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. 2023 May 26;15(11):2470.
doi: 10.3390/polym15112470.

Biomimetic Hydroxyapatite Crystals Growth on Phosphorylated Chitosan Films by In Vitro Mineralization Used as Dental Substitute Materials

Fathia Rahmani  1   2 , Omar Larbi Bouamrane  3   4 , Amina Ben Bouabdallah  2   5   6 , Leonard I Atanase  7   8 , Abdelkader Hellal  4   9 , Aurelian Nichita Apintiliesei  7
Affiliations

Biomimetic Hydroxyapatite Crystals Growth on Phosphorylated Chitosan Films by In Vitro Mineralization Used as Dental Substitute Materials

Fathia Rahmani et al. Polymers (Basel). .

Abstract

Chitosan (CS) films exhibit great potential as a substrate for the in vitro mineralization process. In this study, to mimic the formation of nanohydroxyapatite (HAP) as natural tissue, CS films coated with a porous calcium phosphate were investigated using scanning electron microscopy (SEM), Energy dispersive X-ray spectroscopy (EDX), Fourier transforms infrared spectroscopy (FTIR), X-ray diffractometry (XRD) and X-ray photoelectron spectroscopy (XPS). Calcium phosphate coating deposited on phosphorylated derivatives of CS was obtained by a process based on phosphorylation, Ca(OH)2 treatment and artificial saliva solution (ASS) immersion. The phosphorylated CS films (PCS) were obtained by partial hydrolysis of the PO4 functionalities. It was demonstrated that this precursor phase could induce the growth and the nucleation of the porous calcium phosphate coating when immersed in ASS. Moreover, oriented crystals and qualitative control of calcium phosphate phases on CS matrices are obtained in a biomimetic mode. Furthermore, in vitro antimicrobial activity of PCS was evaluated against three species of oral bacteria and fungi. It revealed an increase in antimicrobial activity with minimum inhibition concentration (MIC) values of 0.10% (Candida albicans), 0.05% (Staphylococcus aureus) and 0.025% (Escherichia coli) which proves their possible use as dental substitute materials.

Keywords: biomineralization; calcium phosphate; chitosan film; dental material; hydroxyapatite; phosphorylated chitosan.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
FTIR spectrum of (a) CS film, (b) PCS film and (c) Ca(OH)2 treated PCS film.
Figure 2
Figure 2
FTIR spectra of Ca(OH)2 treated phosphorylated chitosan film after soaking in ASS for (a) 4 days, (b) 15 days, (c) 21 days and (d) 28 days.
Figure 3
Figure 3
Schematic illustration of the mineralization on PCS film. (a) Scheme of phosphorylated and mineralized CS, (b) Interaction between CS and phosphate to form phosphorylated chitosan films, (c) Formation of OCP phase after addition of Ca2+ ions, and (d) HAP crystals grow.
Figure 4
Figure 4
XRD spectra of samples of (a) Ca(OH)2-treated PCS films. Soaking time in ASS: (b) 4 days, (c) 15 days and (d) 28 days.
Figure 5
Figure 5
Scanning electron micrograph of a sample of PCS films after immersion in saturated Ca(OH)2 for eight days at room temperature. Scale bars: (a) 2 μm, (b) 1 μm.
Figure 5
Figure 5
Scanning electron micrograph of a sample of PCS films after immersion in saturated Ca(OH)2 for eight days at room temperature. Scale bars: (a) 2 μm, (b) 1 μm.
Figure 6
Figure 6
Scanning electron micrograph of samples of Ca(OH)2 treated PCS films after soaking in ASS for 15 days. Scale bars: (a) 20 μm, (b) 2 μm, (c) 1 μm.
Figure 6
Figure 6
Scanning electron micrograph of samples of Ca(OH)2 treated PCS films after soaking in ASS for 15 days. Scale bars: (a) 20 μm, (b) 2 μm, (c) 1 μm.
Figure 7
Figure 7
Scanning electron micrograph of samples of Ca(OH)2 treated PCS films after 28 days immersion in ASS. Scale bars: (a) 3 μm, (b) 1 μm.
Figure 8
Figure 8
Ca/P ratio for the CaP coating formed on Ca(OH)2 treated PCS films as a function of immersion time in ASS solution.
Figure 9
Figure 9
Typical XPS spectra of (a) CS films (b) PCS films (c) PCS-Ca(OH)2 treated films (d) PCS-Ca(OH)2 treated four days biomineralized (e) PCS-Ca(OH)2 treated 15 days biomineralized (f) PCS-Ca(OH)2 treated 28 days biomineralized.
Figure 10
Figure 10
N1S, O1S and P2P XPS high-resolution spectra of: (a,c) unmodified CS films, (b,d,e) PCS films.
Figure 11
Figure 11
Ca2P and P2P XPS high-resolution spectra of (a) PCS-Ca(OH)2 treated (b) PCS-Ca(OH)2 treated four days biomineralized (c) PCS-Ca(OH)2 treated 15 days biomineralized (d) PCS-Ca(OH)2 treated 28 days biomineralized.
Figure 12
Figure 12
C1S XPS high-resolution spectra of (a) unmodified CS films, (b) PCS films.
Figure 13
Figure 13
Weight variation of biomineralized films in ASS as a time function.
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