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Histochemical evaluation of enzymes in the rabbit corneal endothelium after short-term storage
Abstract:
Isolated rabbit corneae were stored at a temperature of 4 degrees C in McCoy's 5a medium (modified) containing 5% dextran T 40 and gentamycin sulphate (250 mug/ml) for a varied number of days. The endothelium of the corneae was examined by light- and electron-microscopic histochemical methods for ATPase, TPPase, and SDH. The activity of each enzyme in the endothelia, stored for 6 days, was found to resemble the enzyme activities in the fresh corneal endothelium. By 8 days, the SDH activity was lowered and specific staining for ATPase and TPPase was decreased. In the 12 day-stored endothelium, the SDH and TPPase activities could not be detected. From these results, it was gathered that the isolated cornea stored in the medium for 6 days may be used in keratoplasty.
Insights
Rabbit corneas stored for 6 days in a special medium maintained enzyme activity, suggesting suitability for keratoplasty. Longer storage periods (8-12 days) showed decreased or absent enzyme function.
Area of Science:
- Ophthalmology
- Histochemistry
- Tissue Preservation
Background:
- Corneal storage is crucial for keratoplasty success.
- Enzyme activity in corneal endothelium reflects tissue viability.
- Optimizing storage media and duration is essential for preserving corneal health.
Purpose of the Study:
- To evaluate the impact of storage duration on corneal endothelium enzyme activity.
- To determine the optimal storage period for isolated rabbit corneas for potential use in keratoplasty.
- To assess the viability of corneal endothelium using histochemical markers after cold storage.
Main Methods:
- Isolated rabbit corneae were stored at 4°C in a modified McCoy's 5a medium with dextran T 40 and gentamycin.
- Corneal endothelium was analyzed using light and electron microscopy.
- Histochemical staining was performed to detect Adenosine Triphosphatase (ATPase), Triphosphatase (TPPase), and Succinate Dehydrogenase (SDH) enzyme activities.
Main Results:
- Corneal endothelium enzyme activity (ATPase, TPPase, SDH) closely resembled fresh tissue after 6 days of storage.
- By 8 days, SDH activity decreased, and ATPase and TPPase staining diminished.
- After 12 days, SDH and TPPase activities were undetectable in the stored corneal endothelium.
Conclusions:
- Isolated rabbit corneas stored for up to 6 days in the tested medium retain significant endothelial enzyme activity.
- The findings suggest that 6-day stored corneas may be viable for keratoplasty.
- Extended storage beyond 6 days leads to a notable decline in critical endothelial enzyme functions, impacting potential graft survival.