A Novel Nano-Hydroxyapatite Agarose-Based Hydrogel for Biomimetic Remineralization of Demineralized Human Enamel: An in-vitro Study

Abstract

Purpose: This study aims to investigate the biomimetic effect of agarose hydrogel loaded with enamel matrix derivative (EMD-agarose) alone or in combination with nano-hydroxyapatite (n-HA-EMD-agarose) on the remineralization of human demineralized enamel. Methods: Extracted human mandibular third molars were sectioned into 54 buccal and lingual halves. Acid-resistant nail varnish was applied to each half, except for two enamel windows. Enamel surface microhardness, energy-dispersive X-ray spectroscopy (EDX), and scanning electron microscopy (SEM) analyses were conducted to evaluate enamel surfaces at baseline, following demineralization with 37% phosphoric acid, and after each hydrogel application and remineralization for two, four, and six days. Remineralization was performed using a phosphate solution at 37°C. Results: At day 6 following remineralization, a statistically significant higher mean microhardness was recorded in n-HA-EMD-agarose hydrogel (260.87 ± 3.52) as compared to EMD-agarose hydrogel (244.63 ± 2.76) (p = 0.027). Similarly, n-HA-EMD-agarose hydrogel showed a higher mean calcium (46.31 ± 2.78), phosphorous (24.92 ± 0.826), and fluoride (0.909 ± 0.053) weight percentage compared to EMD-agarose hydrogel calcium (19.64 ± 1.092), phosphorous (19.64 ± 1.092), and fluoride (0.7033 ± 0.0624) weight percentage (p < 0.05). Further, SEM analysis revealed a substantial deposition of n-HA following the application of the n-HA-EMD-agarose hydrogel, whereas the EMD-agarose exhibited a relatively smooth enamel surface with less visible enamel rods due to mineral deposition. Conclusion: The combined n-HA-EMD-agarose hydrogel demonstrated improved surface microhardness of the remineralized enamel and enhanced mineral content deposition, indicating its potential as a biomimetic approach for dental enamel repair.