Khangura Amberpreet Kaur, Gupta Shally, Gulati Anubha, Singh Simranjit
Department of Oral Pathology & Microbiology, Dr. Harvansh Singh Judge Institute of Dental Sciences and Hospital, Panjab University, Chandigarh, India.
J Oral Maxillofac Pathol. 2021 Sep-Dec;25(3):463-469. doi: 10.4103/jomfp.jomfp_203_21. Epub 2022 Jan 11.
In microscopic assessment of mineralized tissue, decalcification is an important step during tissue processing. The present study was attempted to compare the efficacy of various decalcifying agents and to evaluate the most efficacious decalcifying agent.
The aim was to study and compare the time taken for complete decalcification of the specimen by six different chemical agents; to study and compare the effect of various decalcifying agents on cellular and nuclear changes of hard and soft tissues; to study and compare the effect of various decalcifying agents used on the staining intensity with Ehrlich's Hematoxylin and Eosin stain and to determine the ideal decalcification technique.
The six decalcifying agents, namely 5% nitric acid, 8% formic acid, formalin-nitric acid, 5% trichloroacetic acid, neutral ethylenediaminetetraacetic acid (EDTA) and Perenyi's fluid were used to decalcify 30 human permanent teeth (5 teeth in each solution). The endpoint of decalcification was evaluated by chemical (calcium oxalate test) as well as radiographic methods. The specimens were then subjected to processing, sectioning and staining with hematoxylin and eosin. The stained sections were observed under a light microscope and grading was done.
The results in the present study confirmed the fact that the time required for complete decalcification process was least in Perenyi's fluid, 5% trichloroacetic acid and highest in 14% EDTA. Teeth decalcified in 5% trichloroacetic acid, 8% formic acid, formalin-nitric acid and 5% nitric acid were easy to section. Sectioning was most difficult for teeth decalcified in Perenyi's fluid and 14% EDTA. The overall structure details as well as staining characteristics were best in teeth decalcified by 5% trichloroacetic acid and neutral EDTA and worst in teeth decalcified by Perenyi's fluid.
Five percent trichloroacetic acid was showing the most efficient result as it balances both tissue integrity and time factor suggesting that it can be used as a stable decalcifying agent for routine histopathological diagnosis.
在矿化组织的显微镜评估中,脱钙是组织处理过程中的重要步骤。本研究旨在比较各种脱钙剂的效果,并评估最有效的脱钙剂。
研究并比较六种不同化学试剂使标本完全脱钙所需的时间;研究并比较各种脱钙剂对硬组织和软组织细胞及细胞核变化的影响;研究并比较使用各种脱钙剂后用伊红苏木精染色的染色强度,并确定理想的脱钙技术。
使用六种脱钙剂,即5%硝酸、8%甲酸、甲醛 - 硝酸、5%三氯乙酸、中性乙二胺四乙酸(EDTA)和佩雷尼氏液对30颗人类恒牙进行脱钙处理(每种溶液中5颗牙齿)。通过化学方法(草酸钙试验)以及放射学方法评估脱钙终点。然后对标本进行处理、切片并用苏木精和伊红染色。在光学显微镜下观察染色切片并进行分级。
本研究结果证实,完全脱钙过程所需时间在佩雷尼氏液、5%三氯乙酸中最短,在14% EDTA中最长。用5%三氯乙酸、8%甲酸、甲醛 - 硝酸和5%硝酸脱钙的牙齿易于切片。用佩雷尼氏液和14% EDTA脱钙的牙齿切片最困难。5%三氯乙酸和中性EDTA脱钙的牙齿整体结构细节以及染色特征最佳,佩雷尼氏液脱钙的牙齿最差。
5%三氯乙酸显示出最有效的结果,因为它平衡了组织完整性和时间因素,表明它可作为常规组织病理学诊断的稳定脱钙剂。