==== Front Dent Res J (Isfahan)Dent Res J (Isfahan)DRJDental Research Journal1735-33272008-0255Medknow Publications & Media Pvt Ltd India DRJ-15-276Original ArticleThe effect of fluoride varnish and chlorhexidine gel on white spots and gingival and plaque indices in fixed orthodontic patients: A placebo-controlled study Alavi Shiva 1Yaraghi Navid 21 Department of Orthodontics, Dental Materials Research Center, School of Dentistry, Isfahan University of Medical Sciences, Isfahan, Iran2 Department of Orthodontics, School of Dentistry, Isfahan University of Medical Sciences, Isfahan, IranAddress for correspondence: Dr. Navid Yaraghi, Department of Orthodontics, School of Dentistry, Isfahan University of Medical Sciences, Isfahan, Iran. E-mail: navidyaraghi@yahoo.comJul-Aug 2018 15 4 276 282 10 2017 3 2018 Copyright: © 2018 Dental Research Journal2018This is an open access journal, and articles are distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 License, which allows others to remix, tweak, and build upon the work non-commercially, as long as appropriate credit is given and the new creations are licensed under the identical terms.Background: Difficulties to maintain good oral hygiene during orthodontic treatment can cause prolonged accumulation of dental plaque, thereby increasing the risk of developing gingival inflammation and periodontal disease as well as enamel demineralization and caries. The purpose of this study was to determine the effect of chlorhexidine (CHX) gel and fluoride varnish versus placebo and control groups on plaque and gingival indices as well as their influence on enamel demineralization prevention in orthodontic patients with fixed appliances. Materials and Methods: In this clinical trial, forty patients with fixed orthodontic appliances were participated and were divided into control, CHX gel (0.2%), fluoride varnish (5%), and placebo groups. The parameters evaluated in this research were dental plaque index (DPI), index of gingival inflammation, and white spot lesion (WSL) index. For all the patients, each index was scored at the beginning of orthodontic treatment, then 3 months, 6 months, and 9 months afterward. Data were analyzed using Kruskal–Wallis and Mann–Whitney tests. The level of significance was set at 0.05. Results: All the scores showed statistically significant differences (P < 0.05) between the four groups for each of the three time points. A statistically significant decrease of DPI, WSL index, and gingival index was observed 9 months after treatment in fluoride and CHX groups. Conclusion: Adding CHX gel and fluoride varnish to the patients' oral hygiene regimen can reduce the development of plaque and gingivitis and decrease WSLs in orthodontic patients. Key Words: Chlorhexidinedental plaque indexorthodontic applianceperiodontal indexwhite spot ==== Body INTRODUCTION Dental caries is one of the most important oral cavity diseases, which initially appears as a white spot lesion (WSL) due to the loss of mineral content.[1] After the introduction of orthodontic fixed appliances into the oral cavity, some rapid ecologic changes such as increase of Streptococcus mutans have been reported in the saliva and plaque of patients.[2] Prevention of demineralization and occurrence of new carious lesions in orthodontic patients is rather difficult. Besides, good oral hygiene and regular supplementation techniques are often needed.[34] After bracket placement, WSLs can be identified within 1 month although it takes at least 6 months before caries becomes notable.[4] These lesions are predominantly in sites adjacent to brackets and are usually formed at the buccal surfaces, especially in the gingival region.[56] Plaque also harbors the cariogenic bacteria, potentially capable of hard tissue damage, especially at the bracket margins.[7] Mechanical and chemical cleaning of tooth surface with regular brushing and mouth rinses can help reduce plaque formation and accumulation; therefore, it prevents gingival and dental diseases during orthodontic appliance therapy.[8] Good plaque control is very difficult in patients with fixed orthodontic appliances. To improve mechanical plaque removal, incorporation of a chemotherapeutic agent such as an antibacterial mouth rinse into the oral hygiene regimen can be helpful.[910] Chlorhexidine (CHX) is the most potentially documented antimicrobial agent against mutans streptococci (MS) and dental caries. Different modes of administration are recommended for caries prevention.[111213] The form that CHX is delivered such as varnishes, gels, and rinsing solutions determines the mode of its effect. The contact time of CHX with intraoral tissues, its concentration and rate of release, are factors affecting its persistence of bacterial suppression. Depending on these particular factors, CHX varnishes establish the most persistent reduction in MS, followed by gels and mouthwashes.[1415] Studies have shown that a long-lasting suppression of MS concentrations can be achieved by application of CHX varnishes and gels compared with other forms of application.[1617] High and low concentrations have been reported to reduce the number of MS in plaque and saliva for considerable periods of time.[1819] The safety of CHX has been reportedly confirmed although a drawback of CHX is the associated staining of pellicle. The effect of subgingival irrigation with CHX on gingivitis in adolescents with fixed orthodontics has been reported by Morrow et al.[20] Topical application of fluoride varnish at least around anterior teeth is suggested in high-risk patients, for example, for orthodontic patients at every orthodontic adjustment appointment.[21] However, the most recommended type of fluoride by orthodontists is daily use of 0.05% sodium fluoride rinse in conjunction with fluoridated dentifrice.[22] Perhaps, this recommendation was based on the research done on nonorthodontic adolescent patients, suggesting that this approach may significantly help reduce caries rates. Further, the efficacy of fluoride rinses in WSL prevention in patients undergoing orthodontic treatment is questionable, and there is weak and mixed evidence on this topic. Moreover, it should be noted that patients' compliance with such rinses is poor.[23] There is evidence regarding the ability of amorphous calcium phosphate (ACP) in remineralization of WSL after orthodontic treatment. Some studies have reported no significant advantage for the use of ACP along with regular oral hygiene regimen including 1,000 ppm fluoride toothpaste.[2425] Plaque accumulation which can lead to WSL formation and gingival inflammation represent a significant challenge to excellence in clinical orthodontics. The risk assessment methods and finding the best preventive materials may help to minimize this common problem in your patients; however, no matter how hard we strive, it is unlikely that this problem can be completely eliminated. For this reason, it is imperative for both orthodontists and general practitioners to have consistent protocols for caries risk assessment, prevention, intratreatment management, and treatment of WSL. The purpose of this study was to evaluate the effect of CHX and fluoride varnishes on plaque and gingival indices as well as their efficiency in preventing demineralization in orthodontic patients with fixed appliances. An attempt was also made to compare the results of these groups with control and placebo groups. MATERIALS AND METHODS This clinical trial included 40 patients (20 females and 20 males), who were treated in the Department of Orthodontics of Dental School, Isfahan University of Medical Sciences. This trial is registered with IRCT registry ID: IRCT2016122531558N1. The participants were selected based on simple sampling and were randomly divided into four groups. The participants were qualified on the basis of the following criteria: Having orthodontic appliances in both jaws and undergoing a nonextraction orthodontic therapy Aged over 18 Showing no allergic reaction to the materials used No history of previous orthodontic therapies No systematic diseases affecting teeth such as amelogenesis and dentinogenesis imperfecta No systematic diseases affecting gum inflammation such as lichen planus and pemphigus No clinical evidence of periodontal diseases No medical problems or evidence of current antibiotic therapy or using drugs affecting salivary secretion. Patients who did not follow the given oral hygiene instructions were excluded from this study (To exclude the patients who did not follow the oral hygiene program, participants were questioned each session). The study population had a mean age of 23 years (range: 18–34). The patients were divided into control group (n = 10), CHX group (n = 10), fluoride group (n = 10), and placebo group (n = 10). Clinical procedures Before the beginning of examination, all of the selected volunteers were instructed on toothbrushing technique (bass technique) and flossing. As shown, teeth were selected based on proposed method for plaque and gingival index (GI) and white spot by Silness and Loe: Preparations Prophylaxis For all groups before varnish administration, plaque removal was done using toothbrush and toothpaste. Tested materials were applied to teeth surfaces around brackets, and patients were asked not to drink or eat anything for 2 h. Bonding All the brackets were bonded with nonfluoride adhesives according to standard procedures. Clinical measurements The materials used were CHX varnish (0.2%, Periokin, Spain), sodium fluoride varnish alcohol free (5%, Pascal, America) and 70% Wt% sorbitol solution for placebo group. They were applied in every 3 months. The following parameters, including dental plaque index (DPI) of Silness-Löe and GI and index of gingival inflammation of Löe and Silness, were recorded at baseline (day 0), and after 3, 6, and 9 months.[26] The WSL index was assessed using the International Caries Detection and Assessment System (ICDAS).[27] ICDAS was also recorded through visual evaluation of selected tooth at baseline (day 0) and after 3, 6, and 9 months' surfaces. The scoring was performed as follows: (0) no visible WSL or surface disruption (no demineralization); (1) visible WSL covering less than one-third of the surface, without surface disruption (mild demineralization); (2) visible WSL covering more than one-third of the surface, with a roughened surface but not requiring restoration (moderate demineralization), and (3) visible cavitation, requiring restoration (severe demineralization).[28] Statistical analysis Continuous variables were expressed as mean ± standard deviation (SD), and t-test was used to compare the means among groups. Data were analyzed using Kruskal–Wallis and Mann–Whitney tests. SPSS version 21 (SPSS Inc., Chicago, Illinois, USA) was used for data analysis. P ≤ 0.05 was considered statistically significant for all analyses. RESULTS A total of 40 (20 male, 20 female) orthodontic patients aged 18–34 years (mean ± SD = 23.53 ± 4.5) were included in this study and assigned into four subgroups of 10 (5 males, 5 females). Descriptive statistics for DPI, GI, and WSL index in control and experimental groups (placebo, CHX, and fluoride) at each of the four time intervals are illustrated in Table 1. Table 1 Descriptive statistics for dental plaque index, gingival index, and white spot lesion index in control and examined groups (placebo, chlorhexidine, and fluoride) at the beginning, after 3, 6, and 9 months The frequency rates of patients with WSLs at the beginning and after 3, 6, and 9 months of treatment are presented in Table 2. Table 2 Frequency of patients with white spot lesions at the beginning, 3, 6, and 9 months after treatment Figures 1–3 display significant differences between the examined and control groups for the WSL index, DPI, and GI scores in the four time points (baseline, after 3, 6, and 9 months). Figure 1 Mean white spot lesion values in control and examined groups (placebo, chlorhexidine, and fluoride) at time intervals. Figure 2 Mean plaque index values in control and examined groups (placebo, chlorhexidine, and fluoride) at time intervals. Figure 3 Mean gingival index in control and examined groups (placebo, chlorhexidine, and fluoride) at time intervals. The results of Kruskal–Wallis test showed significant changes in all three indices and all groups after the 9-month period (P < 0.05). After using fluoride and CHX for 9 months, GI decreases significantly (P < 0.05) while the changes in placebo and control groups were not statistically significant. Based on the results of Mann–Whitney test, a significant decrease in plaque was observed when comparing CHX with control (P = 0.00) and CHX with placebo (P = 0.03). CHX effectiveness in WSL reduction was also significant compared to control group (P = 0.001) and placebo group (P = 0.009). Furthermore, decrease in WSLs was significantly more in CHX group compared to fluoride group (P = 0.001). Fluoride was significantly more effective in plaque reduction compared to control group although its difference with placebo group was not significant. The only difference between control and placebo groups was observed in plaque index, being significantly lower in placebo group (P = 0.01). DISCUSSION High prevalence of dental caries, especially in high-risk patients, has made them the target of many interventions. The effect of using fluoridated toothpastes,[29] other topically applied fluorides,[30] fluoridated municipal water,[31] and pit and fissure sealants[27] in the prevention of plaque and caries still remains vague. There is controversy regarding the use of CHX for caries prevention. It has been reported that the most persistent reduction of MS can be achieved by CHX varnishes, followed by gels and mouth rinses. This evidence is rather suggestive but incomplete. Variable study designs and lack of data in high-risk patients such as orthodontic patients support the need to continue conducting randomized, well-controlled clinical trials and search for a practical, effective mode of antimicrobial treatment.[3233] Distinct difference has been reported in the caries-preventive effects of concentrated fluoride solutions, gels, or varnishes. Thus, several factors such as costs, convenience, patient acceptance, and safety can affect the choice of method. Fluoride varnishes are proven to be a feasible and safe approach for fluoride application. With fluoride varnishes, the amounts of fluoride exposure can be better controlled, and less chair time is required, compared with conventional solutions and gels. No dose-response effect on concentrated fluoride agents is apparent, and the benefit of frequent application is not clearly established.[789] In a systematic review done by Chen et al., it was noted that there is lack of reliable evidence to support the effectiveness of remineralizing agents for the treatment of orthodontic WSLs.[34] In a systematic review done by Migliorati et al.,[35] the efficacy of professional hygiene and prophylaxis on preventing plaque increase in orthodontic patients with multibracket appliances was assessed and it was found that an optimal oral health maintenance during orthodontic treatment should be a gold standard in today's practice; nevertheless, literature on this topic is lacking as well as high-quality studies like randomized controlled trial (RCT). Future high-quality research is recommended. In a systematic review in 2016, the effects of CHX varnish on caries during orthodontic treatment were assessed and it was found that high-quality clinical trials to examine CHX varnish effectiveness on caries prevention during orthodontic treatment are clearly needed.[36] In a RCT done by Singh et al.,[37] in 2016, the effects of various remineralizing agents on the outcome of postorthodontic WSLs was evaluated. It was found that the use of 5% NaF varnish in addition to twice daily use of 1000 ppm of fluoride toothpaste had no additional beneficial effect in the remineralization of orthodontic WSLs, which was in contrast with the result of our study. The results of our study demonstrated a statistically significant decrease for DPI and GI in both groups of fluoride and CHX in all three times of follow-up. CHX has known antiplaque and antigingivitis effect and it may be employed as an adjunct to other preventive measures such as professional care and patient-oriented instruction on an intermittent basis to reduce the plaque-induced iatrogenic side effects and to enhance the efficacy of oral hygiene measures in connection with orthodontic therapy with fixed appliances.[1538] Several studies in the literature are in agreement with the results of our study, confirming that CHX and fluoride varnishes can decrease dental plaque and WSLs of orthodontic patients when added to routine oral hygiene treatment (brushing + flossing).[394041] CHX and fluoride groups demonstrated significantly better plaque index and GI scores at all treatment intervals after baseline measurements. In Jayaprakash et al.[42] study, it was concluded that a mouth rinse containing CHX and sodium fluoride could elevate dental hygiene and decrease plaque and gingivitis, which is in agreement with the findings of our study. In Esfahanian et al.[43] study, they reported that CHX has a positive effect in plaque control and it was better when compared to Irsha mouth rinses. Early detection of WSLs during orthodontic treatment is also very important as it would allow clinicians to implement preventive measures to control the demineralization process before progression of the lesions. In this study, we decided to use fluoride varnishes for two reasons; first, because we wanted to use it at 3-month intervals and second, based on the literature, it seems that this type of application of fluoride can provide the patients with the best possible outcome.[44] In this study, a statistically significant decrease of WSLs was registered in the participants of CHX and fluoride groups in all three follow-ups but not in the control or placebo groups. No significant differences were observed between the CHX and fluoride groups for demineralization. To date, clinical detection of WSLs has been carried out primarily by means of traditional methods such as visual inspection after air drying and tactile examination by dental probing. We chose to use this approach in our study. However, the subjectivity and lack of reproducibility of these approaches, together with the prerequisite presence of a significantly advanced lesion, have led to the introduction of several optical techniques during recent decades, including optical caries monitor, use of quantitative laser and light-induced fluorescence, digital imaging with fiber-optic transillumination, laser fluorescence, and computer analysis of digital photographs.[45] Enamel surface roughness affects its visual properties, plaque retention, abrasion, and bond strength.[46] Due to simple sampling, the unification of DMFS (Decay-missing-filling-surfaces) was difficult and one of the limitations of our study. CONCLUSION Adding CHX and fluoride to daily oral hygiene program reduces bacterial plaque accumulation and improves GI. Orthodontists are recommended to enhance their patients' oral hygiene by requiring the use of CHX and fluoride in addition to daily brushing. Financial support and sponsorship Nil. Conflicts of interest The authors of this manuscript declare that they have no conflicts of interest, real or perceived, financial or nonfinancial in this article. ==== Refs REFERENCES 1 Shirani F Sakhaiemanesh M Effect of remineralizing agents on the color change of sound enamel and white spot lesions Journal of Isfahan dental school 2013 9 358 68 2 Lundström F Krasse B Streptococcus mutans and lactobacilli frequency in orthodontic patients; the effect of chlorhexidine treatments Eur J Orthod 1987 9 109 16 3472888 3 Ahn SJ Lim BS Lee SJ Prevalence of cariogenic streptococci on incisor brackets detected by polymerase chain reaction Am J Orthod Dentofacial Orthop 2007 131 736 41 17561051 4 Øgaard B White Spot Lesions during Orthodontic Treatment: Mechanisms and Fluoride Preventive Aspects Seminars in Orthodontics 2008 University of oslo, Norway Elsevier 5 Geiger AM Gorelick L Gwinnett AJ Griswold PG The effect of a fluoride program on white spot formation during orthodontic treatment Am J Orthod Dentofacial Orthop 1988 93 29 37 3276146 6 Ahn SJ Lim BS Yang HC Chang YI Quantitative analysis of the adhesion of cariogenic streptococci to orthodontic metal brackets Angle Orthod 2005 75 666 71 16097239 7 Hickman J Millett DT Sander L Brown E Love J Powered vs. manual tooth brushing in fixed appliance patients: A short term randomized clinical trial Angle Orthod 2002 72 135 40 11999936 8 Vizitiu T Ecaterina I In vitro evaluation of effectiveness in reducing bacterial plaque of antimicrobial substances in patients treated with orthodontic appliances Therapeutics, Pharmacol Clin Toxicol 2011 15 245 48 9 Tufekci E Casagrande ZA Lindauer SJ Fowler CE Williams KT Effectiveness of an essential oil mouthrinse in improving oral health in orthodontic patients Angle Orthod 2008 78 294 8 18251611 10 Sekino S Ramberg P Uzel NG Socransky S Lindhe J Effect of various chlorhexidine regimens on salivary bacteria and de novo plaque formation J Clin Periodontol 2003 30 919 25 14710772 11 Axelsson S Söder B Nordenram G Petersson LG Dahlgren H Norlund A Effect of combined caries-preventive methods: A systematic review of controlled clinical trials Acta Odontol Scand 2004 62 163 9 15370637 12 Botelho MA dos Santos RA Martins JG Carvalho CO Paz MC Azenha C Efficacy of a mouthrinse based on leaves of the neem tree (Azadirachta indica ) in the treatment of patients with chronic gingivitis: A double-blind, randomized, controlled trial J Med Plants Res 2008 2 341 6 13 Gomez SS Basili CP Emilson CG A 2-year clinical evaluation of sealed noncavitated approximal posterior carious lesions in adolescents Clin Oral Investig 2005 9 239 43 14 Chapman JA Roberts WE Eckert GJ Kula KS González-Cabezas C Risk factors for incidence and severity of white spot lesions during treatment with fixed orthodontic appliances Am J Orthod Dentofacial Orthop 2010 138 188 94 20691360 15 Featherstone JD Delivery challenges for fluoride, chlorhexidine and xylitol BMC Oral Health 2006 6 Suppl 1 S8 16934125 16 Schaeken MJ De Haan P Effects of sustained-release chlorhexidine acetate on the human dental plaque flora J Dent Res 1989 68 119 23 2918132 17 Maxfield BJ Hamdan AM Tüfekçi E Shroff B Best AM Lindauer SJ Development of white spot lesions during orthodontic treatment: Perceptions of patients, parents, orthodontists, and general dentists Am J Orthod Dentofacial Orthop 2012 141 337 44 22381494 18 Sandham HJ Brown J Chan KH Phillips HI Burgess RC Stokl AJ Clinical trial in adults of an antimicrobial varnish for reducing mutans streptococci J Dent Res 1991 70 1401 8 1960249 19 Brunner E Langer F Nonparametric analysis of ordered categorical data in designs with longitudinal observations and small sample sizes Biom J 2000 42 663 75 20 Morrow D Wood DP Speechley M Clinical effect of subgingival chlorhexidine irrigation on gingivitis in adolescent orthodontic patients Am J Orthod Dentofacial Orthop 1992 101 408 13 1590289 21 Stecksén-Blicks C Renfors G Oscarson ND Bergstrand F Twetman S Caries-preventive effectiveness of a fluoride varnish: A randomized controlled trial in adolescents with fixed orthodontic appliances Caries Res 2007 41 455 9 17827963 22 Derks A Kuijpers-Jagtman AM Frencken JE Van't Hof MA Katsaros C Caries preventive measures used in orthodontic practices: An evidence-based decision? 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