Goldberg M, Lécolle S
Laboratoire de Biologie et Biomatériaux du Milieu Buccal et Osseux, Faculté de Chirurgie Dentaire, Université Paris V, Montrouge, France.
Histochem J. 1995 May;27(5):401-10.
At neutral pH, poly-L-lysine-gold complexes labelled the predentine extensively, whereas in dentine the number of gold complexes was reduced by half. Hyaluronidase pretreatment of the section at pH 6.8, prior to labelling, suppressed most of the staining in predentine and did not affect dentine. In contrast, alkaline phosphatase pretreatment at pH 9 enhanced the gold complex labelling in predentine and removed most of the labelling in dentine. This proves that at pH 7.2, the polyanions which are stained include a heterogeneous population of glycosaminoglycans, located in predentine, and phosphoproteins, visualized in dentine. At acidic pH levels (2.9 and 1.1), the number of scored gold complexes decreased, but the ratio between predentine and dentine labelling remained constant. Hyaluronidase pretreatment removed or firmly reduced the gold complex labelling both in predentine and dentine, whereas alkaline phosphatase pretreatment of the sections at pH 9 prior to labelling did not induce any change. This argues in favour of an increased specificity of polylysine-gold complex staining for glycosaminoglycans, stained at low pH in both predentine and dentine. Differential staining of glycosaminoglycans and phosphoproteins according to the pH provides a useful tool for studying the role played respectively by the two matrix components in dentine mineralization.
在中性pH值条件下,聚-L-赖氨酸-金复合物广泛标记前期牙本质,而在牙本质中,金复合物的数量减少了一半。在标记前于pH 6.8对切片进行透明质酸酶预处理,可抑制前期牙本质中的大部分染色,且不影响牙本质。相反,在pH 9进行碱性磷酸酶预处理可增强前期牙本质中的金复合物标记,并去除牙本质中的大部分标记。这证明在pH 7.2时,被染色的聚阴离子包括位于前期牙本质中的多种糖胺聚糖和在牙本质中可见的磷蛋白。在酸性pH值水平(2.9和1.1)下,计分的金复合物数量减少,但前期牙本质和牙本质标记之间的比率保持不变。透明质酸酶预处理可去除或显著减少前期牙本质和牙本质中的金复合物标记,而在标记前于pH 9对切片进行碱性磷酸酶预处理则未引起任何变化。这表明聚赖氨酸-金复合物染色对在低pH值下染色的前期牙本质和牙本质中的糖胺聚糖具有更高的特异性。根据pH值对糖胺聚糖和磷蛋白进行差异染色,为研究这两种基质成分在牙本质矿化中分别发挥的作用提供了一个有用的工具。