Ultrastructural distribution at the electron microscopic level of sulfated glycosaminoglycan in human fetal tooth germs
It is well known that glycoproteins, glycosaminoglycans and proteoglycans are of fllndamental importance to the processes of morphogenesis and cytodifferentiation dllring the teeth development. With HID-TCH-SP(High-iron diamine-thiocarbohydrazide-silver proteinate), slllfated glycosaminoglycans sllch as chondroitin slllfate and heparan slllfate have been localized at the 1I1trastructurallevel in a wide variety of tisslles. 까le pllrpose of this study were to characterize slllfated glycosaminoglycan profiles of hllman fetal tooth genns at 비trastructurallevel for the phase of morphogenesis and cytodifferentiation, and to detect the protein expression of sulfated glycosaminoglycan by immunoslot blot. Human tooth germs from the alveolar bone of twenty still born fetuses were f1xed in a mixture of 2% glutaraldehyde/ l% fonnaldehyde. The ultrathin sections were stained with HIDTCH- SP, and some sections were σ'eated with 0.05% solution of testicular hyaluronidase to identify the histochemical properties of the HID-TCH-SP stain deposits. For semiquantitative protein assay, immunoslot blot was done. The obtained results were as follows 1. HID-TCH-SP staining showed sulfated glycocongugated deposits in DEJ, peritublllar dentin, and mantle dentin matrix, enamel prism sheath, interrod area, and enamel matrix. 2. Heparan sulfate deposits in DEJ resisted to testicular hyalllronidase treatment prior to HID-TCH-SP staining 3. In immunoslot blot, chondroitin slllfate was detected higher in enamel and dentin extraα , while heparan slllfate was relatively expressed in enamel and dentin extract, but rarely expressed in enamel or dentin extract. From the aboving results, it was suggested that chondroitin and heparan sulfate would play an important role in the formation of D티, while chondroitin sulfate would in the development of enamel prism sheath, enamel matrix, and mantle or peritllblllar dentin of human fetal t∞th germs.