A methodology to evaluate different histological preparations of soft tissues: Intervertebral disc tissues study

Area of Science:

• Biomaterials Science
• Histology
• Biophysics

Background:

• Tissue preparation is crucial for accurate analysis but can alter sample properties.
• Understanding these alterations is vital for reliable soft tissue research.
• Intervertebral disc (IVD) tissue analysis requires careful consideration of preparation methods.

Purpose of the Study:

• To evaluate the effects of common histological preparation methods on IVD tissue.
• To assess changes in morphology, biomechanics, and chemical composition.
• To identify optimal preparation techniques for microscale soft tissue analysis.

Main Methods:

• Raman spectroscopy and nanoindentation were used to analyze IVD samples.
• Three preparation methods were compared: fresh cryosection, fixed cryosection, and paraffin-embedding.
• Morphological, biomechanical, and chemical properties were quantified for each method.

Main Results:

• Paraffin-embedding provided the most detailed morphology, revealing AF lamellar structure.
• No significant biomechanical difference was found between fresh and fixed cryosections (p>0.05).
• Significant biomechanical differences were observed between fresh cryosections and paraffin-embedded samples (p<0.05).
• Tissue preparation methods altered chemical content and tissue structure, impacting biomechanical properties.

Conclusions:

• Fresh and fixed cryosections are more suitable for biomechanical assessments of histological soft tissues.
• Preparation methods significantly influence microscale tissue characteristics.
• Findings provide a reference for optimizing soft tissue analysis methods.