Standardization of the protocol for oral cavity examination and collecting of the biological samples for microbiome research using the next-generation sequencing (NGS): own experience with the COVID-19 patients.

To date, publications have shown that compositions of oral microbiota differ depending on their habitats (e.g. tongue, tonsils, pharynx). The absence of set standards for the choice of the areas and conditions of material collection makes the oral microbiome one of the most difficult environments for a comparative analysis with other researchers, which is a meaningful limitation during an assessment of the potential effects of microorganisms as biomarkers in the courses of various human diseases. Therefore, standardisation of basic conditions of a dental examination and collection of material for the next generation sequencing (NGS) is worth attempting. The standardisation of the dental exam and collection of the clinical materials: saliva, swab from the tongue ridge, hard palate, palatine tonsils and oropharynx, supragingival plaque and subgingival plaque. Protocol involved the patients (n = 60), assigned to 3 groups: I-COVID-19 convalescents who received antibiotics, n = 17, II-COVID-19 convalescents, n = 23 and III-healthy individuals, n = 20. The collected biological samples were used to conduct NGS (16S rRNA). The conditions of patient preparation for collecting biological materials as well as the schedule of dental examination, were proposed. Based on the research conducted, we have indicated the dental indicators that best differentiate the group of COVID-19 patients (groups I and II) from healthy people (group III). These include the DMFT, D and BOP indices. The use of alpha and beta diversity analysis provided an overall insight into the diversity of microbial communities between specific niches and patient groups. The most different diversity between the studied group of patients (group II) and healthy people (group III) was noted in relation to the supragingival plaque. The order of activities during the dental exam as well as while collecting and securing clinical materials is particularly important to avoid technical errors and material contamination which may result in erroneous conclusions from the analyses of the results of sensitive tests such as the NGS. It has been shown that the dental indices: DMFT, D number, PI and BOP are the best prognostic parameters to assess the oral health. Based on beta diversity the most sensitive niche and susceptible to changes in the composition of the microbiota is the supragingival plaque. The procedures developed by our team can be applied as ready-to-use forms in studies conducted by other researchers.

Oral microbiota study of the patients after hospitalisation for COVID-19, considering selected dental indices and antibiotic therapy using the next generation sequencing method (NGS).

Background: Poor oral hygiene and the increased incidence and severity of periodontitis may exacerbate SARS-CoV-2 infection. The aim was to evaluate the oral microbiota of 60 participants divided into groups: COVID-19 convalescents who received antibiotics during hospitalization (I), COVID-19 convalescents without antibiotic therapy (II) and healthy individuals (III). Materials and Methods: Dental examination was conducted, and oral health status was evaluated using selected dental indexes. Clinical samples (saliva, dorsal swabs, supragingival and subgingival plaque) were collected and used for metagenomic library to the next-generation sequencing (NGS) preparation. Results: Each of the clinical materials in particular groups of patients showed a statistically significant and quantitatively different bacterial composition. Patients from group I showed significantly worse oral health, reflected by higher average values of dental indexes and also a higher percentage of Veillonella, Tannerella, Capnocytophaga and Selenomonas genera in comparison to other groups. Additionally, a statistically significant decrease in the amount of Akkermansia type in both groups with COVID-19 was observed for all materials. Conclusions: The primary factor affecting the composition of oral microbiota was not the SARS-CoV-2 infection itself, but the use of antibiotic therapy. The increased percentage of pro-inflammatory pathogens observed in COVID-19 patients underscores the importance of preventing periodontal disease and improving oral hygiene in the future.

Oral and Mucosal Complaints among Institutionalized Care Seniors in Malopolska Voivodeship-The Utility of the Mirror Sliding Test in an Assessment of Dry Mouth.

The purpose of this article was to evaluate reported oral and mucosal complaints among seniors residing in institutionalized 24-h care of the Municipal Center for Older and Dependent People (MHCOD) and the 3-month rehabilitation program of the Daily Medical Care House (DMCH). We evaluated the feasibility of using the dental mirror slidding test to assess dry mouth of seniors. Patients underwent a questionnaire, and clinical examination. The Visual Analogue Scale (VAS) was used to assess pain, Fox's questionnaire and Challacombe's scale with Clinical Oral Dryness Score to assess dryness of the mouth, dental mirror slidding test to assess buccal mucosal resistance. Dryness, mucosal burning, impaired taste, food intake are symptoms associated with seniors, and their frequency does not depend on the type of care. The incidence of mucosal burning (Mdn = 4.0, IQR = 4.75, p = 0.032) and difficulty in using dental prosthetics (Mdn = 3.0, IQR = 4.00, p = 0.010) increase with the length of stay at MHCOD. Seniors are at risk of side effects of polypharmacy, which cause dryness (p = 0.036), complaints of lack of saliva (p = 0.009) and taste disorders (p = 0.041. Seniors with higher levels of dry mouth are more likely to exhibit mucosal burning (p = 0.026) and difficulty in taking food (p = 0.037). The implementation of the dental mirror slidding test in the scope of the primary care of geriatric examination should be discussed.

Physiotherapeutic rehabilitation of adolescent patients with temporomandibular disorders.

TMD is a group of pathological changes including increased tension in the masticatory muscles, pain in the muscles and/or the temporomandibular joints, abnormal range of the mandibular movement or the presence of acoustic symptoms in the joints in the form of clicking or poping. The aim of the project was to compare the effectiveness of two methods of physiotherapeutic rehabilitation, used in adolescent patients with temporomandibular disorders. MATERIAL AND METHODS: The material consisted of 68 patients, aged 14 to 17, of both sexes, generally healthy, who came to the Dental Prosthetics Department for treatment due to pain in the masticatory muscles. Dental examination and diagnostics revealed Ia in all cases in accordance with RDC/TMD protocol. Manual therapy was performed in group I (34 people) and kinesitherapy with massage was performed in group II (34 people). Patients were allocated randomly to both groups. Contraindications were considered for both methods. RESULTS: Within the first study, mean values of pain intensity between group I and group II were not significantly different (6.12 and 6.24 respectively). Within the second study significantly lower VAS scores in both groups have been revealed (0.92 and 0.74 respectively). Results of the first and second study differed significantly in both groups. Similar results were obtained for the maximum abduction of the mandible. CONCLUSIONS: The results of the conducted studies indicate a beneficial effect of the assessed physiotherapeutic procedures in terms of functional rehabilitation of adolescent patients with temporomandibular disorders.

The Influence of Preventive Activities on Stress Perception among Dentistry Students in the Period of the COVID-19 Pandemic.

BACKGROUND: The COVID-19 pandemic resulted in the strengthening of the earlier stressors and the appearance of new pandemic-related stressors. Many students of dentistry fit the profile of a group who are particularly susceptible to stress related to the pandemic. Thus, it was necessary to implement preventive activities, reducing their stress perception. This was understood as a means of significantly influencing the student's well-being, thus improving the quality of education. Therefore, the aim of the study was to assess the impacts of implemented preventive activities on stress perception among students of dentistry during the pandemic, as well as their influence on this assessment of the selected students' personal experiences regarding the pandemic. METHODS: The study was performed one year after the outbreak of the pandemic on students of dentistry at Jagiellonian University Medical College, Krakow, Poland. It consisted of the completion of a self-designed, voluntary, anonymous, online questionnaire. The respondent's task was to assess the influences of implemented preventive activities on stress perception using a five-point scale. The activities were divided into external (national, global) and internal (institutional). The material was statistically analysed for all students, including pre-clinical and clinical groups. Additionally, the impact of the selected students' personal experiences regarding the COVID-19 pandemic on the assessment was studied. RESULTS: All preventive activities (in total) significantly reduced stress perception (p ≤ 0.001), but the intensity of their impacts (mean rank) was different. The most highly assessed activities were all external preventive activities, with the greatest intensity belonging to the prospect of receiving vaccination against SARS-CoV-2. The remaining external activities were the prospect of developing an effective COVID-19 treatment and the increase in knowledge about SARS-CoV-2. The internal activities were assessed and ascribed lower positions, with the exception of the theoretical classes held online. This activity was placed slightly lower than the highest-rated activity, the prospect of receiving vaccination. CONCLUSIONS: The studied preventive activities reduced stress perception with different levels of intensity. The highest-ranked activities were external activities. One exception was the theoretical classes held online, an internal activity. Due to the lower impact of the internal activities and the ability to modify them by the educators, there is a need to strengthen their effectiveness. The possibility of monitoring and tailoring some preventive activities to the students' needs was the practical aspect of the conducted study. Students' personal experiences related to the COVID-19 pandemic influenced their assessment of the preventive activities, in some cases significantly.

Oral Health of Elderly People in Institutionalized Care and Three-Month Rehabilitation Programme in Southern Poland: A Case-Control Study.

Demographic ageing is a global growing process and the quality of ageing is an important parameter in this process. The aim of the study was to analyse the distribution of remaining dentition in relation to oral hygiene indicators among elderly people remaining in institutional care and those who participated in a 3-month rehabilitation program, aimed at increasing time of independent functioning, in southern Poland. The patients underwent a questionnaire and clinical examination. An analysis of missing teeth was performed, plaque index, and gingival index were measured. Residents of the Municipal Health Centre for Older and Dependent People (n = 50) had a higher incidence of missing teeth in the maxilla (88.4%), mandible (77.6%), as well as in the maxilla and mandible combined (83%) than residents of the Daily Medical Care House (n = 30). The distribution of the remaining teeth, in both groups, corresponds to the outlets of the large salivary glands. The group of 53.8% of patients with dry mouth had PI scored 3. Residents with dry mouth were more likely to have plaque deposits and gingival inflammation. It is necessary to develop and implement an oral care program for patients with reduced saliva secretion, with a particular focus on dependents.

Dental environmental stress during the COVID­19 pandemic at the Jagiellonian University Medical College, Kraków, Poland.

OBJECTIVES: The burden of stress experienced by dentistry students has been the subject of numerous studies aimed at defining the strongest stressors, and at reducing their intensity to improve the conditions and, consequently, the quality of education. Such studies have never been performed in the conditions of a pandemic caused by a new, unknown coronavirus. Therefore, the objective of the study was to identify the sources of their stress in the course of the studies, with reference to the COVID­19 pandemic. MATERIAL AND METHODS: A total of 334 dentistry students of the Institute of Dentistry at the Jagiellonian University Medical College were invited to take part in the study. They represented a range from the first to the fifth year of studies in the 2019/2020 academic year. The tool employed in the study was the Dental Environmental Stress (DES) questionnaire and 11 supplementary questions related to the pandemic. The study was carried out on June 1-25, 2020. RESULTS: The participation rate was 72%. Academic work presented the highest level of stress. A comparison of the overall levels of stress in particular domains as well as questions included in the DES survey revealed significant differences between students of individual years of studies in the 2019/2020 academic year. The strongest positive relationships between the supplementary questions and the DES domains appeared in the area of clinical factors. These concerned returning to clinical classes and contact with patients with regard to SARS-CoV-2 (rs = 0.50, p < 0.001), contact with patients in connection with SARS-CoV-2 (rs = 0.47, p < 0.001), and a lack of practical classes with patients in connection with the COVID­19 pandemic (rs = 0.42, p < 0.001). CONCLUSIONS: Dentistry students were subjected to stress resulting from a conflict between the perceived risk of returning to clinical classes and contact with patients due to SARS-CoV-2, and disruptions in the course of clinical education, which they perceived negatively. Int J Occup Med Environ Health. 2021;34(2):211-22.

The influence of a selected desensitizer on marginal microleakage of a class V composite restoration subjected to thermocycles: an in vitro study.

INTRODUCTION: Dentin hypersensitivity is a painful clinical condition. The frequency of its occurrence varies from 8 to 57%, depending on tested group and different methods of investigations. Recommended desensitizing agents have different mechanism of action and effectiveness. We are still looking for solutions that will improve their effectiveness and simultaneously allow for wider use of e.g. as a base material, counteracting postoperative hypersensitivity, reducing marginal microleakage. The aim of the study was to assess the effect of a selected desensitizing agent occluding dentin tubules with calcium hydroxyapatite on marginal microleakage formation of a class V composite restorations subjected to thermocycles. MATERIALS AND METHODS: In study it was used 40 molars and premolars, which were alternately assigned into two groups. In both groups standardized cavities were prepared. In the study group (study group - SG) before application of bonding agent Teethmate Desensitizer (f. Kuraray, Noritake Dental Inc., Okayama, Japan) was used. In the control group (control group - CG) OptiBond All-in-one (f. Kerr, Bioggio, Switzerland) bonding agent was used and cavities were filled using composite material Gradia Direct (f. GC Europe N.V., Leuven, Belgium). After storage in saline, teeth were subjected to 600 thermocycles, passive dye penetration test was done, teeth were cut in the area of filling, according to its long axis. Under light microscope magnification value of microleakage was measured and marginal microleakage rate (M) was counted. The results of the tests were statistically analyzed using the package STATISTICA 12.0 (StatSoft, USA). RESULTS: The average value of M for the SG group was 0.46 (min 0.05, max 0.76, SD 0.226) and for CG was 0.22 (min 0, max 0.74, SD 0.235). The differences between M values were statistically significant (p = 0.0094). CONCLUSION: A reduction in the number of retention sites for the bonding system, facilitates the formation of microleakage in the experimental conditions and reduces the degree of adhesion of the composite material to the hard tissues of the tooth.