Background: Pomegranate (Punica granatum) fruits are commonly regarded as medicinal plant in Indonesia, and the polyphenols found in pomegranate juice (punicalagin and ellagic acid) have been shown to have antibacterial properties. Objectives: Using monospecies and multispecies biofilms, we sought to examine the effects of pomegranate juice on the viability of three periodontal pathogens: Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Treponema denticola. Methods: Biofilm assays were performed using crystal violet. Pomegranate juice was obtained from pomegranates using a juicer, and the juice was then diluted into different concentrations with phosphate saline buffer. The three pathogens were cultured in both monospecies and multispecies plates. Pomegranate juice was then added to each biofilm well. These were then incubated for 1h, 6h, or 24h, after which the optical density (OD) of the biofilm mass was measured using a microplate-reader (490 nm). Biofilm without treatment was used as a negative control and 0.2% chlorhexidine gluconate as a positive control. Data were analyzed with a one-way ANOVA; the level of significance was set at p<0.05. Results: Compared to the negative control, biofilm mass was significantly decreased after treatment with pomegranate juice across all concentrations and incubation times, for both monospecies and multispecies abiofilm (p<0.05). The best results were achieved with P. gingivalis biofilm, with 100% concentration (OD 0.34 ± 0.03); A. actinomycetemcomitans, 50% concentration (OD 0.22 ± 0.01); and T. denticola, with 25% concentration (OD 0.87 ± 0.08), as well as with a multispecies biofilm with a 50% concentration in 1h incubation time (OD 0.09 ± 0.02). Conclusion: Pomegranate juice effectively inhibited the biofilm formation of P. gingivalis, A. actinomycetemcomitans, and T. denticola. Pomegranate juice may therefore be used as an alternative therapy in preventing periodontal disease. Additional research should explore this effect in an environment that mimics oral cavities.

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Background: Periodontal disease is common chronic adult condition. Antioxidants are present in the body fluid as protection against free radical. Uric acid is one of antioxidants that can be found in saliva. Moreover, the relationship among the antioxidant enzymes activities and clinical periodontal status were investigated. Objectives: The aim of the study was to observe uric acid level activities in the saliva of gingivitis and periodontitis patients. Methods: Six patients with gingivitis and six patients with periodontitis in Dental Hospital Trisakti University were included in the study. Clinical condition of each subject, the plaque index, and probing depth were determined. The salivary uric acid level was measured using the Folin-Wu method. Result: Salivary uric acid levels in the periodontitis patients with a mean ± SD 7.40 ± 0.31 (p = 0.004) were found to be higher compared to the gingivitis patients (mean ± SD = 6.84 ± 0.19). In addition, there were no significant differences in salivary uric acid levels between gender (p = 0.641). Conclusion: Uric acid levels in periodontitis patients were found to be higher than in gingivitis patients. Moreover, uric acid has more role on periodontitis than in gingivitis as an antioxidant agent.

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Background: Probiotic bacteria is commonly used as a food supplement intended to benefit the host by improving intestinal bacterial balance. Probiotics have also been investigated from the perspective of oral health. Objectives: The purpose of this study was to investigate the effect of daily intake of yogurt containing probiotic Bifidobacterium animalis subsp. lactis BB-12 (B. lactis) on salivary Streptococcus mutans (S. mutans) counts in patients undergoing fixed orthodontic treatment. Methods: Saliva samples were collected from each subject (n = 7; mean age, 21 years) using spitting method in centrifuge tubes at baseline and two weeks after daily probiotic yogurt consumption. B. lactis BB-12 and S. mutans ATCC 25175 were cultured in BHI-broth (37°C, anaerobic conditions). After 48 hours incubation, the number of colonies on each dilution plate was used to extrapolate a standard curve. The total number of target DNA molecules were identified using Real-Time PCR followed by SYBR Green reagents and 16S rRNA gene specific primers S. mutans and B. lactis BB-12. Data were analyzed statistically using paired-sample t-tests. Results: Statistical evaluation indicated that there was a significant reduction in the presence of S. mutans before probiotic yogurt consumption, (4.73 ± 1.43) log10 CFU/mL and after two weeks of daily consumption of probiotic yogurt, (4.03 ± 0.77) log10 CFU/mL, p = 0.001. Moreover, no B. lactis was found in the saliva of any of the subjects before probiotic consumption, but after two weeks of consumption, B. lactis was found in the saliva of four subjects. Conclusions: Consuming probiotic yogurt containing B. lactis reduced the quantity of S. mutans in the saliva of subjects during fixed orthodontic treatment. Thus, the probiotic bacteria could be beneficial in improving oral health.

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Background: Enterococcus faecalis is the frequent cause of oral infections, such as periodontitis, infected root canals, and peri-radicular abscesses. Pineapple (Ananas comosus) fruit contains bromelain, one of proteolytic enzymes associated with several health benefits. Bromelain has been shown to promote healthy digestion, stimulate the immune system, improve cardiovascular conditions, and accelerate wound healing. Bromelain compounds possess antiinflammatory and anticancer properties and exhibit antibacterial activity. Objectives: To analyze the enzymatic activity of bromelain extracted from pineapple hump and investigate the antibacterial effect of bromelain against E. faecalis. Methods: Pineapple hump was dried and extracted with maceration technique. Further purification was obtained by ammonium sulfate fractionation, dialysis and ion exchange chromatography. Minimum inhibitory concentration (MIC) test using diffusion and dilution techniques tested the antibacterial activity of the bromelain extract towards E. faecalis. A one-way analysis of variance (ANOVA) test analyzed the significance of the differences in the E. faecalis inhibition zones after treatment with a range of bromelain extract concentrations. Differences were considered statistically significant if p < 0.05. Results: The specific activity of bromelain in the crude extract was 62.89 U/mg. Furthermore, bromelain activity using ammonium sulfate fractionation was 50.99 U/mg, dialysis was 54.59 U/mg, and ion exchange chromatography was 152.38 U/mg. The bromelain extract showed effective inhibitory and bactericidal activity against E. faecalis. The results of the inhibition test using a bromelain extract purified by ion exchange chromatography demonstrated that a concentration as small as 12.5% was effective in inhibiting the growth of E. faecalis (p < 0.05). Conclusion: The highest enzymatic activity of bromelain was found after purification with ion exchange chromatography. Bromelain exerted an antibacterial effect against a potent endodontic pathogen, but further studies are needed to explore this effect.

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Age estimation is a fundamental part in forensic, criminal, legal, and anthropological investigations. The biomolecular analysis is considered to have a good capability in estimating age because it can describe a person's biological age. According to previous studies, DNA methylation has the best effectiveness for estimating biological age, compared to other biomolecular analysis. Although DNA methylation is influenced by a number of factors such as heredity, environment, lifestyle and systemic diseases, DNA methylation still has accuracy that accountable to estimate age. A literature review search of PubMed, ScienceDirect, Scopus, and EBSCO was conducted to get all studies that published before February 2020. The review was then performed on 22 papers that selected based on inclusion and exclusion criteria. The purpose of this reviewed paper was to identify all gene markers that have been used to estimate age using DNA methylation analysis; and to find out the subject, age range, tissue taken for DNA methylation analysis, and the effectiveness of the analysis.

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Background: Chitosan has been shown to inhibit free radicals that cause tooth enamel demineralization. Nano-sized chitosan can penetrate cell membranes that larger particles cannot penetrate. Objective: This study aimed to determine the difference between chitosan nanoparticles, chitosan microparticles, and casein phosphopeptide–amorphous calcium phosphate (CPP-ACP) in preventing tooth enamel demineralization. Methods: We used 0.2% chitosan nanoparticles. A total of 50 mL of chitosan solution was stirred and added in tripolyphosphate to prepare a nanoparticle suspension. It was then stirred for 1 h to generate crosslinking. The nanoparticles' size was 57.6 nm. The demineralization solution consisted of 2.2 mM/L CaCl₂, 2.2 mM/L KH₂PO₄, and 50 mM of acetate buffer. Its acidity was regulated to a pH of 4.06. The sample consisted of 27 maxillary first premolar teeth post extraction due to orthodontic treatment needs divided into three groups: a chitosan microparticle treatment group, a chitosan nanoparticle treatment group, and a CPP-ACP treatment group used as a positive control. A scanning electron microscope with ×5000 magnification was used to observe the enamel surface morphology and mineral release. Results: The mean value of enamel surface microhardness in the chitosan nanoparticle group (233.39 HV) was significantly greater than those in the chitosan microparticle (153.192 HV) and CPP-ACP groups (152.626 HV) (P < 0.05). Moreover, the chitosan nanoparticle treatment resulted in the lowest enamel porosity. Conclusions: Chitosan nanoparticles are more effective than chitosan microparticles and CPP-APP in preventing enamel demineralization.

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Background: Biofilms are involved in a wide variety of microbial infections, including dental caries and periodontitis. The use of probiotics has been a promising prevention and treatment modality with which to combat biofilm-related diseases in the oral cavity. The probiotic Lactobacillus reuteri has been proven to reduce gingivitis and plaque index inside the oral cavity. These bacteria can be found in the digestive system and also in the human oral cavity. Objective: The aim of this study was to identify L. reuteri in the saliva of Indonesian young adults. Methods: Forty saliva samples were collected from 18–24 year-old Indonesian subjects. DNA extraction was performed, and then, the identification of L. reuteri was accomplished using PCR. Six subjects showed positive results. The positive samples were cultured in Rogosa Agar for 24 hours at 37°C anaerobically. Several single colonies were further cultured separately in broth medium before DNA extraction and PCR identification were performed. The four thickest bands were selected for DNA sequencing. Results: An analysis performed using BLAST showed that two of the L. reuteri strains obtained from the Indonesian saliva isolates had 96% (isolate 3.11) and 95% (isolate 5.14) values. This confirmed the presence of new strains based on average nucleotide identity (ANI). The isolate strains of 3.11 and 5.14 have been registered at DDBJ/EMBL/GenBank under the accession number LC382415 and LC382416, respectively. Conclusion: L. reuteri novel strain can be isolated from the saliva of Indonesian young adults. Further studies involving biochemical tests and phenotypic analysis are needed to better understand these new L. reuteri strains.

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Background: The aim of endodontic treatment is to eliminate bacteria from the root canal. Bacterial removal from the root canal can be achieved with a mechanical approach using an instrument and a disinfecting irrigation agent. Enterococcus faecalis is the most prevalent bacteria found in root canals. Research studies have also been conducted to examine irrigation systems using herbal products such as drumstick tree leaf (Moringa oleifera) instead of NaOCl. Active compounds in M. oleifera, such as flavonoids, tannins, saponins, alkaloids, phenolics, and triterpenoids, possess antibacterial effects. Objectives: The aim of this study is to assess the antibacterial effect of drumstick tree extract (M. oleifera) in several concentrations (25%, 50%, 75%, and 100%) as an irrigation solution against E. faecalis through in the root canal ex vivo. Methods: This study used 24 mandibular premolars, divided into six category groups. Group 1 received 5.25% NaOCl as a positive control, Group 2 received 25% M. oleifera extract, Group 3 received 50% M. oleifera extract, Group 4 received 75% M. oleifera extract, Group 5 received 100% M. oleifera extract, and Group 6 received irrigation using phosphate-buffered saline as a negative control. Quantitative polymerase chain reaction methods used to analyze the E. faecalis number in the root canal after treatment with M. oleifera extract. Results: A one-way ANOVA showed significant differences (P = 0.05) between the three types of irrigation solutions against E. faecalis. Among the six study groups, the most prominent efficacy was found in Groups 1, 4, and 5. Conclusion: M. oleifera extract solution at concentrations of 75% and 100% is as effective as 5.25% NaOCl. This extract may be used as an alternative irrigation agent for root canal treatment. However, further studies are warranted to examine the toxicity effect.

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Background: The management of immature central incisors with nonvital pulp is a challenge for dentists because the goals and criteria for successful root canal treatment are difficult to achieve. A necrotic immature root canal system accompanied by apical periodontitis cannot be disinfected by standard protocols with the aggressive use of endodontic files. This case report demonstrated the advantages of pulp revascularization as a treatment method for necrotic immature teeth based on clinical and radiographic outcomes. Case Report: The patient was an 11-year-old boy with a history of trauma to the upper left central incisor who was treated with a revascularization procedure. Clinical and radiographic evaluations showed nonvital pulp, asymptomatic apical periodontitis, and mucogingival swelling before treatment. At the 1-, 3-, and 6-month follow-up visits, the patient's teeth remained asymptomatic and well-functioning. There was a slight thickening of the tooth wall and loss of lesions, although total closure of the apical foramen was observable. Longer clinical and radiographic controls are needed to evaluate the success of this case.Conclusion: Revascularization is a procedure that promotes thickening of the dentinal wall and closure of the apical foramen, thereby preventing teeth from becoming weak. The minimum expectation of revascularization treatment is the absence of undesirable signs and symptoms, but the ultimate goal is full regeneration of the pulp complex and ideal root development.

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Background: Einstein's photoelectric amplification theory provided the template for the development of lasers. With recent technological advancements, the increasing use of lasers in dentistry has had a meaningful impact on the diagnosis and management of oral mucosal lesions. This case series highlights lasers' impact. Case Reports: Five patients with diagnoses of arteriovenous malformation, homogeneous leukoplakia, mucocele, traumatic fibroma, and erosive lichen planus were treated with a diode laser (1,200 J/s; wavelength of 940 nm; 1.5 W; pulse mode). The patients were followed up to evaluate lesion healing and complete healing of the lesion was rapidly achieved with minimal discomfort. Conclusion: Soft-tissue lasers are trending in the management of various oral mucosal lesions due to their advantages of providing higher precision, a clean surgical field with minimal blood loss, accelerated wound healing, and fewer postoperative complications.

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Background: Advances in nanotechnology research make the use of cellulose nanocrystals (CNCs) attractive for improving the mechanical properties of glass ionomer cement (GIC). Sugarcane bagasse (Saccharum officinarum L.) is a CNCs source with a high CNC content (72.5%). Objective: This study aimed to determine the effect of the addition of sugarcane bagasse CNCs on the mechanical properties of GIC. Methods: In total, 42 GIC (Fuji IX, GC, Japan) samples were divided into six groups, with various concentrations of CNCs, added to the samples. After 24 h immersion in distilled water at 37°C, the samples were analyzed using the Vickers hardness test. The samples were also characterized by transmission electron microscopy (TEM). For statistical analysis, a one-way analysis of variance, followed by Tukey's post hoc test, was applied. A value of P < 0.05 denoted statistical significance. Results: The TEM revealed crystalline particles in the form of nanocrystals, with varying particle sizes (lengths of 100–200 nm and diameters of 4–19 nm). The addition of 0.4% of CNCS from bagasse fiber to GIC increased the Vickers hardness of the material by 38.89% (P < 0.05). Conclusion: The addition of 0.4% of sugarcane bagasse can improve the hardness of GIC.

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Background: Angiosarcoma is a rare malignant tumor, which most often occurs in the skin and soft tissue. Its occurrence in the head and neck region, and the intraoral region in particular, is uncommon. The aim of this case report is to highlight the importance of histopathological examination and to underscore the characteristic features of angiosarcoma. Case report: A 52-year-old female presented with a growth in the right maxillary portion of the interdental region of her molar teeth. The growth had been present for approximately one month. A computerized tomography (CT) scan showed metastatic tumors in the lungs, liver, bone, and soft tissue of maxillary gingiva. Intraoral examination found a soft and mobile pedunculated ovoid mass at the right palatal gingiva; the mass was reddish-blue in color. An incisional biopsy was performed to confirm the diagnosis and plan for further treatment. Histopathologic examination revealed malignant endothelial cells in a vasoformative growth pattern, with numerous vascular channels lined with neoplastic endotheliumforming intraluminal buds, projections, and papillae. These projections were interspersed with the tumor cells. Immunohistochemistry analysis showed these endothelial cells demonstrated strong positive immunoreactivity with CD31, factor VIII, and FLi-1. A diagnosis of angiosarcoma was made. Discussion: Because intraoral angiosarcoma is rare and its clinical presentation can resemble other vascular lesions, it can be a challenging diagnosis for clinicians and pathologists. In the present case, the diagnosis of intraoral angiosarcoma was made based on clinical and histopathology findings. The microscopic features of malignant endothelial cells and the presence of intraluminal projections, supported by positive immunoreactivity with CD31, factor VIII, and FLi-1, confirmed the diagnosis. Conclusion: Angiosarcoma poses a diagnostic challenge to dental practitioners and oral pathologists due to the rarity of occurrence in the oral cavity. Histopathological examination is critical for an accurate diagnosis.

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Background: Pain is a common problem encountered after fixed orthodontic appliance placement. Pain from orthodontic treatment can come from ischemia, inflammation, and edema in the periodontal ligaments that were depressed. Objectives: To determine whether there are differences in the pain reduction associated with chewing gum, taking acetaminophen, and gargling green tea after fixed orthodontic appliance placement. Methods: This study is a quasiexperimentwith a controlled randomized post-treatment approach using 40 subjects. The subjects were consecutively selected and then randomly assigned into four equal groups: a group assigned to chewing gum, a group assigned to take acetaminophen, a group assigned to gargle green tea, and a control group. The level of pain was measured using a Visual Analogue Scale (VAS). A one-way ANOVA test was used to analyse differences between the groups. Results: The VAS scores of the chewing gum and acetaminophen groups were lower than the green tea and control groups. At 24 hours after fixed orthodontic appliance placement, only the chewing gum group reported a decrease in the amount of pain experienced. Overall, the chewing gum group reported the lowest pain scores compared to other groups. However, there was no overall statistically significant difference between the chewing gum and acetaminophen group (p>0.05). Additionally, there was no statistically significant difference in the amount of pain reduction experienced by the green tea group and the control group (p>0.05). There was a significant difference in the amount of pain reduction reported between the chewing gum or acetaminophen groups and the green tea group (p<0.05). Conclusion: There was no significant difference between chewing gum and taking acetaminophen in the amount of pain reduction experienced after fixed orthodontic appliance placement, but gargling with green tea was found to be significantly less effective in reducing pain. Because it has fewer side effects, chewing gum may be a more preferable alternative to taking acetaminophen to reduce pain after fixed orthodontic appliance placement.

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Background: Fractured instruments, especially endodontic files, are a common problem in daily practice. A broken file causes a canal blockage that impedes the cleaning and shaping process. Therefore, an attempt to remove the broken file should be considered in most cases. Nowadays, with advances in technology, such as ultrasonic tips, fractured instruments can be easily retrievable. Case Report: This case report presented a 15-year-old female patient with a complaint of a large cavity in the left mandibular region who presented to the department of conservative dentistry and endodontics. The clinical diagnosis was pulp necrosis, and a root canal treatment was performed. During the shaping procedure, a file was broken in the mesiolingual canal. Conclusion: The broken file was removed using ultrasonic tips with a dental operating microscope. After the instrument was retrieved, the obturation was performed successfully.

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Background: Tooth loss in individuals more commonly occurs with increasing age. In a dental practice, many patients present with torus palatinus, which is usually caused by problems and complications resulting from denture fabrication. Therefore, a nonsurgical approach in which horseshoe-shaped complete dentures are fabricated is considered to be a viable option to address torus palatinus. Case Report: A 59-year-old edentulous male came to the Prosthodontic Department, Faculty of Dentistry, Trisakti University, with a chief complaint of the instability of his maxillary complete dentures, and he requested new dentures. The patient had been wearing the dentures for approximately 5 years. Based on the intraoral examination, the patient had a large, single lobule torus palatinus that extended posteriorly through the junction between the hard palate and the soft palate (AH line). It was covered with thin mucosal tissue, and it did not interfere with his speech, his ability to chew, or other oral functions without the dentures. After the problems were thoroughly diagnosed and corrected, horseshoe-shaped complete dentures were chosen as the appropriate solution, and the dentures were fabricated. Conclusion: Torus palatinus tends to have very thin mucosa that causes discomfort and irritation during routine usage of acrylic dentures. Therefore, the horseshoe-shaped dentures facilitated good retention and stability, and they did not irritate the torus palatinus.

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Background: Deep bites represent one of the most difficult cases in orthodontic treatment. In growing patients, strategies for deep bite correction include extrusion of posterior teeth and intrusion of incisors, which can be achieved using anterior bite plates. This case report presents the treatment of a deep bite in a growing patient using an anterior bite plate. Case Report: A 13-year-old female presented with a convex profile, short lower facial height, a Class II skeletal relationship, a deep bite (6 mm overbite), a 6 mm overjet, and severe crowding on both arches. The patient was treated with a removable anterior bite plate in conjunction with a fixed appliance. A normal overbite (2 mm) was achieved, severe crowding was corrected on both arches, the lower facial height was increased, the interincisal angle remained favorable, the mandibular incisors were well positioned in the basal bone, and the smile esthetics improved. Conclusion: The use of an anterior bite plate in combination with fixed appliances is effective in treating deep bites in a growing patient.

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Background: Maldistribution of health workers, including dentists, is a global issue faced by not only poor and developing countries but also developed countries due to the mobility of health workers, who prefer working in urban areas. The World Health Organization (2012) released global policy recommendations that cover education, regulation, incentives, and personal and professional support to overcome this issue. However, they have not been able to completely solve the issue, as is shown by a description of Indonesia's Health Profile (2018), which highlights that 43.53% of public health center lack of dentists. The number of studies concerning the real condition experienced by dentists working in rural and remote areas of Indonesia is still limited. Objectives: To analyze job satisfaction and factors affecting dentists' retention in rural and remote areas of Indonesia. Methods: This study used questionnaire to collect data which was designed based on seven major themes and inserted in Google Form. The link was distributed to dentists who work in rural and remote areas of Indonesia. The sample was selected based on random sampling techniques. Result: The study showed that 45.90% of respondents were not satisfied working in rural and remote areas. Most of the respondents claimed that working conditions, incentives, career and professional development, living conditions, and personal characteristics influenced their interest in working permanently in remote and rural areas. However, political and cultural factors did not give a high impact. Conclusion: The description of job satisfaction and factors affecting dentists' retention in rural and remote areas of Indonesia is expected to have a positive impact on formulating policy regarding dentists' maldistribution.

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Background: Home bleaching with carbamide peroxide is commonly used due to its high success rate and minimum side effects. Although home bleaching is safe, it causes structural changes in surface enamel, thereby facilitating Streptococcus mutans attachment on the enamel surface. Objectives: This study aimed to determine the effect of carbamide peroxide concentration on surface enamel structural changes and S. mutans attachment. Methods: Healthy, caries-free, and calculus-free upper first premolars were divided into 10%, 15%, and 35% carbamide peroxide concentration groups, with five samples in each group. Structural changes in the surface enamel of the teeth before and after the bleaching procedure were assessed subjectively based on observations using a stereomicroscope at × 40. The samples were inoculated in S. mutans culture and incubated for 24 h (37°C) in anaerobic conditions. The samples were then washed with phosphate-buffered saline, and bacterial attachment was released by vortexing for 1 min. Bacterial attachment was assessed using a turbidimetry test and total plate count test. Result: Structural changes in enamel were observed in the samples exposed to carbamide peroxide 35%. The results of an analysis of variance test revealed a significant difference (P < 0.05) in the bacterial attachment test. The samples exposed to carbamide peroxide 35% exhibited the highest amount of bacterial attachment (3 × 10^[6] CFU/mL, optical density: 0.06). Conclusion: The concentration of carbamide peroxide plays a role in structural changes in enamel and S. mutans attachment.

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Background: Periodontitis has been referred to as the sixth complication of diabetes. A number of studies found a higher prevalence of periodontal disease among diabetes patients than among healthy controls. Objective: The objective of the study was to determine the periodontal status and periodontal treatment needs (TN) of diabetes mellitus Type 2 patients attending Talaga Bodas Community Health Center, Bandung City, Indonesia. Methods: This descriptive study included 30 subjects with diabetes mellitus Type 2. The periodontal tissue status and periodontal TN were determined using the Community Periodontal Index of TN. Data on periodontal status and TN are presented in percentages based on subject, sextant, and duration of diabetes mellitus. Results: Periodontitis (53.3%) was more common than gingivitis (43.3%) among diabetes mellitus Type 2 patients. Scaling and root planing were the treatments most commonly required in the diabetes mellitus patients (n = 21.70%) and sextants (n = 81.65.9%). A longer period of diabetes mellitus was associated with a tendency toward more severe periodontal disease and more complex periodontal TN. Conclusion: The prevalence of periodontitis was higher than gingivitis among patients with diabetes mellitus Type 2. The TN of diabetes mellitus Type 2 patients comprised mostly scaling and root planing.

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Background: Mineral trioxide aggregate (MTA) is the most widely used material in endodontics, and recently, it has been used as an endodontic sealer. Bioceramic (BC) is a newly developed material based on calcium silicate, which is already used as a biocompatible root canal obturation material to overcome the weakness of MTA. However, there have been no studies comparing the materials used to seal the root canal. Objective: This study aimed to determine the apical leakage differences between BC and MTA sealers with BC-coated gutta-percha (GP) and conventional GP. Methods: In total, 40 mandibular premolars were cut coronally to 14 mm of working length. All samples were randomized and sorted into four groups: BC sealer with BC-coated GP, bioceramic sealer with conventional GP, MTA sealer with BC-coated GP, and MTA sealer with conventional GP. The samples were stored in an incubator, and nail varnish was applied to all root surfaces except for a 1 mm area from the apex. The samples were also soaked in methylene blue for 72 h before undergoing diaphanization. A stereomicroscope was used to measure the methylene blue penetration. All data were analyzed using analysis of variance. Results: There were significant differences in apical leakage between the BC and MTA sealers (P < 0.05). Conclusion: The BC sealer prevented apical leakage better than MTA. The BC-coated GP had better results than the conventional GP, but the differences were not significant, indicating that the choice of sealer is more important in preventing the apical leakage.

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Background: Lactobacillus reuteri and Lactobacillus casei have been proposed as probiotic bacteria that promote oral health. Objectives: The present study aimed to evaluate the in vitro effects of L. reuteri and L. casei on the biofilm formation of major oral pathogens, Streptococcus mutans and Porphyromonas gingivalis. Materials and Methods: L. casei strain Shirota and L. reuteri ATCC 55730 were isolated from the commercial products and cultured in de Man, Rogosa, and Sharpe broth. Polymerase chain reaction was used to confirm the identity of the species. S. mutans ATCC 25175 and P. gingivalis ATCC 33277 were cultured in brain–heart infusion broth and used for biofilm formation on 96-well microplate platform. The biofilms were treated with the probiotics and appropriate controls in a time-dependent experiment from 15 min to 24 h. The biofilm biomass was evaluated using crystal violet and safranin. Results: The statistical analysis showed a significant reduction in the S. mutans and P. gingivalis biofilms after treatment with the L. reuteri and L. casei probiotics at all incubation times (P < 0.05). Conclusion: The present study demonstrated the potential antibiofilm activity of L. casei strain Shirota and L. reuteri ATCC 55730 against S. mutans and P. gingivalis biofilms in vitro. The foregoing data have formed a basis for future clinical studies to evaluate the beneficial oral health effect of probiotic Lactobacilli strains.

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Background: Toothpaste is essential in the process of oral care. One of the components of toothpaste, Eugenia caryophyllus, might have an anti-inflammatory and antibacterial effect. Objectives: The objective of this study is to analyze the effectiveness of E. caryophyllus in the toothpaste against Streptococcus mutans, Lactobacillus, and total microbial load in the saliva of healthy subjects after toothbrushing using E. caryophyllus toothpaste. Methods: Saliva (n = 10, aged 18–25 years) were collected before and 2 weeks after toothbrushing using toothpaste-containing E. caryophyllus (Antiplaque, Triple-Ace, Indonesia). The total microbial load of saliva was determined by the measurement of colony-forming unit (CFU) number on brain–heart infusion agar, at 37°C, for 24 h, in anaerobic-condition. Real-time polymerase chain reaction technique was used to quantify the S. mutans and Lactobacillus deoxyribonucleic acid using SYBR green and 16S-rRNA gene-specific primers for S. mutans and Lactobacillus. Primers were 5'-ATTCCCGCCGTTGGACCATTCC-3' (fwd); 5'-CCGACAAAGACCATTCCATCTC-3' (rvs) and 5'-CTTGTACACACCGCCC GT CA-3' (fwd); 5'-CTCAAAACTAAACAAAGTTTC-3' (rvs) for S. mutans and Lactobacillus, respectively. Data were analyzed using t-pair test with P < 0.05 set as the level of significance. Results: The result showed that there was a significant reduction of total microbial load, S. mutans, and Lactobacillus number in the saliva after toothbrushing. The total microbial number in the saliva was significantly decreased before (5.84 ± 0.43 log CFU/mL) and 2 weeks after toothbrushing (5.27 ± 0.61 log CFU/mL) (P < 0.05). The number of S. mutans and Lactobacillus was also significantly decreased before (8.27 ± 0.11 and 2.34 ± 0.71 log CFU/mL) and after toothbrushing (8.18 ± 0.11 and 1.91 ± 0.25 log CFU/mL) (P < 0.05). Conclusion: E. caryophyllus toothpaste may reduce the number of total microbial, S. mutans, and Lactobacillus in the saliva of healthy subjects. Further studies are needed to explore this result.

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Background: The storage condition of a toothbrush can influence the growth of bacteria. Indonesia as a developing country has a low awareness of the importance of toothbrush hygiene. Thus, an efficient and economic toothbrush storage solution is needed. Objective: The aim of this study was to evaluate the effect of soaking toothbrushes in mouthwash solution on the total number of Streptococcus mutans and Fusobacterium on the toothbrush. Methods: Eighteen toothbrushes were provided to healthy individuals aged 19–23-years. For 1 month, each toothbrush was soaked in disinfectant solution (25 mL) for 20 min after brushing, with Group 1 using mouthwash and Group 2 using sterile tap water. The real-time polymerase chain reaction (PCR) method was used to evaluate the number of microorganisms on each toothbrush. The total number of DNA targets was identified using real-time PCR followed by SYBR Green reagents and 16S rRNA-gene specific primers for S. mutans and Fusobacterium. A paired sample t-test was used for statistical analysis, and the level of significance was set at P < 0.05. Results: There were significant differences in the total bacterial numbers of S. mutans and Fusobacterium on toothbrushes after soaking with mouthwash solution. The average scores (Log10-CFU/mL ± standard deviation) of the total bacteria (2.66 ± 0.39), S. mutans (1.21 ± 0.18), and Fusobacterium (10.35 ± 6.02) on toothbrushes in Group 1 were significantly decreased compared to the average scores of the total bacterial load (5.19 ± 0.41), S. mutans (2.71 ± 1.59), and Fusobacterium (18.96 ± 4.26) in Group 2. Statistical evaluation brought statistically significant difference of total bacteria numbers and S. mutans between Group 1 and Group 2 (P < 0.05). Conclusion: Soaking toothbrushes in mouthrinse solution reduce the total number of bacterial load, S. mutans, and Fusobacterium. Further studies are needed to explore the effects of mouthwash solution against other oral pathogens.

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Background: Tooth extraction is the most frequently conducted dental care procedure. In Indonesia, there is an extremely high utilization of dental and oral health services for tooth extraction, reaching 79.6%. Pain is a side effect of extraction. Pain due to extraction wounds can be treated with analgesic drugs, but alternative drugs with minimal or no side effects are still being researched. An herbal active beluntas leaf substance can reduce pain from extraction wounds. The beluntas plant not only aids in healing wounds but also exhibits anti-inflammatory and antipyretic effects. Objectives: In this study, the aims were to determine the 50% inhibitory concentration (IC₅₀value) and examine the viability effect of an ethanol extract of beluntas leaves on fibroblast cell cultures in vitro. Methods: Laboratory experiments were carried out. Beluntas leaves were obtained; their leaf extracts were prepared using ethanol as the solvent; phytochemical tests were performed. Triplicate measurements for the viability of 3T3 BALB/c fibroblast cells were carried out using the Methylthiazol sulfophenyl (MTS Assay) method. The extract concentrations were 500, 250, 125, 62.50, 31.25, 15.63, and 7.81 μg/mL. Results: Data analysis was carried out by one-way analysis of variance statistical test. Analysis results revealed that extract concentrations of 500, 31.25, 15.63, and 7.81 μg/mL exhibit a significant difference in the effect of cytotoxicity (P < 0.05) on fibroblast cells, and the IC₅₀value is 265.388 μg/mL. Conclusion: A significant difference in the cytotoxicity effect between the concentrations of the ethanol extract of beluntas (P. indica) leaves on the fibroblast cell cultures in vitro was observed. The beluntas leaf extract at an IC₅₀value of 7.81 μg/mL did not affect cell viability; hence, it is considered safe.

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Background: Tea tree (Melaleuca alternifolia) oil (TTO) is known to have anti-inflammatory, antibacterial, antifungal, and antiviral properties. Objectives: The aim of this study was to determine the effects of TTO on the ability of Porphyromonas gingivalis and Aggregatibacter actinomycetemcomitans biofilms to adhere to enamel surfaces in vitro. Methods: P. gingivalis and A. actinomycetemcomitans were cultured in brain heart infusion (BHI) broth at 37°C for 24 h in anaerobic conditions. Eighteen premolar teeth were inoculated and incubated for 48 h to form biofilms on enamel surfaces. Subsequently, TTO in 6.25%, 12.5%, 25%, and 50% concentrations was added and incubated for 1 and 3 h. Chlorhexidine (0.2%) and BHI broth were used as positive and negative controls, respectively. The remaining biofilm colonies were counted using an enzyme-linked immunosorbent assay reader (490 nm). The teeth were placed in microtubes containing phosphate-buffered saline and vortexed for 20 s. Subsequently, biofilms were cultured in BHI agar for 24 h. The colonies in each concentration were estimated as colony-forming units per milliliter. Statistical analysis was performed using one-way analysis of variance. The level of statistical significance was set to P < 0.05. Results: Treatment with all concentrations of TTO significantly reduces biofilm adhesion compared to the negative control after both incubation periods (P < 0.05). The concentration that most effectively inhibited the adhesion of P. gingivalis was 12.5% after 1 h incubation. The concentration that most effectively inhibited the adhesion of A. actinomycetemcomitans was 25% after 1 h incubation. Conclusion: TTO inhibits the adhesion of P. gingivalis and A. actinomycetemcomitans biofilms to enamel surfaces and may be useful as a treatment for oral diseases. Further studies should examine its efficacy in vivo.

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