Three-dimensional characteristics of submandibular salivary gland of ageing rats: An HRSEM study

Abstract

The characteristics of the submandibular glands of ageing Wistar rats were studied using light and high-resolution scanning electron microscopy (HRSEM) methods. For light microscopy, the samples were fixed in Bouin solution and embedded in paraffin. Sections were stained with hematoxilin-eosin and Azo-Carmin. For conventional scanning electron microscopy, the tissues were fixed in modified Karnovsky solution, and treated with NaOH solution for 3-4 days. The O-D-O method was used for HRSEM. The samples were fixed in 2% osmium tetroxide, macerated in diluted osmium and dehydrated in an increasing series of ethanol. The samples were dried in a critical point dryer, coated with gold-palladium and examined in a Hitachi high-resolution scanning electron microscope, S-900. The results showed that submandibular glands with lobules are separated by connective tissue septum. The acinar formations and the ducts, revealing the serous and mucous cells were observed. After fracture in liquid nitrogen and treatment with NaOH solution to remove the cellular components, the original disposition of the collagen bundles fibers were revealed corresponding to the round, oval or irregular acinar and ductal structures. In the cytoplasm, organelles such as mitochondria, rough endoplasmic reticulum, Golgi apparatus and serous and mucous secretory granules were observed localized in the apical portion in three-dimensional HRSEM images. The serous secretory granules presented different sizes and shapes showing the modifications which occurred in the ageing rats. The striated duct cells revealed the presence of the secretory cells and mitochondria in parallel disposition. The mitochondrial cristae were noted in three-dimensional aspects. The lumen presented numerous cytoplasmic microprojections. The lumen of excretor ducts are covered by polygonal epithelial cells containing numerous microplicae.