Three-Dimensional Assessment of Cervical Vertebrae Anomalies in Patients With Cleft Lip and Palate.

OBJECTIVE: To evaluate the occurrence of cervical vertebrae anomalies (CVA) in patients with unilateral (UCLP) and bilateral cleft lip and palate (BCLP) using cone beam computed tomography (CBCT) examinations. DESIGN: Retrospective assessment of CBCT images. Descriptive statistics were calculated. Fisher exact test or χ2 test was performed to evaluate the differences among each CVA between sex and type of cleft. SETTING: School of Dentistry. PARTICIPANTS: One hundred fifty-one patients with cleft lip and palate (103 UCLP/48 BCLP). INTERVENTIONS: No relevant intervention. MAIN OUTCOME MEASURES: Cone beam computed tomography images were assessed for the presence or absence of 12 most commonly observed CVA: spina bifida, dehiscence, cleft of the posterior arch, cleft of the anterior arch, fusion between cervical vertebrae, block fusion, occipitalization, narrowing of the intervertebral space, posterior ponticle, os odontoideum, ossiculum terminale, and subdental cartilaginous remnants. RESULTS: The presence of subdental cartilaginous remnants was the most frequently observed alteration-found in 81.45% of the sample-and it was the only CVA with statistically significant frequencies in the individuals with BCLP. Considering only the other CVA, 22.51% presented 1 and 5.29% presented 2 or more CVA. CONCLUSIONS: Patients with cleft lip and palate may present an overall high incidence of CVAs. However, when comparing the distribution of the CVAs among sex and types of cleft, the only significant difference noted was a higher incidence of subdental cartilaginous remnants among patients with BCLP.

Craniofacial characterization of Marfan Syndrome.

OBJECTIVE: The morbidity and mortality associated with the Marfan Syndrome (MFS) warrant timely diagnosis and intervention that can improve long-term prognosis. The aim of this study was to test the hypothesis that a distinct craniofacial morphology exists for patients with MFS that can be described quantitatively and qualitatively. METHODS: Subjects with a positive diagnosis of MFS were recruited for this study (N = 36). Craniofacial anthropometric measurements were made on each subject and compared to established norms of age- and sex-matched controls using z-scores calculated for measurements of MFS patients. Lateral and frontal photographs were obtained to make qualitative assessments and describe facial features of subjects, and a clinical examination was completed to document occlusal relationships. RESULTS: The subjects were primarily female (58%) ranging in age between 4 and 57 years (mean age 10.7 ± 6.0 years). Comparison of craniofacial measurements revealed that for 10 of the 12 measurements, ≥65% of the study population had a z-score of ± 2 and fell within the normal range for facial dimension. For 2 of the 12 measurements, over half of the subjects fell outside of the normal range (z-score < -2 or > 2) for facial dimension. Specifically, the majority of participants resided in the supernormal category for biocular width and the subnormal category for width of the face. Photographic assessment revealed retrognathia (54%) and down-slanting palpebral fissures (62%) were most prevalent in MFS patients. CONCLUSION: Our data suggest there are quantitative differences in the facial morphology of patients with MFS when compared to a control population.

Issues involved in the phenotypic classification of orofacial clefts ascertained through a state birth defects registry for the North Carolina Cleft Outcomes Study.

BACKGROUND: Epidemiologic studies involving birth defects are extremely sensitive to phenotype accuracy and precision. We devised a case review and classification protocol for a project to study school achievement in children with idiopathic, nonsyndromic orofacial clefts to improve the reliability of phenotypic classification from the statewide birth defects registry. METHODS: Surveillance-program abstraction data and medical records at the birth or treating hospitals were used when available. Exclusion criteria included: median cleft lip; Tessier cleft; premaxillary agenesis; presence of a recognizable syndrome, phenotype, association, or sequence (other than Robin sequence); clefts with other malformations not considered to be normal or common variants in the newborn; and cases with documented or suspected genetic or teratogenic causes. RESULTS: Of 712 children identified with orofacial clefts, 153 were excluded, leaving 559 nonsyndromic orofacial cleft cases of unknown cause in the final study. These cases were grouped into the following clinically meaningful types: cleft lip with or without cleft alveolus; cleft lip and cleft palate; and cleft palate only. This review and classification process resulted in the elimination of 21.5% of the original cohort of identified cases, with most exclusions being due to suspected syndromic associations. CONCLUSION: Verbatim descriptions of the clinical findings are critical for accurate classification of diagnoses. This review process improved the precision of orofacial cleft phenotype classification for our study. Precision would have been further improved if all of the cases had verbatim descriptions of diagnoses and all medical records could have been reviewed by the classification team.

Evaluation of ICD-9-CM codes for craniofacial microsomia.

BACKGROUND: Craniofacial microsomia (CFM) is a congenital condition characterized by microtia and mandibular underdevelopment. Healthcare databases and birth defects surveillance programs could be used to improve knowledge of CFM. However, no specific International Classification of Diseases, 9th Revision, Clinical Modification (ICD-9-CM) code exists for this condition, which makes standardized data collection challenging. Our aim was to evaluate the validity of existing ICD-9-CM codes to identify individuals with CFM. METHODS: Study sample eligibility criteria were developed by an expert panel and matched to 11 ICD-9-CM codes. We queried hospital discharge data from two craniofacial centers and identified a total of 12,254 individuals who had ≥1 potentially CFM-related code(s). We reviewed all (n = 799) medical records identified at the University of North Carolina (UNC) and 500 randomly selected records at Seattle Children's Hospital (SCH). Individuals were classified as a CFM case or non-case. RESULTS: Thirty-two individuals (6%) at SCH and 93 (12%) at UNC met the CFM eligibility criteria. At both centers, 59% of cases and 95% of non-cases had only one code assigned. At both centers, the most frequent codes were 744.23 (microtia), 754.0 and 756.0 (nonspecific codes), and the code 744.23 had a positive predictive value (PPV) >80% and sensitivity >70%. The code 754.0 had a sensitivity of 3% (PPV <1%) at SCH and 36% (PPV = 5%) at UNC, whereas 756.0 had a sensitivity of 38% (PPV = 5%) at SCH and 18% (PPV = 26%) at UNC. CONCLUSIONS: These findings suggest the need for a specific CFM code to facilitate CFM surveillance and research.

Effect of finishing instrumentation on the marginal integrity of resin-based composite restorations.

PURPOSE: This study evaluated the effect of the use of different finishing instruments on the marginal integrity of resin composite restorations. MATERIALS AND METHODS: Bovine incisors (N = 75) embedded in epoxy resin had the facial enamel ground and polished to 1200-grit. A standardized cavity (3 x 3 mm, 2 mm deep) was prepared on each specimen and restored with a 2-step total-etch adhesive (Single Bond, 3M ESPE, St. Paul, MN, USA) and a hybrid resin composite (Filtek Z250, 3M ESPE) in a single increment. The restorations were mechanically polished to 1200-grit. Specimens were randomized into different groups (N = 5) according to finishing technique: positive control (1200-grit paper), negative control (regular-grit diamond), fine cross-cut laminated burs, straight-cut laminated burs, spiral-cut laminated burs, and finishing diamonds. The straight-cut burs, spiral-cut burs, and finishing diamonds were tested individually as fine, extra-fine, and ultra-fine, as well as sequentially as a series. A high-speed, water-cooled handpiece under standardized pressure (0.5 N) and time (40 seconds) was used for all finishing procedures. Specimens were processed for scanning electron microscope, and margin gaps were systematically measured. Data were analyzed with one-way analysis of variance and Duncan test. RESULTS: The negative control specimens (course diamond) presented the largest gaps, whereas the positive control specimens (mechanically polished) generated the smallest gaps. No statistically significant difference was noted between the finishing diamonds and the positive control. The negative control exhibited significantly larger gaps when compared with the other finishing instruments. Intermediate results were observed for cross-cut, straight-cut, and spiral-cut laminated burs. Fine, extra-fine and ultra-fine finishing diamonds generated smaller gaps compared with laminated burs, but the differences were not always statistically significant. CONCLUSION: Fine, extra-fine and ultra-fine finishing diamonds used to finish composite restorations generated better marginal integrity when compared with carbides and regular-grit diamonds. CLINICAL SIGNIFICANCE When finishing composite restorations, finishing diamond burs result in better composite margins than carbide laminated burs.

Dentin and enamel bond strengths of dual-cure composite luting agents used with dual-cure dental adhesives.

OBJECTIVE: The purpose of this in vitro study was to evaluate and compare dentin and enamel bond strengths obtained with dual-cure composite luting agents when used with dual-cure dental adhesives. METHODS: Human molars were ground flat to expose enamel (n=80) or dentin (n=80). Specimens in each substrate group were randomly assigned to eight treatment sub-groups, according to four adhesive-luting agent combinations and two test conditions (with or without thermocycling). Pre-polymerized composite resin posts (TPH Spectrum) were luted to either the enamel or dentin surfaces with one of the following adhesive-luting agent combinations: (1) Xeno IV Dual Cure (dual-cure self-etch adhesive) and Calibra (dual-cure luting agent); (2) Prime & Bond NT Dual Cure (dual-cure total-etch adhesive) and Calibra; (3) OptiBond All-in-One Dual Cure (dual-cure self-etch adhesive) and Nexus 2 Dual Syringe (dual-cure luting agent); (4) OptiBond Solo Plus Dual Cure (dual-cure total-etch adhesive) and Nexus 2 Dual Syringe. For each treatment sub-group, half the specimens (n=10) were tested after 24h storage in water at 37 degrees C, and the other half (n=10) were tested after thermocycling for 1800 cycles between water baths held at 5 and 55 degrees C, with a dwell time in each bath of 30s, and a transfer time of 10s. Bond strengths were measured in shear mode, and expressed in MPa. The fracture mode (adhesive, cohesive, mixed) was examined. Data were analyzed for statistical significance with a factorial ANOVA and post hoc tests. RESULTS: Mean enamel bond strengths ranged from 8.4MPa for non-thermocycled OptiBond All-in-One|Nexus 2 to 35.5MPa for non-thermocycled Prime & Bond NT|Calibra. Mean dentin bond strengths ranged from 14.5MPa for non-thermocycled OptiBond Solo Plus|Nexus 2 to 30.9MPa for thermocycled Xeno IV|Calibra. The fracture mode was predominantly adhesive for all groups. CONCLUSIONS: On enamel, the total-etch adhesives performed better than their self-etch counterparts, while in dentin, the opposite was found, i.e., the self-etch adhesives performed better than their total-etch counterparts. Thermocycling for 1800 cycles did not affect the SBS of the materials tested to dentin and enamel.

Effect of nanofillers' size on surface properties after toothbrush abrasion.

PURPOSE: To investigate the effect of filler-particle size of experimental and commercial resin composites, undergoing toothbrush abrasion, on three surface properties: surface roughness (SR), surface gloss (G) and color stability (CS). METHODS: Four model (Ivoclar/Vivadent) and one commercial resin composite (Tokuyama) with varying filler-size from 100-1000 nm were examined. Six discs (10 mm x 2 mm) from each product were prepared and mechanically polished. The samples were then submitted to 20,000 brushing strokes in a toothbrush abrasion machine. SR parameters (Ra, Rt and RSm), G, and CS were measured before and after toothbrush abrasion. Changes in SR and G were analyzed by 2-way ANOVA, with Bonferroni post hoc test. CS values were submitted to one-way ANOVA and Bonferroni post hoc test (alpha=0.05). RESULTS: Initial G values ranged between 73-87 gloss units (GU) and were reduced after toothbrush abrasion to a range of 8-64 GU. Toothbrush abrasion resulted in significant modifications in SR and G amongst the materials tested, attributed to filler sizes. There was statistically significant difference in color (delta E* ranged from 0.38-0.88). Filler size did not affect color stability. Toothbrush abrasion resulted in rougher and matte surfaces for all materials tested. Although the individual differences in surface roughness among filler sizes were not always significant, the correlation showed a trend that larger filler sizes resulted in higher surface roughness after abrasion for the SR parameters Ra and Rt (r = 0.95; r = 0.93, respectively). RSm showed an increase after toothbrush abrasion for all resin composites, however no significant correlation was detected (r = 0.21).There was a significant correlation between G and Ra ratios (r = - 0.95).

Using extracted teeth for research: the effect of storage medium and sterilization on dentin bond strengths.

BACKGROUND: The Centers for Disease Control and Prevention has adopted guidelines for infection control of extracted teeth used for research and teaching, requiring that teeth be sterilized before use. The authors conducted a study to test the null hypothesis that the storage medium and sterilization method have no effect on composite-to-dentin bond strengths. MATERIALS AND METHODS: The authors collected 170 bovine incisors, cleaned them and placed them randomly into one of six storage media at 37 degrees C for 60 days: distilled water (dH(2)O), 0.9 percent sodium chloride, 0.5 percent chloramine-T, 5.25 percent sodium hypochlorite (NaClO), 2 percent glutaraldehyde and 10 percent formalin. For sterilization, they autoclaved a subset of 10 specimens from every sample, while they stored another subset of 10 specimens from every sample (except for the 10 percent formalin sample) in 10 percent formalin for 14 days. The authors then embedded the specimens in epoxy and ground flat the facial surface to expose middle-depth dentin, which they polished to 600 grit. They used a dental adhesive to apply composite to the exposed dentin. The authors tested the composite-to-dentin shear bond strength 24 hours after bonding. They analyzed the data using global analysis of variance and, when appropriate, multiple post hoc tests (P = .05). RESULTS: Storage in NaClO resulted in significantly lower bond strength than that of the other treatment specimens. Sterilization with the autoclave negatively affected the bond strength of specimens stored initially in dH(2)O or 10 percent formalin, while sterilization with formalin alone had no significant effect on bond strengths. CONCLUSIONS: Storing bovine teeth in 5.25 percent NaClO may negatively affect composite-to-dentin bond strengths. Immersion in 10 percent formalin might be the best option for storage and sterilization of bovine teeth that are to be used in dental bonding studies in vitro.

Effect of light-curing method on marginal adaptation, microleakage, and microhardness of composite restorations.

The objective of this study was to investigate the effects of different light-curing methods on microleakage, marginal adaptation, and microhardness of composite restorations. Slot-type preparations were made in bovine teeth, with gingival margins on dentin. Specimens were divided into 12 groups (n = 12) according to composite-light-curing unit (LCU) combinations. Three composites were used: Filtek Supreme, Herculite XRV, and Heliomolar. All restorations were placed using the same adhesive. Four LCUs were used: a quartz-tungsten-halogen (QTH) LCU (Optilux 501), a first-generation light-emitting diode (LED) LCU (FreeLight 1), and two second-generation LED LCUs (FreeLight 2 and Translux Power Blue). After finishing and polishing, specimens were subjected to mechanical load cycling (100,000 cycles). Gingival margin adaptation was determined as a function of gap formation using epoxy replicas. Microleakage was evaluated by measuring dye penetration across the gingival wall in cross-sectioned specimens. Microhardness was measured as Knoop Hardness number (KHN) at different occluso-gingival locations in cross-sectioned specimens. Data were analyzed for statistical significance (p = 0.05) using appropriate statistical tests. Marginal adaptation was affected by load-cycling in most specimens, but no significant differences were observed among composites and LCUs. Microleakage was not affected by LCU, except for Heliomolar specimens which when cured with Optilux 501 resulted in higher microleakage scores than those obtained with the other LCUs. For microhardness, Translux Power Blue generally produced the highest values and the FreeLight 1 produced the lowest. The performance of the second-generation LED LCUs generally was similar to that of the QTH control, and better than that of the first-generation LED unit.

Comparison of different methods to assess alveolar cleft defects in cone beam CT images.

OBJECTIVES: This study aimed to evaluate the accuracy of three different methods for assessing the volume of cleft defects in CBCT images. The influence of field of view (FOV) and voxel sizes was also assessed. METHODS: Using three radio-opaque plastic skulls, unilateral defects were created to mimic alveolar clefts and were filled with wax following the contralateral side contours. They were scanned in a CBCT unit using four different acquisition protocols, varying FOV and voxel sizes. Using three different methods, the defect/wax volume was evaluated on the images by defining: (1) the width, height and facial-palatal length of the defect in maximum intensity projection; (2) the areas of the defect on axial slices; and (3) the threshold and segmentation of the region of interest. The values obtained from each method using different acquisition protocols were compared with the real volume of the wax (gold standard) using ANOVA and Tukey's test. RESULTS: Methods 2 and 3 did not differ from the gold standard (p > 0.05). Conversely, Method 1 presented statistically significant overestimated values (p < 0.01). No differences were found among the different FOV and voxel sizes (p > 0.05). CONCLUSIONS: CBCT volumes proved reliable for the volumetric assessment of alveolar cleft defects, when using Methods 2 and 3 regardless of FOV and voxel sizes. It may be possible to improve surgical planning and outcomes by knowing the exact volume of grafting material needed prior to the surgical intervention.

Volumetric nasal cavity analysis in children with unilateral and bilateral cleft lip and palate.

OBJECTIVES/HYPOTHESIS: Children with cleft lip and palate (CLP) often suffer from nasal obstruction that may be related to effects on nasal volume. The objective of this study was to compare side:side volume ratios and nasal volume in patients with unilateral (UCLP) and bilateral (BCLP) clefts with age-matched controls. STUDY DESIGN: Retrospective case-control study using three-dimensional (3D) nasal airway reconstructions. METHODS: We analyzed 20 subjects (age range = 7-12 years) with UCLP and BCLP from a regional craniofacial center who underwent cone beam computed tomography (CT) prior to alveolar grafting. Ten multislice CT images from age-matched controls were also analyzed. Mimics software (Materialise, Plymouth, MI) was used to create 3D reconstructions of the main nasal cavity and compute total and side-specific nasal volumes. Subjects imaged during active nasal cycling phases were excluded. RESULTS: There was no statistically significant difference in affected:unaffected side volume ratios in UCLP (P = .48) or left:right ratios in BCLP (P = .25) when compared to left:right ratios in controls. Mean overall nasal volumes were 9,932 ± 1,807, 7,097 ± 2,596, and 6,715 ± 2,115 mm(3) for control, UCLP, and BCLP patients, respectively, with statistically significant volume decreases for both UCLP and BCLP subjects from controls (P < .05). CONCLUSIONS: This is the first study to analyze total nasal volumes in BCLP patients. Overall nasal volume is compromised in UCLP and BCLP by approximately 30%. Additionally, our finding of no major difference in side:side ratios in UCLP and BCLP compared to controls conflicts with pre-existing literature, likely due to exclusion of actively cycling scans and our measurement of the functional nasal cavity. LEVEL OF EVIDENCE: 3b. Laryngoscope, 126:1475-1480, 2016.

Nasal Airway Dysfunction in Children with Cleft Lip and Cleft Palate: Results of a Cross-Sectional Population-Based Study, with Anatomical and Surgical Considerations.

BACKGROUND: The aesthetic aspects of the cleft lip nasal deformity have been appreciated for over a century, but the functional implications have remained largely underappreciated or misunderstood. This study describes the frequency and severity of nasal obstructive symptoms among children with cleft lip and/or cleft palate, addressing the hypotheses that age, cleft type, and severity are associated with the development of nasal obstructive symptoms. METHODS: Children with nonsyndromic cleft lip and/or cleft palate and a comparison group of unaffected children born from 1997 to 2003 were identified through the North Carolina Birth Defects Monitoring Program and birth certificates. Nasal airway obstruction was measured using the validated Nasal Obstruction Symptom Evaluation scale. RESULTS: The survey was completed by parental proxy for 176 children with cleft lip and/or cleft palate and 333 unaffected children. Nasal obstructive symptoms were more frequently reported in cleft lip with cleft palate compared with unaffected children (p < 0.0001); children who had isolated cleft lip with or without alveolus and isolated cleft palate were not statistically different from unaffected children. Patients with unilateral cleft lip with cleft palate were found to be more severely affected than bilateral cases. Nasal obstruction was observed in early childhood, although severity worsened in adolescence. CONCLUSIONS: This population-based study reports a high prevalence of nasal obstructive symptoms in children with cleft lip and/or cleft palate based on type and severity of the cleft. The authors encourage cleft teams to consider using this or similar screening methods to identify which children may benefit from functional rhinoplasty. CLINICAL QUESTION/LEVEL OF EVIDENCE: Risk, I.

Microhardness evaluation of in situ vital bleaching on human dental enamel using a novel study design.

OBJECTIVES: The aim of this 'in situ' study was to evaluate the microhardness of dental enamel following treatment with an in-office and an at-home vital bleaching agent through a novel approach using samples temporarily bonded 'in vivo'. METHODS: Human dental enamel slabs (n=88) were subjected to sequential polishing and initial Knoop microhardness tests were performed. The slabs were fixed to the facial surfaces of the maxillary first molars of 44 human volunteers. They were divided into four groups (n=11) according to the treatment group: G1- in-office-CP37+ at-home-CP10; G2- in-office-CP37+ at-home-PLA; G3- in-office-PLA and at-home-CP10; G4- in-office and at-home-PLA. After 3 weeks of treatment, final microhardness measurements were performed. RESULTS AND SIGNIFICANCE: ANOVA and Tukey's HSD hoc analysis (alpha=0.05) revealed no differences among initial or final microhardness values (p>0.05); however, significant differences occurred between initial and final values for each group (p<0.01), The evaluation of microhardness revealed a reduction of 6.8% for G1, 4.1% for G2, 3.4% for G3 and 3.5% for G4. In-office bleaching with 37% carbamide peroxide, an at-home bleaching with 10% carbamide peroxide, and a combination of both resulted in lower enamel microhardness when measured immediately post-treatment. However, long-term effects of these treatments are not known, and are believed to be clinically insignificant due to the relatively small reductions observed in enamel microhardness. Interestingly, the carrier, Cabopol 934P, also resulted in similar reduction in enamel microhardness even when administrated alone as a placebo.

Three-dimensional evaluation of nasopharyngeal airways of unilateral cleft lip and palate patients.

OBJECTIVES/HYPOTHESIS: The aim of this study was to compare the volume of nasopharyngeal airways of patients with unilateral cleft lip and palate (UCLP) with a control (noncleft) group of subjects. STUDY DESIGN: Retrospective case-control study. METHODS: The sample consisted of 45 subjects, 30 with UCLP who underwent cone beam computed tomography (CBCT) exams as part of their workups for alveolar bone graft and 15 noncleft subjects seeking orthodontic care who obtained CBCT for diagnostic purposes. The three-dimensional volumes of nasal (NV) and oropharyngeal (PV) airways of these subjects were assessed separately using Mimics software by one well-calibrated evaluator. The images were rendered, and the volumes were measured in cm(3). RESULTS: The mean PV of the UCLP subjects was 6.7 cm(3) and the mean NV was 10.3 cm(3). The control group presented with a mean PV of 8.1 cm(3) and a mean NV of 11.6 cm(3). Student t test showed no statistically significant difference for PV (P = 0.10) or NV (P = 0.23) when comparing cleft and noncleft subjects. CONCLUSION: Evaluation of the nasopharyngeal airways revealed that the NV and PV airways showed no difference between the cleft and noncleft subjects. It is interesting to postulate that the reported nasal abnormalities of the cleft patients do not produce a significant measurable effect on the overall volume, at least as shown in our study sample. In addition, the nasopharyngeal airways of patients are not larger than those of age-matched controls, at least at the ages that were sampled, after cleft palate repair.

3D of Brain Shape and Volume After Cranial Vault Remodeling Surgery for Craniosynostosis Correction in Infants.

The skull of young children is made up of bony plates that enable growth. Craniosynostosis is a birth defect that causes one or more sutures on an infant's skull to close prematurely. Corrective surgery focuses on cranial and orbital rim shaping to return the skull to a more normal shape. Functional problems caused by craniosynostosis such as speech and motor delay can improve after surgical correction, but a post-surgical analysis of brain development in comparison with age-matched healthy controls is necessary to assess surgical outcome. Full brain segmentations obtained from pre- and post-operative computed tomography (CT) scans of 8 patients with single suture sagittal (n=5) and metopic (n=3), non-syndromic craniosynostosis from 41 to 452 days-of-age were included in this study. Age-matched controls obtained via 4D acceleration-based regression of a cohort of 402 full brain segmentations from healthy controls magnetic resonance images (MRI) were also used for comparison (ages 38 to 825 days). 3D point-based models of patient and control cohorts were obtained using SPHARM-PDM shape analysis tool. From a full dataset of regressed shapes, 240 healthy regressed shapes between 30 and 588 days-of-age (time step = 2.34 days) were selected. Volumes and shape metrics were obtained for craniosynostosis and healthy age-matched subjects. Volumes and shape metrics in single suture craniosynostosis patients were larger than age-matched controls for pre- and post-surgery. The use of 3D shape and volumetric measurements show that brain growth is not normal in patients with single suture craniosynostosis.

Photographic protocol for image acquisition in craniofacial microsomia.

Craniofacial microsomia (CFM) is a congenital condition associated with orbital, mandibular, ear, nerve, and soft tissue anomalies. We present a standardized, two-dimensional, digital photographic protocol designed to capture the common craniofacial features associated with CFM.

Photosensitization of in vitro biofilms by toluidine blue O combined with a light-emitting diode.

In natural ecosystems, micro-organisms grow preferentially attached to surfaces, forming matrix-enclosed biofilms. The aim of this study was to determine photodestruction levels in biofilms after subjecting them to photodynamic therapy. Biofilms of Streptococcus mutans, S. sobrinus, and S. sanguinis were grown on enamel slabs for 3, 5 or 7 d. Both the number of viable micro-organisms and the concentration of water-insoluble polysaccharides were analysed, and mineral loss (DeltaZ) analyses were performed on the enamel slabs. The antimicrobial potential of toluidine blue O (0.1 mg ml(-1)), associated with 85.7 J cm(-2) of a light-emission diode, was evaluated on the viability of 5-d biofilms. Both the number of micro-organisms and the concentration of water-insoluble polysaccharide increased with the age of the biofilms. A significant reduction ( approximately 95%) in viability was observed for S. mutans and S. sobrinus biofilms following photosensitization, with a > 99.9% reduction in the viability of S. sanguinis biofilms. In conclusion, a biofilm model was shown to be suitable for studying changes in bacterial numbers and enamel mineralization and for demonstrating the potential value of photosensitization in the control of in vitro biofilms.

An in vitro microbial model for producing caries-like lesions on enamel

This study aimed to develop a low cost in vitro viable microbiological model to produce biofilms to be used in dental researches. Single and multi-species biofilms of S. mutans, S. sobrinus, S. mitis, S. salivarius, S. cricetus and S. sanguinis were grown on bovine enamel slabs during 10 days, in a sterile brain-heart infusion broth, containing 5% sucrose and incubated at 37ºC in an atmosphere of 10% CO2. The slabs were transferred to a fresh medium at every 6, 12 or 24 hours. After the experimental period, enamel volume percent mineral was determined by cross-sectional microhardness. Caries-like lesions were found in all bacterial groups when compared with the control group. No statistical significant differences were found between S. mutans and S. sobrinus with respect of their cariogenicity or among the periods of medium change. However, it was found a statistical significant difference among the cariogenicity of S. salivarius and S. sanguinis (ANOVA followed by Tukey test). This model has successfully developed caries-like lesion on enamel and the medium can be changed at every 24 hours utilizing either S. mutans or S. sobrinus.