In this report, we demonstrated the formation of a biomimetic mineralizing layer obtained on the surface of dental enamel (biotemplate) using bioinspired nanocrystalline carbonate-substituted calcium hydroxyapatite (ncHAp), whose physical and chemical properties are closest to the natural apatite dental matrix, together with a complex of polyfunctional organic and polar amino acids. Using a set of structural, spectroscopy, and advanced microscopy techniques, we confirmed the formation of a nanosized ncHAp-based mineralized layer, as well as studying its chemical, substructural, and morphological features by means of various methods for the pretreatment of dental enamel. The pretreatment of a biotemplate in an alkaline solution of Ca(OH)₂ and an amino acid booster, together with the executed subsequent mineralization with ncHAp, led to the formation of a mineralized layer with homogeneous micromorphology and the preferential orientation of the ncHAp nanocrystals. It was shown that the homogeneous crystallization of hydroxyapatite on the biotemplate surface and binding of individual nanocrystals and agglomerates into a single complex by an amino acid booster resulted in an increase (~15%) in the nanohardness value in the enamel rods area, compared to that of healthy natural enamel. Obtaining a similar hierarchy and cleavage characteristics as natural enamel in the mineralized layer, taking into account the micromorphological features of dental tissue, is an urgent problem for future research.

The aim of this work is to develop a biomimetic interface between the natural tooth tissue and the restorative composite and to study it on the basis of synchrotron micro-FTIR mapping and multidimensional processing of the spectral data array. Using hierarchical cluster analysis of 3D FTIR data revealed marked improvements in the formation of the dentine/adhesive/dental hybrid interface using a biomimetic approach. The use of a biomimetic strategy (application of an amino acid–modified primer, alkaline calcium and a nano-c-HAp–modified adhesive) allowed the formation of a matrix that can be structurally integrated with natural dentine and dental composite. The biomimetic hybrid layer was characterised by homogeneous chemical composition and a higher degree of conversion of the adhesive during polymerisation, which should provide optimal integration of the dental composite with the dentine.

In our paper, we discuss the results of a comprehensive structural-spectroscopic and microscopic analysis of non-stoichiometric nanocrystalline hydroxyapatite (CHAp) with low carbonate anion content and biomimetic hybrid nanomaterials produced on its basis. It was shown that hydroxyapatite nanocrystals synthesized by chemical precipitation and biogenic calcium source mimic the properties of biogenic apatite and also have a morphological organization of “core–shell” type. The “core” of the CHAp nanocrystal is characterized by an overabundance of calcium Ca/P~1.9. Thus “a shell” with thickness of ~3–5 nm is formed from intermediate apatite-like phases where the most probable are octocalcium phosphate, dicalcium phosphate dihydrate and tricalcium phosphate. The multimode model of the Raman profile of samples CHAp and biomimetic composites for spectral region 900–1100 cm⁻¹ proposed in our work has allowed to allocate precise contribution of B-type carbonate substitution, taking into account the presence on a surface of “core” HAp nanocrystal of various third-party intermediate apatite-like phases. The calibration function constructed on the basis of the described model makes it possible to reliably determine small concentrations of carbonate in the structure of hydroxyapatite with the application of Raman express method of diagnostics. The results of our work can inspire researchers to study the processes of induced biomineralization in mineralized tissues of the human body, using non-destructive methods of control with simultaneous analysis of chemical bonding, as well as determining the role of impurity atoms in the functions exhibited by biotissue.

In this work, for the first time, the influence of the coordination environment as well as Ca and P atomic states on biomimetic composites integrated with dental tissue was investigated. Bioinspired dental composites were synthesised based on nanocrystalline calcium carbonate-substituted hydroxyapatite $C{a}_4^{\mathrm{I}}C{a}_6^{\mathrm{II}}{\left(P{O}_4\right)}_{6-x}{\left(C{O}_3\right)}_{x+y}{\left(OH\right)}_{2-y}$ (nano-cHAp) obtained from a biogenic source and a set of polar amino acids that modelled the organic matrix. Biomimetic composites, as well as natural dental tissue samples, were investigated using Raman spectromicroscopy and synchrotron X-ray absorption near edge structure (XANES) spectroscopy. Molecular structure and energy structure studies revealed several important features related to the different calcium atomic environments. It was shown that biomimetic composites created in order to reproduce the physicochemical properties of dental tissue provide good imitation of molecular and electron energetic properties, including the carbonate anion CO₃²⁻ and the atomic Ca/P ratio in nanocrystals. The features of the molecular structure of biomimetic composites are inherited from the nano-cHAp (to a greater extent) and the amino acid cocktail used for their creation, and are caused by the ratio between the mineral and organic components, which is similar to the composition of natural enamel and dentine. In this case, violation of the nano-cHAp stoichiometry, which is the mineral basis of the natural and bioinspired composites, as well as the inclusion of different molecular groups in the nano-cHAp lattice, do not affect the coordination environment of phosphorus atoms. The differences observed in the molecular and electron energetic structures of the natural enamel and dentine and the imitation of their properties by biomimetic materials are caused by rearrangement in the local environment of the calcium atoms in the HAp crystal lattice. The surface of the nano-cHAp crystals in the natural enamel and dentine involved in the formation of bonds with the organic matrix is characterised by the coordination environment of the calcium atom, corresponding to its location in the Ca^I position—that is, bound through common oxygen atoms with PO₄ tetrahedrons. At the same time, on the surface of nano-cHAp crystals in bioinspired dental materials, the calcium atom is characteristically located in the Ca^II position, bound to the hydroxyl OH group. The features detected in the atomic and molecular coordination environment in nano-cHAp play a fundamental role in recreating a biomimetic dental composite of the natural organomineral interaction in mineralised tissue and will help to find an optimal way to integrate the dental biocomposite with natural tissue.

(1) Objectives: This study is the first one to investigate the molecular composition of the dental biofilm during the exogenous and endogenous prophylaxis stages (use of dentifrice/drug) of individuals with different cariogenic conditions using molecular spectroscopy methods. (2) Materials and Methods: The study involved 100 participants (50 males and 50 females), aged 18–25 years with different caries conditions. Biofilm samples were collected from the teeth surface of all participants. The molecular composition of biofilms was investigated using synchrotron infrared microspectroscopy. Changes in the molecular composition were studied through calculation and analysis of ratios between organic and mineral components of biofilm samples. (3) Results: Based on the data obtained by synchrotron FTIR, calculations of organic and mineral component ratios, and statistical analysis of the data, we were able to assess changes occurring in the molecular composition of the dental biofilm. Variations in the phosphate/protein/lipid, phosphate/mineral, and phospholipid/lipid ratios and the presence of statistically significant intra- and inter-group differences in these ratios indicate that the mechanisms of ion adsorption, compounds and complexes arriving from oral fluid into dental biofilm during exo/endogenous prophylaxis, differ for patients in norm and caries development. (4) Conclusions: The conformational environment and charge interaction in the microbiota and the electrostatic state of the biofilm protein network in patients with different cariogenic conditions play an important role. (5) Clinical Significance: Understanding the changes that occur in the molecular composition of the dental biofilm in different oral homeostasis conditions will enable successful transition to a personalised approach in dentistry and high-tech healthcare.

In this exploratory study, we searched for associations between the two most common diseases of the oral cavity—dental caries and periodontal diseases—taking into account additional factors, such as personalized clinical pictures (the individual risk factors of the patient), based on the method of a multivariate data analysis of the molecular changes in the composition of human gingival crevicular fluid (GCF). For this purpose, a set of synchrotron Fourier-transform infrared spectroscopy (FTIR) spectra of gingival crevicular fluid samples from patients with different demographics, levels of dental caries development and periodontal diseases, and the presence/absence of concomitant chronic diseases were obtained and analyzed. Using a set of techniques (v-, F-, Chi-square tests; a principal component analysis (PCA); and the hierarchical clustering of principal components (HCPCs)) implemented in the R package FactoMineR allowed us to assess the relationship between the principal components (PCs) and characteristics of the respondents. By identifying the features (vibrational modes in the FTIR spectra) that contribute most to the differentiation of the spectral dataset, and by taking into account the interrelationships between the patients’ characteristics, we were able to match specific biological markers (specific molecular groups) to the two factors of interest—two types of oral pathologies. The results obtained show that the observed changes in the quantitative and qualitative composition of the modes in the infrared (IR) spectra of the GCF samples from patients with different dental caries developments and periodontal diseases present confirm the difficulty of identifying patient-specific spectral information. At the same time, different periodontal pathologies are more closely associated with other characteristics of the patients than the level of their caries development. The multivariate analysis performed on the spectral dataset indicates the need to take into account not only the co-occurrence of oral diseases, but also some other factors. The lack of this consideration (typical in lots of studies in this area) may lead to misinterpretations and consequently to a loss of data when searching for biological markers of certain oral diseases.