Annotation of the results obtained in 2018
In 2018, during the implementation of the third phase of the Project, the following work was completed: –The development of caries of the human tooth in depth has been studied and initial / residual cariogenic factors have been established; –The conditions of reversibility of the emerging enamel caries on the basis of phase formation, structure, stoichiometry, elemental composition of the deposited and the newly formed enamel layers were determined; - Changes in the molecular composition of saliva in people with multiple caries were studied using spectroscopic methods of analysis; –– Potential biomarkers in the human oral fluid were identified to effectively determine the physiological and pathological processes associated with the development of caries; –– proposed for industrial production and preclinical studies of therapeutic and prophylactic agents based on laboratory samples of biomimetic materials developed in the Project, normalizing metabolic processes in the hard tissues of teeth to prevent the development of carious process. - Algorithms of treatment-and-prophylactic measures are formed from conservative therapy, without preparation of hard tissues during the initial manifestations of the carious enamel process and tooth cement, to restoration therapy focused on prevention in clinically determined foci of destruction of hard tooth tissues. As part of the work performed, the following scientific results were obtained: - Using the methods of IR-spectroscopy, including using synchrotron radiation on the equipment of the “megasynes” class installation - Australian synchrotron of Melbourne, Raman and fluorescence spectroscopy to analyze the processes of demineralization / remineralization, as well as the conditions of reversibility of nascent enamel caries that the biocomposites created by us based on carbonate-substituted hydroxyapatite synthesized from a biological source of calcium and an organic component have The group of compounds found in the enamel tubules of the teeth: amino acids L-arginine hydrochloride, L-histidine, L-lysine hydrochloride and important for hyaluronic acid metabolic processes are promising agents for the remineralization of native dental tissue. - It has been experimentally shown that, due to the multistage nature of the ion exchange process, remineralizing agents should be kept in the oral cavity for a long time and come into contact with the tooth enamel. This requires the active remineralization of dental hard tissue using biocomposites based on carbonate-substituted hydroxyapatite, synthesized from a biological source of calcium and an organic component, having in its composition a group of compounds found in the enamel tubules of the teeth. The return of dicalcium phosphate dihydrate formed in the carious region during the development of the disease to a stable phase occurs through the substitution of OH bonds from the organic complex in the carbonate-substituted hydroxyapatite bond structure and the biological process of remineralization of the enamel and stabilization of the buffer system ensuring the presence of her free calcium ions. - Using the FTIR method, including using synchrotron radiation, the features in the IR spectra were determined, mineral-organic, carbon-phosphate, AmidII / AmidI and protein / thiocyanate ratios were calculated and the changes occurring in the molecular composition of saliva in people with multiple caries using spectroscopic methods of analysis. - Based on a comprehensive analysis of experimental data, it was possible to show that the organomineral balance of oral fluid in persons suffering from multiple caries shifts towards reducing the content of mineral groups and complexes in it and increasing the organic component. An integral indicator of these changes in molecular composition is the ratio AmidII / AmidI. In the group with multiple caries, this ratio increases by ~ 120% in relation to the group of healthy patients. - The most significant changes in the composition of the oral fluid of people suffering from multiple caries occur in relation to the number of –N = C = S groups correlated with the presence of thiocyanate in saliva and observed in the IR spectrum in the 2150 - 1950 cm-1 region. In the case of multiple dental caries in the samples of mixed saliva, the content of thiocyanates more than doubles. In addition, carbonic groups of esters, lipids and carbohydrates appear in the mixed saliva, the appearance of which is characteristic of caries development processes. - The complex results obtained by us and the tested methodology can be used as biomarkers and a diagnostic approach in the analysis of samples of mixed saliva of patients to assess the cariogenic situation. - As a result of detection of low molecular weight cationic proteins and hyaluronic acid using histochemical reactions in hard tooth tissues, it was proposed to Raduga-R, Voronezh to include protein substances and carbonate substituted hydroxyapatite from a biogenic source of calcium in the composition of treatment-and-prophylactic means to prevent development initial carious and secondary carious process during restoration therapy. - The results obtained in the Project allowed us to form algorithms for treatment-and-prophylactic measures aimed at preventing the development of the carious process in the hard tissues of the tooth. - The scientific results of our Project have already made a significant contribution to solving the problem of regenerative and regenerative medicine related to the development of effective preventive measures for socially significant dental diseases of a cariogenic nature, which are of the greatest importance due to the impact on all spheres of human social activity. The methods of preventive dental care developed in the Project due to the normalization of metabolic processes in the solid tissues of a human tooth in vivo using biomimetic materials were communicated to health care organizers in the Russian Federation. - Based on the data obtained in our Project in 2019 in the Voronezh Region, a pilot project is planned to correct the eating behavior of schoolchildren in order to prevent the development of caries in them. The plan of preventive measures to prevent the development of dental caries in children of school age is based on the concept of the etiology of the disease, as well as the conditions for the normalization of metabolic processes in the solid tissues of the human tooth using exogenous and endogenous methods of preventing caries, determined during the implementation of the Project. Links to information resources on the Internet (url-addresses) dedicated to the project: http://www.dentistrytoday.com/news/industrynews/item/3421-people-with-caries-have-more-organic-components-in-their-saliva https://www.eurekalert.org/pub_releases/2018-06/arsc-nwt060818.php https://chelorg.com/2018/05/30/the-doctors-learned-to-predict-the-development-of-caries/ https://indicator.ru/news/2018/05/30/kak-predskazat-razvitie-kariesa/ https://chrdk.ru/news/rossiiskie-uchenye-nauchilis-predskazyvat-karies-po-slyune http://rscf.ru/en/node/3126 https://riavrn.ru/news/voronezhskie-uchenye-razrabotali-metod-diagnostiki-kariesa/ https://rg.ru/2018/06/04/reg-cfo/v-voronezhe-pridumali-sposob-predotvratit-karies.html https://www.vrn.kp.ru/online/news/3135559/ http://www.sib-science.info/ru/institutes/veschestva-pomogut-borotsya-30052018 https://ria.ru/science/20180530/1521663341.html https://www.pravda.ru/news/science/30-05-2018/1385135-zyb-0/ http://polit.ru/news/2018/05/31/ps_rnf/ https://iz.ru/749731/2018-05-30/mediki-nauchilis-predskazyvat-razvitie-kariesa https://nation-news.ru/370134-rossiiskie-mediki-nauchilis-po-slyune-predskazyvat-razvitie-kariesa https://health.mail.ru/news/naydennye_vonbsprtu_veschestva_pomogut_borotsya/ http://www.nanonewsnet.ru/news/2018/naidennye-vo-rtu-veshchestva-pomogut-borotsya-s-kariesom http://vestivrn.ru/novosti/voronezhskie-spetsialistyi-sdelali-otkryitie-sposobnoe-izmenit-predstavlenie-o-lechenii-kariesa_2018-7-10_15-5

 

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Annotation of the results obtained in 2016
In 2016, during the execution of the following work carried out the first stage of the project: 1) An analytical review of information sources on the project issues; 2) Experimental studies and the subsequent comparative analysis of clinical and laboratory parameters of hard tissue intact and carious teeth, and oral fluid in patients aged 18 to 45 years old; 3) to study and analyze the state of the macro and micronutrients in the hard tissues of the teeth in combination with bio-organic polymers in remote clinically indicated intact teeth and teeth affected by caries process in patients aged 16 to 60 years; 4) The experimental studies of structural and spectroscopic methods for the complex, including with the involvement of synchrotron radiation, metabolic processes and the development of caries in the hard tissues of the teeth, the emergence of macro- and micronutrient deficiency, a condition of the tissue barrier, the structure of the organic matrix and its range of biopolymers, able to prevent erosion and penetration into the tooth tissue micro-organisms; 5) Experimental studies of phase formation, structure, stoichiometry, elemental composition and thermodynamic properties of the enamel products in the conditions of development of caries process; 6) A method for producing nanocrystalline biomimetic materials based on hydroxyapatite using biological source of calcium, which reproduce the structural and morphological features of enamel remineralization with a high potential; 7) Samples were synthesized laboratory biomimetic materials under different conditions to change the structural and morphological properties with similar characteristics to the tooth enamel; 8) A comparative analysis of data obtained by structural and spectroscopic methods for intaknoy and decayed enamel, as well as biomimetic materials to determine the chain of phase transformation at the initial stage of development of caries process; 9) The most informative diagnostic and prognostic clinical and laboratory parameters of hard tissue of teeth and oral liquid, is characterized by high resistance to the development of oral caries process, as well as risk factors for patients with moderate and low caries; 10) Prepared by the patent application; As part of the work performed following scientific results were obtained: The analysis of clinical and laboratory parameters of hard tissue intact and carious teeth, and oral fluid in patients aged 18 to 45 years have shown that the following symptoms occurred at an average caries: carious lesion only posterior teeth, not every year, the appearance of new cavities lack of teeth already had a root, a small amount of plaque on the teeth. To lower average caries carious lesions were characteristic of the front teeth, the annual appearance of new cavities, rapid "loss" of seals within one or two years, there are several cavities in one tooth, the presence of teeth already had a root, a large amount of plaque on the teeth. In low caries carious process amazed at all the teeth with progressive destruction of hard tissues. In the work of this group of patients, we do not take into account, as preventive measures at the hard tissue of teeth marked destruction were not effective. Patients last group received a set of therapeutic measures, including therapeutic, surgical and orthopedic methods. The most informative and prognostically relevant clinical and laboratory parameters that characterize the resistance of the mouth to the development of caries are: electrical conductivity of hard tissue of teeth, cariogenicity and content of short-chain fatty acids in plaque, microcrystallisation, viscosity and lysozyme content in the oral fluid, which allows in a complex considered their performance risk in deviation from the reference values ​​at high caries. Based on data from the complex structural and spectroscopic methods for the development of caries in the enamel is characterized by the growth of deformation and stretching vibrations of the NCO, NH and C = O, a decrease in the index of crystallinity and teksturrirovannosti, as well as the absence of preferred orientation of the HAP crystals in the affected enamel, which suggests destructive processes in the organic matrix hydroxyapatite crystal. As a result of subsurface demineralization further degraded enamel and crystal itself. In studying the weight percent of the chemical elements contained in the enamel intact and carious teeth, with the help of factor analysis revealed the importance of the group of elements that determine the pathological changes in the lesion. First, the most significant factor is composed of elements such as carbon, phosphorus, calcium, chlorine and sulfur, the second factor - oxygen third - sodium in pathological process. The values ​​of the other elements are less significant. By Raman spektromikroskopii shown that in the fissure caries in early stages of development of caries formation zarakterno dicalcium dihydrate - CaHPO4. Comparative analysis of structural and spectroscopic data for intaknoy and decayed enamel shows that in the area of ​​the wall demineralisation fissure caries significantly changed the composition of its mineral phase component. The observed change in the redistribution of vibration modes and groups of intact enamel and the occurrence of A-type substitution karbanationom OH-group in the carious region corresponds to the assumptions of the occurrence of a soluble phase - carbonate-substituted hydroxyapatite. Based on the technology of waste material able to obtain biomimetic biocomposites with the composition, wherein the ratio of organic and inorganic was 5/95, 25/75, 40/60. This allowed to simulate not only the composition but also the optical characteristics (IR and luminescence spectra) of native human dental tissues (enamel and dentin). Achieved participate in an international seminar in the role of the main guest speaker (visiting professor) with the presentation of the goals, objectives and results of the existing Project Seminar organized by the largest University in Mexico - Benemérita Universidad Autónoma de Puebla. The seminar was organized a round table with stakeholders in the project outcomes foreign (Mexican and American) universities and representatives of the manufacturers of dental pharmacy. Reporting and information on the report within the framework of the project manager Paul Mid visit as a visiting professor at the University of Puebla, Mexico on the official Facebook page of the Institute of Physics and Chemistry of the University of Puebla, Mexico https://www.facebook.com/LISDER/photos/a.304314102916553.94946.235518136462817/1351403984874221/?type=3&theater https://www.facebook.com/media/set/?set=a.1355956751085611.1073741838.235518136462817&type=3

 

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Annotation of the results obtained in 2017
In 2017, during the second phase of the Project, the following work was carried out: -The processes of demineralization / remineralization, as well as conditions of reversibility of nascent caries of enamel; - The peculiarities of fluorescence from micro-regions of hard tooth tissues, including micro-enamel sections, subjected to demineralization and caries affected at the initial stage of formation, are studied; approaches to the excitation of fluorescence have been developed, which can be the basis of the methods for recording the earliest stages of caries formation; - Mechanisms for the formation of weak phosphates in the interaction of enamel apatite with certain organic acids (the products of microbial activity), including those with the help of detected changes in fluorescence, are specified; -work on the synthesis and study of structural and morphological, biomimetic materials based on hydroxyapatite obtained with a number of impurities introduced, to achieve a change in the physicochemical properties of the material and their chemical and structural affinities to hard tooth tissues, followed by their use to restore caries affected areas of the tooth and model analysis on the effects of different nature; -The role of the organic component in the protection of hydroxyapatite crystals of hard tooth tissues from dissolution and / or its promotion of crystal reprecipitation is studied; - an organomineral system was created and investigated, based on nanocrystalline biomimetic materials, imitating processes at the interface between the dental biofilm / surface of the enamel; -The potential of bioremineralization of oral fluid and hard tissues of human teeth during preventive measures with the use of developed biomimetic materials is studied. Within the framework of the work performed the following scientific results were obtained: Using the methods of Raman spectroscopy to analyze the state and phase composition of human teeth, demineralization / remineralization processes, as well as conditions of reversibility of the incipient enamel caries, we were able to show that at the initial stages of development of the carious process (caries fissure) the elemental composition of the affected area, i.e. the degree of substitution in it varies with the depth of penetration of the caries into the enamel. This means that caries is not just a chain of chemical transformations during hydroxyapatite dissolution processes. With the selective dissolution of minerals, there is a possibility of their reprecipitation, which allows us to speak about possible reverse processes of growth of HAP crystals on the affected areas of hard tooth tissues. Analyzing the results obtained, as well as the literature data of similar studies, it is possible to speak with confidence about the underestimated role of organics in the development of caries, which requires careful and thorough study of it, since it is the organic component that can act as an inhibitor of the growth of hydroxyapatite crystals. So partial dissolution of apatite enamel in the interaction with acid can mean a loss, first of all, of OH groups in the enamel apatite lattice, and as the concentration of acids affecting the enamel of the tooth is not constant over time and hydroxyapatite is the most probable final phase, a short-term restoration of the enamel structure with the formation of a carbonate-substituted hydroxyapatite according to the A-type of substitution is quite possible. Investigations of the micro-sections of fissure caries confirmed the assumption of the formation of weak phosphates in the interaction of enamel apatite with organic acids (the products of microorganisms' vital activity). The experimental results obtained by the Raman spectromicroscopy testify to the formation of dicalcium phosphate dihydrate - CaHPO4 in the region of near-wall demineralization of a carious fissure. A comparative analysis of structural and spectroscopic data for intact and caries-infested enamel shows that the appearance of a more soluble phase - carbonate-substituted hydroxyapatite - is characteristic at the stage of wall-mounted dimineralization. Using the technique of excitation of fluorescence from hard tooth tissues, we investigated the enamel regions with a carious fissure An analysis of the obtained experimental results showed that around the areas of the carious fissure, lighter areas of hard tooth tissues with a correspondingly higher intensity of excited fluorescence are well visualized. This intense luminescence around the carious fissure, differs in color and intensity from the radiation of the entire illuminated area of ​​intact enamel, and has lateral dimensions comparable with the size of fissures. Due to the disorientation of hydroxyapatite crystals in them, the micro-sections of the enamel adjacent to the carious fissure have a higher fluorescence yield than the regions of the intact enamel and can be easily detected even before the white spot stage, when the organomineral complex is irreversibly changed and an intensive removal of the mineral component. Besides: - An organomineral system based on nanocrystalline biomimetic materials was created and investigated, imitating the processes at the interface between the dental biofilm / surface of the enamel; The work was carried out on the equipment of the megascins class facility, the Australian Synchrotron, Melbourne, in order to use the unique equipment of the synchrotron scientific center for carrying out precision optical studies of the mechanisms of formation of weak phosphates in the interaction of enamel apatite with certain organic acids. In addition, work was carried out on the equipment of the Indian Institute of Science in order to clarify the features of fluorescence from micro-regions of hard tooth tissues, including micro-enamel sites that had undergone demineralization and caries affected at the initial stage of formation. During the visit to the Indian Institute of Science, he took part in an international seminar as the main invited speaker (guest professor) with a presentation of the goals, objectives and already available results of the work on the Project. The seminar is organized by the largest scientific center in Bangalore - Indian Institute of Science. Within the framework of the seminar, a round table was organized with representatives of universities and manufacturers of dental pharmacy in India who were interested in the results of the Project. The presentation and the report materials (photo) are attached in the relevant section of this report.

 

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