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The external layer of a teleost fish scale is composed of type I collagen, an amorphous matrix substance and hydroxyapatite crystals. Calcification of this layer can be inhibited in the scale-regenerating process under a calcium and phosphate deficient (CaDPD) conditions, and can be facilitated by incubation in physiological saline. The aim of this study was to evaluate this model of calcification using histological and quantitative analysis in order to promote further understanding of the mechanism of calcification in fish scales. We found that the external layer of the scales produced under a CaDPD condition contained more densely-aligned collagen fibrils with a small amount of the amorphous matrix substance. The CaDPD scale contained only one-third of the amount of calcium and phosphate present in the control fish. After 4-hour incubation, a 2-3 fold increase in calcium content and a 1.5-fold increase in phosphate content were observed. Calcification proceeded in the external layer, and mineral deposits grew only in the electron-dense matrix substance. We conclude that this model would be suitable to study the early process of fish scale calcification that occurs in the non-collagenous substance. The electron-dense substance may contain key molecules that promote calcification

Scale calcification in the goldfish in vitro: histological and quantitative analysis