| کلیدواژههای انگلیسی مقاله |
Diabetes mellitus, Saliva, Gingivitis, Children, IntroductionDiabetes mellitus (DM) is a group of metabolic disorders with serious complications reducing the quality of life [ 1,- 2,]. It is caused by the absolute or relative insulin deficiency due to decreased secretion of this hormone from the pancreas (type-1 DM) or insensitivity of environmental receptors to this hormone (type-2 DM) [ 3,]. The global diabetes prevalence in 2019 is estimated to be 9.3% (463 million people) [ 4,]. Type-1 DM usually occurs in childhood, and adolescence includes 5-10% of all diabetes patients [ 3,]. Periodontal diseases and gingivitis were reported as the sixth most common complication of diabetes, which are usually associated with the severity of disease [ 5,- 6,]. Some studies showed more prevalence of gingival inflammation in children and adolescents with type-1 DM compared to the healthy population [ 7,- 9,].The use of saliva instead of serum has recently been preferred as a diagnostic medium. Saliva offers advantages over blood because it is a cost-effective and non-invasive method that can be collected by persons with modest education [ 10,]. One approach to early diagnosis of periodontitis is the salivary biomarkers. Some biomarkers, such as cytokines, were diagnosed and proposed in the literatures [ 11,- 13,].Accumulation of reactive oxygen species, oxidative stress, and interactions between advanced glycation end products (AGEs) in the periodontal tissues and their receptor (RAGE) all contribute to increased inflammation in the periodontal tissues in people with DM [ 14,]. There is also a lipid metabolism disorder in diabetic patients due to impaired glucose metabolism and changes in insulin secretion and activity. As a result of systemic lipid disorders, high concentrations of lipids have been shown in these patients&apos, blood and saliva. Lipids play the role of nuclei in the dental plaque mineralization and accelerate the activity of the enzyme glucosyltransferase, which is responsible for the carcinogenic activity of oral microorganisms. High cholesterol and triglyceride levels in the plaque, delay the release of lactic acid from it. The presence of lipids in the saliva modulates bacterial hydrophobic surfaces and thus helps to bind them to dental surfaces [ 14,- 15,]. Salivary albumin is regarded as a serum ultrafiltrate to the mouth, and it may diffuse into the mucosal secretions. Hormonal balance, nutrition, and osmotic pressure regulate albumin synthesis. High concentrations of salivary albumin have been detected in a medically compromised condition, such as immunosuppression and DM. Both normal and raised salivary albumin levels have been seen in periodontitis [ 16,- 17,]. In association with &,alpha -amylase, some studies have suggested that this salivary enzyme contributes to the microorganisms&,rsquo adhesion and the microbial plaque formation however, other studies have found that &,alpha -amylase secretion is associated with a reduced risk of caries, a decrease in oral bacteria, and a reduced risk of periodontal disease [ 18,- 20,]. DM can affect the composition and flow of saliva. These changes in saliva can be involved in the onset of symptoms and even the severity of oral complications in diabetic patients [ 21,]. Many studies have been done about the correlation between salivary composition and periodontal disease in DM, but the results have not been conclusive. Regarding the few studies conducted in this field concerning quality control of the disease, the present study performed to investigate the relationship between periodontal status and salivary protein, lipid, and antioxidant capacity in healthy individuals and patients with well-controlled and uncontrolled type-I DM.Materials and MethodStudy populationThis cross-sectional study performed on 6- to 16-year-old diabetic and healthy volunteers with normal body mass index (BMI percentile 5-85%) (Figure 1,) [ 22,]. Figure 1. Individual growth chart 3rd, 5th, 10th, 25th, 50th, 75th, 90th, 95th, 97th percentiles, 2 to 20 years, body mass index-for-age [ 22,] Based on previous studies [ 23,- 24,] and considering an alpha coefficient of 0.05 and a statistical power of 0.8, the sample size was determined to be 120 (case group n=60 and control group n=60). The case was divided into two subgroups of well-controlled DM (n=33) and poorly-controlled DM (n=27).The case group has consisted of patients who were diagnosed with type-1 DM by an endocrinologist at the Amirkola Children Hospital for at least three years. The quality of control of disease was determined based on the level of HbA1c. Patients with HbA1c more than 7.5% were considered into poorly-controlled DM group [ 25,]. The patients were selected through a simple sampling method considering the exclusion criteria, such as having other diseases (asthma, cardiovascular disease, epilepsy, and renal deficiency) and reluctance to participate in the study.The control group included healthy individuals who had not taken any medicine over the last month [ 26,]. They were selected from schools of Babol city (north of Iran) using multistage random sampling. According to the different socio-economic situations in various urban districts and their impact on health and nutrition, a multistage sampling method can be generalized to the whole town. However, in the case of patients, because there is only one children&apos,s hospital in the city, all the patients can be found in the same place. The subjects in both groups were matched for age and gender.Ethical considerationsThe study was approved by the Ethical Committee of the Research Council of Babol University of Medical Sciences (MUBABOL.REC.1395.55). The written consent was obtained from all subjects or their parents.Experimental procedurePersonal information and medical history of patients were obtained through interviewing participants/ parents and their medical records. In order to minimize the effect of the circadian rhythm, all saliva samples were collected from 10 to 11 AM, and then oral examination was done. Unstimulated saliva samples were collected in disposable sterile tubes and immediately transferred to the laboratory in a container containing dry ice at -4&,deg C. The samples were centrifuged at 1500 g and 15 minutes, Clement 2000, North Sydney, Australia), the supernatant was collected into Eppendorf microtubes and were stored at -80&,deg C until analyzes. Measurement of a lipid profileSalivary cholesterol and triglyceride levels were assessed based on the colorimetric method using ZiestChem commercial kits (ZiestChem Diagnostics Co. Iran) according to the manufacturer&apos,s protocol [ 27,]. Measurement of &,alpha -amylase activityIn order to measure the &,alpha -amylase activity, 500&,micro l of reagent was poured into the blank and the sample tubes and incubated at 37&,ordm c for 5min. Then 20&,micro l of the sample was added to the sample tube and incubated at 37&,ordm c for 15min. Then immediately after that, the chemical reaction was stopped by adding 500 &,micro l Iodine solution and 1500 &,micro l distilled water. The absorption for the blank and the sample tubes was compared against distilled water at 405nm using UV-visible spectrophotometer, and &,alpha -amylase activity was estimated by this equation [ 28,], Absorbance of blank-Absorbance of sample Absorbance of blank&,times 1470= &,alpha -Amylase activityULMeasurement of total protein and albumin concentrationSalivary total protein was measured in the Biuret method using ZiestChem commercial kits (ZiestChem Diagnostics Co., Iran) according to the manufacturer&apos,s protocol. Salivary albumin levels were measured based on colorimetric assay using the ZiestChem commercial kit (ZiestChem Diagnostics Co., Iran) also [ 29,- 30,]. Measurement of FRAP and DPPH indexesTotal antioxidant capacity (TAC) of salvia was measured according to Ferric Reducing Ability of Plasma (FRAP) assay, and the free radicals scavenger index was measured by DPPH assay (1.1-Diphenyl-2-picryl-hydrazyl) [ 31,- 32,]. L&,ouml e and Silness gingival index (GI) and Silness and L&,ouml e plaque index (PI) were measured [ 33,]. A senior dental student, using a dental mirror and a probe on a chair and in ambient light, did oral examination. Statistical analysisData were statistically analyzed by Kruskal-Wallis followed by the Mann-Whitney U Test, One-way analysis of variance (ANOVA), Tukey Scheffe, t test, and Pearson correlation test in SPSS-22. All statistical tests were performed at the significance level of the p value less than 0.05.ResultsThe mean ages of the subjects in case and control groups were respectively 10.02&,plusmn 1.39 and 10.07&,plusmn 0.82 years. Thirty-three diabetic patients (55%) were diagnosed with well-controlled DM (HbA1c less than 7.5%) approximately 4.45&,plusmn 2.43 years elapsed from the diagnosis of DM. The levels of salivary triglyceride and cholesterol,&,nbsp amylase, and GI&,nbsp in people with DM were significantly higher than that in healthy subjects (p&,lt 0.001, Table 1,).GroupDiabetic subjectsHealthy subjectsp ValueVariableTriglyceride (mg/dL)*15.82&,plusmn 2.587.74&,plusmn 6.98&,lt 0.001Cholesterol (mg/dL)*11.40&,plusmn 6.495.12&,plusmn 3.37&,lt 0.001Amylase (U/L) *65.40&,plusmn 17.7942.44&,plusmn 17.56&,lt 0.001Total protein (g/dL)*91.53&,plusmn 10.82106.56&,plusmn 11.74&,lt 0.001Albumin (g/dL)*2.48&,plusmn 0.973.10&,plusmn 1.09&,lt 0.001DPPH (%)*95.595.90.77FRAP (&,micro mol/mL)*9.77&,plusmn 0.139.78&,plusmn 0.120.083Plaque index**1.63&,plusmn 0.741.59&,plusmn 0.690.84Gingival index**1.51&,plusmn 0.740.91&,plusmn 0.81&,lt 0.001*, Based on t-test **, Based on Mann-Whitney test |
| نویسندگان مقاله |
Fatemeh Tabatabaei |
Dental Student, Student's Research Committee, Babol University of Medical Sciences, Babol, Iran.
Soleiman Mahjoub |
Cellular and Molecular Biology Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran.
Morteza Alijanpour Aghamaleki |
Non-Communicable Pediatric Diseases Research Center, Health Research Institute, Babol University of Medical Sciences, Babol, Iran.
Amene Moslemnejad |
Clinical Biochemistry, Babol University of Medical Sciences, Babol, Iran.
Samane Gharekhani |
Oral Health Research Center, Dept. of Pediatric Dentistry, Faculty of Dentistry, Babol University of Medical Sciences, Babol, Iran.
Forough Yavarzade |
Dental Student, Student's Research Committee, Babol University of Medical Sciences, Babol, Iran.
Soraya Khafri |
Biostatistics & Epidemiology, Medicine Faculty, Babol University of Medical Sciences, Babol, Iran.
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