Journal of Family Medicine and Primary Care

: 2019  |  Volume : 8  |  Issue : 7  |  Page : 2373--2377

Evaluation of levels of Interleukin-1b, intensity of pain and tooth movement during canine retraction using different magnitudes of continuous orthodontic force

Rajeshwar Singh1, Poonam K Jayaprakash2, Ankit Yadav3, Meeta Dawar3, Harpreet Grewal4, Amit Mishra5,  
1 Department of Orthodontics and Dentofacial Orthopedics, Mekelle University, Mekelle, Tigray, Ethiopia
2 Department of Orthodontics and Dentofacial Orthopedics, Kothiwal Dental College and Research Center, Mora Mustaqueem, Moradabad, Uttar Pradesh, India
3 Department of Orthodontics and Dentofacial Orthopedics, Yadav Dental and Orthodontic Centre, New Delhi, India
4 Professor and Head, Department of Dentistry, University College of Medical Sciences and Guruteg Bahadur Hospital, Dilshad Garden, Delhi, India
5 Post Graduate Student, Mithila Minority Dental College, Darbhanga, Bihar, India

Correspondence Address:
Dr. Poonam K Jayaprakash
Department of Orthodontics and Dentofacial Orthopedics, Kothiwal Dental College and Research Center, Mora Mustaqueem, Moradabad - 244 001, Uttar Pradesh


Aim: The present study was conducted for the evaluation of Interleukin (IL)-1b levels in human gingival crevicular fluid (GCF), intensity of pain, and the amount of tooth movement measured during canine retraction using different magnitudes of continuous orthodontic force. Materials and Method: A statistically significant number of subjects were included for the study (n = 16, 6 male subjects and 10 female subjects). The age ranged from 18 to 24 years and all were diagnosed with Class I bimaxillary protrusion. They underwent first premolar extractions prior to participating in the study. The maxillary cuspids were then retracted using a continuous force of either 50 or 150 g. This was executed using nickel–titanium coil springs on segmented archwires. The opposite counterpart, that is, mandibular cuspid was used as control. GCF was then drawn from the distal aspect of each tooth at defined time intervals. This was followed by the assessment of IL-1b concentrations, pain intensity, using the visual analogue scale (VAS), and the amount of tooth movement. ANOVA test, Friedman test, and paired t-tests were used for comparisons of IL-1b in GCF, the plaque and gingival indices, and the efficiency of tooth movement on pain perception, respectively. Results: The 150 g group showed the highest level of IL-1b concentration at 24 h from baseline and at 2 with significant differences compared with the control group (P < 0.05). The mean VAS score of pain intensity from the 150 g force was significantly greater than from the 50 g force at 24 h (P < 0.01). Conclusion: No significant difference in the amount of tooth movement was found between these two different magnitudes of continuous force at 2 months. A 50 g force could effectively induce tooth movement similar to 150 g with less pain and less inflammation.

How to cite this article:
Singh R, Jayaprakash PK, Yadav A, Dawar M, Grewal H, Mishra A. Evaluation of levels of Interleukin-1b, intensity of pain and tooth movement during canine retraction using different magnitudes of continuous orthodontic force.J Family Med Prim Care 2019;8:2373-2377

How to cite this URL:
Singh R, Jayaprakash PK, Yadav A, Dawar M, Grewal H, Mishra A. Evaluation of levels of Interleukin-1b, intensity of pain and tooth movement during canine retraction using different magnitudes of continuous orthodontic force. J Family Med Prim Care [serial online] 2019 [cited 2021 Jun 25 ];8:2373-2377
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Tissue remodeling is facilitated by orthodontic forces which occurs mainly as a reaction of tissues to some type of mechanical stimulation. A plethora of literature is available online reflecting the results of various studies conducted to determine the magnitude of optimal forces or range of force for orthodontic tooth movement.[1],[2],[3],[4] The appropriate forces for tooth movement of human teeth reportedly range from a force as light as 18 g to one as heavy as 1,515 g.[2],[5] This argument still exists, and no evidence-based optimal force level can be recommended in clinical orthodontics.[5],[6] In addition to the forces optimal for the velocity of human tooth movement, the inflammatory response and pain after orthodontic force is applied which need to be studied.

Since teeth must be moved safely as well as efficiently, it is important to determine the possible adverse effects from various magnitudes of force application, cell biology by cytokines, and patient discomfort from pain intensity. The purpose of this study, therefore, was to compare two different magnitudes of orthodontic force used for canine retraction, with regard to IL-1b secretion in GCF, efficiency of tooth movement, and pain perception. The null hypothesis tested was that there is no difference between forces of 50 and 150 g concerning these measured variables.

 Subjects and Methods

Patient selection

Sixteen patients aged 18–24 years (six males, mean age 20.8 ± 1.2 years; 10 females, mean age 20.2 ± 1.6 years) participated in this study. They all met the following criteria: (1) class I molar relationship and bimaxillary protrusion with very mild crowding, especially in the posterior segment; (2) treatment plan involving extraction of all first premolars and distal retraction of the canines; (3) no evidence of periodontal or gingival disease; and (4) no history of antibiotic therapy during the previous 3 months and no anti-inflammatory drug use within 1 month before the start of the study. The reason for excluding patients with a history of recent antibiotic and inflammatory drug use was that they would affect some of the mediators released and immune functions.

Experimental design

After first premolar extractions, all subjects received oral hygiene instruction and were advised to have a soft food diet and to chew on both sides 1 month before and throughout the experimental period. To prevent plaque formation and the development of gingivitis, all subjects started rinsing with chlorhexidine mouthwash twice daily until the end of the experiment. At each appointment, the oral hygiene of each subject was evaluated using the plaque index (PI) as described by Dababneh et al.[7] and the modified gingival index given by Lobene et al.[8] (GI). A transpalatal arch attached on molar bands was inserted at least 1 week before the experimental procedures.

Brackets (0.022 inch slot, Ormco Corp.) and segmented archwires (0.018 × 0.025 inch stainless steel wire) were placed on the upper posterior teeth. The upper right and left canines of the same patient were randomly retracted using a continuous force of 50 or 150 g with nickel--titanium coil springs (Tomy®, Tokyo, Japan). The accuracy of the force was measured before canine retraction with a calibrated orthodontic force gauge (Gram Gauges, Mecmesin Asia Co. Ltd., Bangkok, Thailand). A lower right or left canine with no appliance was used as the control.[9]

GCF sampling

GCF was collected from the distal site of the experimental and control canines before retraction (baseline) and after retraction at 1 and 24 h, 1 week, 1 month, and 2 months without any reactivation of the coil spring. A paper strip (Periopaper; Proflow™ Incorporated, Amityville, New York, USA) was carefully inserted 1 mm into the gingival crevice on the distal side and left there for 30 s [10] [Figure 1]. After an interval of 90 s, a second strip was carefully placed at the same site. The absorbed fluid volume was measured with a Periotron 8000 (Proflow™ Incorporated). The two periopapers of each sample site were pooled into a sealed tube and immediately frozen at −80°C.{Figure 1}

The periopapers in each tube were eluted with 100 ml of 0.05 M TrisHCl buffer (pH 7.5) and centrifuged at 5000 g, 4°C, for 20 min. A further 50 ml of buffer was then applied, and the procedure was repeated. Subsequently, the supernatants were placed in a new tube and prepared for measurement of protein and IL-1b concentrations.

Protein assay and IL-1b determination

Protein concentrations of each sample site were measured by BCA Assay with bovine serum albumin as a standard. IL-1b levels were determined using the enzyme-linked immunosorbent assay. Total IL-1b was calculated in picograms, and IL-1b concentration in each sample site was calculated from the amount of IL-1b divided by the total protein content in GCF samples (picograms/milligrams of total protein).

Intensity of pain

For evaluation of pain intensity, all subjects were instructed to place a mark on a 100 mm VAS, corresponding to their current level of spontaneous pain intensity, including a feeling of discomfort for the right and left experimental canines separately as well as the control tooth at all experimental time periods without any stimulation. The left end of the line was given a VAS score of 0, indicating no pain, and the right end 100, indicating maximum pain. The distance from the left side to the mark indicating pain intensity was measured three times and averaged.

Determination of the amount of tooth movement

Dental models of all subjects taken before and at 2 months were evaluated with a measuring microscope.

Statistical analyses

Data analysis was performed using the Statistical Package for Social Sciences version 14.0 (SPSS Inc., Chicago, Illinois, USA). Means and standard deviations of total protein and IL-1b concentrations from the GCF samples of all groups were calculated. For comparison of the protein or IL-1b concentrations at each observation time point within each group, repeated measures one-way analysis of variance (ANOVA) was performed. One-way ANOVA was used for comparison of concentrations of protein and IL-1b among the groups and Friedman test for comparisons of the PI and modified GI among the groups. A paired t-test was used for comparing VAS scores of pain intensity or the amount of canine movement between the 50 and 150 g force. The significance level was set at P < 0.05.


All subjects showed good gingival and periodontal status at all experimental time points with no significant difference in PI and modified GI scores [Figure 2] and [Figure 3].{Figure 2}{Figure 3}

GCF volumes showed no significant difference among or within groups at any time point [Table 1]. The mean value of total protein concentrations in the GCF samples of all groups was approximately 12 mg/ml at all-time points (data not shown).{Table 1}

IL-1b concentrations in the 50 and 150 g groups increased, with the greatest mean amounts at 24 h, declined to approximately normal levels during 1 week to 1 month, and increased again at 2 months [Table 2]. No significant difference was found between the two experimental groups (P > 0.05) in Canine retraction after 2 months of applicationof ontinuous orthodontic forces of 50 and 150 g [Figure 4]. Significant differences were found between the control and a force of 150 g at 24 h and 2 months (P < 0.05) [Table 3].{Table 2}{Figure 4}{Table 3}


In this study, an attempt was made to evaluate the efficacy of different amounts of orthodontic force (50 and 150 g) for tooth movement in conjunction with levels of IL-1b as well as intensity of pain because a force of 100–200 g has been recommended for canine retraction.[11]

GCF collection, which is a noninvasive method that has been widely used for analysis of human tooth movement, enables easy detection of various biochemical markers [12] (Uematsu et al., 1996). Because the level of IL-1b in GCF increases with plaque accumulation and gingival inflammation, all subjects were instructed to maintain good oral hygiene practices throughout the period of the study. The PI and GI results for all subjects showed no sign of gingival inflammation or significant changes at any time point. Moreover, as there was no change in GCF volume, this demonstrated good gingival health throughout experimental period.

Interestingly, there was no significant difference between the mean amount of canine movement with forces of 50 and 150 g at 2 months, implying that force magnitudes less than 100 g could produce the same rate of tooth movement as a greater force.[3],[13] Iwasaki et al.[2],[14] used continuous average forces of 18 and 60 g for canine retraction and found that effective tooth movement could be produced with lower forces and that the lag phase was eliminated.

The immediate painful response from initial orthodontic force has been reported to be due to the development of an acute inflammatory process and changes in blood flow in the PDL.[15] To evaluate pain intensity, a VAS was used as this method has been found to be valid and reliable in previous research.[16],[17] In this study, because of the well-aligned posterior teeth, canine retraction by continuous coil springs could be performed immediately after placement of brackets and segmented arch wires. The maxillary first molar bands with the transpalatal arch had been placed more than 1 week earlier to ensure that pain from the band phase had subsided.[18] The highest pain intensity was found in the 150 g group at 24 h, similar to other studies,[19],[20] while pain in the 50 g group was significantly less.

In the present study, at 24 h, IL-1b concentration from a force of 150 g showed the highest data, which was consistent with the reported pain. Thus, the concentration of IL-1b was to some extent related to the pain intensity. It could be considered that there might be a concentration of IL-1b, which induced sufficient tooth movement but not strong pain. A force of 50 g could be considered optimum for canine retraction.


A continuous force of 150 g resulted in significantly higher IL-1b levels at 24 h and after 2 months of initial canine tooth movement when compared with the control teeth. A continuous force of 50 g produced significantly less pain intensity at 24 h compared with a 150 g force. Both forces resulted in movement of the canines after 2 months, but without a statistically significant difference. A continuous force of 50 g could effectively induce canine movement similar to a 150 g force, but with less pain.

Financial support and sponsorship


Conflicts of interest

There are no conflicts of interest.


1Boester CH, Johnston LE. A clinical investigation of the concept of differential and optimal force in canine retraction. Angle Orthodontist 1974;44:113-9.
2Iwasaki LR, Haack JE, Nickel JC, Morton J. Human tooth movement in response to continuous stress of low magnitude. Am J Orthod Dentofacial Orthop 2000;117:175-83.
3Reitan K. Some factors determining the evaluation of forces in orthodontics. Am J Orthod 1957;43:32-45.
4Storey E. The nature of tooth movement. Am J Orthod 1973;63:292-314.
5Ren Y, Maltha JC, Kuijpers-Jagtman AM. Optimum force magnitude for orthodontic tooth movement: A systematic literature review. Angle Orthodontist 2003;73:86-92.
6Limsiriwong S, Khemaleelakul W, Sirabanchongkran S, Pothacharoen P, Kongtawelert P, Ongchai S, et al. Biochemical and clinical comparisons of segmental maxillary posterior tooth distal movement between two different force magnitudes. Eur J Orthod 2018;40:496-503.
7Dababneh RH, Khouri AT, Smith RG, Addy M. Correlation and examiner agreement between a new method of plaque scoring and a popular established plaque index, modelled in vitro. J Clin Periodontol 2002;29:1107-11.
8Lobene RR, Weatherford T, Ross NM, Lamm RA, Menaker L. A modified gingival index for use in clinical trials. Clin Prev Dent 1986;8:3-6.
9Lee KJ, Park YC, Yu HS, Choi SH, Yoo YJ. Effects of continuous and interrupted orthodontic force on interleukin-1beta and prostaglandin E2 production in gingival crevicular fluid. Am J Orthod Dentofacial Orthop 2004;125:168-77.
10Offenbacher S, Odle BM, Van Dyke TE. The use of crevicular fluid PGE2 levels as a predictor of periodontal attachment loss. J Periodontal Res 1986;21:101-12.
11Burstone CJ, Baldwin JJ, Lawless DT. The application of continuous forces to orthodontics. Angle Orthodontist 1961;31:1-14.
12Grieve III WG, Johnson GK, Moor RN, Reinhardt RA, DuBois LM. Prostaglandin E (PGE) and interleukin-1b (IL-1b) levels in gingival crevicular fluid during human orthodontic tooth movement. Am J Orthod Dentofacial Orthop 1994;105:369-74.
13Gianelly AA, Goldman HM. Tooth movement. In: Biological Basis of Orthodontics. Philadelphia: Lea and Febiger; 1971. p. 116-204.
14Iwasaki LR, Gibson CS, Crouch LD, Marx DB, Pandey JP, Nickel JC. Speed of tooth movement is related to stress and IL-1 gene polymorphisms. Am J Orthod Dentofacial Orthop 2006;130:698.e1-9.
15Burstone CJ. The biomechanics of tooth movement. In: Kraus BS, Riedel RA, editors. Vistas in Orthodontics. Philadelphia: Lea and Febiger; 1962. p. 197-213.
16Carlsson AM. Assessment of chronic pain. I. Aspects of the reliability and validity of the visual analogue scale. Pain 1983;16:87-101.
17Krebs EE, Carey TS, Weinberger M. Accuracy of the pain numeric rating scale as a screening test in primary care. J Gen Intern Med 2007;22:1453-8.
18Jones ML, Richmond S. Initial tooth movement: Force application and pain-a relationship? Am J Orthod 1985;88:111-6.
19Ngan P, Kess B, Wilson S Perception of discomfort by patients undergoing orthodontic treatment. Am J Orthod Dentofacial Orthop 1989;96:47-53.
20Polat O, Karaman AI. Pain control during fixed appliance therapy. Angle Orthodontist 2005;75:214-9.