Korean J Orthod.  2008 Jun;38(3):187-201. 10.4041/kjod.2008.38.3.187.

The expression of MMP-1, -8, and -13 mRNA in the periodontal ligament of rats during tooth movement with cortical punching

• Gwack C
• Kim SS
• Park SB
• Son WS
• Kim YD
• Jun ES
• Park MH

• KMID: 1975907
• DOI: http://doi.org/10.4041/kjod.2008.38.3.187

Abstract

OBJECTIVE
The aim of this study was to determine whether cortical punching stimulates the expression of matrix metalloproteinase-1, -8, and -13 in orthodontic tooth movement in rats. METHODS: A total of 32 male sprague-dawley rats at 15 weeks old were divided into two groups of 16 rats each, to form the tooth movement with cortical punching (TMC) group and tooth movement only (TM) group. A total of 20 gm of orthodontic force was applied to rat incisors to cause experimental tooth movement. Cortical punching was performed on the palatal side near the central incisor with a 1.0 mm width microscrew in the TMC group. The duration of tooth movement was 1, 4, 7, and 14 days. RESULTS: Measurements of the mRNA expression were selected as the means to determine the identification of expression of MMP-1, -8, and -13. In the TMC group, the expression of collagen type I was greater than that of the TM group from day 4 to day 14. Expression of TIMP-1 in the TM group was greater than that of the TMC group in the pressure side of PDL and alveolar bone cell at day 4. In the TMC group, TIMP-1 was expressed at the osteoclast, but not at the tooth surface of the TM group at day 14. Maximum induction of the mRNA of MMP-1 was observed on day 4 in the TMC group, but it was observed on day 7 in the TM group. MMP-8 mRNA of the TMC group was twice greater than that of the TM group at 4 days. In the TMC group, maximum induction of MMP-13 mRNA was observed on day 1. CONCLUSIONS: These findings suggested that cortical punching can stimulate remodeling of PDL and alveolar bone connective tissues during experimental orthodontic tooth movement in rats.

Keyword

Matrix metalloproteinase; Collagen type I; Cortical punching

MeSH Terms

Animals
Collagen Type I
Connective Tissue
Humans
Incisor
Male
Matrix Metalloproteinase 1
Osteoclasts
Periodontal Ligament
Rats
Rats, Sprague-Dawley
RNA, Messenger
Tissue Inhibitor of Metalloproteinase-1
Tooth
Tooth Movement
Collagen Type I
Matrix Metalloproteinase 1
RNA, Messenger
Tissue Inhibitor of Metalloproteinase-1

Figure

• 

  Fig. 1 Densitometric results of MMP-1 expression from semi-quantitative RT-PCR between TMC and TM groups. There was a slight increase in the TMC group at day 4 than others. *p< 0.05, considered statistically significant, †p< 0.01, considered statistically significant under paired t test.

• 

  Fig. 2 Densitometric results of MMP-8 expression from semi-quantitative RT-PCR between TMC and TM groups. MMP-8 expression of TMC group from day 4 were more intensive than that of TM group. *p< 0.05, considered statistically significant, †p< 0.01, considered statistically significant under paired t test.

• 

  Fig. 3 Densitometric results of MMP-13 expression from semi-quantitative RT-PCR between TMC and TM group. MMP-13 expression of TMC group increased more from day 1 than that of TM group. *p< 0.05, considered statistically significant under paired t test.

• 

  Fig 4. A-D Collagen type I immunohistochemical staining. Light microscopy photographs of the horizontal section of rat incisor area (horizontal section, 4µm thick, original magnification × 400). Compression side of TMC group at day 1 (A), tension side of TMC group at day 1 (B), compression side of TM group at day 1 (C), and tension side of TM group at day 1 (D). Osteoclast (black triangular head indicator), osteoblast (black diamond head indicator). Alv, Alveolar bone; PDL, periodontal ligament; Tooth, tooth. Bar = 100µm. Immunoreactivity is seen in the periodontal ligament stained by anti-Collagen type I antibody (black arrow).

• 

  Fig 4. E-H Compression side of TMC group at day 4 (E), tension side of TMC group at day 4 (F), compression side of TM group at day 4 (G), and tension side of TM group at day 4 (H). Osteoclast (black triangular head indicator), osteoblast (black diamond head indicator). Alv, Alveolar bone; PDL, periodontal ligament; Tooth, tooth. Bar = 100µm. Immunoreactivity is seen in the periodontal ligament stained by anti-Collagen type I antibody (black arrow). Perpendicular orientation of fibrillar collagen from tooth surface were broken at the compression side near bone surface at day 4 (E and G).

• 

  Fig 4. I-L Compression side of TMC group at day 7 (I), tension side of TMC group at day 7 (J), compression side of TM group at day 7 (K), and tension side of TM group at day 7 (L). Osteoclast (black triangular head indicator), osteoblast (black diamond head indicator). Alv, Alveolar bone; PDL, periodontal ligament; Tooth, tooth. L, Bar = 100µm. Immunoreactivity is seen in the periodontal ligament stained by anti-Collagen type I antibody (black arrow).

• 

  Fig 4. M-P Compression side of TMC group at day 14 (M), tension side of TMC group at day 14 (N), compression side of TM group at day 14 (O), and tension side of TM group at day 14 (P). Osteoclast (black triangular head indicator), osteoblast (black diamond head indicator). Alv, Alveolar bone; PDL, periodontal ligament; Tooth, tooth. P, Bar = 100µm. Immunoreactivity is seen in the periodontal ligament stained by anti-Collagen type I antibody (black arrow).

• 

  Fig 5. A-D TIMP-1 immunohistochemical staining photomicrographs of horizontal sections of maxillary incisors from TMC group and TM group (horizontal section, 4µm thick, original magnification × 400). Osteoclast (black triangular head indicator), osteoblast (black diamond head indicator). Alv, Alveolar bone; PDL, periodontal ligament; Tooth, tooth. D, Bar = 100µm. Immunoreactivity is seen in the alveolar bone and periodontal ligament stained by anti-TIMP-1 antibody (black arrow). TIMP-1 stained in the periodontal ligament cells and perivascular cells in both TMC and TM groups.

• 

  Fig 5. E-H Compression side of TMC group at day 4 (E), tension side of TMC group at day 4 (F), compression side of TM group at day 4 (G), and tension side of TM group at day 4 (H). Osteoclast (black triangular head indicator), osteoblast (black diamond head indicator). Alv, Alveolar bone; PDL, periodontal ligament; Tooth, tooth. H, Bar = 100µm. Immunoreactivity is seen in the alveolar bone and periodontal ligament stained by anti-TIMP-1 antibody (black arrow). Expression of TIMP-1 in TM group was more intensive than that of TMC group from day 4 (G and H).

• 

  Fig 5. I-L Compression side of TMC group at day 7 (I), tension side of TMC group at day 7 (J), compression side of TM group at day 7 (K), and tension side of TM group at day 7 (L). Osteoclast (black triangular head indicator), osteoblast (black diamond head indicator). Alv, Alveolar bone; PDL, periodontal ligament; Tooth, tooth. L, Bar = 100µm. Immunoreactivity is seen in the alveolar bone and periodontal ligament stained by anti-TIMP-1 antibody (black arrow). TIMP-1 stained in the periodontal ligament cells and perivascular cells in both TMC and TM groups.

• 

  Fig 5. M-P Compression side of TMC group at day 14 (M), tension side of TMC group at day 14 (N), compression side of TM group at day 14 (O), and tension side of TM group at day 14 (P). Osteoclast (black triangular head indicator), osteoblas (black diamond head indicator). Alv, Alveolar bone; PDL, periodontal ligament; Tooth, tooth. P, Bar = 100µm. Immunoreactivity is seen in the alveolar bone and periodontal ligament stained by anti-TIMP-1 antibody (black arrow). TIMP-1 stained in the periodontal ligament cells and perivascular cells in both TMC and TM groups.

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