NEED FOR STUDY An increasing number of adult patients are seeking orthodontic treatment to enhance both the appearance of their smile and their masticatory function. One of the major challenges in orthodontics, however, is the treatment duration. As patient expectations evolve, factors like time efficiency and aesthetic concerns have become more significant. In recent years, adjunctive therapies including micro-osteoperforations(MOPs) and vibrational frequency devices such as Electric toothbrush have been explored to accelerate orthodontic tooth movement by modulating the biological processes involved.1,2 Previous research has indicated that the levels of various enzymes and proteins fluctuate following the application of orthodontic force.3 This study seeks to evaluate the enzymatic activity in gingival crevicular fluid (GCF) during orthodontic treatment, following MOPs combined with the use of an electric toothbrush. The enzymes of interest are alkaline phosphatase (ALP), acid phosphatase (ACP), aspartate aminotransferase (AST), and dentin sialoprotein (DSP) which are involved in various aspects of bone remodelling4 inflammation5, and root resorption6, making them ideal biomarkers for evaluating the biological changes occurring during orthodontic tooth movement. During the treatment procedure, If the levels of bone remodelling enzymes, such as ALP and ACP are elevated but remain within normal limits after the procedure, it would be considered favourable to proceed with that treatment. On the other hand, enzymes like AST, which indicate tissue damage, should stay within normal limits. An elevation in these levels could result in increased inflammation, potentially causing delayed healing and other complications. This study aims to evaluate the response of enzymes to three different treatments: MOPs, electric toothbrush use, and a combined therapy of both. It will also examine how these treatments correlate with the rate and effectiveness of tooth movement. BACKGROUND: Orthodontic treatment duration remains a major concern for adult patients. Adjunctive methods like micro-osteoperforations and electric toothbrush use may accelerate tooth movement by influencing biological activity. PURPOSE OF TRIAL: To evaluate the effect of MOPs, electric toothbrush use, and their combination on enzymatic activity in gingival crevicular fluid. The study aims to determine their potential in enhancing the rate of orthodontic tooth movement while maintaining periodontal health
CLINICAL SIGNIFICANCE The clinical significance of this study lies in assessing how the use of MOPs, electric toothbrush and a combination of both affect enzymatic markers in gingival crevicular fluid, potentially accelerating orthodontic tooth movement while preserving periodontal health. The findings could lead to more efficient treatment strategies that enhance treatment outcomes for patients. AIM OF THE STUDY The aim of this study is to enzymatically evaluate the changes in gingival crevicular fluid during orthodontic tooth movement in patients undergoing MOPs combined with the use of an electric toothbrush. OBJECTIVES OF THE STUDY To evaluate the enzymatic changes in gingival crevicular fluid during orthodontic tooth movement under following conditions: Following MOPs only. Following MOPs combined with the use of an electric toothbrush. Following use of electric toothbrush only. Following conventional orthodontic treatment. To evaluate the enzymatic changes in gingival crevicular fluid during orthodontic tooth movement for the following enzymes: Alkaline Phosphatase for bone formation. Acid Phosphatase for bone resorption. Aspartate Amino Transferase for tissue inflammation.
Dentin Sialoprotein (a protein) for root resorption. SOURCE OF DATA
This study will be a prospective, comparative design involving 32 patients in total, all undergoing conventional fixed orthodontic treatment at the Department of Orthodontics and Dentofacial Orthopaedics, A.B. Shetty Memorial Institute of Dental Sciences, Mangalore. The biochemical analysis will be performed at the Research Laboratory, KSHEMA, Nitte Deemed to be University.
METHODOLOGY
After receiving approval from the institutional ethics committee, informed consent will be obtained from the patients prior to the commencement of the study. Patients who require the extraction of maxillary first premolars on both sides of the arch will be included in the study. This is a split-mouth study where participants will be randomly assigned to two groups: a) Group A: Conventional orthodontic treatment combined with bilateral MOPs. Participants will use an electric toothbrush on one side, which is randomly assigned. b) Group B: Conventional orthodontic treatment without MOPs. Participants will use an electric toothbrush on one side, which is randomly assigned. The split-mouth technique is selected to minimize biological variability. The method of simple randomization will be followed throughout the study. Patients will be instructed to have their premolars extracted prior to the initial stage of levelling and aligning. Banding/bonding for both the upper and lower arches will be performed using standard techniques. After the initial alignment, maxillary canine retraction will be performed using NiTi closed coil springs with a 0.017×0.025 stainless steel maxillary arch wire. A force of 150gm per side will be applied.12 The necessary anchorage will be established with mini-implants. Participants in groups A will undergo the MOPs procedure bilaterally. MOPs (micro-osteoperforations) will be performed using commercially available orthodontic mini-implant screws with a diameter of 1.2 mm and a length of 8mm. The screws will be inserted to a depth of 6mm using a hand driver. Three MOPs will be created at a depth of 6 mm, spaced 1–3 mm apart, distal to the canine. These will be placed on both the buccal and palatal sides, as close to the roots as possible, within the extraction space, following the administration of local anesthesia.
No flap will be raised during the procedure. Participants in both Group A and Group B will be provided with a standardized 150 Hz electric toothbrush ensuring consistent specifications and obtained from a single manufacturer. They will be instructed to place it against the mesio-labial surface of the experimental canine to deliver mechanical vibrations for at least 5 minutes, three times a day (at an interval of 6 to 8 hours between each application), throughout the study period.13 Participants will be asked to continue their usual orthodontic cleaning routine for the rest of their teeth. A daily usage log will be provided, and participants will be required to submit it during their monthly follow-up visits. SAMPLE COLLECTION
Gingival crevicular fluid (GCF) will be collected from the maxillary canines. Prior to collection, the cheeks will be retracted using a properly sized cheek retractor. The targeted gingival area will be gently dried using an air syringe, and each tooth will be isolated with a cotton roll to prevent contamination. A calibrated microcapillary tube will be used to collect GCF by positioning the volumetric micropipette extra-crevicularly and standardized volume of GCF will be collected at various time points.14
The initial sample will be obtained from both sides of the canine region before retraction (T0). Subsequent samples will be collected on day 1 (T1), day 7 (T2), day 14 (T3), day 30 (T4), day 60 (T5) after the start of canine retraction.
SAMPLE PREPARATION
100 µL of phosphate-buffered saline (pH 7) containing 0.05% bovine serum albumin (Sorensen’s medium) will be transferred into a vial using a 100 µL micropipette. The volume of buffer added will be 100 times the amount of GCF collected. The vial containing the GCF will be sealed, labelled, and immediately sent to the laboratory for centrifugation to remove any bacterial or cellular debris. The centrifugation will be done for 1 minute. After centrifugation, the supernatant will be carefully separated and stored in a deep freezer at -80°C for subsequent analysis. The collected samples will be analysed for the following enzymes and proteins: I. Alkaline Phosphatase (Spectro-photometry method) II. Acid Phosphatase (Spectro-photometry method) III. Aspartate Aminotransferase (Spectro-photometry method) IV. Dentin Sialoprotein (ELISA method) The data will be analysed and compared across the groups to assess differences in outcomes. |