Biomimetic remineralization of demineralized enamel with nano-complexes of phosphorylated chitosan and amorphous calcium phosphate

Remineralization of enamel plays a crucial role in the progression of carious process and the management of early caries lesion. Based on the influence of phosphorylated proteins in biomineralization, the objective of this study was to synthesize nano-complexes of phosphorylated chitosan and amorphous calcium phosphate (Pchi-ACP), and evaluate their ability to remineralize enamel subsurface lesions in vitro. Pchi was synthesized using a previously established chemical method. The biomimetic remineralizing solution containing nano-complexes of Pchi-ACP was prepared by adding CaCl2 and K2HPO4 into Pchi-ACP solution (0.5 % w/v) in sequence. The final concentrations of calcium and phosphate ions were 10 and 6 mM, respectively. The nano-complexes of Pchi-ACP were characterized by Fourier-transform infrared (FTIR), X-ray diffraction (XRD), transmission electron microscopy (TEM), and selected area electron diffraction (SAED). During testing the enamel lesions were treated with Pchi-ACP and fluoridated remineralizing solutions, respectively. The remineralizing of enamel lesions was examined with field emission electron microscope (FE-SEM) and Micro-CT. ACP was stabilized by Pchi to form nano-complexes that were soluble in water. The size of Pchi-ACP nano-complexes particles was determined to be less than 50 nm. XRD and SAED results confirmed their amorphous phases. FE-SEM and Micro-CT results showed that the remineralizing effect of Pchi-ACP on enamel lesions was similar to that of fluoride. However, the remineralizing rate of Pchi-ACP treatment was significantly higher than that of fluoride treatment (P < 0.05). This study highlighted the potential of nanoparticles functionalized with a natural analogue involved in biomineralization, to remineralize early enamel caries.