J Periodontal Implant Sci.  2018 Dec;48(6):383-394. 10.5051/jpis.2018.48.6.383.

Determination of the optimal diabetes duration for bone regeneration experiments in an alloxan-induced diabetic rabbit calvarial defect model

• Jeong SH
• Jung BH
• Yoo KY
• Um HS
• Chang BS
• Lee JK
• Choi WY

• KMID: 2429866
• DOI: http://doi.org/10.5051/jpis.2018.48.6.383

Abstract

PURPOSE
The purpose of this study was to evaluate the optimal diabetes duration for bone regeneration experiments in an alloxan monohydrate (ALX)-induced diabetic rabbit calvarial defect model by evaluating the association between diabetes duration and bone healing capacity.
METHODS
Twenty-four New Zealand white rabbits were used. Twenty-two rabbits were injected with 100 mg/kg of ALX to induce experimental diabetes. These rabbits were divided into 4 groups, including a control group and groups with diabetes durations of 1 week (group 1), 2 weeks (group 2), and 4 weeks (group 3). Calvarial defects were created at 1, 2, and 4 weeks after ALX injection and in the control rabbits. Cone-beam computed tomography (CBCT) scanning was performed on the day of surgery and at 2 and 4 weeks after surgery. The rabbits were sacrificed 4 weeks after surgery, followed by histological and immunofluorescence analysis.
RESULTS
The diabetic state of all diabetic rabbits was well-maintained throughout the experiment. Reconstructed 3-dimensional CBCT imaging showed more rapid and prominent bone regeneration in the control group than in the experimental groups. Histological staining showed notable bone regeneration in the control group, in contrast to scarce bone formation in the experimental groups. The appearance and immunoreactivity of receptor activator of nuclear factor-kappa B and osteoprotegerin did not show notable differences among the groups.
CONCLUSION
ALX administration at 100 mg/kg successfully induced experimental diabetes in rabbits. The effect of diabetes on bone healing was evident when the interval between diabetes induction and the intervention was ≥1 week.

Keyword

Alloxan; Bone regeneration; Cone-beam computed tomography; Experimental diabetes mellitus; Rabbits

MeSH Terms

Alloxan
Animals
Bone Regeneration*
Cone-Beam Computed Tomography
Diabetes Mellitus, Experimental
Fluorescent Antibody Technique
Osteogenesis
Osteoprotegerin
Rabbits
Receptor Activator of Nuclear Factor-kappa B
Alloxan
Osteoprotegerin
Receptor Activator of Nuclear Factor-kappa B

Figure

• 

  Figure 1 FBS levels of diabetic rabbits. FBS levels over 200 mg/dL were classified as indicating diabetes. All rabbits showed stable FBS throughout the experiment. No rabbits showed relapse of FBS. FBS: fasting blood sugar, W: weeks.

• 

  Figure 2 Reconstructed 3-dimensional cone-beam computed tomography image. The control group (A) showed relatively rapid and prominent bone regeneration compared to groups 1 (B), 2 (C), and 3 (D).

• 

  Figure 3 Hematoxylin and eosin staining and Masson trichrome staining of specimens (scale bae=1 mm). The control group (B) showed notable bone regeneration, whereas groups 1 (C), 2 (D), and 3 (E) showed scarce bone regeneration. (A) baseline. Black arrowheads indicate defect margin. CTL: control, DM: diabetes mellitus, W: weeks.

• 

  Figure 4 Immunofluorescence analysis of OPG and RANKL, as well as merged images (Scale bar = 25 um). There were no obvious differences among the groups. The relative optical density was derived from the immunofluorescence analysis. No statistically significant difference was shown among groups. (A) baseline, (B) control group, (C) group 1, (D) group 2, (E) group 3. OPG: osteoprotegerin, RANKL: receptor activator of nuclear factor-kappa B, CTL: control, DM: diabetes mellitus, IR: immunoreactivity, W: weeks.

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