Investigation of the Effects of Bisphosphonate on the Gingival Crevicular Fluid Levels of Sclerostin and the DKK-1 in Individuals With Postmenopausal Osteoporosis With Periodontal Changes
Phase: PHASE4
Conditions: Osteoporosis, Postmenopausal, Periodontitis
Sponsor: Ondokuz Mayıs University

 
 
 
 
 
 
Investigation of the Effects of Bisphosphonate on the 
Gingival Crevicular Fluid Levels of Sclerostin and the 
DKK-1 in Individuals with Postmenopausal 
Osteoporosis with Periodontal Changes 
 
 
 
 
 
 
Document Date: December, 2018 
Uploaded: September, 2020 
 
 
 

Study Protocol 
 
Individuals with chronic periodontitis and osteoporosis between 51 
and 66 years (mean age: 57.15 ± 4.93) (group A, n=12), 52 to 66 
years (mean age: 57.41 ± 4.94) with systemic healthy chronic 
periodontitis (group B, n=10), between the ages of 50-63 (mean 
age: 55.64 ± 3.79) periodontally healthy osteoporotic individuals 
(group C, n=11) and 52-64 years (mean age: 56.55 ± 4.06) 
periodontal and systemically healthy individuals (group D, n=10) 
were included in study. 43 postmenopausal women in total were 
included in the study. This study involved postmenopausal women 
admitted to the Ondokuz Mayıs University Faculty of Medicine, 
Department of Endocrinology and Metabolism between 2016 and 
2018. The study protocol was approved by Ethical Board of 
Ondokuz Mayis University School of Medicine (OMU KAEK 
2016/100) and was carried out in accordance with the ethical 
standards established in the 1964 Declaration of Helsinki, as 
revised in 2008. Written informed consent was obtained from each 
subject before enrollment in the study. 
Bisphosphonates were started in the patients who were diagnosed 
with osteoporosis. Individuals smokes, who had received antibiotic 
treatment in the last 3 months and/or periodontal treatment in the 
last 6 months were excluded from the study. Following medical 

record retrieving, patients were leaded to Ondokuz Mayıs 
University Faculty of Dentistry Periodontology Department for 
periodontal examination, receipt of clinical records and gingival 
crevicular fluid (GCF) samples. Following the receipt of medical 
records; standard periodontal recordings including plaque index 
(PI) (1), gingival index (GI) (2), bleeding on probing index (BOP), 
pocket depth measurement (PD), clinical attachment loss 
measurement (CAL), dental radiographs were received from each 
patient. Clinical measurements were taken from the region under 
each tooth (mesio-vestibule, mid-vestibule, disto-vestibule, mesio-
lingual, mid-lingual, disto- lingual) using Williams probe. In the 
first 
clinical 
and 
radiographic 
evaluation, 
groups 
with 
postmenopausal osteoporosis and healthy groups were divided into 
2 subgroups as chronic periodontitis and healthy control groups. 
The diagnosis of chronic periodontitis was made clinically and 
radiographically according to the criteria determined by the “1999 
Periodontology Workshop” (3). Periodontitis patients consisted of 
patients with 4 or more probes with bleeding index (BOP) positive, 
5 mm or more deeper pocket depth (PD) and ≥6 mm attachment 
loss (CAL) and radiographically determined alveolar bone loss. 
Individuals with <3 mm PD and no radiographically determined 
alveolar bone loss were preferred to form groups with healthy 
periodontium. In the groups with chronic periodontitis gingival 

crevicular fluid (GCF) was collected with the help of periopaper 
(Periopaper®; Oraflow, New York, NY, USA) from the regions with 
the deepest pocket depth with periodontal defect, and in healthy 
groups it was collected by taking the right mandibular molar 
region as reference. Each time, the volume value of the collected 
GCF was determined by the Periotron 8000 device. GCF collected 
paper strips were placed in eppendorf tubes and stored at -80 ° C 
until biochemical analysis was performed (NUAIRE Ultra-Low 
Freezer Model no: Nu-6420E). Following the initial sample 
collection and recording of patient data (medical, periodontal and 
radiographic), patients were placed under phase 1 periodontal 
treatment (scaling and root planing), and oral hygiene habits of 
the patients were corrected and followed up for 1 year. In this 
period, periodontal treatments, controls and sample collections of 
the patients in Group A and Group B who completed phase 1 
treatment were repeated at the end of 6th month and at the end of 
the 12th month after the end of the treatment, their clinical 
changes were recorded. The clinical data and GCF SOST and 
DKK-1 levels of osteoporosis groups (Group A and C) and non-
osteoporotic 
chronic 
periodontitis 
group 
(Group 
B) 
with 
bisphosphonate were reevaluated in the determined times. 
 
 

Biochemical Analysis 
 
SOST and DKK-1 levels in gingival crevicular fluid were 
determined by ELISA (Enzyme-linked immunosorbent assay) kits 
(SunRed Human DKK1 ELISA Kit, Cat.No:201-12-063, and 
SunRedHuman SOST ELISA Kit, Cat No: 201-12 -5418) The 
SOST and DKK1 assay ratios according to the manufacturer's 
instructions are 0.2-60 ng / mL 0.5-78 ng / mL, respectively. 
Sensitivity is 0.412 ng / mL for DKK-1 and 0.17 ng / mL for SOST. 
Color intensity is measured at 450 nm and the results were 
calculated using standard curves for each kit. GCF SOST and 
DKK-1 concentrations were obtained by dividing total amount of 
SOST 
and 
DKK1 
values 
by 
GCF 
volume 
(microliter). 
Concentration values will be presented in ng / mL. 
 
Statistical Analysis 
 
Statistical analysis was performed using SPSS (version 21, SPSS 
Inc. Chicago, IL, USA) program. The Shapiro-Wilk test was used 
to show if the data were consistent with the normal distribution. 
Comparisons of biochemical data between groups were performed 
with non-parametric Kruskal-Wallis test and p <0.05 was 
considered as statistically significant. In determining the 

difference group, p value was divided by the number of comparison 
and p <0.016 was accepted as significance level (Mann- Whitney U 
test with Bonferroni correction). Friedman test was used to 
determine the time difference in groups and p<0.05 was 
considered as statistically significant. 
 
 
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Correlation between oral hygiene and periodontal condition. 
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