Establishment of a femoral critical-size bone defect model in immunodeficient mice

Stefan Zwingenberger, Eik Niederlohmann, Corina Vater, Stefan Rammelt, Romano Matthys, Ricardo Bernhardt, Roberto Daniel Valladares, Stuart Barry Goodman, Maik Stiehler
Journal of Surgical Research 2013 May 1, 181 (1): e7-e14

BACKGROUND: The development of innovative therapies for bone regeneration requires the use of advanced site-specific bone defect small-animal models. The achievement of proper fixation with a murine model is challenging due to the small dimensions of the murine femur. The aim of this investigation was to find the optimal defect size for a murine critical-size bone defect model using external fixation method.

METHODS: An external fixation device was attached to the right femur of 30 mice. Femoral bone defects of 1 mm (n = 10), 2 mm (n = 10), and 3 mm (n = 10) were created. Wounds were closed without any additional treatment. To investigate bone healing during the 12-wk observation period, x-ray analysis, histomorphology, immunohistochemistry, and μCT scans were performed.

RESULTS: MicroCT analyses after 12 wk showed that 3/8 1-mm defects, 5/8 2-mm defects, and 8/8 3-mm defects remained as nonunions. The defect volumes were 0.36 ± 0.42 mm³ (1-mm group), 1.40 ± 0.88 mm³ (2-mm group), and 2.88 ± 0.28 mm³ (3-mm group; P < 0.001, between all groups).

CONCLUSION: Using external fixation, a defect size of 3 mm is necessary to reliably create a persisting femoral bone defect in nude mice.

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