Lab Anim Res.  2010 Sep;26(3):233-239. 10.5625/lar.2010.26.3.233.

The Bone Grinding and Scaffold Grafting Techniques for Guide Bone Regeneration Induce the Stress on the Rat Brain

• Lee YK
• Kim JE
• Nam SH
• Choi SI
• Lee EP
• Choi HW
• Kim HS
• Lee JH
• Jung YJ
• Kang BC
• Cho JS
• Hwang DY

• KMID: 2312072
• DOI: http://doi.org/10.5625/lar.2010.26.3.233

Abstract

Guided bone regeneration (GBR) is a technique that a barrier membrane is placed over the bone defect to prevent the cell growth from the connective tissue and epithelium. In this study, in order to determine whether GBR technique could induce stress in rats, the standardized bone defect in rat calvaria was covered with apatitte membrane. Bone and brain tissues were collected from rats at 3 days, 2, 4, and 16 weeks post-operation, and then alteration of the new bone formation at the defects and stress-related factors were detected with histological examination and Western blot, respectively. From 4 to 16 weeks after the operation, the apatitte membrane was attached to the region of regenerated bone and encapsulated with a thick fibrous layer. Furthermore, the concentration of cortisol, a good indicator of stress, significantly increased 3 days post-operation. However, the increase at 3 days was returned to the basal level in 2 weeks. In Western blot analysis, the highest phosphorylation level of extracellular signal-regulated kinase (ERK) was observed 3 day post-operation, while those of the c-jun N-terminal kinase (JNK) and p38 were detected 4 weeks post-operation. Taken together, the results suggest that GBR technique may induce the serious stress on the brain tissue via the induction of ERK phosphorylation during 2 weeks, and that the stress responses restored in 4 week via JNK and p38 signaling pathway.

Keyword

Guided bone regeneration; cortisol; mitogen-activated protein kinase (MAPK); stress; apatitte membrane

MeSH Terms

Animals
Blotting, Western
Bone Regeneration
Brain
Connective Tissue
Epithelium
Hydrocortisone
JNK Mitogen-Activated Protein Kinases
Membranes
Osteogenesis
Phosphorylation
Phosphotransferases
Rats
Skull
Transplants
Hydrocortisone
JNK Mitogen-Activated Protein Kinases
Phosphotransferases

Figure

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  Figure 1. Experimental scheme and procedure of guided bone regeneration of bone defect. A. Two cavities were created using a drill. A round defect of 3-5 mm in diameter and 0.1-0.2 mm in depth was formed in the calvaria and covered with apatite membrane. B. After the operation of apatite membrane, bone and brain tissue were collected from rats at 3 days and 2, 4 and 16 weeks to examine histological alterations and Western blot analysis. C. Rat cranium showing two full-thickness bone defects with 3-5 mm in diameter.

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  Figure 2. Representative histological findings of control (A) and 3 days (B) and 2 (C), 4 (D) and 16 weeks (E) post-operation. The arrow in Figure 2B indicates regenerative film.

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  Figure 3. Changes of the plasma cortisol concentration in rats with regenerated bone. The serum cortisol level was measured by radioimmunoassay. Values are mean±SD. ∗P<0.05 is the significance level compare to control group.

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  Figure 4. Effects of guided bone regeneration on MAPK signaling pathway. Fifty micrograms of the protein per sample were immunoblotted with the antibodies for ERK, p-ERK, JNK, p-JNK, p38, p-p38 and β-actin, respectively. The samples were assayed in triplicate. The values are mean±SD. ∗P<0.05 is the significance level compare to control group.

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