|Year : 2022 | Volume
| Issue : 3 | Page : 118-121
Smoking effect on the interleukin-1β expression and periodontal status of periodontitis patients
Muhammad Ihsan Rizal1, Redha Putri Wiranda2, Ria Aryani Hayuningtyas1, Fatimah Maria Tadjoedin3, Ferry Sandra1, Melanie Sadono Djamil1
1 Department of Oral Biology, Biochemistry and Molecular Biology Division, Faculty of Dentistry, Universitas Trisakti, Jakarta, Indonesia
2 BioCORE, Faculty of Dentistry, Universitas Trisakti, Jakarta, Indonesia
3 Department of Periodontology, Faculty of Dentistry, Universitas Indonesia, Jakarta, Indonesia
|Date of Submission||05-Jul-2022|
|Date of Decision||02-Aug-2022|
|Date of Acceptance||18-Aug-2022|
|Date of Web Publication||15-Nov-2022|
Muhammad Ihsan Rizal
Department of Oral Biology, Biochemistry and Molecular Biology Division, Faculty of Dentistry, Universitas Trisakti, Jakarta
Source of Support: None, Conflict of Interest: None
Background: The smoking habit is a risk factor for periodontitis. Periodontitis is a multifactorial inflammatory disease associated with biofilm plaque dysbiosis and is characterized by the progressive deterioration of the periodontal tissue. Pro-inflammatory cytokines, such as interleukin-1β (IL-1β), can trigger inflammation of the tissue and become a stimulator to destroy it. Moreover, IL-1β will increase in response to inflammation. Objective: The aim of this study was to evaluate the differences in periodontal status and IL-1β gene expression between smoking and nonsmoking periodontitis subjects. Methods: This research was an analytic observational study with a cross-sectional design. The anthropometric demographic data included name, address, gender, age, plaque index examination, papillary bleeding index (PBI), calculus index, smoking status, and clinical photos to diagnose periodontitis. The collection of gingival crevicular fluid and the analysis of the IL-1β gene expression were conducted using real-time polymerase chain reaction. Results: There were significant differences in periodontal status for plaque index, calculus index, debris index, oral hygiene index, and PBI between smoking and nonsmoking subjects. The IL-1β gene expression was significantly higher (P = 0.047) in smoking subjects than in nonsmoking subjects, with mean ± standard deviation (11.34 ± 9.11) and (0.24 ± 0.42), respectively. Conclusion: This study revealed differences in periodontal status and the IL-1β gene expression between smok ing and nonsmoking periodontitis subjects. The periodontal rate was higher in periodontitis smokers compared to nonsmokers periodontitis. In this study, it was also found that the expression of the IL-1β gene was significantly higher in subjects with smoking periodontitis.
Keywords: Gingival crevicular fluid, IL-1β gene expression, periodontitis, smoking
|How to cite this article:|
Rizal MI, Wiranda RP, Hayuningtyas RA, Tadjoedin FM, Sandra F, Djamil MS. Smoking effect on the interleukin-1β expression and periodontal status of periodontitis patients. Sci Dent J 2022;6:118-21
|How to cite this URL:|
Rizal MI, Wiranda RP, Hayuningtyas RA, Tadjoedin FM, Sandra F, Djamil MS. Smoking effect on the interleukin-1β expression and periodontal status of periodontitis patients. Sci Dent J [serial online] 2022 [cited 2022 Dec 3];6:118-21. Available from: https://www.scidentj.com/text.asp?2022/6/3/118/361159
| Background|| |
Smoking is the most common form of tobacco use. The world’s most numerous cigarette consumers are from developing countries. The number of smokers in the world was estimated to reach approximately 1.3 billion people in 2014., In this regard, Indonesia has the third largest number of active smokers, behind China and India. According to the data released by the Basic Health Research (RISKESDAS) in 2018, the prevalence of smokers aged ten years old or older in Indonesia accounted for 28.8% of all Indonesian aged 10-years old or older. Apart from these figures, smoking has adverse effects on health, particularly on the heart, lungs, teeth, and mouth, with no exception for periodontal tissues.
The 2018 data of RISKESDAS revealed that the number of periodontitis cases in Indonesia amounted to 74.1% of the total population. Periodontitis itself is a multifactorial inflammatory disease associated with biofilm plaque dysbiosis and characterized by a progressive breakdown of the periodontal tissues. Pro-inflammatory cytokines such as interleukin (IL)-1β, tumor necrosis factor (TNF)-α, IL-6, and IL-17 can trigger the inflammation of the periodontal tissues. IL-1β can serve as a stimulator that can damage those tissues.
An increased level of IL-1β is often detected in the gingival crevicular fluid (GCF) of patients with periodontitis. IL-1β will increase in response to the occurrence of inflammation. Furthermore, the nicotine contained in cigarettes can affect immune and inflammatory responses. When inflammation occurs in the body, pro-inflammatory cytokines, such as IL-1β, TNF-α, and IL-6 will respond actively to it by increasing the level of cytokines. According to a previous study conducted by Cheng et al. in a monoculture system, the stimulation of nicotine led to an increase of IL-1β gene expression in the serum of human periodontal ligament cells. In this regard, smoking becomes one of the risk factors in the development of periodontitis. Smoking-induced periodontitis could increase the level of IL-1β since the yielded bacteria produce toxins that will allow for the stimulation of the host response. However, there has been limited investigation into this issue. To the best of the researcher’s knowledge, few studies have examined the periodontal status and IL-1β gene expression of smokers and nonsmokers in the same periodontitis group. Therefore, the purpose of this study was to compare the periodontal status and IL-1β gene expression of smokers and nonsmokers with periodontitis.
| Materials and Methods|| |
This research was an analytical observational study with a cross-sectional design. This research was conducted at BioCore Laboratory Faculty of Dentistry, Universitas Trisakti. This study has been granted research ethics approval with permission number 388/S1/KEPK/FKG/8/2020.
The subjects for the study were obtained from the suburbs of a densely populated neighborhood in Menteng Sukabumi in Central Jakarta from January to March 2020. This study’s inclusion criteria were males and females aged 45–60 years, suffering from periodontitis, and smoking or nonsmoking status. Subjects with a history of systemic disease (diabetes mellitus and hypertension), taking antibiotics in the previous three months, pregnant and lactating women, and those who had received periodontal treatment in the last three months were excluded. Samples from 32 subjects were collected. The GCF was collected with a paper point and stored in a sterilized phosphate buffer saline (PBS). The paper points were centrifuged at 3000 ×g for 5 min at 4°C, and then the supernatant was taken and frozen at -20°C until further analysis. The anthropometric demographic data of the subjects, such as name, address, gender, age, plaque index (PI) examination, papillary bleeding index (PBI), debris index (DI), calculus index (CI), and smoking status, were also recorded.
IL-1β gene expression analysis The RNA was extracted by GENEzol (Geneaid, Taipei, Taiwan) in accordance with the manufacturer’s instruction. RNA concentration was then measured using BioDrop (Biochrom, Cambourne, UK). The extracted RNA was reverse transcribed using ReverTra Ace quantitative polymerase chain reaction (qPCR) reverse transcription (RT) Master Mix with gDNA remover (Toyobo, Osaka, Japan). The IL-1β and GAPDH (a housekeeping gene for internal control) gene expression was analyzed by primer 3plus (IL-1β F 5′-GCATCCAGCTACGAATCTCC-3′; IL-1β R 5′-GAACCAGCATCTTCCTCAGC-3′; GAPDH F 5′-CAACGACCACTTTGTCAAGC-3′; GAPDH R 5′-TTACTCCTTGGAGGCCATGT-3′). qPCR was performed using an SYBR Green Supermix PCR kit (Bio-Rad, CA, USA) and Rotor Gene Q (Qiagen, Hilden, Germany). Initial denaturation at 95°C for 15 min, followed by 35 cycles of denaturation at 95°C for 15 s, annealing at 58°C for 15 s, and extension at 72°C for 30 s. The experimental quantification cycle values were normalized to GAPDH, and the 2−∆∆Ct method was used to discover out the relative gene expression levels.
The data were analyzed using Statistical Package for the Social Sciences (SPSS) software program, version 24.0 for the Shapiro–Wilk, Mann–Whitney, and t tests. The dataset was then transformed into tables and graphs.
| Results|| |
Anthropometric data analysis
Based on the Mann–Whitney and t test results, there was a significant difference between smoking and nonsmoking subjects in each variable. According to this study, the number of smoking subjects was 14 men and two women, whereas the overall number of nonsmoking subjects was one man and 15 women. With regard to age, the value of the smoking subjects was significantly higher (P = 0.004) than that of the nonsmoking ones, with mean ± standard deviation (SD) being (55.8 ± 5.36) and (49.38 ± 5.53), respectively. In terms of the PI, it was significantly higher (P = 0.013) in the subjects with the smoking status with mean ± SD (1.53 ± 0.71) than in those with the nonsmoking status with mean ± SD (0.99 ± 0.41). For the CI, the smoking subjects with mean ± SD (1.37 ± 0.69) had a significantly higher value (P = 0.006) than the nonsmoking ones with mean ± SD (0.78 ± 0.35). In the DI, the value of the smoking subjects with mean ± SD (1.36 ± 0.73) was significantly higher (P = 0.020) than that of the nonsmoking subjects with mean ± SD (0.85 ± 0.35). In relation to the oral hygiene index (OHI), the smoking subjects with mean ± SD (2.68 ± 1.36) had a significantly higher value (P = 0.000) than the nonsmoking ones with mean ± SD (0.84 ± 0.52). For the PBI, the value of the subjects with the smoking status (1.55 ± 0.80) was significantly higher (P = 0.006) than that of those with the nonsmoking status (0.85 ± 0.52). The complete details can be seen in [Table 1].
|Table 1: Mean and standard deviation of smoking status by gender, age, plaque index, calculus index, debris index, OHI, and PBI|
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Interleukin-1β gene expression
The results of the Mann–Whitney test showed that the P-value of the IL-1β was 0.047. The value of IL-1β varied between the smoking subjects and the nonsmoking ones. In addition, the mean ± SD value of IL-1β of the smoking subjects (11.34 ± 9.11) was significantly higher than that of the nonsmoking ones (0.24 ± 0.42). Therefore, the level of IL-1β of the smoking subjects was higher than that of nonsmoking ones, as shown in [Figure 1].
|Figure 1: Mean value of interleukin-1β based on smoking status. The Mann–Whitney test revealed that the level of interleukin-1β between the smoking and the nonsmoking periodontitis subjects was significantly distinct (P = 0.047)|
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| Discussion|| |
Currently, smoking continues to be a dangerous habit. Owing to the nicotine, it contains, which can lead to addiction, cigarettes have become an unavoidable necessity for their consumers. Cigarettes initiate a dose-response effect, which means that the younger the smoker, the greater the impact. The prevalence of smoking in Indonesia is extremely high; people start at the young age of 15 years. This research showed that the number of smoking subjects was higher in males than in females, namely as shown in this study, 14 men and two women. A previous study by Puscasu et al. found that there was a higher number of smoking subjects between the ages of 45 and 55 compared to nonsmoking ones in the same age group.
This research indicated that the PI, CI, DI, OHI, and PBI were significantly higher in smoking subjects than in nonsmoking ones. The increased PI, CI, DI, OHI, and PBI can function as signs of periodontitis. Periodontitis occurs due to the presence of bacterial plaque found in the subgingival area, which typically extends to the gingival groove, causing the loss of tooth attachment to the periodontal tissues. Previous studies proved that periodontitis is two to seven times greater in smokers than in nonsmokers since the former tend to have poor oral hygiene. In this regard, adult smokers have a higher risk of suffering from periodontitis at the age of 30 years old and over. This is adherent to the fact that the level of human immunity decreases over time, thus making them more susceptible to diseases. A study by Leite et al. showed a significant positive association between the habit of smoking and the increased risk of periodontitis in a prospective longitudinal study. In addition, if smoking is eliminated in this population, the risk of periodontitis will be reduced by 14%. This is consistent with the findings of this study, which showed that the occurrence of periodontitis was higher in smokers than in nonsmokers.
A previous study conducted by Moeintaghavi et al. revealed that the level of the IL-1β gene expression in nonsmoking periodontitis subjects was higher than in smoking periodontitis subjects. However, this research showed that the level of the IL-1β gene expression was higher in smoking periodontitis subjects than in nonsmoking periodontitis subjects. This is because Moeintaghavi et al. used periodontitis subjects who had undergone the first phase of periodontal treatment therapy. Therefore, the intensity of tissue inflammation was lower than that of the sample taken when the gingiva was completely inflamed. Meanwhile, in this research, periodontal treatment therapy had not been performed. Another difference lies in the primer used in each study, which may possibly lead to different results. A study by Meenawat et al. indicated that the level of IL-1β gene expression was significantly higher in smoking subjects than in nonsmoking ones. This is in line with the results of this research, which used subjects who had not been treated with periodontal therapy and had not taken antibiotics for the last three months. Furthermore, the chief difference between this study and the previous study by Meenawat et al. is the gender of the subjects. The former study included only male subjects, whereas this study included both male and female subjects. The limitation of this study is that the number of subjects whose interleukin levels can be perceived did not reach the desired sample size. However, based on these results and a post hoc power analysis test that was conducted, it was declared to have the ability to meet the requirements (1-β = 0.95).
| Conclusion|| |
This study discovered a difference in the occurrence of periodontitis between smokers and nonsmokers. The periodontal status (PI, CI, DI, OHI, and PBI) was significantly higher in the smoking periodontitis subjects than in the nonsmoking periodontitis subjects. Furthermore, smokers had significantly higher levels of IL-1β gene expression than nonsmokers.
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Conflicts of interest
There are no conflicts of interest.
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