Takagi M, Kazama T, Shimada K, Hosokawa Y, Hishikawa H
Department of Anatomy, Nihon University School of Dentistry, Tokyo, Japan.
Anat Rec. 1989 Dec;225(4):279-87. doi: 10.1002/ar.1092250404.
We have investigated ultrastructural cytochemical properties of elastic elements in Alligator periodontal ligaments decalcified with EDTA and stained with 1) the tannic acid-uranyl acetate (TA-UA) method for elastin in combination with elastase digestion; 2) the high iron diamine-thiocarbohydrazide-silver proteinate (HID-TCH-SP) method with prior treatment of specimens with either monopersulphate or cupric-sulphite reagent for the localization of disulphide- and/or sulphydryl-containing material (i.e., oxytalan fibers); and 3) HID-TCH-SP alone for sulphated complex carbohydrates. Many microfibrils accumulated to form either large or small bundles. Large bundles having a diameter of 2.50 +/- 1.10 microns (mean +/- SD; n = 50) each showed an apico-occlusal distribution, although small bundles measuring 0.63 +/- 0.13 microns (mean +/- SD; n = 50) in diameter each were exclusively localized in interstitial areas rich in vessels and nerves. The former bundles always lacked TA-UA reactivity and represented oxytalan fibers; the latter bundles frequently contained TA-UA-reactive elastase digestible components and were similar in appearance to immature elastic fibers or elaunin fibers. HID-TCH-SP after oxidation strongly stained both the oxytalan and elastic fiber microfibrils but stained the amorphous elastin very weakly or not all. In nonoxidized specimens, there was no definite HID-TCH-SP staining of microfibrils and the amorphous elastin, although adjacent matrix proteoglycans stained consistently. These results indicate that although there is a marked difference in the distribution and size of oxytalan and elastic fibers in Alligator periodontal ligaments, their associated microfibrils lack stainable sulphate groups but are enriched with disulphide and/or sulphydryl groups, as has been described in mammals.
我们研究了用乙二胺四乙酸(EDTA)脱钙并用以下方法染色的短吻鳄牙周韧带中弹性成分的超微结构细胞化学特性:1)用于弹性蛋白的鞣酸 - 醋酸铀(TA-UA)法结合弹性蛋白酶消化;2)高铁二胺 - 硫代碳酰肼 - 蛋白银(HID-TCH-SP)法,在标本先用过一硫酸盐或亚硫酸铜试剂处理后用于定位含二硫键和/或巯基的物质(即氧化弹力纤维);3)单独使用HID-TCH-SP法用于硫酸化复合碳水化合物。许多微原纤维聚集形成大束或小束。直径为2.50±1.10微米(平均值±标准差;n = 50)的大束均显示出根尖 - 咬合方向的分布,而直径为0.63±0.13微米(平均值±标准差;n = 50)的小束仅局限于富含血管和神经的间质区域。前者的束始终缺乏TA-UA反应性,代表氧化弹力纤维;后者的束经常含有TA-UA反应性的可被弹性蛋白酶消化的成分,外观与未成熟的弹性纤维或弹力素纤维相似。氧化后的HID-TCH-SP强烈染色氧化弹力纤维和弹性纤维微原纤维,但对无定形弹性蛋白染色非常弱或根本不染色。在未氧化的标本中,微原纤维和无定形弹性蛋白没有明确的HID-TCH-SP染色,尽管相邻的基质蛋白聚糖持续染色。这些结果表明,尽管短吻鳄牙周韧带中氧化弹力纤维和弹性纤维在分布和大小上存在明显差异,但它们相关的微原纤维缺乏可染色的硫酸基团,却富含二硫键和/或巯基,这与哺乳动物中的情况一致。