Mehmet GÜL, Semir GÜL, Azibe YILDIZ, Nuray BAYAT, Mustafa HÜZ, Ali OTLU

A Comparison of Three Different Agents of Decalcification for a Histological Examination of Bone Tissues

Amaç: Kemik dekalsifikasyonu histopatoloji laboratuvarlarında hala zahmetli ve zaman alıcı bir süreçtir. Bu çalışmada, kemik dekalsifikasyonu için kullanılan formik asit, Biodec-R ve Decalcifier II dekalsifikasyon ajanları ile kompakt kemik dokularının dekalsifikasyon derecesi ve boyanma özelliklerinin karşılaştırılması amaçlandı. Gereç ve Yöntem: Çalışmada toplam altı adet sağlıklı erkek sıçan (200-220 g) kullanıldı. Sıçanlar servikal dislokasyonla dekapite edildi ve her iki femurları çıkarıldı. Femurlardan alınan 0.5 cm uzunluğundaki parçalar %10luk formaldehitte 36 saat süreyle tespit edildi. Daha sonra kemik dokuları dekalsifikasyon sıvılarında altı gün oda ısısında bekletildi. Dekalsifikasyonun ardından kemik doku örnekleri rutin doku takip işlemlerinden geçirilerek ışık mikroskobik inceleme için hazırlandı. Lamlar üzerine alınan 6 μm kalınlığındaki kesitlere hematoxylin eosin, Gomoris trichrome ve Periodic acid-Schiff boyamaları yapıldı. Kesitler Leica DFC 280 ışık mikroskobu ve Leica Q Win görüntü analiz sisteminde (Leica Micros Imaging Solutions Ltd.; Cambridge, U.K) incelendi. Bulgular: Her üç dekalsifikasyon ajanı aynı süre, aynı deney dizaynıyla uygulandığında; formik asitin kemik dokunun doğal histolojik yapısını en iyi koruduğunu ve boyanma özelliklerinin belirgin şekilde daha kaliteli olduğu gözlendi. Biodec-R ve Decalcifier IInin hücre ve doku detaylarının korunması ve boyanma özellikleri açısından birbirine benzer olduğu saptandı. Sonuç: Kompakt kemik dokunun histolojik incelenmesinde formik asit dekalsifikasyonu, histolojik boyanma ve mikroskopik görüntü kalitesi açısından tercih edilebilir.

Kemik Dokunun Histolojik İncelemesi İçin Üç Farklı Dekalsifikasyon Ajanının Karşılaştırılması

Objectives: Bone-decalsification is still a time consuming and laboring process in histopathology laboratories. In this study, we have aimed a comparision of decalsification degrees and staining properties of compact bone tissue decalcificated by formic acid, Biodec-R, and Decalcifier II as decalcification agents. Materials and Methodology: A total of 6 healthy male rats (200-220 g) were used in this study. Rats were decapitated by cervical dislocation. Femurs were removed and 0.5 cm long pieces from these femurs were fixed in 10% formaldehyde for 36 hours. Subsequently, the bone-tissues were stored in decalcification fluids at room temperature for six days. The bone-tissue samples were processed by routine tissue procedures. They were further processed for light microscopic examination and stained with hematoxylin-eosin, Gomori s trichrome, and Periodic acid-Schiff. We have examined the bone sections under a Leica DFC280 light microscope and Leica Q Win Image Analysis System (Leica Micros Imaging Solutions Ltd.; Cambridge, U.K). Results: When all three decalcification agents were applied for equal time periods and at the same experimental design, it was observed that formic acid is more effectible for the preservation of natural structure of the bone tissue and on the quality of the staining properties. It was observed that Biodec R and Decalcifier II are similar to each other in terms of staining properties and preservation of structural details of cells and tissue. Conclusion: Formic acid decalcification is adviced for histologic staining and for a higher qualitiy of microscopic view during histological examination of compact bone tissues.

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