Ultrastructural and immunohistochemical studies of medullary bone calcification, with special reference to sulphated glycosaminoglycans

Abstract

Histochemical, immunohistochemical and electron energy-loss spectroscopic studies were performed to examine the relationship between sulphated glycosaminoglycans and medullary bone calcification using oestrogen-injected male Japanese quail. Sulphated glycosaminoglycans, detected by high iron diamine (HID) or HID-thiocarbohydrazide-silver protein (HID-TCH-SP) methods, were distributed throughout the matrix of medullary bone, some periphery and extending tips of the trabeculae stained weakly, and the globular structures at osteoid areas were exclusively positive for HID-TCH-SP stain. Immunohistochemistry identified keratan sulphate located in the globular structures at osteoid areas and calcified matrix, but chondroitin-4 sulphate and chondroitin-6 sulphate were not detected in the matrix. Using electron spectroscopic imaging, sulphur was determined to be localized in the globular structures. These results demonstrate that medullary bone matrix accumulates keratan sulphate in the globular structures, which are the foci for calcification, and eventually in the calcified areas. This suggests that keratan sulphate containing sulphur is maintained in the calcified matrix. These results indicate a unique process of calcification exists in medullary bone.

Ultrastructural and Immunocytochemical Analysis of Multilineage Differentiated Human Dental Pulp- and Umbilical Cord-Derived Mesenchymal Stem Cells

“…At the ultrastructural level, DPSC show all stages of osteogenesis, starting with MSCs which develop into mature osteocytes. In addition, the secretory activity of differen- tiated DPSC very closely resembles the globular secretion pattern which is seen during in vivo osteogenesis [Yamamoto et al, 2005]. The presence of hydroxyapatite needles confirms the occurrence of real mineralization in the culture.…”

Assessment of an Efficient Xeno-Free Culture System of Human Periodontal Ligament Stem Cells

“…To our knowledge, the current study showed for the first time the morphological characteristics and gene modulation of xeno-free hPDLSCs cultured in osteogenic inductive medium. Some differentiation steps of osteogenic cells and, in particular, the secretion of ECM resembling the secretion pattern during in vivo osteogenesis 44,45 were documented.…”

Sequential Expression of Osteoblast Phenotypic Genes During Medullary Bone Formation and Resorption in Estrogen‐Treated Male Japanese Quails

“…Organ cultures of medullary bone matrix from female birds and E2‐treated male birds included glycosaminoglycans, such as keratan sulfate and chondroitin sulfate (Fisher and Schraer, ’80; Hunter and Schraer, ’83). Yamamoto et al () demonstrated that these sulfated glycosaminoglycans accumulated in the bone matrix near cortical bone and acted as foci for mineralization, suggesting that they played an important role in the process of medullary bone mineralization. Meanwhile, Wang et al () suggested that the accumulation of keratan sulfate‐containing proteoglycans in the medullary bone matrix contributed to its rapid resorption.…”