Intraoral Exposure Research Articles

Physical and chemical changes of clear aligners after thermoforming and intraoral exposure

Physical and chemical changes of clear aligners after thermoforming and intraoral exposure

Effect of use of antiseptics and fluorides during orthodontic treatment on working properties of NiTi archwires in levelling dental arches : Arandomized controlled trial.

The goal was to investigate whether the application of antiseptics and fluorides during orthodontic treatment affects the biomechanics of the levelling of dental arches by changing the working properties of nickel-titanium (NiTi) archwires. The sample consisted of 60individuals aged 12-22years (53% females). There were 20individuals in each experimental group: (I)individuals conducted regular oral hygiene, (II)individuals used ahigh concentration of fluorides for intensive prophylaxis for the first month; and (III)individuals used chlorhexidine in the same manner. NiTi alloy archwires (dimensions 0.508 × 0.508 mm) were analyzed 3months after intraoral exposure and compared to as-received NiTi wires. Elastic modulus, yield strength, springback ratio, and modulus of resilience were calculated. Dimensions of the dental arches were analyzed when NiTi alloy (T1) was placed intraorally and after 3months (T2). Change was quantified as the difference in dimensions (T2-T1). Anterior width-to-length ratio was used as ameasure for dental arch shape. Intraoral exposure reduced elastic modulus, yield strength, springback ratio, modulus of resilience, loading, and unloading forces of NiTi wires (p ≤ 0.021). Chlorhexidine mouthwash and gel with high concentration of fluorides did not change these properties more than saliva with regular hygiene. The amount of change of dental arch shape in the maxilla and mandible did not differ significantly between the experimental groups. Using antiseptics or ahigh concentration of fluorides during orthodontic treatment does not significantly affect the mechanical properties of NiTi wires and would not have clinical implications in changing orthodontic biomechanics.

Do Oral Antiseptics Affect the Force Degradation of Elastomeric Chains?

Objective: This study aimed to compare the force degradation of elastomeric chains submerged in commonly used mouthrinses. Methods: One hundred samples of elastomeric orthodontic chains from five different brands (Ormco, GC, RMO, Forestandent, and 3M Unitek) were initially activated on double length, and the force was measured with a universal mechanical testing machine. Then, elastomeric modules were thermocycled and immersed into four different mouthrinses: Octenident, Vitis Orthodontic, Perio Plus+, and Listerine through a total number of three cycles which simulated 30 days of intraoral exposure. Force decay was measured after each cycle. Results: All specimens showed statistically significant force degradation over the tested period (p < 0.001). After thermocycling and immersion in oral antiseptics, the lowest measured force was found in Forestadent EOC in Listerine with a median of 70 cN (70–75 cN) and Vitis orthodontic with a median of 70 cN (70–80 cN). On the contrary, the least prone to force reduction was the control group of Ormco 280 cN (275–285 cN) and RMO 280 cN (270–280 cN). Conclusions: Elastomeric chains’ force degradation could be exacerbated by the use of mouth rinses. These data could be beneficial in choosing the appropriate combination of elastomeric chain and mouthrinse for optimal results of orthodontic therapy.

Effect of Real-Time Environment on Mechanical Properties of Preformed Stainless Steel Archwires: An In Vivo Study.

Clinicians should be aware of any effect the oral environment may have on archwires. Laboratory models fail to closely imitate intraoral conditions. The aim was to evaluate the change in mechanical properties of preformed stainless steel archwires after 15 weeks of exposure to the oral environment. Three commercially manufactured 0.019 × 0.025″ stainless steel archwires were evaluated. Young's modulus, yield strength, spring factor, and hardness were studied. The unexposed distal end cuts (control samples) and archwires were tested after 15 weeks of intraoral exposure (test samples). Tension tests, Vickers microhardness tests, and nanoindentation tests were carried out. Normality was tested using the Shapiro-Wilk test. Statistical analyses included the paired t-test for intragroup comparisons and Kruskal-Wallis ANOVA with the post hoc Dunn test for comparison of mean percentage reduction in values. At T15, Young's modulus showed a statistically significant decrease. Changes in yield strength and spring factor were not significant for groups other than American Orthodontics wires. The reduction in hardness was significant in 3M Unitek. Vickers, tension, and nanoindentation tests demonstrated an expansive range between hardness and Young's modulus so determined. 3M Unitek archwires showed the highest difference in Young's modulus. Yield strength values increased in Ortho Organizers archwires. Spring factor decreased only in 3M Unitek archwires. Hardness values obtained from various tests did not produce identical results.

In situ evaluation of microbial profile formed on Ti-6Al-4V additive manufacturing disks: 16S rRNA sequencing

Statement of problemDental implants obtained by additive manufacturing may present changes in the microbiome formed. However, studies profiling the microbial communities formed on Ti-6Al-4V are lacking. PurposeThe purpose of this in situ study was to characterize the profile of the microbial communities formed on Ti-6Al-4V disks produced by additive manufacturing and machining. Material and methodsTitanium disks produced by additive manufacturing (AMD) and machining (UD) were housed in the buccal region of removable intraoral devices. These devices containing both disks were used by eight participants for 96 hours. After every 24 hours of intraoral exposure, the biofilm that had formed on the disks was collected. The 16S rRNA genes from each specimen were amplified and sequenced with the Miseq Illumina instrument and analyzed. Total microbial quantification was evaluated by analysis of variance-type statistics using the nparLD package. The Wilcoxon test was used to evaluate alpha diversity (α=.05). ResultsA difference was found in the microbial communities formed on additively manufactured and machined disks, with a reduction in operational taxonomic units (OTUs) for the AMD group compared with the UD group. Firmicutes and Proteobacteria were the most abundant phyla. Of the 1256 genera sequenced, Streptococcus predominated on both disks. ConclusionsThe microbiome of the biofilm formed on the Ti-6Al-4V disks was significantly influenced by the fabrication method. The AMD disks showed lower total microbial counts than the UD disks.

In vivo aging-induced surface roughness alterations of Invisalign® and 3D-printed aligners.

To assess the surface roughness of in-house 3D-printed orthodontic aligners compared with Invisalign® appliances, both retrieved as well as in the 'as-received' control status. An in vitro study following intra-oral material aging. Twelve clinically used Invisalign® appliances and the same number of 3D-printed aligners, without involvement of attachments, were obtained from a respective number of patients. A similar number of 'as-received' aligners, of each material, were used as control (CON) groups. Four groups of materials were examined: A = Invisalign® CON; B = Invisalign® used; C = 3D-printed CON; and D = 3D-printed used. Optical profilometry was employed to examine the following surface roughness parameters: amplitude parameters Sa, Sq and Sz and functional parameters Sc and Sv. Descriptive statistics and quantile regression modeling were conducted, and the level of statistical significance was set at α = 0.05. Intra-oral exposure of 3D-printed aligners was significantly associated with increase in all tested parameters (P < 0.001 at all occasions). Significant differences were detected in the retrieved 3D-printed aligners compared with Invisalign® retrieved, with the exception of Sz. The respective effect sizes (median differences) were as follows: Sa: 169 nm, 95% confidence interval [CI] = 89-248, P < 0.001; Sq: 315 nm, 95% CI = 152-477, P < 0.001; Sc: 233 nm3/nm2, 95% CI = 131-335, P < 0.001; and Sv: 43 nm3/nm2, 95% CI = 17-68, P = 0.002. Within the limitations of this study, we concluded that surface roughness differences existed between 3D-printed aligners and Invisalign® in the retrieved status, as well as between the control and retrieved 3D-printed groups.

Manual and mechanical stripping-induced enamel roughness and elemental composition in vivo.

Interproximal enamel reduction (IPR) is routinely used in orthodontics to generate small to moderate amounts of space within the dental arch. Aim of this ex vivo study was to evaluate the effect of two different IPR systems on the enamel surface's waviness, roughness, and elemental composition after 6 months of intraoral exposure. Fifteen orthodontic extraction patients were included in the present study. The 39 healthy premolars, which were scheduled to be extracted, were subjected to IPR at least 6 months before their extraction. IPR was performed on their mesial side with two different methods: (1) instrumented method with the Ortho-Strips system (on handpiece) and (2) manually with the Intensiv ProxoStrip (strips)-each with four different grits for contouring, finishing, and polishing. The distal side of each premolar served as its own internal control. Treated and untreated tooth surfaces were evaluated by optical profilometry, Raman, and scanning electron microscope/X-ray energy-dispersive (EDX) analyses. Data were analysed with descriptive statistics and generalized linear models at alpha = 5%. Both IPR methods significantly reduced the waviness of the enamel surface (P < 0.001), with manual IPR leading to smaller waviness reductions than the instrumented IPR (P ≤ 0.001). On the other side, both IPR methods led to a significant increase in enamel surface roughness (P < 0.001), with no significant differences between IPR methods. EDX and Raman analyses did not demonstrate any alterations on elemental composition of enamel after at least 6 months of intraoral exposure. Both stripping systems led to a flatter but rougher enamel surface. Further polishing is needed to restore the initial enamel smoothness. The elemental composition of the stripped enamel returns to the baseline level after 6 months of intraoral exposure.

Ultrastructure of the Dentin Pellicle and the Impact of Erosion

While the ultrastructure of the enamel pellicle and its erosion protective properties are well studied, the dentin pellicle is still neglected in dental research. Therefore, the ultrastructure and erosion protective properties of a pellicle formed on bovine dentin specimens were investigated in the present study. The dentin pellicle was formed in situ for 3, 30, 120, and 360 min at buccal or palatal oral sites of 3 subjects and analyzed by transmission electron microscopy. In order to clarify the impact of an erosive challenge to the ultrastructure of the pellicle and the underlying dentin, specimens were exposed to the oral cavity and eroded in vivo with 0.1% or 1% citric acid either immediately or after 30 min of pellicle formation. Specimens that were eroded without exposure to the oral cavity served as control. In another trial, specimens with a 30-min pellicle were exposed to the oral cavity for a further 60 min after the erosive challenge to investigate the effect of saliva on the impaired pellicle and dentin. Transmission electron micrographs reveal a globular and granular structured pellicle layer, which was thicker when the pellicle was formed buccally or with longer formation times. Erosion with citric acid reduced the thickness of the pellicle and interrupted its continuity. The dentin was also affected by erosion, which was represented by a lower electron density and formation of demineralized lacunae. These were infiltrated by a granular structured material when specimens were exposed to the oral cavity. After further intraoral exposure, the infiltration was more pronounced, indicating a significant impact of saliva on the demineralized dentin. A reformation of the dentin pellicle on the other hand did not occur. In conclusion, the dentin pellicle is neither acid-resistant nor able to effectively protect dentin from erosion.

Leaching from a 3D-printed aligner resin.

To quantitatively assess the degree of conversion and the water-leaching targeted compound from 3D-printed aligners. 3D-printed aligners were made of photopolymerized resin (Tera Harz TC85A). The molecular structure and degree of conversion of the set resin were investigated by ATR-FTIR spectroscopy (n = 5). The aligners (n = 10) were immersed in double distilled water for 1 week at 37°C and the eluents were analysed using liquid chromatography/mass spectrometry methods (LC-ESI-MS/MS for urethane dimethacrylate [UDMA] and LC-APCI-MS/MS for bispenol-A [BPA]). The resin was composed of aliphatic vinyl ester-urethane monomers, with acrylate and/or methacrylate functionalization. The degree of conversion was estimated as to 83%. There was no detection of BPA in any of the assessed samples (0.25 µg/l). Quantifiable amounts of UDMA were detected in all the exposed samples, ranging from 29 to 96 µg/l. Although efficiently polymerized and BPA free, the great variability in the amount of UDMA monomer leached from the examined samples may raise concerns on potential health hazards after repeated intraoral exposure, which is indicated for this class of materials.

The Value of Three-Dimensional Modeling on Repositioning and Soft Tissue Filling in Microsurgical Reconstruction of Secondary Mandibular Defects: A Retrospective Study.

PurposeSecondary mandibular defect reconstruction is a challenging operation. It is performed rather rarely, as in most of the cases a primary reconstruction is preferable. Restoration of function and symmetry, in secondary reconstruction of mandibular defects, requires accurate repositioning of segments and appropriate soft tissue filling. The purpose of this study is to estimate the value of three-dimensional (3D) digital modeling to meet the above requirements, in secondary microsurgical reconstruction of mandibular defects.MethodsFive cases of mandibular defects, with various degrees of asymmetry and dysfunction, were restored secondarily, with fibula flaps, after virtual reconstruction on a 3D model derived from their computed tomography, with the utilization of CAD-CAM software. Software reproduced symmetrical 3D models by mirroring the healthy side. Occlusion was restored by fine adjustments on 3D models and finally a reconstruction plate was pre-bent on them, prior to its sterilization for use in surgery. Three out of the five cases received an osteo-cutaneous flap, while in the other two patients, an osseous flap was used. Lower face symmetry, mouth opening, and main patient’s complaints were evaluated pre- and postoperatively to assess the value of the 3D modeling.ResultsAll flaps survived. Hematoma occurred in two patients and was resolved after evacuation. In two patients, a fistula observed, was attributed to screw loosening, and treated after a surgical debridement and screw replacement. Intraoral exposure in the posterior part of the fibula flap was recorded in one patient, possibly from wound dehiscence due to tension of the intraoral tissue, and successfully covered with an ipsilateral nasolabial flap. The resulting symmetry and function were satisfactory in all the patients.ConclusionSecondary mandibular defects are often related with cosmetic disfigurement, misalignment of mandibular segments, and mandibular malfunction. For the correction of mandibular continuity, symmetry, and restoration of function, preoperative 3D modeling may be an important tool, according to our results.

Differences in Metal Ions Released from Orthodontic Appliances in an In Vitro and In Vivo Setting

The aim was to assess the composition of released metal ions from fixed orthodontic appliances both in an in vitro and in vivo setting and to compare their quantities to evaluate any possible health risks associated with them. For the in vitro setting, a set of 24 as-received sterile SS brackets, 2 SS sterile archwires, and 2 NiTi sterile archwires were aged for 90 days in artificial saliva, and released metal ion concentrations were determined by inductively coupled plasma mass spectrometry (ICP-MS), using matrix-matched standards for calibration. For the in vivo setting, 15 brackets, 15 SS, and 15 NiTi archwires were retrieved after 90 days of intraoral exposure in 15 subjects. Debris composition on each part of the orthodontic appliance was assessed using scanning electron microscopy with energy dispersive spectroscopy (SEM/EDS). The present study evidenced a difference in the composition of released metal ions from fixed orthodontic appliances detected in an in vitro and in vivo setting. Generally, the relative content of metal ions was higher in the retrieved debris than that released in artificial saliva. The concentration of released metal ions from all tested alloys was below the upper recommended limit of daily intake; however, possible local effects in terms of hypersensitivity due to their accumulation in the debris cannot be excluded even with such concentrations.

Bioadhesion on Textured Interfaces in the Human Oral Cavity-An In Situ Study.

Extensive biofilm formation on materials used in restorative dentistry is a common reason for their failure and the development of oral diseases like peri-implantitis or secondary caries. Therefore, novel materials and strategies that result in reduced biofouling capacities are urgently sought. Previous research suggests that surface structures in the range of bacterial cell sizes seem to be a promising approach to modulate bacterial adhesion and biofilm formation. Here we investigated bioadhesion within the oral cavity on a low surface energy material (perfluorpolyether) with different texture types (line-, hole-, pillar-like), feature sizes in a range from 0.7–4.5 µm and graded distances (0.7–130.5 µm). As a model system, the materials were fixed on splints and exposed to the oral cavity. We analyzed the enzymatic activity of amylase and lysozyme, pellicle formation, and bacterial colonization after 8 h intraoral exposure. In opposite to in vitro experiments, these in situ experiments revealed no clear signs of altered bacterial surface colonization regarding structure dimensions and texture types compared to unstructured substrates or natural enamel. In part, there seemed to be a decreasing trend of adherent cells with increasing periodicities and structure sizes, but this pattern was weak and irregular. Pellicle formation took place on all substrates in an unaltered manner. However, pellicle formation was most pronounced within recessed areas thereby partially masking the three-dimensional character of the surfaces. As the natural pellicle layer is obviously the most dominant prerequisite for bacterial adhesion, colonization in the oral environment cannot be easily controlled by structural means.

Influence of periodic milk or cream treatment on the anti-erosive potential of the acquired enamel pellicle

ObjectiveThe purpose of the present study was to assess the anti-erosive potential of the acquired enamel pellicle formed in situ under the influence of periodic milk or cream treatment. MethodsThe pellicle was formed on bovine enamel specimens in the oral cavity at buccal and palatal sites of upper molars in 6 subjects, using removable acrylic splints. During 6-h of intraoral exposure, splints were removed from the oral cavity every 25 min, treated with milk or cream for 5 min, and subsequently re-inserted into the oral cavity. After 6 h, pellicle covered specimens were immersed in citric acid (0.1 or 1.0 %) for 1 min, and processed for measurement of surface microhardness, determination of calcium release by atomic absorption spectroscopy, scanning and transmission electron microscopy. Statistical analysis was performed with SAS. ResultsStatistical analysis did not indicate major differences between erosive surface alterations on enamel specimens covered by pellicles treated with cream or milk, and those covered by control pellicles. In addition, TEM analysis did not reveal any differences concerning the ultrastructure of the different pellicle treatments during acid exposure. All pellicles were dissolved in part after exposure to 0.1 % citric acid and were nearly completely removed after treatment with 1.0% citric acid. ConclusionsIt is concluded that periodic treatment with milk or cream during pellicle formation in situ does not improve the protective potential of the acquired enamel pellicle against erosion. Clinical significanceModification of the pellicle by consumption of milk or cream prior to an acidic challenge cannot sufficiently protect enamel from erosion.

Influence of intraoral application of antiseptics and fluorides during orthodontic treatment on corrosion and mechanical characteristics of nickel-titanium alloy in orthodontic appliances.

To explore whether the commercial agents recommended for controlling dental biofilm formation had a significant effect in vivo on mechanical and corrosion properties of nickel-titanium (NiTi) alloy. NiTi archwires (dimensions 0.508 × 0.508 mm) were collected from 36 orthodontic patients aged 13-42 years after a 3-month intraoral exposure. Three experimental groups were formed: (1) subjects conducting regular oral hygiene, (2) subjects who used fluorides for intensive prophylaxis for the first month, and (3) subjects who used chlorhexidine in the same manner. Corrosion behavior, surface characteristics, stiffness, hardness, and friction were analyzed. Exposure to intraoral conditions significantly reduced the stiffness and hardness of the NiTi alloy (P ≤ .015). Fluoride tended to reduce stiffness and hardness more than did saliva or antiseptic, but not significantly. Roughness and friction were not significantly influenced by oral exposure. Intraoral aging predominantly produced general corrosion independent of the adjuvant prophylactic agent, although localized corrosion may also have occurred. Fluorides and the antiseptic chlorhexidine do not increase corrosion more than saliva itself, nor do they further modify the mechanical properties of the NiTi alloy.

Effects of Experimental Agents Containing Tannic Acid or Chitosan on the Bacterial Biofilm Formation in Situ.

Chitosan and tannic acid are known for their antibacterial properties. In the present in-situ study, their antibacterial and anti-adherent effects on biofilm formation on enamel were investigated. Six subjects carried upper jaw splints with bovine enamel specimens, allowing in-situ biofilm formation. During the two-day trial, subjects rinsed with experimental solutions that contained either chitosan, tannic acid (pH = 2.5), tannic acid (pH = 7) or hydrochloric acid. Water served as the negative and chlorhexidine as the positive control. Rinsing occurred four or five times following two different rinsing protocols to investigate both the immediate and long-lasting effects. After 48 h of intraoral exposure, the dental plaque was stained with LIVE/DEAD® BacLight, and fluorescence micrographs were evaluated by using the software ImageJ. The results were verified by scanning electron microscopy. Rinsing with chitosan resulted in little immediate antibacterial and anti-adherent effects but failed to show any long-lasting effect, while rinsing with tannic acid resulted in strong immediate and long-lasting effects. Except for a slightly lower antibacterial effect, the neutral solution of tannic acid was as good as the acidic solution. Hydrochloric acid showed neither an antibacterial nor an anti-adherent effect on dental biofilm formation. Experimental solutions containing tannic acid are promising anti-biofilm agents, irrespective of the pH values of the solutions. Chitosan, on the other hand, was not able to prevent biofilm formation.

Transbuccal Approach in Management of Mandible Angle Fracture.

With expanding dimensions and an eminent member of trauma team many Ear Nose Throat specialists are exposed to Oro-maxillofacial trauma. Mandibular angle is a difficult region to operate owing to its unique anatomy. Anglefractures form around 30-40% of the mandibular fracture. Open reduction and internal plating is the management of choice in most angle fractures and various approaches have been described in literature. We conducted a study of trans-buccal approach for management of mandibular angle fractures. It was a non-randomised observational study. 57 patients of mandible angle fractures were included in the study. The ORIF was done in these pts under general anaesthesia after careful surgical planning using trans-buccal approach. This approach included an intraoral exposure with stab incision for the trans-buccal passage of drill and screw diver. Out of 57 cases in our study 33 were males and 24 were females. 23/57 patients were in age group 25-35years whereas 19/57 patients were in age group 15-25years All 57 patients had good fracture healing. Occlusion dysfunction was seen in only 1 case. Infection was seen 3 cases while intraoral exposure of plates occurred in 2 cases. Average mouth opening was 43.3mm at 6weeks with progressive improvement on follow up. Extraoral scar healed well with very good cosmesis in all cases. We strongly suggest the use of transbuccal approach for ORIF in mandibular angle fractures owing to the ease of procedure, comfort of surgeon and less complication rate.

Evaluation of Surface Roughness and Microbial Biofilm Adhesion of Different Orthodontic Arch-wires

Objectives: To assess the capability of different orthodontic arch-wires to retain oral biofilm and to correlate the adhesion to surface roughness of the wires. Methods: Four types of orthodontic arch- wires were used for the study, Nickel-titanium (NiTi), Copper nickel-titanium (Cu-NiTi) , Beta Titanium (TMA ) & Beta III Titanium (CNA) new arch wires and 4 weeks after intraoral usage, were examined for Surface roughness (SR) using an atomic force microscope (AFM). Adhesion of Streptococcus mutans (MS), Staphylococcus aureus (SA), and Candida albicans (CA) were performed using colony count method. Statistically, the following tests were done: analysis of variance, Pearson correlation coefficient test, post hoc Tukey test. Results: The four wire types showed significant increases in SR (P˂ 0.05) after 4 weeks of intra-oral usage, TMA wires recorded the highest roughness values while the lowest ones were for NiTi wires. Bacterial adhesion was detected on all wires, ANOVA showed significant differences between the wires concerning MS, SA and CA adhesion. A significant positive correlation (P=.001) was observed between bacterial adhesion and surface roughness after intra-oral exposure. Conclusions: SR of the wires increased after intra-oral use and there was a positive correlation with the biofilm adhesion.

Thickness of orthodontic clear aligners after thermoforming and after 10\u2009days of intraoral exposure: a prospective clinical study

BackgroundClear aligners (CA) are among the most chosen orthodontic therapies for patients who require an invisible treatment. Previous studies showed that the thermoforming process and the complexity of the intraoral environment might alter the properties of these devices. The aim of the current prospective clinical study was to assess the thickness changes of the CA after 10 days of intraoral use. The secondary aim was to assess the reproducibility of the thermoforming process, in terms of aligner thickness.Materials and methodsCA from 18 consecutive patients (13 women, 5 men, mean age 28.8 ± 9.6 years) were investigated. Before intraoral exposure (T0), the thickness of the unused CA was measured at different occlusal points on a 3D model with a dedicated software (Geomagic Qualify 2013; 3D Systems, Rock Hill, SC, USA). Two CA configurations were studied: passive maxillary aligner (P—no tooth movement; no shape for attachments) and active maxillary aligner (A—tooth movement; shape for attachments and divot). The used aligners were returned after 10 days (T1) and the thickness measurements were repeated. A Student’s t test for paired data (T1 vs. T0) was applied to compare the thicknesses of used and unused devices (significance level after Bonferroni correction for multiple comparison was set at p < 0.0014). Furthermore, to study the reproducibility of the thermoforming process, P and A aligners were thermoformed twice, and the thicknesses of the two unused thermoformed devices were compared by means of Student’s t test for paired data (significance level after Bonferroni correction for multiple comparison was set at p < 0.0014) and Dahlberg’s error.ResultsThe thermoforming process showed good reproducibility for both aligner configurations, with a maximum Dahlberg’s error of 0.13 mm. After intraoral use, the thickness of P showed some statistically significant, but not clinically relevant, thickness changes as compared to the unused aligners, while A did not show any significant changes.ConclusionConsidering the thickness changes, the thermoforming process is reliable both with active and passive aligner configurations. Also, the CA examined show good thickness stability after physiological intraoral ageing in a population of healthy adults.

Activity and distribution pattern of enzymes in the in-situ pellicle of children

ObjectiveThis study investigated, for the first time, pellicle enzymes with respect to their activity, distribution and fluorescence pattern in children with different caries experience. DesignIn-situ pellicles were collected from 41 children (aged 4–6 years) with different caries status; 17 of them were caries-free (dmf = 0), 12 had dental restorations but no current caries (dmf ≥ 2) and 12 had at least two carious lesions (dmf ≥ 2). Bovine enamel samples were fixed on individual upper jaw braces for pellicle formation. After 30 min of intraoral exposure, the pellicle and saliva samples were analysed for the activities of amylase, lysozyme, peroxidase and glucosyltransferase (GTF). The distribution of these enzymes, including GTF-isoforms B, C and D, and the pellicle ultrastructure were examined by gold-immunolabelling and transmission electron microscopy (TEM). Furthermore, interactions between pellicle enzymes and adherent bacteria were visualised using combined fluorescence and immunofluorescence labelling. ResultsThere were no significant differences in the pellicle enzyme activities between the study groups. TEM analysis revealed the absence of GTF C and D in the pellicle of caries-active children. Amylase, peroxidase and GTF-isoforms showed a random distribution within the pellicle layer; lysozyme was found in the form of clusters. A similar ultrastructural pattern was observed for all subjects. Fluorescence labelling technique enabled visualisation of all enzymes, except for GTF B. ConclusionPellicle enzyme activities and ultrastructure are not associated with children’s caries status. Further investigation is needed to assess the influence of individual GTF-isoforms on caries susceptibility in children.

Oil droplet formation on pellicle covered tooth surfaces studied with environmental scanning electron microscopy.

Lipophilic components are known to modulate the process of bioadhesion on the tooth surface. However, the presence of lipid droplets at the acquired pellicle under oral conditions has not been demonstrated, yet. The purpose of the present study was to establish a method for direct visualisation of lipids on the surface of hydrated, pellicle covered tooth samples by environmental scanning electron microscopy (ESEM), and to use this technique for studying the effects of rinsing with edible oils on the acquired pellicle under in vivo conditions. In situ pellicle formation was performed by 3 min exposure of enamel and dentin specimens in the oral cavity of volunteers. Subsequently, the volunteers rinsed in vivo with safflower oil or linseed oil for 30 s, and the specimens were further carried intraorally for periods from 0 min up to several hours. After intraoral exposure the specimens were treated by osmium tetroxide vapour, and were subsequently analysed by ESEM. This technique was capable to directly visualise the presence of lipid droplets at the pellicle's surface under hydrated conditions. ESEM analyses revealed that surface bound nano- and micro-sized lipid droplets were present at the acquired pellicle's surface even several hours after rinsing with edible oils indicating that these droplets had tightly adhered to the pellicle surface. Pellicle modification by edible oil rinsing as demonstrated in the present study might have the potential to be beneficial as an adjunct in dental prophylaxis.