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Culture of Murine Embryonic Metatarsals: A Physiological Model of Endochondral Ossification
Published on: December 3, 2016
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Structural differences in epiphyseal and physeal hypertrophic chondrocytes
Frederic Shapiro1, Evelyn Flynn1
1Department of Orthopaedic Surgery, Orthopaedic Research Laboratory, Boston Children's Hospital , Boston, MA, USA.
Bonekey Reports
|May 20, 2015
Summary
Epiphyseal and physeal hypertrophic chondrocytes in mice exhibit distinct ultrastructural differences. Optimized tissue preparation techniques enhance cell structure visualization for microscopy, suggesting region-specific chondrocyte functions.
Area of Science:
- Cell Biology
- Histology
- Developmental Biology
Background:
- Hypertrophic chondrocytes are crucial for endochondral ossification.
- Previous studies have not fully detailed the ultrastructural differences between epiphyseal and physeal hypertrophic chondrocytes.
- Variations in chondrocyte morphology may indicate region-specific functional adaptations.
Purpose of the Study:
- To investigate and compare the light microscopic and ultrastructural characteristics of epiphyseal and physeal hypertrophic chondrocytes in BALB/c mice.
- To identify optimal tissue preparation methods for preserving chondrocyte morphology for light microscopy (LM) and electron microscopy (EM).
Main Methods:
- BALB/c mice were used for tissue collection.
- Tissue preparation involved controlled osmolarity and membrane stabilization using ruthenium hexammine trichloride (RHT) or lanthanum nitrate.
- Optimal fixation protocols were developed for LM (glutaraldehyde, paraformaldehyde, RHT, JB-4 plastic, toluidine blue) and EM (glutaraldehyde, paraformaldehyde, RHT, cacodylate buffer, osmium tetroxide).
Main Results:
- Significant differences in light microscopic and ultrastructural appearance were observed between epiphyseal and physeal hypertrophic chondrocytes.
- Physeal chondrocytes showed size increase and elongation towards the metaphysis, with a central nucleus and dispersed rough endoplasmic reticulum (RER).
- Epiphyseal chondrocytes remained circular to oval, featuring a transcellular nucleocytoplasmic column or bud with peripherally located RER, often associated with fat deposits.
Conclusions:
- Epiphyseal and physeal hypertrophic chondrocytes display distinct morphological features, even at similar differentiation stages.
- Optimized fixation and preparation techniques are essential for accurate ultrastructural analysis of chondrocytes.
- These morphological differences suggest potentially different functional roles for hypertrophic chondrocytes in different regions of developing bone.
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