End-Point Determination In Acid Decalcification Of Bone: A New Novel Method
Abstract
Background: Calcium oxalate test is currently the chemical method employed in testing endpoint of decalcification in acid decalcification. However, there are challenges with this existing method used; firstly, the method takes a long duration of 15-30 minutes to complete; secondly, the method uses ammonia which is a highly irritating gas with a sharp suffocating odor necessitating its use in a fume hood; thirdly, exposure to ammonia gas through breathing, swallowing or skin contact causes burning and irritation of the contact surfaces, coughing and can result in blindness, lung damage or death; fourthly, ammonia loses potency over time and lastly, containers of ammonia may explode when exposed to high heat. Additionally, when a high concentration of ammonia is mixed with air, it can explode when ignited. Accordingly, there is a need to device a new chemical method for testing endpoint of decalcification in acid decalcification.
Objective: The purpose of this study was to introduce a new novel chemical method involving optimized chemical reagents and procedure, allowing the determination of endpoint of decalcification of bone in acid decalcification method.
Design: Experimental research design was conducted.
Data analysis: asymmetric measures of agreement for binary data was used to analyze the results of endpoint of determination using two chemical methods, calcium oxalate test and a new novel method, sodium carbonate test.
Setting: The research study was conducted in a teaching histology/histopathology laboratory at Murang’a University of Technology.
Method: Bone samples that were harvested from the radius bone of a rabbit and fixed in 10% neutral buffered formalin were used in this study. The fixed bone samples were decalcified in 5% aqueous nitric acid solution for a period of 2-7 days and tested for end-point of decalcification using both calcium oxalate test method and the new novel method, sodium carbonate test method. Formation of a cloudy precipitate and clear solution in both methods indicated incomplete and complete decalcification respectively.
Results: there was agreement between the results of calcium oxalate and sodium carbonate test methods in determination of endpoint of decalcification in acid decalcification of bone, in that, the bone samples that showed incomplete and complete decalcification in the two test methods were similar.
Conclusion: According to this study, the findings suggests that the new novel method, sodium carbonate test method is a suitable alternative to the existing calcium oxalate test method in routine histopathological determination of endpoint of decalcification in acid decalcification. Additionally, the new novel method proves to be the best with quick time of getting results and overall safety.
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