Tissue integrity, costs and time associated with different agents for histological bone preparation

Area of Science:

• Histopathology
• Biomedical Research
• Laboratory Medicine

Background:

• Effective decalcification is crucial for bone sample analysis in pathology.
• Selection criteria include cellularity, diagnostic urgency, and cost-effectiveness.
• Optimizing demineralization balances tissue integrity with diagnostic needs.

Purpose of the Study:

• To evaluate various decalcification solutions for formalin-fixed, paraffin-embedded bone samples.
• To determine the optimal demineralization procedure based on cellular preservation, time, and cost.
• To assess the impact of different solutions on bone tissue, collagen, and adjacent muscle.

Main Methods:

• Rat femurs were sectioned into bone disks and divided into five groups.
• Groups were demineralized using nitric acid, formic acid, acetic acid, EDTA, or Morsés solution.
• Sections were stained (hematoxylin-eosin, Masson's trichrome, picrosirius red) to assess osteocyte preservation, collagen integrity, and muscle attachment.

Main Results:

• 10% acetic acid yielded the highest osteocyte preservation (95.64%).
• Nitric acid (5%) demonstrated the shortest demineralization time (0.8-5.7 days) and lowest cost.
• EDTA (pH 7.4) showed superior preservation of collagen types I and III and adjacent muscle tissue.

Conclusions:

• 10% acetic acid is most effective for preserving bone tissue structure.
• 5% EDTA best maintains collagen and adjacent muscle integrity.
• 5% nitric acid provides the most efficient demineralization, balancing time and cost.