Determination of the decalcification time of rat sternum, knee joint and nasal cavity specimens

J.Yu. Ustenko, Pathomorphologist, Department of Histology and Pathomorphology, ORCID 0000-0003-1299-0200;
D.A. Vikulina, Senior Laboratory Assistant, ORCID 0000-0002-4819-5519;
Ya.A. Gushchin, Head of the Department of Histology and Pathomorphology, ORCID 0000-0002-7656-991X

Research and manufacturing company «Home оf Pharmacy»,
188663, Russia, Leningrad oblast, Vsevolozhskiy district, Kuzmolovskiy t.s., Zavodskaya st. 3-245
Е-mail: ustenko.ju@doclinika.ru

Abstract

Вone tissue and calcified tissue need preliminary decalcification before histological processing. Decalcification is the technique for removing minerals from pre-fixed tissue samples. Different solutions are used for decalcification depending on the type of study, the time allotted to work and the type of sample.

It is important to determine the "end point" of decalcification.  The end point is the moment when all inorganic material is removed from the tissue, leaving the remainder of the organic matrix intact. There are a number of methods for determining the end point. For example, manipulation (probing and bending), weighing, radiography. In hospitals end point control is usually performed for each batch of samples. In preclinical studies, there are many samples containing bone tissue, they are a routine practice, the procedure for their selection is standardized within a specific laboratory, as a rule. In preclinical studies, there are many samples containing bone tissue, they are a routine practice, the procedure for their selection is standardized within a specific laboratory as a rule. These features allow standardizing the decalcification method and determining control points for typical samples using one of the methods listed above.

This article presents a comparative characteristic of methods for determining the end point of decalcification and establishes the decalcification time of rat sternum, knee joint and nasal cavity specimens. The weighing is most suitable method for monitoring end point in routine preclinical studies. It is accurate, simple to perform, fast, and available to most laboratories. The optimal decalcification period for samples of the sternum, knee joint and nasal cavity – 6 days – was established when using a solution based on hydrochloric and formic acids and in accordance with the described sampling procedure - the optimal decalcification period for samples of the sternum, knee joint and nasal cavity was established when using a solution based on hydrochloric and formic acids and in accordance with the described sampling procedure. The shorter time of the samples in the decalcifying solution results in thick sections with more artifacts. A longer time leads to the loss of normal cell morphology and their ability to be stain. It is necessary to validate the decalcification method in the laboratory, taking into account the reagents, the type of samples and the method of their selection.

Full text avaliable in Russain only 

Authors contribution 

Zh. Ustenko – idea, concept and design of the research, conducting research, systematization materials, writing the article, editing the article, providing of the photo materials.

D. Vikulina – conducting research, systematization materials, writing the article, editing the article.

Ya. Gushchin – idea, concept and design of the research, editing the article.

Conflict of interest

The authors declare no conflict of interest requiring disclosure in this article.

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