How does the early life environment influence the oral microbiome and determine oral health outcomes in childhood?

The first 1000 days of life, from conception to 2 years, are a critical window for the influence of environmental exposures on the assembly of the oral microbiome, which is the precursor to dental caries (decay), one of the most prevalent microbially induced disorders worldwide. While it is known that the human microbiome is susceptible to environmental exposures, there is limited understanding of the impact of prenatal and early childhood exposures on the oral microbiome trajectory and oral health. A barrier has been the lack of technology to directly measure the foetal "exposome", which includes nutritional and toxic exposures crossing the placenta. Another barrier has been the lack of statistical methods to account for the high dimensional data generated by-omic assays. Through identifying which early life exposures influence the oral microbiome and modify oral health, these findings can be translated into interventions to reduce dental decay prevalence.

What is the Risk of Developing Osteonecrosis Following Dental Extractions for Patients on Denosumab for Osteoporosis?

PURPOSE: Osteonecrosis of the jaws (ONJ) occurs in patients on antiresorptive drugs for osteoporosis with the risk with oral bisphosphonates being known to be of the order of 0.1 to 0.3% while the risk for patients on denosumab for osteoporosis is not known. The aim of this study was to determine the risk of developing ONJ in a consecutive series of patients on denosumab for osteoporosis having dental extractions. MATERIAL AND METHODS: A prospective cohort study of patients on denosumab for osteoporosis having dental extractions in the period January 1, 2017 and June 30, 2021 were compared to a control group not on antiresorptives. Detailed demographic records including length of time on antiresorptives and CTX values were obtained. Comparison to further define risk factors was made between those patients developing ONJ to those who didn't. RESULTS: The treatment group included 427 patients who were on denosumab for osteoporosis; they collectively underwent 561 episodes of dental treatment involving extractions for a total of 1081 extractions, with 10 developing ONJ (risk 2.3%). The control group consisted of 299 patients who were not taking denosumab; they collectively underwent 315 episodes of dental treatment for a total of 669 extractions, and none of them developed ONJ. There were significant differences in age and sex, but not medical comorbidities between the treatment and control groups. Within the treatment group, there were no significant differences in any of these characteristics between those who did, and those who didn't, develop ONJ. Within the treatment group, the number of extractions modified the risk of developing ONJ (odds ratio, 1.35; confidence interval, 1.1-1.7). Of the 76 patients who had extractions between 6 and 7 months after the last denosumab injection, none developed ONJ. CONCLUSIONS: The risk of ONJ in patients on denosumab for osteoporosis is a magnitude greater than for patients on the oral bisphosphonates 2.3% v 0 - 0.3%, which is 7.7 times more likely. Number of extractions and early resumption of the next dose of denosumab increases the risk of ONJ.

The genetic and environmental contributions to variation in the permanent dental arch form: a twin study.

OBJECTIVE: The objective of this study was to examine the relative contributions of genetic and environmental influences on variation in dental arch form in individuals who have largely completed their craniofacial growth. MATERIAL AND METHODS: The subjects of this study comprised dental casts of 50 monozygotic twins and 24 dizygotic twins from the collection of records of twins housed at the Adelaide Dental School, Australia. The subjects were of Western European descent, with an average age of 20.93 ±â€…5.58 years. Dental casts were scanned using a 3D scanner to analyse the dental arch form. Landmark-based inter-arch and intra-arch measurements were performed. Structural equation modelling was employed to analyse the quantitative data using the normal assumptions of the twin model. RESULTS: Genetic modelling revealed that additive genetic and unique environmental factors best explained the observed variation for all occlusal traits measured, except for mandibular intercanine width. High heritability was observed for most intra-arch occlusal variables (0.61-0.85) including the maxillary and mandibular intercanine and intermolar widths, arch depth and perimeter. In contrast, moderate heritability was found for inter-arch occlusal variables (0.52-0.59) such as overjet and overbite. Sexual dimorphism was evident, with males displaying larger posterior arch width than females (P < 0.05). LIMITATIONS: Our sample was limited to individuals of Western European ancestry. CONCLUSION: The predominant source of occlusal variation within this group of Australian twins of Western European descent was controlled by genetic effects, and most were highly heritable. Generally, intra-arch occlusal variables showed greater heritability compared with inter-arch occlusal variables.

Heritability of dental arches and occlusal characteristics: a systematic review and meta-analysis.

BACKGROUND AND OBJECTIVE: The genetic basis of dentoalveolar characteristics has been investigated by several studies, however, the findings are equivocal. The objective of this systematic review and meta-analysis was to evaluate the heritability of dental arches and occlusal parameters in different stages of human dentition. SEARCH METHODS: Electronic databases PubMed, Embase, Scopus, Web of Science, and Dentistry and Oral Science Source were searched up to August 2023 without the restriction of language or publication date. SELECTION CRITERIA: Empirical studies investigating the heritability of dentoalveolar parameters among twins and siblings were included in the review. DATA COLLECTION AND ANALYSIS: Study selection, data extraction, and risk of bias assessment were performed independently and in duplicate by two authors and a third author resolved conflicts if needed. Joanna Briggs Institute's critical appraisal tool was used to evaluate the risk of bias among studies and the certainty of evidence was assessed using the Grading of Recommendation, Assessment, Development, and Evaluation (GRADE) criteria. RESULTS: Twenty-eight studies were included in the systematic review, of which 15 studies reporting heritability coefficients in the permanent dentition stages were deemed suitable for the meta-analysis. Random-effects meta-analyses showed high heritability estimates for maxillary intermolar width (0.52), maxillary intercanine width (0.54), mandibular intermolar width (0.55), mandibular intercanine width (0.55), maxillary arch length (0.76), mandibular arch length (0.57), and palatal depth (0.56). The heritability estimates for the occlusal parameters varied considerably, with relatively moderate values for crossbite (0.46) and overbite (0.44) and low values for buccal segment relationship (0.32), overjet (0.22), and rotation and displacement of teeth (0.16). However, the certainty of evidence for most of the outcomes was low according to the GRADE criteria. CONCLUSIONS: Based on the available evidence, it can be concluded that the dental arch dimensions have a high heritability while the occlusal parameters demonstrate a moderate to low heritability. REGISTRATION: PROSPERO (CRD42022358442).

Patterns of heritability across the human diphyodont dental complex: Crown morphology of Australian twins and families.

OBJECTIVES: This study generates a series of narrow-sense heritability estimates for crown morphology of the deciduous and permanent dentition with two overarching aims. The first is to test the hypothesis that deciduous teeth provide a more faithful reflection of genetic information than their permanent successors. The second is to use quantitative genetic methods to evaluate assumptions underlying common data collection and analysis practices in biodistance research. MATERIALS AND METHODS: Dental morphology data were collected from longitudinal dental casts representing Australian twins and families using Arizona State Dental Anthropology System standards. Polygenic models and estimates of narrow-sense heritability were generated using SOLAR v.8.1.1. Each model considered age, sex, and age/sex interaction as covariates. RESULTS: Heritability estimates significantly differed from zero for the majority of morphological crown characters. Most estimates fell within the 0.4-0.8 range typically observed for crown morphology. Mean heritability was stable across the dental complex, but for paired homologues, permanent traits often yielded higher estimates than their deciduous counterparts. Results indicate directional asymmetry in environmental influence for crown morphology and inform biodistance "best practices" related to data collection and treatment. CONCLUSIONS: Overall, results for this sample support the use of crown morphology as a proxy for genetic variation in evolutionary research. This includes the deciduous dentition, which justifies the expansion of efforts to incorporate subadults into reconstructions of past microevolutionary processes. Results do not indicate that deciduous phenotypes more closely approximate underlying genotype, at least for deciduous/permanent homologues.

Technical note: The use of 3D printing in dental anthropology collections.

OBJECTIVES: Rapid prototyping (RP) technology is becoming more affordable, faster, and is now capable of building models with a high resolution and accuracy. Due to technological limitations, 3D printing in biological anthropology has been mostly limited to museum displays and forensic reconstructions. In this study, we compared the accuracy of different 3D printers to establish whether RP can be used effectively to reproduce anthropological dental collections, potentially replacing access to oftentimes fragile and irreplaceable original material. METHODS: We digitized specimens from the Yuendumu collection of Australian Aboriginal dental casts using a high-resolution white-light scanning system and reproduced them using four different 3D printing technologies: stereolithography (SLA); fused deposition modeling (FDM); binder-jetting; and material-jetting. We compared the deviations between the original 3D surface models with 3D print scans using color maps generated from a 3D metric deviation analysis. RESULTS: The 3D printed models reproduced both the detail and discrete morphology of the scanned dental casts. The results of the metric deviation analysis demonstrate that all 3D print models were accurate, with only a few small areas of high deviations. The material-jetting and SLA printers were found to perform better than the other two printing machines. CONCLUSIONS: The quality of current commercial 3D printers has reached a good level of accuracy and detail reproduction. However, the costs and printing times limit its application to produce large sample numbers for use in most anthropological studies. Nonetheless, RP offers a viable option to preserve numerically constraint fragile skeletal and dental material in paleoanthropological collections.

The Influence of Chorion Type on Health Measures at Birth and Dental Development in Australian and Dutch Twins: A Comparative Study.

Chorion type may significantly influence the prenatal environment of twins. This study explored the associations between chorion type and gestational age, birth weight, birth length, and the timing of emergence of the first primary tooth in two populations of twins, Australian and Dutch. Additionally, we investigated the relationship between chorion type and birth weight discordance (BWD) in order to determine whether a significant relationship existed between discordance in birth weight and discordance in the timing of emergence of the first primary tooth. The two study samples consisted of 409 Australian twin pairs and 301 Dutch twin pairs, all of European ancestry. Data were collected through a combination of questionnaires and recording charts administered to the parents and through linkage with biological databases. In the Australian sample, monozygotic monochorionic (MZMC) twins experienced the shortest mean gestation time (35 weeks), the lowest mean birth length (46 cm) and the lowest mean birth weight (2.3 kg) compared with other twin groups. For the same variables in the Dutch sample, these trends with MZMC twinning were not observed. Chorion type did not significantly affect the mean timing of emergence of the first primary tooth in either sample. Monochorionicity was found to be significantly associated with BWD in both samples, but there was a significant association between BWD in MZMC twin pairs and timing of emergence of the first primary tooth only in the Australian sample. Results from this study support previous findings that the timing of emergence of the first primary tooth is influenced strongly by genetic factors and is well protected from environmental disturbances.

Mirror, mirror? An evaluation of identical twin mirroring in tooth crown morphology.

It has been estimated that 25% of monozygotic ("identical") twin pairs exhibit reverse asymmetry (RA) or "mirroring" of minor anatomical features as a result of delayed zygote division. Here, we examine whether identical twin mirroring accounts for patterns of dental asymmetry in a sample of monozygotic and dizygotic ("fraternal") twins. We focus on crown morphology to approach the following question: is there an association between dental RA frequency and twin type suggestive of the presence of mirror image twins in our sample? Data were collected from 208 deciduous and 196 permanent dentitions of participants of the University of Adelaide Twin Study using Arizona State University Dental Anthropology System standards. RA frequencies were compared across morphological complexes (deciduous, permanent), twin types (monozygotic, dizygotic), and traits. Fisher's exact tests were performed to formally evaluate the association between twin type and dental RA. Across the entire dataset, RA rates failed to exceed 8% for any twin type. In monozygotic twins, deciduous mirroring totaled 5.3% of observed cases, while permanent mirroring totaled 7.8% of observed cases. We found no statistically significant association between RA and twin type for any morphological character (p-value range: 0.07-1.00). Our results suggest the timing of monozygotic twin division does not explain the structure of asymmetry for our morphology dataset and that published estimates of identical twin mirroring rates may be inflated or contingent upon phenotype. Instead, rates reported for this sample more closely align with the proposed etiology of this condition.

Genetic, epigenetic, and environmental influences on dentofacial structures and oral health: ongoing studies of Australian twins and their families.

The Craniofacial Biology Research Group in the School of Dentistry at The University of Adelaide is entering an exciting new phase of its studies of dental development and oral health in twins and their families. Studies of the teeth and faces of Australian twins have been continuing for nearly 30 years, with three major cohorts of twins recruited over that time, and currently we are working with twins aged 2 years old to adults. Cross-sectional data and records relating to teeth and faces of twins are available for around 300 pairs of teenage twins, as well as longitudinal data for 300 pairs of twins examined at three different stages of development, once with primary teeth, once at the mixed dentition stage, and then again when the permanent teeth had emerged. The third cohort of twins comprises over 600 pairs of twins recruited at around birth, together with other family members. The emphasis in this third group of twins has been to record the timing of emergence of the primary teeth and also to sample saliva and dental plaque to establish the timing of colonization of decay-forming bacteria in the mouth. Analyses have confirmed that genetic factors strongly influence variation in timing of primary tooth emergence. The research team is now beginning to carry out clinical examinations of the twins to see whether those who become colonized earlier with decay-forming bacteria develop dental decay at an earlier age. By making comparisons within and between monozygotic (MZ) and dizygotic (DZ) twin pairs and applying modern molecular approaches, we are now teasing out how genetic, epigenetic, and environmental factors interact to influence dental development and also oral health.

Influence of Intraoral Scanners, Operators, and Data Processing on Dimensional Accuracy of Dental Casts for Unsupervised Clinical Machine Learning: An In Vitro Comparative Study.

Purpose: This study assessed the impact of intraoral scanner type, operator, and data augmentation on the dimensional accuracy of in vitro dental cast digital scans. It also evaluated the validation accuracy of an unsupervised machine-learning model trained with these scans. Methods: Twenty-two dental casts were scanned using two handheld intraoral scanners and one laboratory scanner, resulting in 110 3D cast scans across five independent groups. The scans underwent uniform augmentation and were validated using Hausdorff's distance (HD) and root mean squared error (RMSE), with the laboratory scanner as reference. A 3-factor analysis of variance examined interactions between scanners, operators, and augmentation methods. Scans were divided into training and validation sets and processed through a pretrained 3D visual transformer, and validation accuracy was assessed for each of the five groups. Results: No significant differences in HD and RMSE were found across handheld scanners and operators. However, significant changes in RMSE were observed between native and augmented scans with no specific interaction between scanner or operator. The 3D visual transformer achieved 96.2% validation accuracy for differentiating upper and lower scans in the augmented dataset. Native scans lacked volumetric depth, preventing their use for deep learning. Conclusion: Scanner, operator, and processing method did not significantly affect the dimensional accuracy of 3D scans for unsupervised deep learning. However, data augmentation was crucial for processing intraoral scans in deep learning algorithms, introducing structural differences in the 3D scans. Clinical Significance. The specific type of intraoral scanner or the operator has no substantial influence on the quality of the generated 3D scans, but controlled data augmentation of the native scans is necessary to obtain reliable results with unsupervised deep learning.

Development of the oral resistome during the first decade of life.

Antibiotic overuse has promoted the spread of antimicrobial resistance (AMR) with significant health and economic consequences. Genome sequencing reveals the widespread presence of antimicrobial resistance genes (ARGs) in diverse microbial environments. Hence, surveillance of resistance reservoirs, like the rarely explored oral microbiome, is necessary to combat AMR. Here, we characterise the development of the paediatric oral resistome and investigate its role in dental caries in 221 twin children (124 females and 97 males) sampled at three time points over the first decade of life. From 530 oral metagenomes, we identify 309 ARGs, which significantly cluster by age, with host genetic effects detected from infancy onwards. Our results suggest potential mobilisation of ARGs increases with age as the AMR associated mobile genetic element, Tn916 transposase was co-located with more species and ARGs in older children. We find a depletion of ARGs and species in dental caries compared to health. This trend reverses in restored teeth. Here we show the paediatric oral resistome is an inherent and dynamic component of the oral microbiome, with a potential role in transmission of AMR and dysbiosis.

Genetic, environmental and epigenetic influences on variation in human tooth number, size and shape.

The aim of this review is to highlight some key recent developments in studies of tooth number, size and shape that are providing better insights into the roles of genetic, environmental and epigenetic factors in the process of dental development. Advances in molecular genetics are helping to clarify how epigenetic factors influence the spatial and temporal regulation of the complex processes involved in odontogenesis. At the phenotypic level, the development of sophisticated systems for image analysis is enabling new dental phenotypes to be defined. The 2D and 3D data that are generated by these imaging systems can then be analysed with mathematical approaches, such as geometric morphometric analysis. By gathering phenotypic data and DNA from twins, it is now possible to use 'genome-wide' association studies and the monozygotic co-twin design to identify important genes in odontogenesis and also to clarify how epigenetic and environmental factors can affect this process. Given that many of the common dental anomalies affecting the human dentition are interrelated, apparently reflecting pleiotropic genetic effects, the discoveries and new directions described in this paper should have important implications for clinical dental practice in the future.

Genetic Correlation, Pleiotropy, and Molar Morphology in a Longitudinal Sample of Australian Twins and Families.

This study aims to expand our understanding of the genetic architecture of crown morphology in the human diphyodont dentition. Here, we present bivariate genetic correlation estimates for deciduous and permanent molar traits and evaluate the patterns of pleiotropy within (e.g., m1-m2) and between (e.g., m2-M1) dentitions. Morphology was observed and scored from dental models representing participants of an Australian twin and family study (deciduous n = 290, permanent n = 339). Data collection followed Arizona State University Dental Anthropology System standards. Genetic correlation estimates were generated using maximum likelihood variance components analysis in SOLAR v.8.1.1. Approximately 23% of deciduous variance components models and 30% of permanent variance components models yielded significant genetic correlation estimates. By comparison, over half (56%) of deciduous-permanent homologues (e.g., m2 hypocone-M1 hypocone) were significantly genetically correlated. It is generally assumed that the deciduous and permanent molars represent members of a meristic molar field emerging from the primary dental lamina. However, stronger genetic integration among m2-M1/M2 homologues than among paired deciduous traits suggests the m2 represents the anterior-most member of a "true" molar field. The results indicate genetic factors act at distinct points throughout development to generate homologous molar form, starting with the m2, which is later replaced by a permanent premolariform crown.

Differential network analysis of oral microbiome metatranscriptomes identifies community scale metabolic restructuring in dental caries.

Dental caries is a microbial disease and the most common chronic health condition, affecting nearly 3.5 billion people worldwide. In this study, we used a multiomics approach to characterize the supragingival plaque microbiome of 91 Australian children, generating 658 bacterial and 189 viral metagenome-assembled genomes with transcriptional profiling and gene-expression network analysis. We developed a reproducible pipeline for clustering sample-specific genomes to integrate metagenomics and metatranscriptomics analyses regardless of biosample overlap. We introduce novel feature engineering and compositionally-aware ensemble network frameworks while demonstrating their utility for investigating regime shifts associated with caries dysbiosis. These methods can be applied when differential abundance modeling does not capture statistical enrichments or the results from such analysis are not adequate for providing deeper insight into disease. We identified which organisms and metabolic pathways were central in a coexpression network as well as how these networks were rewired between caries and caries-free phenotypes. Our findings provide evidence of a core bacterial microbiome that was transcriptionally active in the supragingival plaque of all participants regardless of phenotype, but also show highly diagnostic changes in the ways that organisms interact. Specifically, many organisms exhibit high connectedness with central carbon metabolism to Cardiobacterium and this shift serves a bridge between phenotypes. Our evidence supports the hypothesis that caries is a multifactorial ecological disease.

The genetic architecture of anterior tooth morphology in a longitudinal sample of Australian twins and families.

OBJECTIVE: This study presents a quantitative genetic analysis of human anterior dental morphology in a longitudinal sample of known genealogy. The primary aim of this work is to generate a suite of genetic correlations within and between deciduous and permanent characters to access patterns of integration across the diphyodont dental complex. DESIGN: Data were recorded from casted tooth crowns representing participants of a long-term Australian twin and family study (deciduous n = 290, permanent n = 339). Morphological trait expression was observed and scored following Arizona State University Dental Anthropology System standards. Bivariate genetic correlations were estimated using maximum likelihood variance decomposition models in SOLAR v.8.1.1. RESULTS: Genetic correlation estimates indicate high levels of integration between antimeres but low to moderate levels among traits within a tooth row. Only 9% of deciduous model comparisons were significant, while pleiotropy was indicated for one third of permanent trait pairs. Canine characters stood out as strongly integrated, especially in the deciduous dentition. For homologous characters across dentitions (e.g., deciduous i1 shoveling and permanent I1 shoveling), ∼70% of model comparisons yielded significant genetic correlations. CONCLUSIONS: Patterns of genetic correlation suggest a morphological canine module that spans the primary and secondary dentition. Results also point to the existence of a genetic mechanism conserving morphology across the diphyodont dental complex, such that paired deciduous and permanent traits are more strongly integrated than characters within individual tooth rows/teeth.

Genetic modeling of primary tooth emergence: a study of Australian twins.

The aim of this study was to quantify contributions of genetic and environmental factors to variation in timing of emergence of the primary teeth in a sample of monozygotic and dizygotic twins, using univariate model-fitting approaches. The sample comprised 94 pairs of monozygotic twins and 125 pairs of dizygous twins, all of European ancestry, aged from 2-6 years. Tooth emergence timing was based on parental report, with a subset of data validated by clinical assessment. Heritability estimates for tooth emergence timing were generally high, around 90%, however estimates for the lower right lateral incisor and the lower canines were around 50%. These findings confirm a strong genetic influence on observed variation in the timing of emergence of the human primary teeth.

Genetic contribution to palatal morphology variation using three-dimensional analysis in Australian twins.

OBJECTIVE: This study aimed to provide insight into the relative contributions of genetic and environmental factors to palatal morphology variation in a cohort of Australian twins. METHODS: Healthy Australian twins, aged 12-15 years (45 monozygotic, 46 same sex dizygotic, and 32 opposite-sex dizygotic) were included in the study groups. A scanner was used to obtain three-dimensional data of the maxillary arch. Palatal depth was defined by a line to the deepest point measured from the reference plane at the mid-point of the inter-pre-molar or inter-molar line. This line was then divided into 10 equal sections in order to created 10 different depths for each palatal width. Each palatal width was divided into anterior and posterior areas. Univariate genetic analysis, using the OpenMx structural equation modelling package in R, was carried out on the quantitative data using the normal assumptions of a twin model. RESULTS: Heritability estimates for anterior palatal width ranged from 0.75 to 0.80, and from 0.78 to 0.86 for posterior palatal width. Estimates for anterior and posterior palatal depth were 0.72 and 0.86, respectively. CONCLUSIONS: Palatal morphology tends to have a moderate to relatively high genetic contribution overall. Palate height has a higher genetic contribution posteriorly than anteriorly. The width of the deep palate is under marginally less stringent genetic regulation than the width of the shallow palate.

Longitudinal Study of Oral Microbiome Variation in Twins.

Humans are host to a multitude of microorganisms that rapidly populate the body at birth, subject to a complex interplay that is dependent on host genetics, lifestyle, and environment. The host-associated microbiome, including the oral microbiome, presents itself in a complex ecosystem important to health and disease. As the most common chronic disease globally, dental caries is induced by host-microbial dysbiosis in children and adults. Multiple biological and environmental factors are likely to impact disease predisposition, onset, progression, and severity, yet longitudinal studies able to capture these influences are missing. To investigate how host genetics and environment influenced the oral microbial communities over time, we profiled supragingival plaque microbiomes of dizygotic and monozygotic twins during 3 visits over 12-months. Dental plaque DNA samples were amplified by targeting the 16S rRNA gene V4 region, and microbial findings were correlated with clinical, diet and genetic metadata. We observed that the oral microbiome variances were shaped primarily by the environment when compared to host genetics. Among the environmental factors shaping microbial changes of our subjects, significant metadata included age of the subject, and the age by which subjects initiated brushing habits, and the types of actions post-brushing. Relevant heritability of the microbiome included Actinomyces and Capnocytophaga in monozygotic twins and Kingella in dizygotic twins. Corynebacterium and Veillonella abundances were associated with age, whereas Aggregatibacter was associated with younger subjects. Streptococcus abundance showed an inverse association over time, and Selenomonas abundances increased with brushing frequency per day. Unraveling the exact biological mechanisms in caries has the potential to reveal novel host-microbial biomarkers, pathways, and targets important to effective preventive measures, and early disease control in children.

Clinical investigation of C-terminal cross-linking telopeptide test in prevention and management of bisphosphonate-associated osteonecrosis of the jaws.

PURPOSE: The aim of this study was to determine, in a clinical setting, the effectiveness of the C-terminal cross-linking telopeptide test (CTX) test in the prevention and management of osteonecrosis of the jaws (ONJ) in patients taking bisphosphonates. PATIENTS AND METHODS: A total of 348 patients underwent a fasted morning CTX test. Of these, 222 were patients at risk of ONJ who had been referred for extractions, 15 had ONJ, and 113 were controls. RESULTS: The 215 patients taking long-term oral bisphosphonates were older (71 +/- 11.6 years), were predominantly women with osteoporosis, and were medically compromised. The average CTX value was 238 +/- 144 pg/mL, with 98 having a value less than 200 pg/mL. One patient with a CTX value of 126 pg/mL developed ONJ after an extraction. Seven intravenous bisphosphonate patients underwent extractions with no cases of ONJ developing. The CTX value was 329 +/- 354, with 4 less than 200 pg/mL. Fifteen patients developed ONJ, 12 after extractions and 3 spontaneously. Of these, 7, who were still taking a bisphosphonate at presentation, had a CTX value of 116 pg/mL. A CTX value of less than 150 pg/mL did not correlate with the clinical risk factors of age, gender, comorbidities, bone disease, or bisphosphonate duration. A statistically significant difference in the CTX value was found for those taking alendronate compared with those taking risedronate (P < .0001). If the bisphosphonate was ceased, the CTX value increased at approximately 25 pg/mL per month. CONCLUSIONS: The CTX test is not predictive of the development of ONJ for an individual patient but does identify those in the "risk zone," which is a value of less than 150 pg/mL to 200 pg/mL. If medically appropriate, the bisphosphonate can be ceased so that the CTX value increases to bring the patient out of the "risk zone."