Postembedding Decalcification of Mineralized Tissue Sections Preserves the Integrity of Implanted Biomaterials and Minimizes Number of Experimental Animals

Figure 4

Postembedding decalcification of rat femur preserves the integrity of calcium-containing materials and allows investigating osteocytes. The SrCPC cement in the fracture defect was embedded in PMMA T9100. (a-b) von Kossa/van Gieson staining of undecalcified bone sample differentiates soft tissue around the fracture defect (red) and stains ossified tissue and materials in black. (c-d) Silver staining of postembedding decalcified sections visualizes osteocytes within the matrix and differentiates the implant from the surrounding tissue. Images (a) and (c) were acquired by 5x magnification objective; then individual tiles were stitched together by Leica application suite (LASX) software. Images (b) and (d) were acquired by 40x magnification objective while inset of image (d) was acquired by 100x magnification objective.

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