J Periodontal Implant Sci.  2017 Oct;47(5):339-350. 10.5051/jpis.2017.47.5.339.

Determination of the critical diabetes duration in a streptozotocin-induced diabetic rat calvarial defect model for experimentation regarding bone regeneration

• Kim HJ
• Jung BH
• Yoo KY
• Han JW
• Um HS
• Chang BS
• Lee JK

• KMID: 2392720
• DOI: http://doi.org/10.5051/jpis.2017.47.5.339

Abstract

PURPOSE
The purpose of this study was to determine the critical diabetes duration in a streptozotocin (STZ)-induced diabetic rat calvarial defect model for experimentation regarding bone regeneration by evaluating the association between diabetes duration and bone healing capacity through histological and radiographic analyses.
METHODS
Experimental diabetes was induced in 50 of 60 rats by an STZ injection. The rats were divided into 5 groups, including a control group (group 1), according to diabetes durations of 0, 2, 4, 6, and 8 weeks, respectively. Eighteen rats survived: 4 in group 1, 4 in group 2, 4 in group 3, 5 in group 4, and 1 in group 5. Calvarial defects were created at 0, 2, 4, 6, and 8 weeks after STZ injection in groups 1-5. Cone-beam computed tomography scanning was performed at baseline and at 5 and 7 weeks after surgery. The rats were sacrificed 7 weeks after surgery, followed by histological evaluation.
RESULTS
The voxel gray values (VGVs) of group 1 and group 2 increased, whereas the VGVs of group 3 and group 4 decreased starting 5 weeks after surgery, although this trend did not reach statistical significance between groups. On the reconstructed 3-dimensional images and based on an analysis of histological features, groups 1 and 2 showed apparent bone regeneration, while groups 3-5 showed very limited bone regeneration.
CONCLUSIONS
The critical diabetes duration in an STZ-induced diabetic rat calvarial defect model for experimentation regarding bone regeneration was between 2 and 4 weeks. It is suggested that researchers who use STZ-induced diabetic rats wait for more than 2 weeks following diabetes induction before placing implants or conducting bone regeneration studies to allow definite disturbances in bone healing to emerge.

Keyword

Bone regeneration; Cone-beam computed tomography; Diabetes mellitus; Streptozocin

MeSH Terms

Animals
Bone Regeneration*
Cone-Beam Computed Tomography
Diabetes Mellitus
Rats*
Streptozocin
Streptozocin

Figure

• 

  Figure 1 (A) Axial view of CBCT showing the VGV of the ROI. (B) Sagittal view of CBCT indicating 3 different reference planes (red lines) for measurements.CBCT: cone-beam computed tomography, VGV: voxel gray value, ROI: region of interest, Min: minimum, Max: maximum, Avg: average, Std: standard deviation.

• 

  Figure 2 Changes in blood glucose levels after STZ injection in the surviving experimental rats. Diabetes induction was confirmed by the measurement of a blood glucose concentration greater than 300 mg/dL (red line). The diabetic state of all diabetic rats was well maintained throughout the experimental period.STZ: streptozotocin.

• 

  Figure 3 Reconstructed 3D images based on CBCT. Group 1 (A) and group 2 (B) showed apparent bone regeneration interconnecting the bone defect margin, while group 3 (C), group 4 (D), and group 5 (E) showed reduced bone healing capacity, limited to the defect margin area.3D: 3-dimensional, CBCT: cone-beam computed tomography, W: week.

• 

  Figure 4 Histological features of defect healing 7 weeks after defect creation (Masson trichrome staining, scale bar=1 mm). Group 1 (A) showed notable bone regeneration filling most of the defect area. There was also an apparent bone regeneration pattern in the central part of the defect in group 2 (B). Group 3 (C), group 4 (D), and group 5 (E) showed very limited bone regeneration. White asterisks indicate newly formed bone. Black arrowheads indicate defect margin.

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