Study design and patients’ selection
PD, allergic, bleeding, diabetes
BLEEDING, PERIODONTITIS, STERILE, SEVERE PERIODONTAL DISEASE, PERIODONTAL DISEASE, PLAQUE, DIABETES
To evaluate the use of the RTCA system in antibiotic selection for the individualized treatment of patients with periodontal disease, and to compare the Required sample size was calculated by a power estimation based on the differences in clinical attachment level (CAL) of 1 mm between the groups with variation in the population of 0.5, a confidence level of 80% and a power of 95%, which suggested a sample size of 20 individuals per group. Due to the length of the study and the multiple visits, a high dropout rate (>15%) was expected. Therefore, a final sample size of over 60 volunteers fulfilling the inclusion/exclusion criteria was estimated.Inclusion criteria included: adults between 40 and 70 years old, non-smokers or smokers of less than 10 cigarettes a day, with stage III and IV grade A-B periodontitis, presence of at least 20 teeth in the mouth, including at least one molar per quadrant and good general health. Exclusion criteria included: smokers of more than 10 cigarettes a day, patients who have been treated with periodontal therapy in the last 12 months, patients who have been taking antibiotics in the last three months or who routinely used antiseptics in the previous month, patients who due to their systemic condition take antibiotics, pregnant or lactating woman, patients who take medication that might alter the immune system or bone metabolism, patients who are allergic to the antibiotics used in the study and people with diabetes with more than 7% glycosylated hemoglobin. After signing the informed consent form, clinical periodontal indexes such as bleeding on probing (BOP), probing depth (PD), clinical attachment level (CAL) and plaque presence (PLAQUE) were evaluated as previously described (After the evaluation of clinical parameters, the subgingival plaque samples of 64 volunteers were collected into VMG III transport media, as described above, for both biofilm growth monitoring in real-time (Real-Time culture; RT-Culture) using the xCELLigence impedance monitoring system (Agilent Technologies) and for bacterial composition determination before treatment. In parallel, subgingival plaque samples from the same volunteers were collected into sterile tubes using micro-IDentAfter four and eight weeks of systemic antibiotic treatment, the patient’s oral health was re-evaluated, including all clinical parameters described above (Diagram of the clinical study protocol: subgingival plaque samples from 64 individuals with severe periodontal disease were collected in VMGIII transport media and grown in an impedance system for monitoring periodontal biofilm growth in real time in the presence of antibiotics (Real-Time Culture). Derived biofilms were collected at 8h of growth for DNA extraction and 16S rRNA gene sequencing. In parallel, subgingival plaque samples from these 64 patients were used for standard hybridization-based periodontal pathogen quantification and associated antibiotic recommendations by the German Society of Periodontology (