Genetic information such as DNA sequences has been limited to fully explain mechanisms of gene regulation and disease process. Epigenetic mechanisms, which include DNA methylation, histone modification and non-coding RNAs, can regulate gene expression and affect progression of disease. Although studies focused on epigenetics are being actively investigated in the field of medicine and biology, epigenetics in dental research is at the early stages. However, studies on epigenetics in dentistry deserve attention because epigenetic mechanisms play important roles in gene expression during tooth development and may affect oral diseases. In addition, understanding of epigenetic alteration is important for developing new therapeutic methods. This review article aims to outline the general features of epigenetic mechanisms and describe its future implications in the field of dentistry.

While it is reasonably well known that certain dental procedures increase the temperature of the tooth's surface, of greater interest is their potential damaging effect on the pulp and tooth-supporting tissues. Previous studies have investigated the responses of the pulp, periodontal ligament, and alveolar bone to thermal irritation and the temperature at which thermal damage is initiated. There are also many in vitro studies that have measured the temperature increase of the pulp and tooth-supporting tissues during restorative and endodontic procedures. This review article provides an overview of studies measuring temperature increases in tooth structures during several restorative and endodontic procedures, and proposes clinical guidelines for reducing potential thermal hazards to the pulp and supporting tissues.

Early detection of carious lesions increases the possibility of treatment without the need for surgical intervention. Optical coherence tomography (OCT) is an emerging three-dimensional imaging technique that has been successfully used in other medical fields, such as ophthalmology for optical biopsy, and is a prospective candidate for early caries detection. The technique is based on low coherence interferometry and is advantageous in that it is non-invasive, does not use ionizing radiation, and can render three-dimensional images. A brief history of the development of this technique and its principles are discussed in this paper. There have been numerous studies on caries detection, which were mostly in vitro or ex vivo experiments. Through these studies, the feasibility of OCT for caries detection was confirmed. However, further research should be performed, including in vivo studies of OCT applications, in order to prove the clinical usefulness of this technique. In addition, some technological problems must be resolved in the near future to allow for the use of OCT in everyday practice.

During a composite resin restoration, an anticipating contraction gap is usually tried to seal with low-viscosity resin after successive polishing, etching, rinsing and drying steps, which as a whole is called rebonding procedure. However, the gap might already have been filled with water or debris before applying the sealing resin. We hypothesized that microleakage would decrease if the rebonding agent was applied before the polishing step, i.e., immediately after curing composite resin. On the buccal and lingual surfaces of 35 extracted human molar teeth, class V cavities were prepared withthe occlusal margin in enamel and the gingival margin in dentin. They were restored with a hybrid composite resin Z250 (3M ESPE, USA) using an adhesive AdperTM Single Bond 2 (3M ESPE). As rebonding agents, BisCover LV (Bisco, USA), ScotchBond Multi-Purpose adhesive (3M ESPE) and an experimental adhesive were applied on the restoration margins before polishing step or after successive polishing and etching steps. The infiltration depth of 2% methylene blue into the margin was measured using an optical stereomicroscope. The correlation between viscosity of rebonding agents and mciroleakage was also evaluated. There were no statistically significant differences in the microleakage within the rebonding procedures, within the rebonding agents, and within the margins. However, when the restorations were not rebonded, the microleakage at gingival margin was significantly higher than those groups rebonded with 3 agents (p < 0.05). The difference was not observed at the occlusal margin. No significant correlation was found between viscosity of rebonding agents and microleakage, except very weak correlation in case of rebonding after polishing and etching at gingival margin (r = -0.326, p = 0.041).

This study evaluated the influence of the type of restoration and the amount of interdental spacing on the stress distribution in maxillary central incisors restored by means of porcelain laminate veneers and direct composite resin restorations.

Three-dimensional finite element models were fabricated to represent different types of restorations. Four clinical situations were considered. Type I, closing diastema using composite resin. Labial border of composite resin was extended just enough to cover the interdental space; Type II, closing diastema using composite resin without reduction of labial surface. Labial border of composite resin was extended distally to cover the half of the total labial surface; Type III, closing diastema using composite resin with reduction of labial surface. Labial border of the preparation and restored composite resin was extended distally two-thirds of the total labial surface; Type IV, closing diastema using porcelain laminate veneer with a feathered-edge preparation technique. Four different interdental spaces (1.0, 2.0, 3.0, 4.0 mm) were applied for each type of restorations.

For all types of restoration, adding the width of free extension of the porcelain laminate veneer and composite resin increased the stress occurred at the bonding layer. The maximum stress values observed at the bonding layer of Type IV were higher than that of Type I, II and III. However, the increasing rate of maximum stress value of Type IV was lower than that of Type I, II and III.

The purpose of this study was to evaluate the pulp tissue reaction to direct pulp capping of mechanically exposed beagle dogs'pulp with several capping materials. A total of 36 teeth of 2 healthy beagle dongs were used. The mechanically exposed pulps were capped with one of the followings: (1) Mineral Trioxide Aggregate (MTA: ProRoot® MTA, Dentsply, Tulsa, USA), (2) Clearfil SE Bond (Dentin adhesive system: Kuraray, Osaka, Japan), (3) Ultra-Blend (Photo-polymerized Calcium hydroxide: Ultradent, South Jordan, USA), (4) Dycal (Quick setting Calcium hydroxide: LD Caulk Co., Milford, USA) at 7, 30, and 90 days before sacrificing. The cavities were restored with Z350 flowable composite resin (3M ESPE, St. Paul. MN, USA). After the beagle dogs were sacrificed, the extracted teeth were fixed, decalcified, prepared for histological examination and stained with HE stain. The pulpal tissue responses to direct pulp capping materials were assessed.

In MTA, calcium hydroxide, and photo-polymerized calcium hydroxide groups, initial mild inflammatory cell infiltration, newly formed odontoblast-like cell layer and hard tissue bridge formation were observed. Compared with dentin adhesive system, these materials were biocompatible and good for pulp tissue regeneration.

In dentin adhesive system group, severe inflammatory cell infiltration, pulp tissue degeneration and pulp tissue necrosis were observed. It seemed evident that application of dentin adhesive system in direct pulp capping of beagle dog teeth cannot lead to acceptable repair of the pulp tissue with dentine bridge formation.

The aim of this study was to measure the dentinal tubular fluid flow (DFF) during and after amalgam and composite restorations. A newly designed fluid flow measurement instrument was made. A third molar cut at 3 mm apical from the CEJ was connected to the flow measuring device under a hydrostatic pressure of 15 cmH₂O. Class I cavity was prepared and restored with either amalgam (Copalite varnish and Bestaloy) or composite (Z-250 with ScotchBond MultiPurpose: MP, Single Bond 2: SB, Clearfil SE Bond: CE and Easy Bond: EB as bonding systems). The DFF was measured from the intact tooth state through restoration procedures to 30 minutes after restoration, and re-measured at 3 and 7days after restoration.

Inward fluid flow (IF) during cavity preparation was followed by outward flow (OF) after preparation. In amalgam restoration, the OF changed to IF during amalgam filling and slight OF followed after finishing.

In composite restoration, application CE and EB showed a continuous OF and air-dry increased rapidly the OF until light-curing, whereas in MP and SB, rinse and dry caused IF and OF, respectively. Application of hydrophobic bonding resin in MP and CE caused a decrease in flow rate or even slight IF. Light-curing of adhesive and composite showed an abrupt IF. There was no statistically significant difference in the reduction of DFF among the materials at 30 min, 3 and 7 days after restoration (P > 0.05).

The purpose of this study was to investigate the effect of rigidity of post core systems on stress distribution by the theoretical technique, finite element stress-analysis method. Three-dimensional finite element models simulating an endodontically treated maxillary central incisor restored with a zirconia ceramic crown were prepared and 1.5 mm ferrule height was provided. Each model contained cortical bone, trabecular bone, periodontal ligament, 4 mm apical root canal filling, and post-and-core. Six combinations of three parallel type post (zirconia ceramic, glass fiber, and stainless steel) and two core (Paracore and Tetric ceram) materials were evaluated, respectively. A 50 N static occlusal load was applied to the palatal surface of the crown with a 60° angle to the long axis of the tooth. The differences in stress transfer characteristics of the models were analyzed. von Mises stresses were chosen for presentation of results and maximum displacement and hydrostatic pressure were also calculated. An increase of the elastic modulus of the post material increased the stress, but shifted the maximum stress location from the dentin surface to the post material. Buccal side of cervical region (junction of core and crown) of the glass fiber post restored tooth was subjected to the highest stress concentration. Maximum von Mises stress in the remaining radicular tooth structure for low elastic modulus resin core (29.21 MPa) was slightly higher than that for high elastic modulus resin core (29.14 MPa) in case of glass fiber post. Maximum displacement of glass fiber post restored tooth was higher than that of zirconia ceramic or stainless steel post restored tooth.

The purpose of this study was to investigate the pulpal response to direct pulp capping with dentin sialo-protein (DSP) -derived synthetic peptide in teeth of dogs, and to compare its efficacy to capping substances Ca(OH)₂ and white mineral trioxide aggregate (WMTA). A total of 72 teeth of 6 healthy male beagle dogs were used. The mechanically exposed pulps were capped with one of the following: (1) DSP-derived synthetic peptide (PEP group); (2) Ca(OH)₂ (CH group); (3) a mixture paste of peptide and Ca(OH)₂ (PEP+CH group); or (4) white MTA (WMTA group). The access cavity was restored with a reinforced glass ionomer cement. Two dogs were sacrificed at each pre-determined intervals (2 weeks, 1 month, and 3 months). After the specimens were prepared for standard histological processing, sections were stained with hematoxylin and eosin. Under a light microscope, inflammatory response and hard tissue formation were evaluated in a blind manner by 2 observers. In the PEP group, only 3 of 17 specimens showed hard tissue formation, indication that the DSP-derived synthetic peptide did not induce proper healing of the pulp. Compared with the CH group, the PEP group demonstrated an increased inflammatory response and poor hard tissue formation. The CH and WMTA groups showed similar results for direct pulp capping in mechanically exposed teeth of dogs.

The purpose of this study was to assess the current materials, methods and difficulties according to the year of licence and educational background of Korean dentists in Class II direct composite resin restorations.

Total 17 questions were included in the questionnaire. Questions were broadly divided into two parts; first, operator's information, and second, the materials and methods used in Class II posterior composite restoration. The questionnaire was sent to dentists enrolled in Korean Dental Association via e-mail. Total 12,193 e-mails were distributed to dentists, 2,612 e-mails were opened, and 840 mails (32.2%) were received from respondents. The data was statically analyzed by chi-square test using SPSS(v. 12.0.1, SPSS Inc, Chicago, IL, USA).

Male dentists among respondents was 79%. 60.3% of the respondents acquired their licences recently (1998-2007), and 77% practiced in private offices. 83.4% have acquired their knowledge through school lectures, conferences and seminars.

For the Class II restorations, gold inlays were preferred by 65.7% of respondents, while direct composite resin restorations were used by 12.1% amalgam users were only 4.4% of respondents.

For the restorative technique, 74.4% of respondents didn't use rubber dam as needed. For the matrix, mylar strip (53.4%), metal matrix (33.8%) and Palodent system (6.5%) were used. 99.6% of respondents restored the Class II cavity by incremental layering.

Obtaining of the tight interproximal contact was considered as the most difficult procedure (57.2%) followed by field isolation (21%).

Among various bonding systems, 22.6% of respondents preferred SE Bond and 20.2% used Single Bond. Z-250 was used most frequently among a variety of composite resins.

Deterioration of long-term dentin adhesion durability is thought to occur by hydrolytic degradation within hydrophilic domains of the adhesive and hybrid layers. This study investigated the hypothesis that priming the collagen network with an organic solvent displace water without collapse and thereby obtain good bond strength with an adhesive made of hydrophobic monomers and organic solvents. Three experimental adhesives were prepared by dissolving two hydrophobic monomers, bisphenol-A-glycidylmethacrylate (Bis-GMA) and triethylenegly-col dimethacrylate (TEGDMA), into acetone, ethanol or methanol. After an etching and rinsing procedure, the adhesives were applied onto either wet dentin surfaces (wet bonding) or dentin surfaces primed with the same solvent (solvent-primed bonding). Microtensile bond strength (MTBS) was measured at 48 hrs, 1 month and after 10,000 times of thermocycles. The bonded interfaces were evaluated using a scanning electron microscope (SEM). Regardless of bonding protocols, well-developed hybrid layers were observed at the bonded interface in most specimens. The highest mean MTBS was observed in the adhesive containing ethanol at 48 hrs. With solvent-primed bonding, increased MTBS tendencies were seen with thermocycling in the adhesives containing ethanol or methanol. However, in the case of wet bonding, no increase in MTBS was observed with aging.

This study was aimed to investigate whether an oxygen inhibition layer (OIL) is essential for the interfacial bonding between resin composite layers or not.

A composite (Z-250, 3M ESPE) was filled in two layers using two aluminum plate molds with a hole of 3.7 mm diameter. The surface of first layer of cured composite was prepared by one of five methods as followings, thereafter second layer of composite was filled and cured: Group 1 - OIL is allowed to remain on the surface of cured composite; Group 2 - OIL was removed by rubbing with acetone-soaked cotton; Group 3 - formation of the OIL was inhibited using a Mylar strip; Group 4 - OIL was covered with glycerin and light-cured; Group 5 (control) - composite was bulk-filled in a layer. The interfacial shear bond strength between two layers was tested and the fracture modes were observed. To investigate the propagation of polymerization reaction from active area having a photo-initiator to inactive area without the initiator, a flowable composite (Aelite Flow) or an adhesive resin (Adhesive of ScotchBond Multipurpose) was placed over an experimental composite (Exp_Com) which does not include a photoinitiator and light-cured. After sectioning the specimen, the cured thickness of the Exp_Com was measured.

The bond strength of group 2, 3 and 4 did not show statistically significant difference with group 1. Groups 3 and 4 were not statistically significant different with control group 5. The cured thicknesses of Exp_Com under the flowable resin and adhesive resin were 20.95 (0.90) um and 42.13 (2.09), respectively.

The objective of this study was to compare dentin shear bond strength (DSBS) of dentin bonding agents (DBAs) cured with a plasma arc (PAC) light curing unit (LCU) and those cured with a light emitting diode (LED) LCU. Optical properties were also analyzed for Elipar freelight 2 (3M ESPE); LED LCU, Apollo 95E (DMT Systems); PAC LCU and VIP Junior (Bisco); Halogen LCU. The DBAs used for DSBS test were Scotchbond Multipurpose (3M ESPE), Singlebond 2 (3M ESPE) and Clearfil SE Bond (Kuraray). After DSBS testing, fractured specimens were analyzed for failure modes with SEM.

The total irradiance and irradiance between 450 nm and 490 nm of the LCUs were different. LED LCU showed narrow spectral distribution around its peak at 462 nm whereas PAC and Halogen LCU showed a broad spectrum. There were no significant differences in mean shear bond strength among different LCUs (P > 0.05) but were significant differences among different DBAs (P < 0.001)

The purpose of this study was: (1) to compare nanoleakage patterns of a conventional 3-step etch and rinse adhesive system and two experimental hydrophobic adhesive systems and (2) to investigate the change of the nanoleakage patterns after load cycling. Two kinds of hydrophobic experimental adhesives, ethanol containing adhesive (EA) and methanol containing adhesive (MA), were prepared. Thirty extracted human molars were embedded in resin blocks and occlusal thirds of the crowns were removed. The polished dentin surfaces were etched with a 35% phosphoric acid etching gel and rinsed with water. Scotchbond Multi-Purpose (MP), EA and MA were used for bonding procedure. Z-250 composite resin was built-up on the adhesive-treated surfaces. Five teeth of each dentin adhesive group were subjected to mechanical load cycling. The teeth were sectioned into 2 mm thick slabs and then stained with 50% ammoniacal silver nitrate. Ten specimens for each group were examined under scanning electron microscope in backscattering electron mode. All photographs were analyzed using image analysis software. Three regions of each specimen were used for evaluation of the silver uptake within the hybrid layer. The area of silver deposition was calculated and expressed in gray value. Data were statistically analyzed by two-way ANOVA and post-hoc testing of multiple comparisons was done with the Scheffe's test. Silver particles were observed in all the groups. However, silver particles were more sparsely distributed in the EA group and the MA group than in the MP group (p < .0001). There were no changes in nanoleakage patterns after load cycling.

This study evaluated the effectiveness and safety of an experimental bleaching strip (Medison dental whitening strip, Samsung medical Co., Anyang, Korea) containing 2.9% hydrogen peroxide. Twenty-three volunteers used the bleaching strips for one and a half hour daily for 2 weeks. As control group, the same strips in which hydrogen peroxide was not included were used by 24 volunteers with the same protocol. The shade change (ΔE*, color difference) of twelve anterior teeth was measured using Shade Vision (X-Rite Inc., S.W. Grandville, MI, USA), Chroma Meter (Minolta Co., Ltd. Osaka, Japan) and Vitapan classical shade guide (Vita Zahnfabrik, Germany). The shade change of overall teeth in the experimental group was significantly greater than that in the control group (p < 0.05) and was easily perceivable. The change resulted from the increase of lightness (CIE L* value) and the decrease of redness (CIE a* value) and yellowness (CIE b* value). The shade change of individual tooth was greatest in canine, and smallest in central incisor. The safety of the bleaching strip was also confirmed.

The purpose of this study is to evaluate prospectively the effect of different bonding systems and retention grooves on the clinical performance of resin restorations in non-carious cervical lesions (NCCLs). Thirty-nine healthy adults who had at least 2 NCCLs in their premolar areas were included in this study. One hundred and fifty teeth were equally assigned to six groups: (A) Scotchbond Multi-Purpose (SBMP, 3M ESPE, St. Paul, MN, USA, 4th generation bonding system) without retention grooves; (B) SBMP with retention grooves; (C) BC Plus (Vericom Co., Anyang, Gyeonggido, Korea, 5th generation bonding system) without retention grooves; (D) BC Plus with retention grooves; (E) Adper Prompt (3M ESPE, Seefeld, Germany, 6th generation bonding system) without retention grooves; (F) Adper Prompt with retention grooves. All cavities were filled with a hybrid composite resin, Denfil (Vericom Co., Anyang, Gyeonggido, Korea) by one operator. Restorations were evaluated at baseline and at 6-month recall, according to the modified USPHS (United States Public Health Service) criteria. Additionally, clinical photographs were taken and epoxy resin replicas were made for SEM evaluation. At 6-month recall, there were some differences in the number of alpha ratings among the experimental groups. But, despite the differences in the number of alpha ratings, there was no significant difference among the 3 adhesive systems (p > 0.05). There was also no significant difference between the groups with or without mechanical retention (p > 0.05). Follow-ups for longer periods than 6 months are needed to verify the clinical performance of different bonding systems and retention grooves.

This study investigated the hypothesis that the dentin bond strength of self-etching adhesive (SEA) might be improved by applying additional layer of bonding resin that might alleviate the pH difference between the SEA and the restorative composite resin. Two SEAs were used in this study; Experimental SEA (Exp, pH: 1.96) and Adper Prompt (AP, 3M ESPE, USA, pH: 1.0). In the control groups, they were applied with two sequential coats. In the experimental groups, after applying the first coat of assigned SEAs, the D/E bonding resin of All-Bond 2 (Bisco Inc., USA, pH: 6.9) was applied as the intermediate adhesive. Z-250 (3M ESPE, USA) composite resin was built-up in order to prepare hourglass-shaped specimens. The microtensile bond strength (MTBS) was measured and the effect of the intermediate layer on the bond strength was analyzed for each SEA using t-test. The fracture mode of each specimen was inspected using stereomicroscope and Field Emission Scanning Electron Microscope (FE-SEM). When D/E bonding resin was applied as the second coat, MTBS was significantly higher than that of the control groups. The incidence of the failure between the adhesive and the composite or between the adhesive and dentin decreased and that of the failure within the adhesive layer increased. According to the results, applying the bonding resin of neutral pH can increase the bond strength of SEAs by alleviating the difference in acidity between the SEA and restorative composite resin.

The aim of this study was to evaluate the effect of cavity shape, bond quality of bonding agent and volume of resin composite on shrinkage stress developed at the cavity floor. This was done by measuring the shear bond strength with respect to iris materials (cavity shape; adhesive-coated dentin as a high C-factor and Teflon-coated metal as a low C-factor), bonding agents (bond quality; Scotchbond™ Multi-purpose and Xeno®III) and iris hole diameters (volume; 1 mm or 3 mm in diameter × 1.5 mm in thickness). Ninety-six molars were randomly divided into 8 groups (2 × 2 × 2 experimental setup). In order to simulate a Class I cavity, shear bond strength was measured on the flat occlusal dentin surface with irises. The iris hole was filled with Z250 restorative resin composite in a bulk-filling manner. The data was analyzed using three-way ANOVA and the Tukey test. Fracture mode analysis was also done. When the cavity had high C-factor, good bond quality and large volume, the bond strength decreased significantly. The volume of resin composite restricted within the well-bonded cavity walls is also be suggested to be included in the concept of C-factor, as well as the cavity shape and bond quality. Since the bond quality and volume can exaggerate the effect of cavity shape on the shrinkage stress developed at the resin-dentin bond, resin composites must be filled in a method, which minimizes the volume that can increase the C-factor.

This study compared the dentin shear bond strengths of currently used dentin bonding agents that were irradiated with an LED (Elipar FreeLight, 3M-ESPE) and a halogen light (VIP, BISCO). The optical characteristics of two light curing units were evaluated. Extracted human third molars were prepared to expose the occlusal dentin and the bonding procedures were performed under the irradiation with each light curing unit. The dentin bonding agents used in this study were Scotchbond Multipurpose (3M ESPE), Single Bond (3M ESPE), One-Step (Bisco), Clearfil SE bond (Kuraray), and Adper Prompt (3M ESPE). The shear test was performed by employing the design of a chisel-on-iris supported with a Teflon wall. The fractured dentin surface was observed with SEM to determine the failure mode.

The spectral appearance of the LED light curing unit was different from that of the halogen light curing unit in terms of maximum peak and distribution. The LED LCU (maximum peak in 465 ㎚) shows a narrower spectral distribution than the halogen LCU (maximum peak in 487 ㎚). With the exception of the Clearfil SE bond (P < 0.05), each 4 dentin bonding agents showed no significant difference between the halogen light-cured group and the LED light-cured group in the mean shear bond strength (P > 0.05).

The results can be explained by the strong correlation between the absorption spectrum of cam-phoroquinone and the narrow emission spectrum of LED.

The purpose of this study is to monitor the secretion of matrix metalloproteinase-1 (MMP-1) and tissue inhibitor of metalloproteinase-1 (TIMP-1) produced by human osteosarcoma cell line (MG63) stimulated with Prevotella nigrescens lipopolysaccharides (LPS), and to compare the level of secretion before and after the treatment of calcium hydroxide on P. nigrescens LPS.

LPS was extracted and purified from anaerobically cultured P. nigrescens. MG63 cells were stimulated by the LPS (0, 1, 10 µg/ml) or LPS (10 µg/ml) pretreated with 12.5 mg/ml of Ca(OH)₂ for 3 days.

Total RNA was isolated from the cell, and real-time quantitative polymerase chain reaction (PCR) was performed for quantification of MMP-1 and TIMP-1.

The results were as follows.

1. MMP-1 mRNA expression at 48 hr was highly increased by stimulation with P. nigrescens LPS. The increase was dose-dependent.

2. When stimulated with 1 µg/ml of LPS, TIMP-1 mRNA expression was highly increased at 24 hr and 48 hr. However, TIMP-1 expression was suppressed at higher concentration (10 µg/ml).

3. When P. nigrescens LPS was pretreated with Ca(OH)₂, MMP-1 and TIMP-1 gene expression was downregulated.

The results of this study suggest that transcriptional regulation of MMP-1 and TIMP-1 by P. nigrescens LPS could be one of the important mechanisms in bone resorption of periapical inflammation. The result of calcium hydroxide on MMP-1 and TIMP-1 gene expression suppression shows that calcium hydroxide detoxified bacterial LPS and thus should be used the medication of choice for intracanal dressings in root canal infected with black-pigmented bacteria.

Low-viscosity composite resins may produce better sealed margins than stiffer compositions (Kemp-Scholte and Davidson, 1988; Crim, 1989). Flowable composites have been recommended for use in Class V cavities but it is also controversial because of its high rates of shrinkage. On the other hand, in the study comparing elastic moduli and leakage, the microfill had the least leakage (Rundle et al. 1997). Furthermore, in the 1996 survey of the Reality Editorial Team, microfills were the clear choice for abfraction lesions.

The purpose of this study was to evaluate the microleakage of 6 compostite resins (2 hybrids, 2 microfills, and 2 flowable composites) with and without load cycling.

Notch-shaped Class V cavities were prepared on buccal surface of 180 extracted human upper premolars on cementum margin. The teeth were randomly divided into non-load cycling group (group 1) and load cycling group (group 2) of 90 teeth each. The experimental teeth of each group were randomly divided into 6 subgroups of 15 samples. All preparations were etched, and Single bond was applied. Preparations were restored with the following materials (n=15): hybrid composite resin [Z250(3M Dental Products Inc. St.Paul, USA), Denfil(Vericom, Ahnyang, Korea)], microfill [Heliomolar RO(Vivadent, Schaan, Liechtenstein), Micronew(Bisco Inc. Schaumburg, IL, USA)], and flowable composite [AeliteFlo(Bisco Inc. Schaumburg, IL, USA), Revolution(Kerr Corp. Orange, CA, USA)]. Teeth of group 2 were subjected to occlusal load (100N for 50,000 cycles) using chewing simulator(MTS 858 Mini Bionix II system, MTS Systems Corp., Minn. USA). All samples were coated with nail polish 1mm short of the restoration, placed in 2% methylene blue for 24 hours, and sectioned with a diamond wheel. Enamel and dentin/cementum margins were analyzed for microleakage on a sclale of 0 (no leakage) to 3 (3/3 of wall). Results were statistically analyzed by Kruscal-Wallis One way analysis, Mann-Whitney U-test, and Student-Newmann-Keuls method. (p=0.05)