三种染色方法在小鼠下颌第一磨牙牙根不同发育时期的染色效果比较

Application of three staining methods during root development of mandibular first molar in mice

摘要:

目的 比较3种染色方法在小鼠下颌第一磨牙牙根不同发育时期的染色效果，为选择合适的染色方法提供参考。方法 选取出生后第5、7、14、21天的ICR小鼠，制备小鼠下颌第一磨牙冠状切片，采用H-E染色、Masson染色法和Gomori染色法进行组织形态学研究，比较染色结果。结果 在小鼠出生后第5天（PN5）至PN7为牙根发育起始阶段，H-E染色显示： 成牙本质细胞、成釉细胞及上皮根鞘细胞形态最为清晰；Masson染色法显示： 图片色彩鲜艳，牙釉质、牙本质及前期牙本质间层次分明；Gomori染色方法显示： 细胞核呈单一的蓝黑色，但软、硬组织对比度不佳。从PN14小鼠牙齿开始萌出到PN21牙根发育基本完成阶段，H-E染色显示： 成釉细胞、成牙本质细胞及牙周膜成纤维细胞形态最为清晰；Masson染色法显示： 牙骨质与牙槽骨间的胶原纤维排列规则，牙槽骨形态清晰；而Gomori染色效果总体不佳。结论 在小鼠下颌第一磨牙牙根不同发育时期，3种染色方法各有优势。H-E染色适用于各类细胞形态的观察，Masson染色适用于牙体硬组织及骨组织的观察，Gomori染色法总体效果不佳。

Abstract:

Objective To compare the effects of three different staining methods during the root developmental stages of mandibular first molar in mice. Methods The coronal sections of mandibular first molars were prepared from ICR mice at the postnatal day5(PN5), PN7, PN14 and PN21. H-E staining, Masson trichrome staining and Gomori staining were used to analyze the root development, and the staining results were compared among three methods. Results From PN5 to PN7, the root development was at the initial stage, the morphology of odontoblast, ameloblast and Hertwigs root sheath(HERS) were stained clearly in H-E staining. The results of Masson trichrome staining were brightly colored with distinct layers of enamel, dentin and predentin. In Gomori staining, the nucleus were in a single blue-black color, but the contrast of soft and hard tissue was poor. From PN14(tooth eruption stage) to PN21(root development completed stage), the morphology of odontoblast, ameloblast and periodontal fibroblast stained clearly in H-E staining. Masson staining was the most effective method to investigate periodontal tissues as the collagen fiber was arranged regularly between the cementum and alveolar bone. Gomori staining was generally inferior to the other two methods. Conclusion The three different staining methods in the root development of the mandibular first molar in mice have their own advantages. H-E staining is appropriate for the observation of all kinds of cell morphology. Masson trichrome staining is suitable for the tooth hard tissue and bone tissue. Gomori staining is generally inferior to other two methods.