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 Table of Contents  
ORIGINAL ARTICLE
Year : 2018  |  Volume : 9  |  Issue : 2  |  Page : 49-54

Analysis of salivary biomarkers during orthodontic tooth movement with conventional bracket and self-ligating brackets: An in vivo study


Department of Orthodontics, JSS Dental College and Hospital, Jagadguru Sri Shivarathreeshwara University, Mysore, Karnataka, India

Date of Web Publication30-May-2018

Correspondence Address:
Dr. Samson Thomas
Department of Orthodontics, JSS Dental College and Hospital, Jagadguru Sri Shivarathreeshwara University, Mysore, Karnataka
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/ijor.ijor_46_17

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  Abstract 

Objective: The aim of this study is to evaluate and compare salivary enzyme levels during orthodontic tooth movement with conventional brackets and self-ligating brackets.
Materials and Methods: Twenty patients (15–25 years of age) where 10 patients treated with mechanical biological treatment prescription and 10 patients were treated with Damon prescription requiring after first premolar extraction participated in the study. The canine retraction was started with nickel-titanium (NiTi) coil spring with 0.019 × 0.025” stainless steel wire. Saliva sampling was done after initial alignment before retraction and at 1, 2, 3, 4, and 5 weeks after the application of orthodontic force. A volume of 5 ml of unstimulated whole saliva will be collected from the subject for each prescription. Aspartate aminotransferase (AST), alkaline phosphatase (ALP), and lactate dehydrogenase (LDH) enzyme samples will be analyzed with fully automated clinical chemistry analyzer model TOSHIBA 120R from Agappe Diagnostics. The salivary sample for tartrate-resistant acid phosphatase (TRAP) will be analyzed with the enzyme-linked immune sorbent assay (ELISA) technique ELISA.
Results and Discussion: During canine retraction with NiTi coil spring the salivary enzyme levels for LDH and TRAP showed a significant difference from baseline to week 5 with Group A (conventional bracket) after the initiation of compressive orthodontic. The salivary enzyme levels for LDH, AST, TRAP, and ALP showed no significant difference from baseline to week 5 with Group B (self-ligating bracket) after the initiation of compressive orthodontic force. When compared between Group A and Group B at different time intervals for LDH, AST, TRAP, and ALP salivary enzyme levels, Group B showed a significant difference. The significant difference was seen with LDH at week 0 to week 2, AST at week 5, and TRAP at week 4, whereas ALP showed no significant difference. A significant difference with Group A was only seen with TRAP enzyme at week 1.
Conclusion: The LDH, AST, TRAP, and ALP level in Group A showed a significant increase whereas Group B showed no significant difference after the initiation of orthodontic.

Keywords: Alkaline phosphatase, aspartate aminotransferase, conventional bracket, lactate dehydrogenase, nickel-titanium coil spring, self-ligating bracket, tartrate-resistant acid phosphatase


How to cite this article:
Thomas S, Raghunath N. Analysis of salivary biomarkers during orthodontic tooth movement with conventional bracket and self-ligating brackets: An in vivo study. Int J Orthod Rehabil 2018;9:49-54

How to cite this URL:
Thomas S, Raghunath N. Analysis of salivary biomarkers during orthodontic tooth movement with conventional bracket and self-ligating brackets: An in vivo study. Int J Orthod Rehabil [serial online] 2018 [cited 2018 Jun 17];9:49-54. Available from: http://www.orthodrehab.org/text.asp?2018/9/2/49/233544


  Introduction Top


Tooth movement can be classified into physiological tooth movement and orthodontic tooth movement (OTM).[1] The application of orthodontic force can change the dental and paradental tissues. Previous studies have shown that several enzymes are expressed during these phases. These enzymes have been described as biomarkers during bone remodeling.[2],[3],[4],[5] Biomarkers are biologically active substances which are classified as biomarkers of inflammation, bone resorption, cell necrosis, bone deposition, and mineralization. The enzymatic biomarkers include lactate dehydrogenase (LDH), aspartate aminotransferase (AST), Tartrate-resistant Acid Phosphatase (TRAP), and alkaline phosphatase (ALP).[6],[7] There is a need to evaluate the levels of biomarkers during the phase of tooth movement with force delivering systems to understand the biomechanics of tooth movement. The present study is aimed to measure the level of LDH, AST, ALP, and TRAP of young patients undergoing orthodontic canine retraction with nickel-titanium (NiTi) coil spring. The study is also aimed to compare the levels of LDH, AST, ALP, and TRAP between conventional brackets and self-ligating brackets.

Aims and objectives

  1. To evaluate salivary enzyme levels during OTM with conventional ligating bracket
  2. To evaluate salivary enzyme levels during OTM with self-ligating bracket
  3. To compare the salivary enzyme levels during OTM between conventional and self-ligating bracket at different time intervals.



  Materials and Methods Top


In this study, a total of 20 Orthodontic patients aged between 15 and 25 years, irrespective of gender were selected from the outpatients in the Department of Orthodontics and Dentofacial Orthopedics, JSS Dental College and Hospital, JSS University, Mysuru. A total of 10 Orthodontic patients were treated with mechanical, biological treatment prescription [Figure 1], and 10 orthodontic patients were treated with Damon prescription [Figure 2]. The selected patients underwent orthodontic treatment with sequential wire changes until 0.019 × 0.025” [Figure 3] stainless steel wire were placed. Saliva sampling was done after initial alignment before retraction and at 1 week, 2 weeks, 3 weeks, 4 weeks, and 5 weeks after the application of orthodontic force. The retraction was done with NiTi closed coil spring, size 0.010 and length 9 mm [Figure 4]. A total of 5 ml of unstimulated whole saliva was collected from the subject for each prescription. The subject will be asked to salivate directly into sterile containers [Figure 5]. AST, ALP, and LDH enzyme samples will be analyzed with fully automated clinical chemistry analyzer model TOSHIBA 120R from Agappe Diagnostics [Figure 6]. The salivary sample for TRAP will be analyzed with the enzyme-linked immune sorbent assay (ELISA) technique ELISA [Figure 7].
Figure 1: 3M Unitek

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Figure 2: Damon Q (Damon prescription)

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Figure 3: Arch wire

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Figure 4: NiTi closed coil spring

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Figure 5: Sterile containers

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Figure 6: Chemical Analyser - Toshiba 120FR

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Figure 7: Elisa kit

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  Results Top


Statistical methods applied

Descriptive statistics

The descriptives procedure displays univariate summary statistics for several variables in a single table and calculates standardized values (z scores). Variables can be ordered by the size of their means (in ascending or descending order), alphabetically, or by the order in which the researcher specifies.

Following descriptive statistics were employed in the present study – mean, standard deviation, frequency, and percent.

Paired-samples t-test

The Paired-Samples t- test procedure compares the means of two variables for a single group. It computes the differences between values of the two variables for each case and tests whether the average differs from 0.

Repeated measure ANOVA

General linear model (GLM) repeated measures analyze groups of related dependent variables that represent different measurements of the same attribute. This dialog box lets you define one or more within-subjects factors for use in GLM repeated measures. Note that, the order in which you specify within-subjects factors is important. Each factor constitutes a level within the previous factor.

Independent-samples t-test

The independent-samples t-test procedure compares means for two groups of cases. Ideally, for this test, the patients should be randomly assigned into two groups and hence that any difference in response is due to the treatment (or lack of treatment) and not to other factors.

When groups were compared across different time intervals for changes in LDH values, nonsignificant differences were observed, indicating the similarity in the pattern of changes across 2 groups (F = 1.041, P = 0.399).

When groups were compared across different time intervals for changes in AST values, nonsignificant differences were observed, indicating the similarity in the pattern of changes across 2 groups (F = 1.604, P = 0.167).

When groups were compared across different time intervals for changes in TRAP values, nonsignificant differences were observed, indicating the similarity in the pattern of changes across 2 groups (F = 4.372, P = 0.001).

When groups were compared across different time intervals for changes in ALP values, nonsignificant differences were observed, indicating the similarity in the pattern of changes across 2 groups (F = 1.817, P = 0.117).


  Discussion Top


OTM is a process characterized by bone remodeling with bone deposition on tension side and bone resorption on the compression side.[8] An optimal force is important for adequate biological response in the periodontal ligament. A force level of 150–250 g is considered as the optimum force range for retraction of the canine tooth.[2],[3],[9]

A biomarker is defined as an indicator of normal biological, pathogenic processes, pharmacological responses to a therapeutic and other healthcare intervention.[4]

LDH, AST, TRAP and ALP in human saliva and gingival crevicular fluid (GCF) have the potential to serve as a biological marker for OTM monitoring.[5],[7],[10]

Enzyme levels of LDH in Group A (Conventional Bracket) found a significant increase (P ≤ 0.05) from baseline to week 5 [Graph 1]. This phenomenon shows the inflammatory process takes place 7–42 days after the start of treatment. During orthodontic treatment, inflammatory processes occur as a result of mechanical pressure imposed on the teeth. LDH will be released when lysis cells during inflammatory processes occur.[11] The inflammatory process that occurs during this orthodontic treatment will cause the death of part of a cell resulting from mechanical stress which, in turn, causes LDH enzymes to be released. The same increased LDH level is observed in GCF studies when inflammation occurs when orthodontic treatment is administered.[11]



LDH enzyme levels of Group B (self-ligating bracket) showed significant increase (P ≤ 0.05) at week 0 and week 1 [Graph 1] resulting inflammation taking place at 7 and 14 days, this minimal response of LDH in the saliva might have been caused by southern leaf blight, which has the potential to reduce the frictional resistance produced by the archwire and the bracket. Moreover, saliva itself is one of the factors that can affect frictional resistance. This finding is in agreement with the previous studies by Rohaya et al. and Shahrul Hisham et al.[5],[6],[7],[10]

AST is an intracellular enzyme that is normally confined to the cell cytoplasm but is released into the extracellular environment on cell death.[12] We found a significant difference (P ≤ 0.05) of enzyme levels of AST with Group A (conventional bracket) between week 1 and week 5 and week 2 to week 5 [Graph 2], this shows that there is cell necrosis taking place during 14–42 days, after inflammation.



According to von Böhl et al.,[13] necrotic tissue was formed during the second phase of tooth movement (after approximately 2 days of force application). However, the level gradually decreased over the next 3 weeks of treatment. We observed no significant difference (P > 0.05) with enzyme levels of AST of Group B (self-ligating bracket) between baseline to week 5 in saliva during this study.

Meanwhile, TRAP enzyme levels for Group A (conventional bracket) shows a significant increase (P ≤ 0.05) on 28, 35, and 42 days and is followed by bone formation from week 1 to week 3 and week 1 to week 4 [Graph 3].



Therefore, the profile of TRAP enzyme levels obtained for Group A (Conventional Bracket) indicates that the resorption process by osteoclastic cells is active on days 28, 35, and 42 days. In this phase, the resorption process by osteoclasts involves the degradation of organic and mineral elements in the bone matrix (Hill 1998). Next, the bone remodeling process will take over the role of completing the bone modeling cycle during orthodontic treatment. This phenomenon can be seen when ALP activity used as an indicator of the presence of active osteoblast cells where a significant increase (P ≤ 0.05) on 28 and 35 days after the increase of TRAP or osteoclast activity. Similar studies by Perinetti et al. (2002) showed ALP activity increased in GCF after 7 days of orthodontic treatment was imposed on patients.

There was a significant increase (P ≤ 0.05) of TRAP enzyme levels of Group B (Self-ligating bracket) at week 4, meaning bone resorption taking place. ALP enzyme levels showed no significant difference. These enzyme biomarker profiles showed that the remodeling cycle in Group B (self-ligating bracket) might be completed earlier, thus shortening the treatment time. However, the enzyme levels in saliva were very low and thus did not present a clear picture of the remodeling. Therefore, saliva samples would require more sensitive detection methods to obtain a clearer picture of the remodeling cycle. The use of self-ligating bracket tends to induce less bone resorption and bone formation, with desired tooth movement.[14]


  Conclusion Top


The conclusions drawn from the present study were

  • During canine retraction with NiTi coil spring, the LDH salivary enzyme levels in Group A (conventional bracket) were increased at baseline to week 5 after the initiation of compressive orthodontic force
  • During canine retraction with NiTi coil spring, the LDH salivary enzyme levels in Group B (self-ligating bracket) showed no significant difference from baseline to week 5 after the initiation of compressive orthodontic force
  • During canine retraction with NiTi coil spring, the AST salivary enzyme levels in Group A (conventional bracket) showed no significant difference from baseline to week 5 after the initiation of compressive orthodontic force
  • During canine retraction with NiTi coil spring, the AST salivary enzyme levels in Group B (self-ligating bracket) showed no significant difference from baseline to week 5 after the initiation of compressive orthodontic force
  • During canine retraction with NiTi coil spring, the TRAP salivary enzyme levels in Group A (conventional bracket) showed a significant difference from baseline to week 5 after the initiation of compressive orthodontic force
  • During canine retraction with NiTi coil spring, the TRAP salivary enzyme levels in Group B (self-ligating bracket) showed no significant difference from baseline to week 5 after the initiation of compressive orthodontic force
  • During canine retraction with NiTi coil spring, the ALP salivary enzyme levels in Group A (conventional bracket) showed no significant difference from baseline to week 5 after the initiation of compressive orthodontic force
  • During canine retraction with NiTi coil spring, the ALP salivary enzyme levels in Group B (self-ligating bracket) showed no significant difference from baseline to week 5 after the initiation of compressive orthodontic force
  • Comparison of LDH salivary enzyme levels between Group A and Group B at different time intervals where Group B (self-ligating bracket) shows a significant difference at week 0, week 1, and week 2 after the initiation of compressive orthodontic force
  • Comparison of AST salivary enzyme levels between Group A (conventional bracket) and Group B (self-ligating bracket) at different time intervals where Group A shows a significant difference at week 5 after the initiation of compressive orthodontic force
  • Comparison of TRAP salivary enzyme levels between Group A (conventional bracket) and Group B (self-ligating bracket) at different time intervals where Group A shows a significant difference at week 1 and Group B at week 4 after the initiation of compressive orthodontic force
  • Comparison of ALP salivary enzyme levels between Group A (conventional bracket) and Group B (self-ligating bracket) at different time intervals where Group A and Group B showed no significant difference after the initiation of compressive orthodontic force [Graph 4].



Generalized conclusion from the present study is

  • The LDH, AST, TRAP, and ALP level in Group A (Conventional Bracket) showed a significant increase after the initiation of orthodontic force
  • Where the salivary enzyme levels of LDH, AST, TRAP and ALP in Group B (Self-ligating Bracket) showed no significant difference after the initiation of orthodontic force.


Financial support and sponsorship

Nil.

Conflicts of interest

There are no conflicts of interest.

 
  References Top

1.
Abdul Wahab RM, Abu Kasim N, Senafi S, Jemain AA, Zainol Abidin IZ, Shahidan MA, et al. Enzyme activity profiles and ELISA analysis of biomarkers from human saliva and gingival crevicular fluid during orthodontic tooth movement using self-ligating brackets. Oral Health Dent Manag 2014;13:194-9.  Back to cited text no. 1
[PUBMED]    
2.
Krishnan V, Davidovitch Z. Cellular, molecular, and tissue-level reactions to orthodontic force. Am J Orthod Dentofacial Orthop 2006;129:469.e1-32.  Back to cited text no. 2
    
3.
Perinetti G, Paolantonio M, D'Attilio M, D'Archivio D, Tripodi D, Femminella B, et al. Alkaline phosphatase activity in gingival crevicular fluid during human orthodontic tooth movement. Am J Orthod Dentofacial Orthop 2002;122:548-56.  Back to cited text no. 3
    
4.
Atkinson AJ, Colburn WA, De Gruttola VG, et al. Biomarkers and surrogate endpoints: Preferred definitions and conceptual framework. Clin Pharmacol Therap2001;69:89-95.  Back to cited text no. 4
    
5.
Rohaya MA, Shahrul Hisham ZA, Khazlina K. The activity of aspartate aminotransferase during canine retraction (bodily tooth movement) in orthodontic treatment. J Med Sci 2008;8:553-8.  Back to cited text no. 5
    
6.
Shahrul Hisham ZA, Mohd Faiz E, Rohaya MA, Yosni B, Sahidan S. Profile of lactate dehydrogenase, tartrate resistant acid phosphatase and alkaline phosphatase in saliva during orthodontic tooth movement. Sains Malaysiana 2010;39:405-12.  Back to cited text no. 6
    
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Rohaya MA, Shahrul Hisham ZA, Khazlina K. Preliminary study of aspartate aminotransferase in gingival crevicular fluids during orthodontic treatment. J Appl Sci 2009;9:1393-6.  Back to cited text no. 7
    
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d'Apuzzo F, Cappabianca S, Ciavarella D, Monsurrò A, Silvestrini-Biavati A, Perillo L, et al. Biomarkers of periodontal tissue remodeling during orthodontic tooth movement in mice and men: Overview and clinical relevance. ScientificWorldJournal 2013;2013:105873.  Back to cited text no. 8
    
9.
Angolkar PV, Arnold JV, Nanda RS, Duncanson MG Jr. Force degradation of closed coil springs: An in vitro evaluation. Am J Orthod Dentofacial Orthop 1992;102:127-33.  Back to cited text no. 9
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Rohaya MA, Maryati MD, Sahidan S, Asma AA, Abdul Aziz J, Nurfathiha AK, et al. Crevicular tartrate resistant acid phosphatase activity and rate of tooth movement under different continuous force applications. Afr J Pharm Pharmacol 2011;5:2213-9.  Back to cited text no. 10
    
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Serra E, Perinetti G, D'Attilio M, Cordella C, Paolantonio M, Festa F, et al. Lactate dehydrogenase activity in gingival crevicular fluid during orthodontic treatment. Am J Orthod Dentofacial Orthop 2003;124:206-11.  Back to cited text no. 11
    
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Meeran NA. Biological response at the cellular level within the periodontal ligament on application of orthodontic force – An update. J Orthod Sci 2012;1:2-10.  Back to cited text no. 12
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von Böhl M, Maltha JC, Von Den Hoff JW, Kuijpers-Jagtman AM. Focal hyalinization during experimental tooth movement in beagle dogs. Am J Orthod Dentofacial Orthop 2004;125:615-23.  Back to cited text no. 13
    
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Kawashima-Ichinomiya R, Yamaguchi M, Tanimoto Y, Asano M, Yamada K, et al. External apical root resorption and the release of interleukin-6 in the gingival crevicular fluid induced by a self-ligating system. Open J Stomatol 2012;2:116-21.  Back to cited text no. 14
    


    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7]



 

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