Time-series expression profiles of mRNAs and lncRNAs during mammalian palatogenesis.

OBJECTIVES: Mammalian palatogenesis is a highly regulated morphogenetic process to form the intact roof of the oral cavity. Long noncoding RNAs (lncRNAs) and mRNAs participate in numerous biological and pathological processes, but their roles in palatal development and causing orofacial clefts (OFC) remain to be clarified. METHODS: Palatal tissues were separated from ICR mouse embryos at four stages (E10.5, E13.5, E15, and E17). Then, RNA sequencing (RNA-seq) was used. Various analyses were performed to explore the results. Finally, hub genes were validated via qPCR and in situ hybridization. RESULTS: Starting from E10.5, the expression of cell adhesion genes escalated in the following stages. Cilium assembly and ossification genes were both upregulated at E15 compared with E13.5. Besides, the expression of cilium assembly genes was also increased at E17 compared with E15. Expression patterns of three lncRNAs (H19, Malat1, and Miat) and four mRNAs (Cdh1, Irf6, Grhl3, Efnb1) detected in RNA-seq were validated. CONCLUSIONS: This study provides a time-series expression landscape of mRNAs and lncRNAs during palatogenesis, which highlights the importance of processes such as cell adhesion and ossification. Our results will facilitate a deeper understanding of the complexity of gene expression and regulation during palatogenesis.

Two rare variants reveal the significance of Grainyhead-like 3 Arginine 391 underlying non-syndromic cleft palate only.

OBJECTIVES: Non-syndromic cleft palate only (NSCPO) is one of the most common craniofacial birth defects with largely undetermined genetic etiology. It has been established that Grainyhead-like 3 (GRHL3) plays an essential role in the pathogenesis of NSCPO. This study aimed to identify and verify the first-reported GRHL3 variant underlying NSCPO among the Chinese cohort. METHODS: We performed whole-exome sequencing (WES) on a Chinese NSCPO patient and identified a rare variant of GRHL3 (p.Arg391His). A validated deleterious variant p.Arg391Cys was introduced as a positive control. Zebrafish embryos injection, reporter assays, live-cell imaging, and RNA sequencing were conducted to test the pathogenicity of the variants. RESULTS: Zebrafish embryos microinjection demonstrated that overexpression of the variants could disrupt the normal development of zebrafish embryos. Reporter assays showed that Arg391His disturbed transcriptional activity of GRHL3 and exerted a dominant-negative effect. Interestingly, Arg391His and Arg391Cys displayed distinct nuclear localization patterns from that of wild-type GRHL3 in live-cell imaging. Bulk RNA sequencing suggested that the two variants changed the pattern of gene expression. CONCLUSIONS: In aggregate, this study identified and characterized a rare GRHL3 variant in NSCPO, revealing the critical role of Arginine 391 in GRHL3. Our findings will help facilitate understanding and genetic counseling of NSCPO.

Accuracy of integration of dental cast and cephalograms compared with cone-beam computed tomography: a comparative study.

This study proposes a method that integrates maxillary dental cast and cephalograms and evaluates its accuracy compared with cone-beam computed tomography (CBCT) scans. The study sample comprised 20 adult patients with records of dental casts, cephalograms, and craniofacial CBCT scans. The maxillary dental cast was integrated with lateral and frontal cephalograms based on best-fit registration of palatal and dental outline curves from dental cast with cephalogram tracings. Linear measurement was conducted to assess the intra- and inter-examiner reproducibility of the proposed integration method using intraclass correlation coefficients; linear and angular measurements were conducted to assess its accuracy with CBCT scans as a standard reference. Paired t test, one sample t test, and mean ± standard deviation of the absolute value of difference were used to compare the integrated images and CBCT. The integration method showed good intra- and inter-examiner reproducibility (intraclass correlation coefficients > 0.98). The differences in linear and angular measurements between the integrated images and CBCT were not statistically significant but with a large deviation. When absolute value of difference was computed, the linear distance error was 0.51 ± 0.34 mm, the tooth point coordinate errors in X, Y and Z axes were 0.22 ± 0.22, 0.38 ± 0.32 and 0.21 ± 0.21 mm, respectively; the angular error in pitch, roll and yaw of the dental cast was 0.82 ± 0.51, 0.92 ± 0.59 and 0.80 ± 0.41 degree, respectively. The proposed method for integration of dental cast and cephalograms showed good reproducibility and acceptable accuracy compared with CBCT. It could be helpful for researchers to study three-dimensional tooth growth changes using the existing craniofacial growth data especially cephalograms.

Dynamic Change in Oral Microbiota of Children With Cleft Lip and Palate After Alveolar Bone Grafting.

To investigate the longitudinal influence of alveolar bone grafting on the oral microbiota of children with cleft lip and palate (CLP).Twenty-eight children with nonsyndromic CLP were recruited and underwent secondary alveolar bone grafting at the first time. Unstimulated saliva and plaque samples were collected from the subjects preoperatively and at 2 days, 1 month, and 3 months postoperatively. The v3-v4 hypervariable regions of the 16S rRNA gene from bacterial DNA were sequenced using the Illumina MiSeq sequencing platform.The alpha diversity of the saliva and plaque microbiota was significantly decreased at 2 days postoperatively and then increased at 1 and 3 months postoperatively. The saliva and plaque microbiota compositions at 2 days postoperatively differed from those at the other time points, and the microbiota compositions at 1 and 3 months postoperatively showed a gradual shift toward the preoperative composition. The saliva, but not plaque, microbiota composition 3 months postoperatively was similar to that preoperatively.The effect of secondary alveolar bone grafting on the plaque microbiota in children with CLP lasted longer than the saliva microbiota. Alveolar bone grafting altered the saliva microbiota in children with CLP within 3 months postoperatively.

Apolipoprotein E is an effective biomarker for orthodontic tooth movement in patients treated with transmission straight wire appliances.

INTRODUCTION: Orthodontic tooth movement (OTM) is the core component of orthodontic treatment and is increasingly popular for treating malocclusions. In this study, we aimed to investigate the role of apolipoprotein E (ApoE) in OTM. METHODS: Thirty patients treated with transmission straight wire technology were selected and longitudinally tracked at 2 different stages of orthodontic treatment (initial 2 months and 12 months of orthodontic treatment). Total saliva was collected and analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. Western blotting was used to detect the difference in ApoE expression in the saliva samples of the 2 groups. The expression of ApoE was further verified by immunohistochemical staining in a mouse model of tooth movement. RESULTS: The results of matrix-assisted laser desorption/ionization time-of-flight mass spectrometry showed significant differences in the components of the salivary peptides in the 2 groups and peptides with a molecular weight of 2010.7 Da were predicted to be ApoE by database analysis. Western blotting further verified a significant difference in the expression of salivary ApoE in the 2 groups. In addition, an OTM model was successfully constructed in mice. The immunohistochemical staining results showed that ApoE expression significantly increased after force loading in the OTM model. CONCLUSIONS: This study indicated that ApoE participated in and played a role during OTM in patients treated with transmission straight wire technology. This relationship might be related to alveolar bone reconstruction and root resorption. The results provide new ideas for research on the mechanism of tooth movement using precision medicine based on saliva detection.

Analysis of facial features and prediction of lip position in skeletal class III malocclusion adult patients undergoing surgical-orthodontic treatment.

OBJECTIVES: This study presents a retrospective study aimed to analyze the facial features at each stage of surgical-orthodontic treatment for skeletal class III malocclusion, and predict the changes in the lips after treatment. MATERIALS AND METHODS: There were 49 skeletal class III malocclusion patients treated with bimaxillary surgery and orthodontic treatment enrolled in this study. Lateral cephalograms were obtained before treatment (T0), 1 month before surgery (T1), 1 month after surgery (T2), and after debonding (T3) for cephalometric measurements. After the measurement of the required variables, paired t-test, Pearson's correlation analysis, and multiple linear regression were performed using SPSS 19.0. RESULTS: The main factors associated with changes in the upper lip included ΔUIE-V, ΔA-V, ΔU1A-V, and ΔL1A-V, and those associated with changes in the lower lip included ΔLIE-V, ΔL1A-V, ΔB-V, ΔPog-V, and Δfacial angle. The predicted regression equation for the horizontal change in the upper lip was represented as ΔUL-vertical reference line (VRL) = 9.430 + 0.779 (ΔUIE-VRL) - 0.542(VULT) (P < 0.05) with a mean error of 1.04 mm; the corresponding equation for the lower lip was ΔLL-VRL = -1.670 + 0.530 (ΔB-VRL) + 0.360 (Ls-E) + 0.393 (ΔLIE-VRL) (P < 0.05), with a mean error of 1.51 mm. CONCLUSIONS: This study explored the relationship between orthognathic surgery and changes in the lips and obtained the predictive equations of lip position after treatment by using multiple linear regression, which likely offers a reference for prediction of soft tissue changes before surgical-orthodontic treatment in patients with skeletal class III malocclusion. CLINICAL RELEVANCE: The findings can help dentists to rapidly predict the lip changes after surgical-orthodontic treatment in patients with skeletal class III malocclusion. The study has been registered with the Chinese Clinical Trial Registration (No: ChiCTR1800017694).

Nell-1-ΔE, a novel transcript of Nell-1, inhibits cell migration by interacting with enolase-1.

NELL-1 is a secreted protein that was originally found to be upregulated in pathologically fusing and fused sutures in non-syndromic unilateral coronal synostosis patients. Apart from the ability of NELL-1 to promote osteogenesis in long and craniofacial bones, NELL-1 reportedly inhibits the formation of several benign and malignant tumors. We previously identified a novel transcript of Nell-1 that lacked a calcium-binding epidermal growth factor (EGF)-like domain compared with full-length Nell-1; this new transcript was named Nell-1-ΔE. Three obvious structural differences between these two isoforms were revealed by homology modeling. Furthermore, the recombinant Nell-1-ΔE protein, but not the full-length Nell-1 protein, inhibited cell migration in vitro. However, full-length Nell-1 and Nell-1-ΔE proteins were present in similar subcellular locations and displayed similar expression patterns in both the intracellular and extracellular spaces. The results from the co-immunoprecipitation and liquid chromatography/tandem mass spectrometry analyses using two cell lines demonstrated that Nell-1-ΔE but not full-length Nell-1 interacted with enolase-1 in the extracellular spaces of both cell lines. The results of wound healing assays using ENO-1-overexpressing cells treated with full-length Nell-1/Nell-1-ΔE suggested that Nell-1-ΔE inhibited cell migration by interacting with ENO-1. Our study indicated that the novel transcript Nell-1-ΔE, but not full-length Nell-1, might be a candidate tumor suppressor factor for basic research and clinical practice.

A novel IRF6 mutation causing non-syndromic cleft lip with or without cleft palate in a pedigree.

Non-syndromic cleft lip with or without cleft palate (NSCLP) is the most common congenital craniofacial malformation, and its harmful influence on affected individuals is apparent. Despite many studies, the causative genes and their mechanisms are not completely clear. We recruited a Han Chinese NSCLP family and explored the causative variant in this pedigree. We performed whole-exome sequencing on two patients. Bioinformatics screening and analysis were used to identify the mutation. We also performed species conservation analysis, mutation function predictions, and homology protein modelling to evaluate the influence of the mutation. We identified a rare mutation in interferon regulatory factor 6 (IRF6) (c.26G>A; p.Arg9Gln) as a candidate of causative mutation. This mutation was predicted to be deleterious. The codon is conserved in many species. The residue change caused by this mutation would affect the structure of IRF6 to a degree. Our study suggested that the rare IRF6 variant is probably the pathogenic mutation in this family. Our result adds evidence that IRF6 variants play a role in the aetiology of orofacial clefts.

A novel PTCH1 mutation underlies nonsyndromic cleft lip and/or palate in a Han Chinese family.

OBJECTIVES: Cleft lip and/or palate (CL/P) is the most common craniofacial congenital disease, and it has a complex aetiology. This study aimed to identify the causative gene mutation of a Han Chinese family with CL/P. SUBJECTS AND METHODS: Whole exome sequencing was conducted on the proband and her mother, who exhibited the same phenotype. A Mendelian dominant inheritance model, allele frequency, mutation regions, functional prediction and literature review were used to screen and filter the variants. The candidate was validated by Sanger sequencing. Conservation analysis and homology modelling were conducted. RESULTS: A heterozygous missense mutation c.1175C>T in the PTCH1 gene predicting p.Ala392Val was identified. This variant has not been reported and was predicted to be deleterious. Sanger sequencing verified the variant and the dominant inheritance model in the family. The missense alteration affects an amino acid that is evolutionarily conserved in the first extracellular loop of the PTCH1 protein. The local structure of the mutant protein was significantly altered according to homology modelling. CONCLUSIONS: Our findings suggest that c.1175C>T in PTCH1 (NM_000264) may be the causative mutation of this pedigree. Our results add to the evidence that PTCH1 variants play a role in the pathogenesis of orofacial clefts.

Exploring the Genomic Diversity and Cariogenic Differences of Streptococcus mutans Strains Through Pan-Genome and Comparative Genome Analysis.

Pan-genome refers to the sum of genes that can be found in a given bacterial species, including the core-genome and the dispensable genome. In this study, the genomes from 183 Streptococcus mutans (S. mutans) isolates were analyzed from the pan-genome perspective. This analysis revealed that S. mutans has an "open" pan-genome, implying that there are plenty of new genes to be found as more genomes are sequenced. Additionally, S. mutans has a limited core-genome, which is composed of genes related to vital activities within the bacterium, such as metabolism and hereditary information storage or processing, occupying 35.6 and 26.6% of the core genes, respectively. We estimate the theoretical core-genome size to be about 1083 genes, which are fewer than other Streptococcus species. In addition, core genes suffer larger selection pressures in comparison to those that are less widely distributed. Not surprisingly, the distribution of putative virulence genes in S. mutans strains does not correlate with caries status, indicating that other factors are also responsible for cariogenesis. These results contribute to a more understanding of the evolutionary characteristics and dynamic changes within the genome components of the species. This also helps to form a new theoretical foundation for preventing dental caries. Furthermore, this study sets an example for analyzing large genomic datasets of pathogens from the pan-genome perspective.

Assessing the Interdental Septal Thickness in Alveolar Bone Grafting Using Cone Beam Computed Tomography.

OBJECTIVE: To assess the interdental septal thickness of grafted bone bridges using cone beam computed tomography (CBCT). PATIENTS: Of 71 patients with cleft lip and/or palate having undergone alveolar bone grafting for the first time at least 6 months previously, 52 patients with 57 grafted sites rated type I or II based on the Bergland scale using occlusal radiographs were selected. INTERVENTIONS: CBCT was performed for each bone-grafted alveolar cleft within 1 week after the occlusal radiographs were taken. MAIN OUTCOME MEASURES: The thickness of the grafted bone bridge was evaluated using CBCT according to the relationship between crest thickness and the root width of cleft-adjacent teeth, and the results were classified into four categories, with scores of 1 to 4 indicating that the thickness of the bony bridge was ≥100%, ≥75%, ≥50%, and <50% of the root width of the cleft-adjacent teeth, respectively. RESULTS: Of the 34 grafted sites rated type I on the Bergland scale, 15 (44.12%), 10 (29.41%), 4 (11.76%), and 5 (14.71%) clefts were scored 1 to 4 on interdental septal thickness using CBCT, respectively. Of the 23 cases of type II, 3 (13.04%), 9 (39.13%), 1 (3.45%), and 10 (43.48%) clefts were scored 1 to 4, respectively. CONCLUSIONS: The interdental septal thickness of grafted bone bridges with clinically successful heights based on the Bergland scale (type I or II) using occlusal radiographs varied significantly in the evaluation using CBCT.

Exosome analysis: a promising biomarker system with special attention to saliva.

Today, exosome-related studies have become a focus in science and technology. Recently, three scientists won the Nobel Prize for determining the mechanisms of exosomal transport, making exosomes a promising biomarker system for disease diagnosis and treatment. This review provides a general introduction of exosomes and explores the recent progress on the function, application, isolation, and identification of exosomes as biomarkers in blood and other body fluids, especially in saliva. Detailed information of exosomal proteins and RNAs is discussed in the paper because of their ability to determine the function of exosomes. Due to their noninvasive assessment for quick and convenient diagnosis of diseases, salivary exosomes may well be promising biomarkers.

Rare loss-of-function variants in FLNB cause non-syndromic orofacial clefts.

Orofacial clefts (OFCs) are the most common congenital craniofacial disorders, of which the etiology is closely related to rare coding variants. Filamin B (FLNB) is an actin-binding protein implicated in bone formation. FLNB mutations have been identified in several types of syndromic OFCs and previous studies suggest a role of FLNB in the onset of non-syndromic OFCs (NSOFCs). Here, we report two rare heterozygous variants (p.P441T and p.G565R) in FLNB in two unrelated hereditary families with NSOFCs. Bioinformatics analysis suggests that both variants may disrupt the function of FLNB. In mammalian cells, p.P441T and p.G565R variants are less potent to induce cell stretches than wild type FLNB, suggesting that they are loss-of-function mutations. Immunohistochemistry analysis demonstrates that FLNB is abundantly expressed during palatal development. Importantly, Flnb-/- embryos display cleft palates and previously defined skeletal defects. Taken together, our findings reveal that FLNB is required for development of palates in mice and FLNB is a bona fide causal gene for NSOFCs in humans.

Tooth lengths of the permanent upper incisors in patients with cleft lip and palate determined with cone beam computed tomography.

Objective : To measure the tooth lengths of fully developed permanent upper incisors and to qualitatively evaluate the root shapes of the incisors in patients with cleft lip and palate (CLP). Design : Cross-sectional, noninterventional, case-control imaging study. Setting : Hospital and Stomatology Unit of Peking University, Beijing, China (institutional tertiary care). Participants : Sixty consecutive nonsyndromic CLP patients (including 40 unilateral [UCLP] and 20 bilateral [BCLP]), and 53 age- and sex-matched controls were selected for this study. Main Outcome Measure : Crown heights and root lengths of permanent upper incisors were measured from cone beam computed tomography scans, and the root shapes of upper incisors were evaluated. Results : Compared with controls, the crown heights of upper incisors in CLP patients were reduced by 9.7% to 22.5% (p < .05), and the root lengths were reduced by 15.8% to 31.7% (p < .05). BCLP patients had greater reductions than the UCLP cases (p < .05). There were no significant differences between incisors and their antimeres in controls and BCLP patients. However, measurements on the cleft side in UCLP patients were lower than those of the noncleft side (p < .05). The prevalence of atypical root shape was higher in CLP than in the control group (p  =  .002); of these, 83.3% (30/36) occurred in central incisors near the cleft. Conclusion : The permanent upper incisors in nonsyndromic CLP patients are underdeveloped. Incisor developmental deficiency was greater in teeth adjacent to the cleft.

[A novel TaqMan® MGB probe for specifically detecting Streptococcus mutans].

OBJECTIVE: To design a new TaqMan® MGB probe for improving the specificity of Streptococcus mutans's detection. METHODS: We extracted six DNA samples from different streptococcal strains for PCR reaction. Conventional nested PCR and TaqMan® MGB real-time PCR were applied independently. The first round of nested PCR was carried out with the bacterial universal primers, while a second PCR was conducted by using primers specific for the 16S rRNA gene of Streptococcus mutans. The TaqMan® MGB probe for Streptococcus mutans was designed from sequence analyses, and the primers were the same as nested PCR. Streptococcus mutans DNA with 2.5 mg/L was sequentially diluted at 5-fold intervals to 0.16 µg/L. Standard DNA samples were used to generate standard curves by TaqMan® MGB real-time PCR. RESULTS: In the nested PCR, the primers specific for Streptococcus mutans also detected Streptococcus gordonii with visible band of 282 bp, giving false-positive results. In the TaqMan® MGB real-time PCR reaction, only Streptococcus mutans was detected. The detection limitation of TaqMan® MGB real-time PCR for Streptococcus mutans 16S rRNA gene was 20 µg/L. CONCLUSION: We designed a new TaqMan® MGB probe, and successfully set up a PCR based method for detecting oral Streptococcus mutans. TaqMan® MGB real-time PCR is a both specific and sensitive bacterial detection method.

Inclination of mandibular incisors and symphysis in severe skeletal class III malocclusion.

OBJECTIVE: The aim of this study was to systematically explore the inclination of the lower central incisor and symphysis in alveolar bone in severe skeletal class III patients. MATERIALS AND METHODS: A total of 198 severe skeletal class III patients (ANB ≤ -4°) who underwent combined orthodontic and orthognathic treatment were divided into three groups based on the mandibular plane angle (MP-SN). Pretreatment lateral cephalograms were analysed and compared among the three groups. We also assessed cone-beam computed tomography (CBCT) images of 11 samples to investigate the reliability of the cephalometric analysis. RESULTS: ANOVA showed no statistically significant differences in the angle between the long axis of the mandibular symphysis and the long axis of the lower central incisor (MIA) among the low-angle, normal-angle and high-angle groups (P > 0.05), while significant differences were found in the angle between the axis of the lower incisor and the mandibular plane (IMPA) among the three groups (P < 0.001). The mean IMPA decreased with increasing MP-SN in the 198 patients. The mean MIA in the low-angle and normal-angle groups was 3.70° and 3.52°, respectively, while the value (2.33°) was smaller in the high-angle group. Paired-samples t test showed no statistically significant differences between the cephalometric and CBCT measurements of the MP-SN, the angle between the mandibular plane and the Frankfort plane (FH-MP) and the MIA (P > 0.05). CONCLUSIONS: In severe skeletal class III patients, the long axis of the lower central incisor was highly consistent with the long axis of the mandibular symphysis, which was more obvious in the high-angle subjects. The MIA reflects the physiological inclination of the lower central incisor better than the IMPA.

Magnetic bead-based salivary peptidome profiling analysis during orthodontic treatment durations.

Orthodontic treatment induces various biological responses, including tooth movement and remodeling of alveolar bone. Although some studies have investigated the contribution of orthodontic procedures to changes in saliva conditions, little is known about the effects of different treatment durations on the saliva proteome. To identify the discriminating protein profiles in unstimulated whole saliva of orthodontic patients with different treatment durations, we used matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) combined with magnetic bead, and peptide mass fingerprints were created by scanning MS signals. Saliva samples from 40 patients (10 in each of four groups: the group without an appliance and groups under treatment for 2, 7, and 12 months) were analyzed. The results showed eight mass peaks with significant differences. Furthermore, mass peak intensities at proteins 1817.7, 2010.7, 2744 and 2710.2 Da represented a steady time-dependent increasing trend, whereas protein 4134 Da exhibited a decreasing tendency. Differential expression of the peptidome profile also occurred in the multiple comparisons, and we established a fitting model. Thus, the potential discriminating biomarkers investigated in this study reflected the complicated changes in periodontal tissues during orthodontic treatment and indicated dynamic interactions between orthodontic treatment and the saliva proteome. The results provide novel insights into alterations in salivary proteins due to different orthodontic treatment durations and may lead to the development of a therapeutic monitoring strategy for orthodontics.

Magnetic bead-based salivary peptidome profiling for periodontal-orthodontic treatment.

BACKGROUND: Patients with periodontitis seek periodontal-orthodontic treatment to address certain functional and aesthetic problems. However, little is known of the effect of periodontitis on orthodontic treatment. Thus, we compared the differences in peptide mass fingerprints of orthodontic patients with and without periodontitis by MALDI-TOF MS using a magnetic bead-based peptidome analysis of saliva samples. In this way, we aimed to identify and explore a panel of differentially-expressed specific peptides. RESULTS: Saliva samples from 24 patients (eight orthodontic patients without periodontitis, eight with periodontitis and another eight with periodontitis but no orthodontic treatment) were analyzed, and peptide mass fingerprints were created by scanning MS signals using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) combined with magnetic beads. Nine mass peaks showed significant differences. Orthodontic patients in the group without periodontal disease showed higher mass peaks for seven peptides of the nine, whereas the mass peaks for the other two peptides were higher in the periodontal-orthodontic patients. Besides, these differentially-expressed peptides were sequenced. CONCLUSIONS: The elucidated candidate biomarkers indicated interactions between periodontal condition and orthodontic treatment and their contributions to the changes of saliva protein profiles. Our results provide novel insight into the altered salivary protein profile during periodontal-orthodontic treatment, and may lead to the development of a therapeutic monitoring strategy for periodontics and orthodontics.

Local osteoprotegerin gene transfer inhibits relapse of orthodontic tooth movement.

INTRODUCTION: In orthodontic treatment, teeth can relapse after tooth movement without retention. The aim of this study was to evaluate the inhibition effects of local osteoprotegerin (OPG) gene transfer on orthodontic relapse. METHODS: Eighteen male Wistar rats were divided into 3 groups. The maxillary right first molars of all animals were subjected to orthodontic force and moved mesially. Three weeks later, the force was removed, and the teeth relapsed. During the 2-week relapse period, the 3 groups of rats received local OPG gene transfer (experimental group), mock vector transfer (mock group), and no injections (control group). Tooth movement and relapse were measured by using palatal superimpositions of 3-dimensional digital models. Histomorphometric analysis was used to quantify osteoclasts, and microcomputed tomography analysis was done to quantify the alveolar bone and the tibia. RESULTS: Relapse was significantly inhibited and the number of osteoclasts was reduced in the experimental group. On the other hand, bone mineral density and bone volume fraction of alveolar bone were significantly increased. Bone mineral density and bone volume fraction of the tibia showed no significant difference between the groups. CONCLUSIONS: Local OPG gene transfer to periodontal tissues could inhibit relapse after orthodontic tooth movement, through the inhibition of osteoclastogenesis.