BACKGROUND: Amelogenesis imperfecta is a hereditary disorder affecting dental enamel. Among its phenotypes, hypocalcified AI is characterized by mineral deficiency, leading to tissue wear and, consequently, dental sensitivity. Excessive fluoride intake (through drinking water, fluoride supplements, toothpaste, or by ingesting products such as pesticides or insecticides) can lead to a condition known as dental fluorosis, which manifests as stains and teeth discoloration affecting their structure. Our recent studies have shown that extracts from Colombian native plants, Ilex guayusa and Piper marginatum, deposit mineral ions such as phosphate and orthophosphate into the dental enamel structure; however, it is unknown whether these extracts produce toxic effects on the dental pulp.
OBJECTIVE: To assess cytotoxicity effects on human dental pulp stem cells (hDPSCs) exposed to extracts isolated from I. guayusa and P. marginatum and, hence, their safety for clinical use.
METHODS: Raman spectroscopy, fluorescence microscopy, and flow cytometry techniques were employed. For Raman spectroscopy, hDPSCs were seeded onto nanobiochips designed to provide surface-enhanced Raman spectroscopy (SERS effect), which enhances their Raman signal by several orders of magnitude. After eight days in culture, I. guayusa and P. marginatum extracts at different concentrations (10, 50, and 100 ppm) were added. Raman measurements were performed at 0, 12, and 24 h following extract application. Fluorescence microscopy was conducted using an OLIMPUS fv1000 microscope, a live-dead assay was performed using a kit employing a BD FACS Canto TM II flow cytometer, and data analysis was determined using a FlowJo program.
RESULTS: The Raman spectroscopy results showed spectra consistent with viable cells. These findings were corroborated using fluorescence microscopy and flow cytometry techniques, confirming high cellular viability.
CONCLUSIONS: The analyzed extracts exhibited low cytotoxicity, suggesting that they could be safely applied on enamel for remineralization purposes. The use of nanobiochips for SERS effect improved the cell viability assessment.

For almost three decades from its discovery, amorphous calcium phosphate (ACP) was not considered a suitable biomaterial due to its structural instability. Thanks to its unique properties in respect to crystalline calcium phosphate phases, nowadays ACP is used in promising devices for hard tissue regeneration. Here we have highlighted the features of ACP that were harnessed to create excellent biomaterials for dental remineralization, self-setting bone cements, drug delivery, and coatings of prostheses. Its current limitations as well as future perspectives of development were concisely described. Despite more research works are needed, we envisage that the future of ACP is bright.

White spot lesions (WSL) and incipient caries on enamel surfaces are the earliest clinical outcomes for demineralization and caries. If left untreated, the caries can progress and may cause complex restorative procedures or even tooth extraction which destroys soft and hard tissue architecture as a consequence of connective tissue and bone loss. Current clinical practices are insufficient in treating dental caries. A long-standing practical challenge associated with demineralization related to dental diseases is incorporating a functional mineral microlayer which is fully integrated into the molecular structure of the tooth in repairing damaged enamel. This study demonstrates that small peptide domains derived from native protein amelogenin can be utilized to construct a mineral layer on damaged human enamel in vitro. Six groups were prepared to carry out remineralization on artificially created lesions on enamel: (1) no treatment, (2) Ca2+ and PO43- only, (3) 1100 ppm fluoride (F), (4) 20 000 ppm F, (5) 1100 ppm F and peptide, and (6) peptide alone. While the 1100 ppm F sample (indicative of common F content of toothpaste for homecare) did not deliver F to the thinly deposited mineral layer, high F test sample (indicative of clinical varnish treatment) formed mainly CaF2 nanoparticles on the surface. Fluoride, however, was deposited in the presence of the peptide, which also formed a thin mineral layer which was partially crystallized as fluorapatite. Among the test groups, only the peptide-alone sample resulted in remineralization of fairly thick (10 μm) dense mineralized layer containing HAp mineral, resembling the structure of the healthy enamel. The newly formed mineralized layer exhibited integration with the underlying enamel as evident by cross-sectional imaging. The peptide-guided remineralization approach sets the foundation for future development of biomimetic products and treatments for dental health care.

Analyze the effect of varnishes containing xylitol compared to commercial fluoridated varnishes on the remineralization of artificial enamel caries lesions in situ.
Twenty subjects took part in this crossover, double-blind study performed in four phases of 5days each. Each subject worn palatal appliances containing four predemineralized bovine enamel specimens. Artificial caries lesions were produced by immersion in 30ml of lactic acid buffer containing 3mM CaCl2·2H2O, 3mM KH2PO4, 6μM tetraetil metil diphosphanate (pH 5.0) for 6days. The specimens in each subject were treated once with the following varnishes: 20% xylitol (experimental); Duofluorid™ (6% NaF, 6% CaF2), Duraphat™ (5% NaF, positive control) and placebo (no-F/xylitol, negative control). The varnishes were applied in a thin layer and removed after 6h. Fifteen subjects were able to finish all phases. The enamel alterations were quantified by surface hardness and transversal microradiography. The percentage of surface hardness recovery (%SHR), the integrated mineral loss and lesion depth were statistically analyzed by Friedmann and Dunn\'s tests test (p<0.05).
Enamel surface remineralization was significantly increased by Duraphat™, Duofluorid™ and 20% xylitol formulations. Significant subsurface mineral remineralization could also be seen for the experimental and commercial varnishes, except for Duraphat™, when the parameter \"lesion depth\" was considered.
20% xylitol varnish seem to be a promising alternative to increase surface and subsurface remineralization of artificial caries lesions in situ.
effective vehicles are desirable for caries control. Xylitol varnishes seem to be promising alternatives to increase enamel remineralization in situ, which should be confirmed by clinical studies.

OBJECTIVE: Analyse the effect of varnishes containing xylitol alone or combined with fluoride on the remineralization of artificial enamel caries lesions in vitro.
METHODS: Bovine enamel specimens were randomly allocated to 7 groups (n=15/group). Artificial caries lesions were produced by immersion in 30 mL of lactic acid buffer containing 3mM CaCl2·2H2O, 3mM KH2PO4, 6 μM tetraetil metil diphosphanate (pH 5.0) for 6 days. The enamel blocks were treated with the following varnishes: 10% xylitol; 20% xylitol; 10% xylitol plus F (5% NaF); 20% xylitol plus F (5% NaF); Duofluorid™ (6% NaF, 2.71% F+6% CaF2), Duraphat™ (5% NaF, positive control) and placebo (no-F/xylitol, negative control). The varnishes were applied in a thin layer and removed after 6h. The blocks were subjected to pH-cycles (demineralization-2h/remineralization-22 h during 8 days) and enamel alterations were quantified by surface hardness and transversal microradiography. The percentage of surface hardness recovery (%SHR), the integrated mineral loss and lesion depth were statistically analysed by ANOVA/Tukey\'s test or Kruskal-Wallis/Dunn\'s test (p<0.05).
RESULTS: Enamel surface remineralization was significantly increased by Duraphat™, 10% xylitol plus F and 20% xylitol plus F formulations, while significant subsurface mineral remineralization could be seen only for enamel treated with Duraphat™, Duofluorid™ and 20% xylitol formulations.
CONCLUSIONS: 20% xylitol varnishes seem to be promising alternatives to increase remineralization of artificial caries lesions.
CONCLUSIONS: effective vehicles are desirable for caries control. Xylitol varnishes seem to be promising alternatives to increase enamel remineralization in vitro, which should be confirmed by in situ and clinical studies.