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Fig 1.

Quantitative evaluation of new bone formation in X-ray and μCT images for grades.

Values refer to the percentage of the given defect border that is bridged by bone tissue of each side of the defect. A score of zero was used when no bone bridging or bone formation was found.

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Fig 1 Expand

Fig 2.

Gross observation of different radius defects after euthanasia at 12 weeks.

Both the borders and center of the defects were covered by a newly formed and irregularly-shaped callus, which declined gradually with the defect size increasing (a: 1.0cm, b: 1.2cm, c: 1.4cm, d: 1.7cm, e: 2.0 cm).

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Fig 2 Expand

Fig 3.

Measurement of defects on the X-ray just after operation.

Size of different bone defects on the X-ray just after operation showed no statistical differences with the measurement of bone defects during operation (P >0.05).

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Fig 3 Expand

Fig 4.

Radiograph of different radius defects just after operation and 12 weeks postoperatively.

The size of the central bone defect gradually increased with respect to an increase in created defect size (a,a’: 1.0cm, b,b’: 1.2cm, c,c’: 1.4cm, d,d’: 1.7cm, e,e’: 2.0 cm, a-e: just after operation, a’-e’: 12 weeks postoperative, scale bar: 10mm).

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Fig 4 Expand

Fig 5.

Radiographic grades of 1.0-cm and 1.2-cm groups showed more new bone formation on the X-ray.

At 12 weeks postoperative, the score from X-ray images of 1.4-cm,1.7cm and 2.0-cm groups showed significantly lower new bone formation compared with the 1.0cm and 1.2cm groups (* P <0.05 compared with 1.0-cm groups, and # P <0.05 compared with 1.2-cm groups, Grade: value of score).

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Fig 5 Expand

Fig 6.

Radiographic grades of 1.0-cm and 1.2-cm groups showed more new bone formation on the Computer tomograph (CT).

CT scans of different radial defects 12 weeks postoperatively. * P <0.05 compared with 1.0-cm groups, and # P <0.05 compared with 1.2-cm groups. Column graph b (1.2-cm group) showed no statistically significant difference with column graph a (1.0-cm group) (P >0.05). (a: 1.0cm, b: 1.2cm, c: 1.4cm, d: 1.7cm, e: 2.0 cm, scale bar: 10mm, Grade: value of score).

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Fig 6 Expand

Fig 7.

Histological assay at 12 weeks postoperative showed large amount of newly formed callus in most samples of the 1.0-cm and 1.2-cm groups.

Bone tissue histology after decalcification with H&E staining at 12 weeks postoperative. The percentage of newly formed bone for the 1.0-cm and 1.2-cm groups were significantly higher than those of other groups (* P <0.05 compared with 1.0-cm groups, and # P <0.05 compared with 1.2-cm groups) (a: 1.0cm, b: 1.2cm, c: 1.4cm, d: 1.7cm, e: 2.0 cm, F: fibrous or fat tissue, C: connection of new bone formation between the two borders of bone defect, red arrow: osteoblasts, blue arrow: chondroblasts, black arrow: mature Haversian system, scale bar: 25μm).

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Fig 7 Expand

Fig 8.

Gross observation of different radius defects cut through longitudinal section after decalcification.

The central defect size was reduced with the increase of created defect size (a: 1.0cm, b: 1.2cm, c: 1.4cm, d: 1.7cm, e: 2.0 cm).

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Fig 8 Expand