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. 2021 May 13;26(10):2899.
doi: 10.3390/molecules26102899.

Mineralization of Phosphorylated Fish Skin Collagen/Mangosteen Scaffolds as Potential Materials for Bone Tissue Regeneration

Eduardo P Milan  1   2 , Murilo Á V Rodrigues  3 , Virginia C A Martins  3 , Ana M G Plepis  1   3 , Thomas Fuhrmann-Lieker  2 , Marilia M Horn  2
Affiliations

Mineralization of Phosphorylated Fish Skin Collagen/Mangosteen Scaffolds as Potential Materials for Bone Tissue Regeneration

Eduardo P Milan et al. Molecules. .

Abstract

In this study, a potential hard tissue substitute was mimicked using collagen/mangosteen porous scaffolds. Collagen was extracted from Tilapia fish skin and mangosteen from the waste peel of the respective fruit. Sodium trimetaphosphate was used for the phosphorylation of these scaffolds to improve the nucleation sites for the mineralization process. Phosphate groups were incorporated in the collagen structure as confirmed by their attenuated total reflection Fourier transform infrared (ATR-FTIR) bands. The phosphorylation and mangosteen addition increased the thermal stability of the collagen triple helix structure, as demonstrated by differential scanning calorimetry (DSC) and thermogravimetry (TGA) characterizations. Mineralization was successfully achieved, and the presence of calcium phosphate was visualized by scanning electron microscopy (SEM). Nevertheless, the porous structure was maintained, which is an essential characteristic for the desired application. The deposited mineral was amorphous calcium phosphate, as confirmed by energy dispersive X-ray spectroscopy (EDX) results.

Keywords: fish skin collagen; mangosteen; mineralization; phosphorylation.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
SDS-PAGE of (A) a protein marker; and (B) the extracted collagen from Tilapia skin.
Figure 2
Figure 2
Quantitative comparison of phosphorous contents in phosphorylated scaffolds. (Same letter in the graphs means no significant difference; significance level of 5%).
Figure 3
Figure 3
FTIR spectra of (A) collagen and phosphorylated collagen scaffolds: (a) C; (b) CP; (c) C10P; and (d) C30P, and (B) mineralized scaffolds: (a) CP25; (b) C10P25; (c) C10P37; (d) C30P25 and (e) C30P37.
Figure 4
Figure 4
FTIR spectrum of the extract obtained from the mangosteen peel.
Figure 5
Figure 5
DSC curves of the scaffolds. In (a) C; (b) CP; (c) C10P; and (d) C30P.
Figure 6
Figure 6
TGA curves for (a) CP25, (b) C10P25; (c) C10P37; (d) CP; and (e) C10P.
Figure 7
Figure 7
TGA curves for (a) C30P37; (b) C30P25; (c) C30P; and (d) C.
Figure 8
Figure 8
SEM Micrographs of collagen scaffold before and after phosphorylation. (A) C, (B) CP, (C) C10P, (D) C30P. Magnification of 200×.
Figure 9
Figure 9
SEM Micrographs of collagen scaffold and calcium phosphate mineralization. (A) CP25; (B) C10P25, with respective EDX insert; (C) C30P25 (Magnification of 700×) and (D) C30P25 (Magnification of 12,000×).
See this image and copyright information in PMC

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