|Year : 2021 | Volume
| Issue : 1 | Page : 90-93
Comparative evaluation of interleukin-17 in gingival crevicular fluid of patients with aggressive periodontitis and healthy gingival sites
Anand Narayanrao Wankhede1, Prasad Vijayrao Dhadse2, Priyanka Ganesh Jaiswal2, Vidya Sudhindra Baliga2
1 Department of Periodontology, V.Y.W.S. Dental College and Hospital, Amravati, Maharashtra, India
2 Department of Periodontics, Sharad Pawar Dental College and Hospital, Datta Meghe Institute of Medical Sciences University, Wardha, Maharashtra, India
|Date of Submission||16-Jul-2019|
|Date of Decision||28-Jan-2020|
|Date of Acceptance||04-Feb-2020|
|Date of Web Publication||29-Jul-2021|
Dr. Anand Narayanrao Wankhede
Department of Periodontology, V.Y.W.S. Dental College and Hospital, Amravati, Maharashtra
Source of Support: None, Conflict of Interest: None
Background: Aggressive periodontitis (AgP) is a severe form of periodontitis with rapid bone loss and clinical attachment loss. Many investigations aimed to understand its etiology and pathogenesis, however there is inadequate literature available regarding the level of interleukin-17 (IL-17) at sites of periodontal inflammation in AgP. Aim: The purpose of this study was to compare the levels of IL-17 in gingival crevicular fluid (GCF) of AgP patients with healthy sites of gingiva. Materials and Methods: GCF samples were obtained from individuals with clinically healthy gingiva (n = 15) and from AgP patients (n = 15). Clinical parameters including probing pocket depth (PPD), clinical attachment level (CAL), Papillary Bleeding Index (PBI), and Plaque index by Turkesy– Gilmore–Glickman Modification Of Quigley-Hein (PI) were measured. The level of IL 17 was measured by using enzyme linked immunosorbent assay (ELISA). Results: The level of GCF IL-17 was found to be statistically significantly increase in AgP (1.12 ± 0.29) as compared to healthy gingiva (0.64 ± 0.23). Clinical parameters such as modified PI, PPD, and PBI were significantly higher in AgP as compared to healthy clinical sites. Conclusion: The result of this study had shown that GCF IL-17 level was found to be elevated in AgP compared to healthy gingiva.
Keywords: Aggressive, cytokines, enzyme-linked immunosorbent assay, gingival crevicular fluid, interleukin-17, periodontitis
|How to cite this article:|
Wankhede AN, Dhadse PV, Jaiswal PG, Baliga VS. Comparative evaluation of interleukin-17 in gingival crevicular fluid of patients with aggressive periodontitis and healthy gingival sites. J Datta Meghe Inst Med Sci Univ 2021;16:90-3
|How to cite this URL:|
Wankhede AN, Dhadse PV, Jaiswal PG, Baliga VS. Comparative evaluation of interleukin-17 in gingival crevicular fluid of patients with aggressive periodontitis and healthy gingival sites. J Datta Meghe Inst Med Sci Univ [serial online] 2021 [cited 2021 Sep 16];16:90-3. Available from: http://www.journaldmims.com/text.asp?2021/16/1/90/322590
| Introduction|| |
Periodontitis is an inflammatory condition of the periodontium. It is complex and multifactorial in nature and is characterized by gingival inflammation and alveolar bone resorption.,, Periodontitis is further classified into chronic periodontitis and aggressive periodontitis (AgP). AgP differs from chronic periodontitis by the rapid rate of disease progression seen in an otherwise healthy individual, absence of large accumulations of plaque and calculus, and family history suggestive of a genetic trait in AgP.,
The balance between anti-inflammatory and pro-inflammatory condition plays a crucial role in any disease progression. In periodontitis, disruption between T-helper (Th) (Th1 and Th2) subsets and cytokine synthesis by them play an important role in the initiation and progression of periodontal inflammation. In recent years, another subset of Th cells, called Th17 cells which produce pro-inflammatory cytokines, has received great attention in progression of periodontitis.
Interleukin-17 (IL-17) is a pro-inflammatory cytokine and was first described as a T-cell product. IL-17 was discovered in 1993. The family consists of six members: IL-17A, IL-17B, IL-17C, IL-17D, IL-17E (IL-25), and IL-17F. IL-17 is also responsible to stimulate other pro-inflammatory cytokines such as IL-1β, IL-6, IL-8, PGE2, and matrix metalloproteinases through the stimulation of epithelial, endothelial, and fibroblastic cells.,, IL-17 aggravates periodontal disease by activating gingival fibroblasts to produce inflammatory cytokines. There is abundant documentation that suggest major tissue destruction in periodontitis results from the recruitment of host cells through the activation of monocytes/macrophages, lymphocytes, and fibroblast cell.,,
IL-17 may play a significant role in AgP because of the association of IL-17 pathways with the recruitment of neutrophils. There is no adequate literature available regarding the level of IL-17 at sites of periodontal inflammation in AgP. Therefore, the aim of this study was to determine the level of gingival crevicular fluid (GCF) IL-17 in healthy gingiva and generalized AgP.
| Materials and Methods|| |
Sixty samples from thirty individuals aged 25–55 years (mean age = 26.53 years) were selected from the outpatient department of periodontology. The samples were divided into two groups as follows:
[TAG:2]Group A (Healthy) [/TAG:2]
It consisted of 15 individuals with clinically healthy periodontium with no evidence of disease, showing no clinical attachment loss (CAL), probing pocket depth (PPD) <1.
Group B (aggressive periodontitis)
AgP patients were <35 years of age and had a minimum of six teeth other than the first molars and incisors with PPD and CAL ≥5 mm.
Exclusion criteria were (1) systemic diseases that could affect the periodontium, (2) tobacco in any form and alcoholics, (3) pregnant or lactating female, (4) periodontal treatment received for the past year, and (5) use of anti-inflammatory and antibiotic drugs in the past 6 months.
Information regarding dietary status, mouth-cleaning habits, and gingival and periodontal status along with other routine clinical data was recorded in a specially designed chart. An informed consent was signed by the patients agreeing for the study. The study protocol was approved by the institutional ethics committee.
The evaluation of the clinical parameters were determined by using Papillary bleeding index (PBI) by Muhlemann, Plaque index by Turkesy– Gilmore–Glickman Modification Of Quigley-Hein (PI), Probing pocket depth (PPD), and Clinical attachment level (CAL). PPD and CAL measurements were performed at four sites per tooth (mesial, distal, buccal, and palatal/ lingual aspects). CAL was measured from the cemento-enamel junction to the base of the probable pocket.
Laboratory analysis of gingival crevicular fluid
Gingival crevicular fluid collection and sampling
GCF samples were collected from the first maxillary molar in the right and left quadrants in each individual. Maxillary molar sites were considered for GCF sampling to prevent saliva contamination. In all cases, before the collection of GCF, all supragingival plaque was removed in the sampling sites with the help of an ultrasonic scaler. The sampling areas were isolated with cotton rolls to prevent the samples from being contaminated by saliva, and these sites were delicately dried using air spray. GCF samples from the sites of each individual were collected using calibrated volumetric microcapillary pipettes by putting the tip of the pipette extracrevicularly until the consistent volume of 1-μl GCF was collected and placed into Eppendorf tubes. It was stored at −70°C until laboratory analyses.
Laboratory analysis of the gingival crevicular fluid
IL-17 levels in GCF samples were estimated by enzyme-linked immunosorbent assay (ELISA) method by using IL-17 ELISA kit (Ç: Diaclone IL-17 ELISA kit, France (Catalog No.850.940.096) [Figure 1], as per the guidelines prescribed by the manufacturer which were followed very cautiously. The absorbance of each well was calibrated on the ELISA reader with 450 nm serving as the primary wavelength [Figure 2]. The attained concentration of IL-17 in each sample was determined by extrapolating optical density (OD) values against IL-17 standard concentration using the standard curve.
The Institutional Ethics Committee of DMIMSDU has approved the Research work proposed to be carried out at Sharad Pawar Dental College, Sawangi(M), Wardha. Date: 3rd July 2017 with Reference no DMIMS (DU)/IEC/2017-18/6427.
| Results|| |
Statistical analysis was done by using descriptive and inferential statistics. For all clinical parameters and IL-17 levels, the mean and standard deviation were calculated by using the Student's unpaired t-test. Correlation between PPD and CAL with IL-17 in AgP group was done by using Pearson's correlation coefficient. Software used in the analysis was SPSS 22.0 version (IBM, Armonk, NY, USA) and GraphPad Prism 6.0 version (San Diego, California, USA), and P < 0.05 was considered as the level of significance.
The concentration of IL-17 in GCF (pg/ml) in two groups along with the clinical parameters is viewed in [Table 1]. The calculated concentration of IL-17 was highest (statistically significant, P < 0.0001) in AgP as compared to healthy gingival site. Clinical parameters such as PI, PBI, and PPD were found to be statistically significant in AgP as compared to healthy gingival site. A positive correlation was found between PPD and IL-17 and also with clinical attachment level [Table 2].
|Table 1: Comparison of clinical parameters between healthy and aggressive periodontitis|
Click here to view
|Table 2: Correlation between probing pocket depth, clinical attachment level, and interlukin-17 in aggressive periodontitis group|
Click here to view
| Discussion|| |
Cytokines are considered a key component in the pathogenesis of inflammatory condition, including periodontitis., GCF is an exudate that flows into the oral cavity from periodontal pockets. As a result of gingival inflammation, GCF is formed when fluids leak from dilated blood vessels within the gingival connective tissue. As this fluid flows through the inflamed connective tissue, it picks up enzymes and other mediators involved in the immune response.
Recent studies had shown that IL-17 plays an important role in the progression of periodontitis. There are studies that investigated the level of IL-17 in gingival tissues and GCF of chronic periodontitis patients, which have shown significant increase in the level of IL-17 in chronic periodontitis as compared to healthy sites. A positive correlation has been found between PPD and clinical attachment loss.,,,,
In our study, the level of IL-17 in GCF in AgP was compared with healthy gingiva. Very few studies have investigated the total amount and concentration of IL-17 in AgP. The study conducted by Shaker and Ghallab found that the total amount of IL-17 was significantly higher in AgP than that of chronic periodontitis and control group. In contrast, some investigations have revealed lower levels of IL-17 in the GCF of AgP patients as compared to healthy group., In our study, the level of IL-17 was found to be statistically significantly increased (P < 0.0001) as compared to healthy group. Significant correlation was found between IL-17 with PPD and clinical attachment loss, which is in accordance with the study conducted by Lester et al.
| Conclusion|| |
Based on our results, the level of GCF IL-17 was found to increase statistically as compared to healthy gingiva. Investigations in all ethnic groups with larger populations are needed to clarify the specific contribution of IL-17 in the immunopathology of AgP.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Wankhede AN, Sayed AJ, Bhutada GP. Assessment of periodontal disease activity by salivary biochemical markers (enzymes) and its evaluation after non-surgical periodontal therapy. Int J Healthc Bio Res 2017;05:95-104.
Wankhede AN, Wankhede SA, Wasu SP. Role of genetic in periodontal disease. J Int Clin Dent Res Organ 2017;9:53-8. [Full text]
Baer PN. The case for periodontosis as a clinical entity. J Periodontol 1971;42:516-20.
Gemmell E, Seymour GJ. Cytokines and T cell switching. Crit Rev Oral Bio Med 1994;5:249-79.
Honda T, Domon H, Okui T, Kajita K, Amanuma R, Yamazaki K. Balance of inflammatory response in stable gingivitis and progressive periodontitis lesions. Clin Exp Immunol 2006;144:35-40.
Yetkin Ay Z, Sütçü R, Uskun E, Bozkurt FY, Berker E. The impact of the IL-11:IL-17 ratio on the chronic periodontitis pathogenesis: A preliminary report. Oral Dis 2009;15:93-9.
Wong CK, Lit LC, Tam LS, Li EK, Wong PT, Lam CW. Hyperproduction of IL-23 and IL-17 in patients with systemic lupus erythematosus: Implications for Th17-mediated inflammation in auto-immunity. Clin Immunol 2008;127:385-93.
Kuby J. Immunology. New York: WH Freeman; 1992.
Belardelli F, Ferrantini M. Cytokines as a link between innate and adaptive antitumor immunity. Trends Immunol 2002;23:201-8.
Rouvier E, Luciani MF, Mattéi MG, Denizot F, Golstein P. CTLA-8, cloned from an activated T cell, bearing AU-rich messenger RNA instability sequences, and homologous to a herpesvirus saimiri gene. J Immunol 1993;150:5445-56.
Toh ML, Gonzales G, Koenders MI, Tournadre A, Boyle D, Lubberts E, et al
. Role of interleukin 17 in arthritis chronicity through survival of synoviocytes via regulation of synoviolin expression. PLoS One 2010;5:e13416.
Fossiez F, Djossou O, Chomarat P, Flores-Romo L, Ait-Yahia S, Maat C, et al
. T cell interleukin-17 induces stromal cells to produce proinflammatory and hematopoietic cytokines. J Exp Med 1996;183:2593-603.
Van bezooijen RL, Farih-Sips HC, Papapoulos SE, Löwik CW. Interleukin-17: A new bone acting cytokine in vitro
. J Bone Miner Res 1999;14:1513-21.
Shih DQ, Targan SR, McGovern D. Recent advances in IBD pathogenesis: Genetics and immunobiology. Curr Gastroenterol Rep 2008;10:568-75.
Crispín JC, Oukka M, Bayliss G, Cohen RA, Van Beek CA, Stillman IE, et al
. Expanded double negative T cells in patients with systemic lupus erythematosus produce IL-17 and infiltrate the kidneys. J Immunol 2008;181:8761-6.
Jaiswal PG, Bhongade ML, Bhake A, Thakare KT, Pande S, Pavan Bajaj P, et al
. Comparison of the Phagocytic Activity Of Gingival Crevicular Fluid (GCF) Neutrophils At Healthy Sites With Periodontally Diseased Sites. J Appl Dent Med Sci 2017;3:51-59.
Shaker OG, Ghallab NA. IL-17 and IL-11 GCF levels in aggressive and chronic periodontitis patients: Relation to PCR bacterial detection. Mediators Inflamm 2012;17:47-64.
Mühlemann HR. Psychological and chemical mediators of gingival health. J Prev Dent 1977;4:6-17.
Schneider HG, Göbbels E, Apel EM. Simplification of plaque index method of Quigley and Hein. Stomatol DDR 1989;39:91-4.
Vernal R, Dutzan N, Chaparro A, Puente J, Antonieta Valenzuela M, Gamonal J. Levels of interleukin-17 in gingival crevicular fluid and in supernatants of cellular cultures of gingival tissue from patients with chronic periodontitis. J Clin Periodontol 2005;32:383-9.
Chaplin DD. 1. Overview of the human immune response. J Allergy Clin Immunol 2006;117:S430-5.
Orozco A, Gemmell E, Bickel M, Seymour GJ. Interleukin 18 and periodontal disease. J Dent Res 2007;86:586-93.
Lamster IB, Novak MJ. Host mediators in gingival crevicular fluid: Implications for the pathogenesis of periodontal disease. Crit Rev Oral Biol Med 1992;3:31-60.
Cardoso CR, Garlet GP, Crippa GE, Rosa AL, Júnior WM, Rossi MA, et al
. Evidence of the presence of T helper type 17 cells in chronic lesions of human periodontal disease. Oral Microbiol Immunol 2009;24:1-6.
Mitani A, Niedbala W, Fujimura T, Mogi M, Miyamae S, Higuchi N, et al
. Increased expression of interleukin (IL)-35 and IL-17, but not IL-27, in gingival tissues with chronic periodontitis. J Periodontol 2015;86:301-9.
Lester SR, Bain JL, Johnson RB, Serio FG. Gingival concentrations of interleukin-23 and -17 at healthy sites and at sites of clinical attachment loss. J Periodontol 2007;78:1545-50.
Nagireddy RR, Chavan V, Subramanyam MB, Reddy VS, Pasupuleti MK, Avula KK. Estimation of interleukin-17 levels in gingival crevicular fluid from healthy individuals and patients with chronic periodontitis. J NTR Univ Health Sci 2013;2:191-5. [Full text]
Chitrapriya MN, Rao SR, Lavu V. Interleukin-17 and interleukin-18 levels in different stages of inflammatory periodontal disease. J Indian Soc Periodontol 2015;19:14-7.
] [Full text]
Ay ZY, Yılmaz G, Ozdem M, Koçak H, Sütçü R, Uskun E, et al
. The gingival crevicular fluid levels of interleukin-11 and interleukin-17 in patients with aggressive periodontitis. J Periodontol 2012;83:1425-31.
Sadeghi R, Sattari M, Dehghan F, Akbari S. Interleukin-17 and interleukin-23 levels in gingival crevicular fluid of patients with chronic and aggressive periodontitis. Cent Eur J Immunol 2018;43:76-80.
[Figure 1], [Figure 2]
[Table 1], [Table 2]