatment-related adverse event. Caspases are important effector molecules for apoptosis. At present, little is known about the mechanisms underlying ORR and apoptosis in the cementum. The aim of the present in vivo study was to investigate the expression of tartrate-resistant acid phosphatase (TRAP), caspase 3, caspase 8, and receptor activator of nuclear factor kappa-B ligand (RANKL) in the cementum in response to a heavy or an optimum orthodontic force.
METHODS
The maxillary molars of male Wistar rats were subjected to an orthodontic force of 10 g or 50 g using a closed coil spring. The rats were sacrificed each experimental period on days 1, 3, 5, and 7 after orthodontic force application. And the rats were subjected to histopathological and immunohistochemical analyses.
RESULTS
On day 7 for the 50-g group, hematoxylin and eosin staining revealed numerous root resorption lacunae with odontoclasts on the root, while immunohistochemistry showed increased TRAP- and RANKL-positive cells. Caspase 3- and caspase 8-positive cells were increased on the cementum surfaces in the 50-g group on days 3 and 5. Moreover, the number of caspase 3- and caspase 8-positive cells and RANKL-positive cells was significantly higher in the 50-g group than in the 10-g group.
CONCLUSIONS
In our rat model, ORR occurred after apoptosis was induced in the cementum by a heavy orthodontic force. These findings suggest that apoptosis of cementoblasts is involved in ORR.

Keyword

Apoptosis; Caspase; Receptor activator of nuclear factor kappa-B ligand; Root resorption

MeSH Terms

Acid Phosphatase
Animals
Apoptosis
Caspase 3
Caspase 8
Caspases*
Dental Cementum
Eosine Yellowish-(YS)
Hematoxylin
Humans
Immunohistochemistry
Male
Models, Animal
Molar
Osteoclasts
Rats
Rats, Wistar
Root Resorption*
Tooth Movement
Acid Phosphatase
Caspase 3
Caspase 8
Caspases
Eosine Yellowish-(YS)
Hematoxylin

Figure

• 

  Figure 1 The rat model of orthodontic tooth movement used in the present study. Tooth movement is reproduced with a closed coil spring ligated to the maxillary first molar using a 0.008-inch stainless steel ligation wire (wire size, 0.005 inch; diameter, 1/12 inch). The other side of the coil spring is ligated using the same ligation wire to a hole in the maxillary incisor, which is drilled in the cleft just above the gingival papilla using a 1/4 round bur. The maxillary right first molar was moved in the mesial direction via the application of a force of 10 g or 50 g by the sealed coil spring.

• 

  Figure 2 A schematic diagram showing a survey area (shaded box) at the mesial center of the distal root of the maxillary first molar obtained from a rat model of orthodontic tooth movement. The periodontal tissue in the region of compression was defined as the tissue connecting the center of the distobuccal (DB) root and the center of the mesial root (M) of the maxillary first molar, and it comprises one-fourth of the mesial region facing the DB root. The large arrows indicate the direction of force. Root resorption is investigated in 300-µm sections (shaded box) from the area close to the furcation on the mesial surface of the DB root, which was the side of compression during tooth movement. MP, Mesial palatal root; DP, distal palatal root; MB, mesial buccal root.

• 

  Figure 3 Light microscopic images showing the effects of various orthodontic forces (0, 10, and 50 g) on the expression of multinuclear osteoclasts (hematoxylin and eosin staining, ×400) at 1, 3, 5, and 7 days after force application in a rat model of orthodontic tooth movement. The expression of odontoclasts (arrow) on the root surfaces in the 50-g group is greater than that on the root surfaces in the 10-g and control groups on days 5 and 7. The arrows indicate the sites of root resorption. Scale bar = 50 µm. PDL, Periodontal ligament; C, cementum; D, dentin.

• 

  Figure 4 A, Effects of the different orthodontic forces (0, 10, and 50 g) on the expression of tartrate-resistant acid phosphatase (TRAP)-positive odontoclasts at 1, 3, 5, and 7 days after force application in a rat model of orthodontic tooth movement. The arrows indicate TRAP-positive cells (×400). Scale bar = 50 µm. B, A graph showing the quantitative evaluation of cellular changes at 1, 3, 5, and 7 days after force application (0, 10, and 50 g) in a rat model of orthodontic tooth movement. PDL, Periodontal ligament; C, cementum; D, dentin. *p < 0.01, significant difference when compared with the control group; †p < 0.01, significant difference when compared with the 10-g group.

• 

  Figure 5 A, Effects of different orthodontic forces (0, 10, and 50 g) on the expression of caspase 3-positive cells at 1, 3, 5, and 7 days after force application in a rat model of orthodontic tooth movement. The arrows indicate caspase 3-positive cells (×400). Scale bar = 50 µm. B, A graph showing the quantitative evaluation of cellular changes at 1, 3, 5, and 7 days after force application (0, 10, and 50 g) in a rat model of orthodontic tooth movement. PDL, Periodontal ligament; C, cementum; D, dentin. *p < 0.01, significant difference when compared with the control group; †p < 0.01, significant difference when compared with the 10-g group.

• 

  Figure 6 A, Effects of different orthodontic forces (0, 10, and 50 g) on the expression of caspase 8-positive cells at 1, 3, 5, and 7 days after force application in a rat model of orthodontic tooth movement. The arrows indicate caspase 8-positive cells (×400). Scale bar = 50 µm. B, A graph showing the quantitative evaluation of cellular changes at 1, 3, 5, and 7 days after force application (0, 10, and 50 g) in a rat model of orthodontic tooth movement. PDL, Periodontal ligament; C, cementum; D, dentin. *p < 0.05 and **p < 0.01, significant difference when compared with the control group; †p < 0.01, significant difference when compared with the 10-g group.

• 

  Figure 7 A, Effects of different orthodontic forces (0, 10, and 50 g) on the expression of receptor activator of nuclear factor kappa-B ligand (RANKL)-positive cells at 1, 3, 5, and 7 days after force application in a rat model of orthodontic tooth movement. The arrows indicate RANKL-positive cells (×400). Scale bar = 50 µm. B, A graph showing the quantitative evaluation of cellular changes at 1, 3, 5, and 7 days after force application (0, 10, and 50 g) in a rat model of orthodontic tooth movement. PDL, Periodontal ligament; C, cementum; D, dentin. *p < 0.01, significant difference when compared with the control group; †p < 0.05 and ††p < 0.01, significant difference when compared with the 10-g group.

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