Multistability and hysteresis in states of oral microbiota: Is it impacting the dental clinical cohort studies?

INTRODUCTION: Microbiome from a "healthy cohort" is used as a reference for comparison to cases and intervention. However, the studies with cohort-based clinical research have not sufficiently accounted for the multistability in oral microbial community. The screening is limited to phenotypic features with marked variations in microbial genomic markers. Herein, we aimed to assess the stability of the oral microbiome across time from an intervention-free "healthy" cohort. METHODS: We obtained 33 supragingival samples of 11 healthy participants from the biobank. For each participant, we processed one sample as baseline (T0) and two samples spaced at 1-month (T1) and 3-month (T2) intervals for 16S ribosomal RNA gene sequencing analysis. RESULTS: We observed that taxonomic profiling had a similar pattern of dominant genera, namely, Rothia, Prevotella, and Hemophilus, at all time points. Shannon diversity revealed a significant increase from T0 (p < .05). Bray Curtis dissimilarity was significant (R = -.02, p < .01) within the cohort at each time point. Community stability had negative correlation to synchrony (r = -.739; p = .009) and variance (r = -.605; p = .048) of the species. Clustering revealed marked differences in the grouping patterns between the three time points. For all time points, the clusters presented a substantially dissimilar set of differentially abundant taxonomic and functional biomarkers. CONCLUSION: Our observations indicate towards the presence of multistable states within the oral microbiome in an intervention-free healthy cohort. For a conclusive and meaningful long-term reference, dental clinical research should account for multistability in the personalized therapy approach to improve the identification and classification of reliable markers.

Effects of incorporating 45S5 bioactive glass into 30% hydrogen peroxide solution on whitening efficacy and enamel surface properties.

OBJECTIVES: This study aimed to evaluate the effects of 30% hydrogen peroxide (HP) solution containing various contents of 45S5 bioactive glass (BAG) on whitening efficacy and enamel surface properties after simulating the clinical bleaching procedure. MATERIALS AND METHODS: A total of 60 bovine enamel specimens discolored with black tea were divided into five groups treated with distilled water (DW), HP, 0.01 wt.% BAG + HP, 1.0 wt.% BAG + HP, and 20.0 wt.% BAG + HP (n = 12). The pH change was observed for 20 min immediately after mixing the experimental solutions, which were applied for 20 min/week, at 37 °C over 21 days. Color, gloss, roughness, microhardness, and micromorphology measurements were conducted before and after bleaching treatment. RESULTS: All groups containing BAG experienced an increase in pH from 3.5 to 5.5 in less than 1 min, and the final pH increased as the BAG content increased. The ΔE of all experimental groups was significantly higher than that of the DW group (p < 0.05), but there were no significant differences between different BAG contents (p > 0.05). Gloss significantly decreased in all experimental groups compared to the DW group, and the increased BAG content had significantly affected the decrease in gloss (p < 0.05). There was no statistical difference in surface roughness (p > 0.05), but hardness increased significantly with BAG content after bleaching treatment (p < 0.05). CONCLUSIONS: HP containing 45S5 BAG showed efficacy in tooth whitening. Also, the pH value of the HP remained acidic near 3.5 for 20 min, while the HP containing the 45S5 BAG showed an increase in pH, which inhibited the demineralization of the enamel surface, and maintained the surface morphology. CLINICAL RELEVANCE: These novel materials are promising candidates to minimize enamel surface damage caused by HP during bleaching procedure in dental clinic.

Effects of 35% hydrogen peroxide solution containing hydrated calcium silicate on enamel surface.

OBJECTIVES: The objectives of this study were to develop a novel bleaching material containing hydrated calcium silicate (hCS) particles and investigate the effects of hCS on the bleaching efficacy, microhardness, and surface morphology of bovine enamel. MATERIALS AND METHODS: To prepare the hCS particles, white Portland cement was mixed with distilled water and ground into a fine powder. The particles in various proportions were then mixed with 35% hydrogen peroxide solution (HP), while HP without hCS was used as a control (HP), and teeth whitening gel was used as a commercial control (CC). Following the thrice application of experimental and control solutions on the discolored bovine enamel surface for 15 min, color change (n = 10), microhardness (n = 10), and micromorphology (n = 2) of the enamel surface were analyzed. RESULTS: The Δ E* of the enamel surface treated with the experimental solution containing hCS was significantly higher than that of the CC, but there were no significant differences between the different hCS contents. The experimental solution containing hCS reduced the percentage of microhardness loss on the enamel surface, and the percentage of microhardness loss significantly decreased as the content of hCS increased (p < 0.05). The erosion pattern was only observed on enamel surfaces treated with HP and CC. CONCLUSIONS: This study suggests that HP containing hCS is effective in bleaching efficacy. In addition, hCS could also minimize the microhardness loss of tooth structure caused by HP and maintain enamel surface morphology. CLINICAL RELEVANCE: This novel bleaching material is promising for inhibiting demineralization and promoting the remineralization of teeth during bleaching treatment in dental clinics.

Accuracy of 3-dimensional printed bracket transfer tray using an in-office indirect bonding system.

INTRODUCTION: In this study, we aimed to evaluate and compare the bracket positioning accuracy of the indirect bonding (IDB) transfer tray fabricated in-clinic using the tray printing (TP) and marker-model printing methods (MP). METHODS: The TP group was further divided into 2 groups (single-tray printing [STP] and multiple-tray printing [MTP]) depending on the presence of a tray split created using the 3-dimensional (3D) software. Five duplicated plaster models were used for each of the 3 experimental groups, and a total of 180 artificial teeth, except the second molar, were evaluated in the experiment. The dental model was scanned using a model scanner (E3; 3Shape Dental Systems, Copenhagen, Denmark). Virtual brackets were placed on facial axis points, and the IDB trays were designed and fabricated using a 3D printer (VIDA; EnvisionTEC, Mich). The accuracy of bracket positioning was evaluated by comparing the planned bracket positions and the actual bracket positions using 3D analysis on inspection software. The main effects and first-order interaction effects were analyzed together by analysis for the analysis of variance. RESULTS: The mean distance and height errors were significantly lower in the STP group than those in the MP and MTP groups (P <0.05). The mean distance error was 0.06 mm in the STP group and 0.09 mm in the MP and MTP groups. The mean height error was 0.10 mm in the STP group and 0.15 mm and 0.18 mm in MP and MTP groups, respectively. However, no significant differences were observed in the angular errors among the 3 groups. CONCLUSIONS: The in-office-fabricated IDB system with computer-aided design and 3D printer is clinically applicable after considering the linear and angular errors. We recommend IDB trays fabricated using the STP method owing to the lower frequency of bracket positioning errors and ease of fabrication.

Time-dependent effects after enamel fluoride application on an acid etching system in orthodontic bracket bonding.

OBJECTIVES: This study investigated the effectiveness of acid etching and bond strength in an orthodontic bonding system in accordance with various time periods after 1.23% acidulated phosphate fluoride (APF) gel applications. MATERIALS AND METHODS: APF gel was applied on the enamel surface of bovine teeth, which were assigned to group F0. The tooth specimens were then immersed in distilled water for 1 (F1), 7 (F7), 14 (F14), 21 (F21) and 28 (F28) days. A group without fluoride pretreatment was a control group. All groups were etched with 35% phosphoric acid and evaluated using a surface hardness tester and scanning electron microscope (SEM). A shear bond strength (SBS) test was performed using a universal testing machine, and the bond failure modes were also examined. RESULTS: After the 1.23% APF gel treatment, the hardness of the acid-etched enamel surface decreased as the immersion period increased. SEM images of the test groups showed etched patterns similar to control group as the immersion period increased. The SBS increased with increasing immersion period, whereas group F21 was not significantly different from the control group. Relatively high adhesive remnant index scores were identified as the ageing period increased. CONCLUSIONS: The present study indicated that in order to obtain suitable bond strength, it is recommended to use an acid etching-based bonding system for the fixation of orthodontic appliances on the enamel surface 21 days after the use of APF gel. CLINICAL RELEVANCE: Fluoride pre-treatment interferes with the acid etching effects on enamel, causing a reduction in the bond strength with orthodontic brackets.

Accuracy of bracket position using thermoplastic and 3D-printed indirect bonding trays.

AIM: The positional accuracy of bracket placement planned through tooth setup vs actual placement was evaluated by means of conventional thermoplastic indirect bonding trays and customized 3D-printed indirect bonding trays. MATERIALS AND METHODS: A total of 280 bracket positions placed on the crowns of 10 dental plaster models were evaluated. The manual setup method and a thermoplastic indirect bonding tray were used for the manual group. For the CAD/CAM group, the bracket was positioned using a digital setup and a corresponding 3D-printed tray. The positional accuracy of the bracket placement on the duplicated gypsum model using the trays was evaluated by means of 3D software. Six errors of bracket position (height, depth, mesiodistal, torque, rotation, and tip errors), including linear and angular errors, were measured. Differences in variables were compared across subgroups using the independent t test or the Mann-Whitney U test. RESULTS: Only the height error differed significantly (P < 0.05) between groups (manual: 0.2 mm; CAD/CAM: 0.12 mm). For both incisors and molars, the manual group showed significantly greater height errors than the CAD/CAM group (P < 0.05). The analysis of variance of the position error to the whole bracket showed statistically significant differences between tooth positions, linear measurements, and angular measurements (P < 0.05). CONCLUSION: A 3D-printed indirect bonding tray showed accuracy similar to that of conventional methods for bracket placement, with slightly greater bracket height accuracy. Further studies should strive to improve accuracy in terms of tooth positions.

Enhanced Osteogenic Differentiation of Human Mesenchymal Stem Cells on Amine-Functionalized Titanium Using Humidified Ammonia Supplied Nonthermal Atmospheric Pressure Plasma.

The surface molecular chemistry, such as amine functionality, of biomaterials plays a crucial role in the osteogenic activity of relevant cells and tissues during hard tissue regeneration. Here, we examined the possibilities of creating amine functionalities on the surface of titanium by using the nonthermal atmospheric pressure plasma jet (NTAPPJ) method with humidified ammonia, and the effects on human mesenchymal stem cell (hMSC) were investigated. Titanium samples were subjected to NTAPPJ treatments using nitrogen (N-P), air (A-P), or humidified ammonia (NA-P) as the plasma gas, while control (C-P) samples were not subjected to plasma treatment. After plasma exposure, all treatment groups showed increased hydrophilicity and had more attached cells than the C-P. Among the plasma-treated samples, the A-P and NA-P showed surface oxygen functionalities and exhibited greater cell proliferation than the C-P and N-P. The NA-P additionally showed surface amine-related functionalities and exhibited a higher level of alkaline phosphatase activity and osteocalcin expression than the other samples. The results can be explained by increases in fibronectin absorption and focal adhesion kinase gene expression on the NA-P samples. These findings suggest that NTAPPJ technology with humidified ammonia as a gas source has clinical potential for hard tissue generation.

Osteogenic Properties of Novel Methylsulfonylmethane-Coated Hydroxyapatite Scaffold.

Despite numerous advantages of using porous hydroxyapatite (HAp) scaffolds in bone regeneration, the material is limited in terms of osteoinduction. In this study, the porous scaffold made from nanosized HAp was coated with different concentrations of osteoinductive aqueous methylsulfonylmethane (MSM) solution (2.5, 5, 10, and 20%) and the corresponding MH scaffolds were referred to as MH2.5, MH5, MH10, and MH20, respectively. The results showed that all MH scaffolds resulted in burst release of MSM for up to 7 d. Cellular experiments were conducted using MC3T3-E1 preosteoblast cells, which showed no significant difference between the MH2.5 scaffold and the control with respect to the rate of cell proliferation (p > 0.05). There was no significant difference between each group at day 4 for alkaline phosphatase (ALP) activity, though the MH2.5 group showed higher level of activity than other groups at day 10. Calcium deposition, using alizarin red staining, showed that cell mineralization was significantly higher in the MH2.5 scaffold than that in the HAp scaffold (p < 0.0001). This study indicated that the MH2.5 scaffold has potential for both osteoinduction and osteoconduction in bone regeneration.

Effect of Heat and Sonic Vibration on Penetration of a Flowable Resin Composite Used as a Pit and Fissure Sealant.

OBJECTIVE: To evaluate penetration of a flowable resin composite into fissures using three different application methods: (1) conventional, (2) heat, and (3) sonic vibration. STUDY DESIGN: Forty-five sound maxillary third molars were divided randomly into three groups (n=15 per group). The occlusal surfaces of the teeth were etched and flowable resin composites were applied into the fissure using the assigned application method. The crowns were sectioned and examined with an optical microscope to assess penetration. In addition, three-point flexural strength was analyzed. RESULTS: The sonic vibration group exhibited significantly greater penetration into the fissure compared with the other test groups (p<0.001). The heat group exhibited greater penetration into the fissure compared with the conventional group (p=0.003). However, three-point flexural strength was similar among all groups (p>0.05). CONCLUSIONS: Sonic vibration and heat increased penetration into fissures. Notably, sonic vibration exhibited the greatest penetration. We found that the application method did not influence the three-point flexural strength.

Evaluation of the marginal and internal discrepancies of CAD-CAM endocrowns with different cavity depths: An in vitro study.

STATEMENT OF PROBLEM: The marginal and internal discrepancies of computer-aided design and computer-aided manufacturing (CAD-CAM) endocrowns are unknown. PURPOSE: The purpose of this in vitro study was to evaluate the marginal and internal discrepancies of endocrowns with different cavity depths by measuring them with microcomputed tomography (µCT). MATERIAL AND METHODS: Endocrowns (n=48) of 2 different cavity depths (2 mm and 4 mm) were fabricated in 2 different chairside CAD-CAM systems (CEREC AC and E4D). A µCT scan was made before and after cementation. For analysis of the marginal and internal discrepancies, reference points were selected in 2-dimensional views of 3 buccolingual cross-sections and 3 mesiodistal cross-sections. To calculate the total discrepancy volume, the µCT sections were reconstructed 3-dimensional views, and changes in volume and surface area were examined. Statistical analysis was performed using 2-way ANOVA with Bonferroni correction (α=.05). RESULTS: An endocrown with a 4-mm cavity showed a larger marginal and internal volume than one with a 2-mm cavity. Cementation did not show significant differences in total discrepancy thickness. Discrepancies on the pulpal floor were largest in other sites. Both chairside CAD-CAM systems showed similar discrepancy in the endocrowns. CONCLUSIONS: Based on the present study, marginal and internal discrepancies increased depending on cavity depth. Cementation did not increase the dimension of the discrepancy between the restoration and the cavity wall. The discrepancy on the pulpal floor appeared to affect these results.

Titanium-Silver Alloy Miniplates for Mandibular Fixation: In Vitro and In Vivo Study.

PURPOSE: Titanium (Ti) alloys have received considerable attention as materials for oral and maxillofacial surgery, which require high mechanical strength, osteosynthesis, and biocompatibility. The objective was to implant miniplates fabricated from commercially pure Ti (CP Ti) and newly developed Ti-silver (Ag) alloy in fractured mandibles of adult dogs after preliminary mechanical and biological characterization. MATERIALS AND METHODS: The surface characteristics, biocompatibility, and pre-osteoblast adhesion and proliferation of CP Ti (grade 3) and Ti-Ag (2 at% Ag) alloys were evaluated. Next, the bending strength of 6- and 8-hole miniplates fabricated from CP Ti and Ti-Ag was compared according to ISO (International Organization for Standardization) 9585. Six-hole miniplates were implanted for 12 weeks in fractured mandibles of adult dogs. The Ag ion concentration in each alloy and implanted bone block with soft tissue was measured by inductively coupled plasma mass spectroscopy after euthanasia according to ISO 10993-12. RESULTS: Precipitated Ag was detected in Ti-Ag by alpha- and beta-phase Ti in x-ray powder diffraction. The biocompatibility with pre-osteoblasts of Ti-Ag and CP Ti was comparable in terms of cytotoxicity, cell adhesion, and proliferation (P > .05). Ti-Ag miniplates had up to 3-fold greater bending strength than CP Ti miniplates (P < .05). An in vivo study showed that CP Ti and Ti-Ag miniplates had comparable soft and hard tissue regeneration ability (P > .05). Ag ions were detected in Ti-Ag alloys and applied mandible blocks. CONCLUSIONS: The results of this study suggest that Ti-Ag alloys can be used to produce miniplates with high mechanical properties, as well as considerable biocompatibility, osteosynthesis ability, and Ag ion-release properties. Further studies, including preclinical investigations, are required to enable clinical use of Ti-Ag bone plates.

Non-thermal atmospheric pressure plasma increased mRNA expression of growth factors in human gingival fibroblasts.

OBJECTIVES: The aim of this in vitro study was to investigate the effects of a non-thermal atmospheric pressure plasma jet (NTAPPJ) on the cellular activity of human gingival fibroblasts (HGF) for possible non-surgical application of it during gingival wound healing. MATERIALS AND METHODS: HGF cells were exposed with NTAPPJ for 1, 2, and 4 min and were investigated for cellular attachment, cell viability, morphology of attached cells, proliferation rate, and messenger ribonucleic acid (mRNA) expression of various growth factors. Also, scavengers for chemicals produced by NTAPPJ were used to identify the chemical species responsible for the effects. RESULTS: There was no significant change in the number of HGF cells attached or their proliferation following NTAPPJ exposure. Also, high cell viability resulted from exposure of all of HGF cells to NTAPPJ for 1, 2, and 4 min. However, cells were more stretched while the mRNA expressions of transforming growth factor and vascular endothelial growth factor were significantly increased following NTAPPJ exposure. Additionally, the scavenger test showed that nitric oxide is likely to be the chemical responsible for an increase of cellular activity. CONCLUSION: The results demonstrated that the NTAPPJ increased mRNA expressions of growth factors in human gingival fibroblasts. CLINICAL RELEVANCE: Application of NTAPPJ would be useful in gingival wound healing in clinics though additional studies confirming the effects would be needed.

Catechol-Functionalized Synthetic Polymer as a Dental Adhesive to Contaminated Dentin Surface for a Composite Restoration.

This study reports a synthetic polymer functionalized with catechol groups as dental adhesives. We hypothesize that a catechol-functionalized polymer functions as a dental adhesive for wet dentin surfaces, potentially eliminating the complications associated with saliva contamination. We prepared a random copolymer containing catechol and methoxyethyl groups in the side chains. The mechanical and adhesive properties of the polymer to dentin surface in the presence of water and salivary components were determined. It was found that the new polymer combined with an Fe(3+) additive improved bond strength of a commercial dental adhesive to artificial saliva contaminated dentin surface as compared to a control sample without the polymer. Histological analysis of the bonding structures showed no leakage pattern, probably due to the formation of Fe-catechol complexes, which reinforce the bonding structures. Cytotoxicity test showed that the polymers did not inhibit human gingival fibroblast cells proliferation. Results from this study suggest a potential to reduce failure of dental restorations due to saliva contamination using catechol-functionalized polymers as dental adhesives.

An Alternative to Annealing TiO2 Nanotubes for Morphology Preservation: Atmospheric Pressure Plasma Jet Treatment.

Titanium oxide nanotube layer formed by plasma electrolytic oxidation (PEO) is known to be excellent in biomaterial applications. However, the annealing process which is commonly performed on the TiO2 nanotubes cause defects in the nanotubular structure. The purpose of this work was to apply a non-thermal atmospheric pressure plasma jet on diameter-controlled TiO2 nanotubes to mimic the effects of annealing while maintaining the tubular structure for use as biomaterial. Diameter-controlled nanotube samples fabricated by plasma electrolytic oxidation were dried and prepared under three different conditions: untreated, annealed at 450 °C for 1 h in air with a heating rate of 10 °C/min, and treated with an air-based non-thermal atmospheric pressure plasma jet for 5 minutes. The contact angle measurement was investigated to confirm the enhanced hydrophilicity of the TiO2 nanotubes. The chemical composition of the surface was studied using X-ray photoelectron spectroscopy, and the morphology of TiO2 nanotubes was examined by field emission scanning electron microscopy. For the viability of the cell, the attachment of the osteoblastic cell line MC3T3-E1 was determined using the water-soluble tetrazolium salt assay. We found that there are no morphological changes in the TiO2 nanotubular structure after the plasma treatment. Also, we investigated a change in the chemical composition and enhanced hydrophilicity which result in improved cell behavior. The results of this study indicated that the non-thermal atmospheric pressure plasma jet results in osteoblast functionality that is comparable to annealed samples while maintaining the tubular structure of the TiO2 nanotubes. Therefore, this study concluded that the use of a non-thermal atmospheric pressure plasma jet on nanotube surfaces may replace the annealing process following plasma electrolytic oxidation.

Air atmospheric-pressure plasma-jet treatment enhances the attachment of human gingival fibroblasts for early peri-implant soft tissue seals on titanium dental implant abutments.

OBJECTIVE: Although dental implants are commonly used for tooth restoration, there is a lack of studies of treatment regimens for preventing extra-oral infection and decreasing osseointegration failures by establishing early peri-implant soft tissue seals on titanium dental implant abutments. In this study, air atmospheric-pressure plasma-jet (AAPPJ) treatment was applied to titanium disks to assay the potential for early peri-implant soft tissue seals on titanium dental implant abutment. MATERIALS AND METHODS: After titanium disks were treated with AAPPJ for 10 s at 250, 500, 1000 and 1500 sccm, surface analysis was performed; the control group received air only or no treatment. Human gingival fibroblasts (HGF) were seeded onto the specimens for evaluating cell attachment and proliferation and adherent-cell morphology was visualized via confocal microscopy. RESULTS: In AAPPJ-treated specimens, the water contact angle decreased according to increased flow rate. Oxygen composition increased in XPS, but no topographical changes were detected. The effect of AAPPJ treatment at 1000 sccm was apparent 2 mm from the treated spot, with a 20% increase in early cell attachment and proliferation. Adherent HGF on AAPPJ-treated specimens displayed a stretched phenotype with more vinculin formation than the control group. CONCLUSIONS: Within the limitations of this study, the results indicate that AAPPJ treatment may enhance the early attachment and proliferation of HGF for establishing early peri-implant soft tissue seals on titanium dental implant abutments with possible favorable effects of osseointegration of dental implant.

Osteoconduction capacity of human deciduous and permanent teeth ash in a rat calvarial bone defect model.

The aim of this study was to confirm the osteoconduction capacities and determine the potential of permanent teeth ash (PTA), and deciduous teeth ash (DTA) as bone substitutes. Rats (n = 71) were divided randomly into four groups: sham, micro macroporous biphasic calcium phosphate (MBCP), PTA, and DTA. A sample of the each group was transplanted into preformed 8-mm calvarial defects (one per rat). The density of new bone was calculated and the crystallinities of the PTA and DTA were analyzed by X-ray diffraction. The degree of new bone formation was high in the MBCP and DTA groups but low in the PTA groups. The DTA was highly crystalline, whereas the PTA was not. The percentages of ß-tricalcium phosphate in the DTA and PTA were 10.7 and 3.7%, respectively. DTA has a high osteoconduction capacity, suggesting that it is a useful bone substitute.

Strain of bone-implant interface and insertion torque regarding different miniscrew thread designs using an artificial bone model.

OBJECTIVES: To evaluate the initial stability of dual-thread miniscrews by analyzing the strain at the bone-implant interface and insertion torque during implantation in artificial bone models with different cortical bone thicknesses. MATERIALS AND METHODS: Insertion torque, and strain, measured with a five-element strain gauge in 1.0, 1.5, and 2.0-mm artificial cortical bone, during insertion of single- (OAS-T1507) and dual-thread (MPlant-U3) type self-drilling miniscrews were assessed. RESULTS: Both dual- and single-thread miniscrews showed greater than 7790 µstrain for all cortical bone thicknesses, and dual-thread miniscrews reached up to 19580 µstrain in 2.00 m m cortical bone. The strain of dual-thread miniscrews increased with increasing cortical bone thicknesses of 1.0-2.0mm. For single-thread miniscrews, the maximum insertion torque was relatively constant, but maximum insertion torque increased significantly in dual-thread groups with increasing cortical bone thicknesses (P < 0.0001). The maximum insertion torque with all cortical bone thicknesses was significantly lower with single- than dual-thread types (P < 0.0001). CONCLUSIONS: Self-drilling dual-thread miniscrews provide better initial mechanical stability, but may cause strain over the physiological bone remodelling limit at the bone-implant interface in thick cortical bone layers.

A comparative study of three cytotoxicity test methods for nanomaterials using sodium lauryl sulfate.

The biocompatibility evaluation of nanomaterials is essential for their medical diagnostic and therapeutic usage, where a cytotoxicity test is the simplest form of biocompatibility evaluation. Three methods have been commonly used in previous studies for the cytotoxicity testing of nanomaterials: trypan blue exclusion, colorimetric assay using water soluble tetrazolium (WST), and imaging under a microscope following calcein AM/ethidium homodimer-1 staining. However, there has yet to be a study to compare each method. Therefore, in this study three methods were compared using the standard reference material of sodium lauryl sulfate (SLS). Each method of the cytotoxicity test was carried out using mouse fibroblasts of L-929 exposed to different concentrations of SLS. Compared to the gold standard trypan blue exclusion test, both colorimetric assay using water soluble tetrazolium (WST) and imaging under microscope with calcein AM/ethidium homodimer-1 staining showed results that were not statistically different. Also, each method exhibited various advantages and disadvantages, which included the need of equipment, time taken for the experiment, and provision of additional information such as cell morphology. Therefore, this study concludes that all three methods of cytotoxicity testing may be valid, though careful consideration will be needed when selecting tests with regard to time, finances, and the amount of information required by the researcher(s).

Air atmospheric pressure plasma jet pretreatment for drop-wise loading of dexamethasone on hydroxyapatite scaffold for increase of osteoblast attachment.

Periodontal disease affects alveolar bone resorption around the involved teeth. To gain bone height, bone graft materials have been widely used with drug carriers. Application of an atmospheric pressure plasma jet (APPJ) treatment is widely studied due to its ability to change surface characteristics without topographical change. The aim of this study is to identify whether the air APPJ (AAPPJ) treatment before drop-wise loading performance could change loaded amount of dexamethasone, and induce increase of cell attachment and proliferation. The results suggested that AAPPJ treatment decreased the contact angle down to about 13 degrees, which increased gradually but significantly lowered at least 4 days compared to no-treated group. After AAPPJ treatment, hydrocarbon was removed with change of zeta potential into positive charge. However, the AAPPJ treatment did not change the quantity or releasing profile of dexamethasone (p > 0.05). Confocal analysis combined with DNA proliferation analysis showed increase of osteoblast attachment and proliferation. Hence, AAPPJ could be a useful pretreatment method before drop-wise loading on HA scaffold with dexamethasone for increase of osteoblast attachment.

Positive control for cytotoxicity evaluation of dental vinyl polysiloxane impression materials using sodium lauryl sulfate.

OBJECTIVES: Vinyl polysiloxane (VPS) is elastomeric dental impression material which, despite having very few reports of adverse reactions, has shown high levels of cytotoxicity that is difficult to be interpreted without referencing to the positive control material. Therefore, in this study, positive control VPS was developed using sodium lauryl sulfate (SLS) for the reference of cytotoxicity test. MATERIALS AND METHODS: The positive control VPS with SLS was formed with a different proportion of SLS (0, 1, 2, 4, 8 and 16 wt%) added to the base. The cytotoxicity test was then carried out using the extractions or dilutions of the extractions from each of the test samples using murine fibroblast cells (L929). RESULTS: The final product of positive control VPS behaved similar to commercially available VPS; being initially liquid-like and then becoming rubber-like. Ion chromatography showed that the level of SLS released from the product increased as the proportion of added SLS increased, consequently resulting in an increased level of cytotoxicity. Also, the commercially available VPS was less cytotoxic than the positive control VPS with more or equal to 2 wt% of SLS. However, even the VPS with the highest SLS (16 wt%) did not cause oral mucosa irritation during the animal study. CONCLUSIONS: The positive control VPS was successfully produced using SLS, which will be useful in terms of providing references during in vitro cytotoxicity testing.