A retrospective longitudinal cohort study was conducted at St Luke’s International Hospital, a large teaching hospital in Tokyo, Japan. All participants who underwent health examinations at the hospital’s Centre for Preventive Medicine between 2005 and 2011 and had an estimated glomerular filtration rate (eGFR) > 15 mL/min/1.73 m² were included. Participants who required regular dialysis, regardless of whether it was hemo- or peritoneal dialysis, were excluded. The primary outcome was a composite renal outcome that was compared with the frequency of tooth brushing. The St Luke’s Ethics Committee Institutional Review Board approved this study (approval number: 18-R203 – Comprehensive approvals for studies about social habits using this dataset).
Outcomes
The primary outcome was a composite renal outcome consisting of a 25% eGFR reduction from baseline, an eGFR of <15 mL/min/1.73 m², and a requirement for regular dialysis [14-16]. The eGFR for each patient was calculated based on the following formula for Japanese patients [17]:
Male: eGFR = 194 × (serum creatinine)-1.094 × (age)-0.287
Female: eGFR = 194 × (serum creatinine)-1.094 × (age)-0.287 × 0.739
The baseline eGFR for each participant was defined as the eGFR measured at their first visit to the center between 2005 and 2011. The eGFR was measured at every subsequent visit to the center for health examinations. The initiation of regular dialysis was dependent on each physician’s decision; relevant information was obtained from the electronic medical records or participants’ self-reports. Each patient was followed until December 31, 2018.
Frequency of tooth brushing
All participants were asked about their tooth brushing frequency as part of a questionnaire during their health examinations between 2005 and 2011. Each participant was asked to select one of the following responses to the question regarding tooth brushing frequency: less than daily, once a day, once to twice a day, or after every meal. Due to the small number of participants who brushed their teeth less than daily, we combined this group of patients with those that brushed their teeth once a day; this was defined as the reference group. Information about tooth brushing frequency was subsequently obtained at every visit to the center and considered as a time-dependent variable.
Covariates
As a part of the questionnaire provided during health examinations, we obtained information about participants’ demographics, medical histories, laboratory measures and physical examinations as potential covariates for our study. Health habits including alcohol consumption (abstainer, occasional drinker, regular drinker), smoking status (never smoker, former smoker, current smoker) and exercise habits (almost none, 1-2 times a week, 3-5 times a week, almost every day) were also assessed. During the health examinations, trained staff measured participants’ heights and weights and calculated their body mass indexes (BMI). Each participant was categorized into one of three groups based on their BMI and Asian criteria: underweight (BMI < 18.5 kg/m2), normal (BMI 18.5–24.9 kg/m2) or obese/overweight (BMI ≥ 25.0 kg/m2) [18]. Information about comorbidities related to the progression of CKD such as diabetes, hypertension [19], or cancer [20], was also obtained from participants’ self-reports. This information was obtained at every health examination and considered as a time-dependent variable.
Finally, information regarding participants’ baseline kidney function was also evaluated and retained as a potential covariate. The eGFR of each participant was calculated based on the Japanese formula outlined above and used to categorize patients into a baseline CKD risk group (low risk, moderately increased risk, high risk, very high risk) according to The Kidney Disease: Improving Global Outcomes (KDIGO) 2012 Clinical Practice Guideline [21]. The baseline eGFRs and CKD risk group classifications were used as covariates.
Statistical methods
First, we compared participants’ baseline characteristics with their tooth brushing frequency. We then conducted multivariate analyses using a generalized estimating equation with the logit function and a binomial distribution to evaluate the association between frequency of tooth brushing and composite renal outcome, adjusting for potential covariates. We used different potential covariates in each of the models: Model 1 was adjusted for participant age and sex, baseline eGFR, and baseline CKD risk category; Model 2 was adjusted for health habits (alcohol consumption, smoking status, and exercise habits) in addition to the covariates used in Model 1; Model 3 was adjusted for BMI in addition to the covariates used in Model 2; and Model 4 was adjusted for comorbidities (hypertension, diabetes, and any type of cancer) in addition to the covariates used in Model 3. Our final model was model 4, but we also presented the results of other models as part of the sensitivity analysis to show that the results are consistent with varying covariates. We also stratified the data by baseline CKD risk category to evaluate the impact of tooth brushing frequency on composite renal outcomes in sub-analyses. All analyses were performed using STATA 14 (STATA Corp., College Station, TX, USA).