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Persistent Endodontics Pathogens Biofilm Inhibited by Lactobacillus reuteri Indonesian Strain Widyarman, Armelia Sari; Lazaroni, Njoo Kiky Elysia
Journal of Dentistry Indonesia Vol. 26, No. 3
Publisher : UI Scholars Hub

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Abstract

Lactobacillus reuteri is known as probiotics that effective to improve oral health. Objective: To analyze the effect of L. reuteri Indonesian strain, towards Enterococcus faecalis and Candida albicans biofilm growth. Methods: This study was conducted using biofilm assay. Each E. faecalis ATCC-29212 and C. albicans ATCC-10231 were cultured in Brain Heart Infusion Broth, distributed into 96 well-microplate and incubated for 24h, 37°C, anaerobic condition for E. faecalis and aerobic condition for C. albicans. L. reuteri LC382415 (Indonesian strain) was cultured in deMann Rogosa Shar pe Broth and diluted into different concentrations (108,106, and 104CFU/mL) subsequently, each concent ration dist ributed into biofilm well. Biofilm wells without probiotic was used as negative cont rol. Biofilm mass were measured using crystal-violet dye at 490nm using microplate-spectrophotometer. Data was statistically analyzed by one-way ANOVA test, statistical significance set as p<0.05. Results: Significant reduction of biofilm growth of E. faecalis and C. albicans after treatment with L. reuteri. The ideal concentration was found at L. reuteri 108 CFU/mL with 79.2% E. faecalis biofilm reduction and L. reuteri 104CFU/mL with 62.5% C. albicans biofilm reduction compared to control ( p<0.05) set at 100%. The ANOVA test results showed that L. reuteri in all concentrations and all time periods in this study had the ability to inhibit biofilm growth of both species ( p<0.05) compared to negative control. Conclusion: L. reuteri Indonesian strain inhibit the biofilm growth of E. faecalis and C. albicans. This antibiofilm effect may be useful in preventing biofilm growth in root canal.
Effectiveness of Lentinus edodes Mushroom Extract on Eradication of Enterococcus faecalis Biofilm Anggiarini, Putri Lestari; Amin, Meiny F; Gunawan, Juanita A; Widyarman, Armelia Sari
Journal of Dentistry Indonesia Vol. 27, No. 2
Publisher : UI Scholars Hub

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Abstract

Sodium hypochlorite is a commonly used irrigation solution in endodontic procedures, but it irritates tissues and has toxic effects. Lentinus edodes is a mushroom that has antibacterial properties. Enterococcus faecalis is an anaerobic bacterium that can cause root canal treatment failure. Objective: This study aimed to determine the effect of L. edodes extract on the eradication of E. faecalis biofilms. Methods: Phytochemical tests of L. edodes were performed to analyze alkaloids, steroids, triterpenoids, phenolics, tannins, flavonoids, and glycosides from this extract qualitatively. E. faecalis ATCC 29212 was cultured in brain heart infusion broth for 24 h at 37°C in an anaerobic atmosphere. Biofilm assay was performed to analyze the eradication of E. faecalis biofilm after treatment with L. edodes extract. The application times were 5, 15, and 30, and 10%, 20%, 40%, and 80% concentrations were used. Distilled water was used as a negative control, and NaOCl was used as a positive control. Data were statistically analyzed via one-way analysis of variance, where p < 0.05 was set as the level of significance. Results: L. edodes mushroom extract was effective in eradicating E. faecalis biofilms in all concentrations and incubation times compared with the control (p < 0.05). Significant differences were found between the application times of 5 and 15 min compared with 30 min (p < 0.05). The most effective concentration in eradicating E faecalis biofilms was 40% with an application time of 30 min. Conclusion: L. edodes mushroom extract proves its antibiofilm activity against E. faecalis biofilm. Further study is necessary to determine which substances are have the most influence on the effectiveness of L. edodes extract in eradicating E. faecalis biofilm in vivo.
Antibacterial and Antibiofilm Efficacy of Pineapple Hump (Ananas comosus) on Porphyromonas gingivalis in vitro Soulissa, Abdul Gani; Lombardo, Billy; Widyarman, Armelia Sari
Journal of Dentistry Indonesia Vol. 28, No. 3
Publisher : UI Scholars Hub

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Abstract

Background: Periodontal disease is one of the most prevalent oral health problems in Indonesia that affects supporting tissues of the teeth. Porphyromonas gingivalis plays an important role in the pathogenesis of periodontal disease. Alternative therapy with natural plant extracts, including pineapple (Ananas comosus) hump extract may inhibit the growth of bacteria that cause periodontal disease. Objective: To determine the effect of pineapple hump extract on bacterial growth and adhesion of Porphyromonas gingivalis biofilms. Method: The bacterial inhibition test was performed by the agar well diffusion method, and biofilm density measurements were made using the biofilm assay method. Results: Pineapple hump extract can inhibit bacterial growth optimally at a concentration of 100%, with an average zone of inhibition of 7.3 mm. The extract at a 50% concentration can eradicate the biofilms in a 6 h incubation time with an average OD of 0.124. The Shapiro-Wilk method confirmed a normal distribution of the data. Both one-way ANOVA and post hoc test showed a significant difference between the inhibitory ability of pineapple hump extract and between concentrations to restrict formation of Porphyromonas gingivalis biofilms. Conclusion: Pineapple hump extract was able to inhibit bacterial growth and eradicate the adhesion of Porphyromonas gingivalis biofilms in vitro.
The Effect of Parabiotic Reuterin on the Expression of Genes Involved in Candida albicans Biofilm Formation: An Ex vivo Study Riad, Anastasya Muna; Widyarman, Armelia Sari
Journal of Dentistry Indonesia Vol. 28, No. 3
Publisher : UI Scholars Hub

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Abstract

Candida albicans has a number of properties, including resistance to various antimicrobial agents, which allow it to survive in the root canals. Lactobacillus reuteri plays a role in maintaining oral health through interactions with the oral microbiome. L. reuteri has potential as a preventive and therapeutic agent against inflammatory diseases. Objective: The aim of this study was to investigate the effect of irrigation with a reuterin-containing solution on BCR1, ACE2, EFG1, and TEC1 gene expression in C. albicans root canal biofilms. Methods: L. reuteri was cultured in MRS broth and incubated anaerobically for 24 hours at 37°C. C. albicans was cultured in Sabouraud dextrose broth at 37°C for 48 hours. A total of 24 single-rooted premolar teeth were standardized and inoculated with C. albicans before irrigation with 50 μg/mL reuterin as a single, independent variable (Indonesian strain), 50 μg/mL reuterin Prodentis (a strain combination of L. reuteri DSM 17938 and L. reuteri ATCC PTA 5289), 2.5% sodium hypochlorite as positive control, and saline as negative control. A real-time quantitative polymerase chain reaction (RT-qPCR) assay was used to detect the expression of BCR1, ACE2, EFG1, and TEC1 in C. albicans root canal biofilms. Results: Reuterin significantly reduced the expression of BCR1 and ACE2 genes, which play a role in C. albicans biofilm formation, at the biofilm maturation stage (P < 0.05). Reuterin also affected the expression of the EFG1 and TEC1 genes, although the effect was not significant. Conclusion: A reuterin isolate of L. reuteri exhibits antibiofilm activity against the expression of C. albicans genes involved in biofilm formation. Reuterin has potential as an irrigation agent in the treatment of root canals. Further research is needed to shed light on the effectiveness of reuterin against the expression of genes that play important roles in the formation of C. albicans biofilms.
Strawberry Extract’s Effects on Enterococcus faecalis and Porphyromonas gingivalis Biofilms in vitro Armelia Sari Widyarman; Stephanie Brigitta Widjaja; Erik Idrus
Scientific Dental Journal Vol. 1 No. 1 (2017): September
Publisher : Faculty of Dentistry, Trisakti University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26912/sdj.v1i1.1911

Abstract

Background: Enterococcus faecalis (E. faecalis) and Porphyromonas gingivalis (P. gingivalis) are oral bacteria related to root canal infection and periodontal disease pathogenesis. Strawberries (Fragaria x ananassa) fruit are rich in vitamins and minerals, have antibacterial and antioxidant effects. Objective: This study investigated the inhibition effect of strawberry extract on monospecies and multispecies E. faecalis and P. gingivalis bacteria grown as biofilms in vitro. Methods: This study used E. faecalis ATCC 29212 and P. gingivalis ATCC 33277. It analyzed the effect of strawberry extract on bacteria biofilm formation using a biofilm assay on microplate wells. Five concentrations of strawberry extracts were used (100%, 50%, 25%, 12.5%, and 6.25%), and the inhibition effect was observed after a 1h, 3h, 6h, and 24h incubation period. Biofilms without the strawberry extract were used as the negative controls, and crystal violet and safranin (0.5%w/v) were used to count the biofilm mass. The biofilms grown on microplates were counted using an ELISA reader at 450 nm after 200 mL of 90% ethanol was added to attract the absorbed stain. The strawberry extract inhibition effectiveness on the biofilm formation of each bacterium tested was analyzed using one-way Anova, where p<0.05 was defined as a significant difference. Result: The strawberry extract inhibited the tested monospecies and multispecies bacteria biofilm formation. The optimal strawberry extract concentration for the inhibition of either monospecies biofilms was 100%. However, the optimal incubation time for the strawberry extract to inhibit the multispecies biofilm formation was 24h, which was the study’s biofilm maturity phase. Conclusions: The 100% strawberry extract concentration inhibited the formation of both the monospecies and multispecies E. faecalis and P. gingivalis biofilms. Future studies are needed to evaluate the potential of strawberry extract as an alternative dental therapy.
Consumption of Yogurt Containing Probiotic Bifidobacterium Lactis Reduces Streptococcus mutans in Orthodontic Patients Armelia Sari Widyarman; Shirley Trisna Yunita; Tjokro Prasetyadi
Scientific Dental Journal Vol. 2 No. 1 (2018): January
Publisher : Faculty of Dentistry, Trisakti University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26912/sdj.v2i1.1913

Abstract

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 (37ºC, 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.
Comparing the Effectivities of Chitosan Citrate and Chitosan Acetate in Eradicating Enterococcus faecalis Biofilm Uppalavanna Witedja; Tien Suwartini; Anastasia E Prahasti; Armelia Sari Widyarman
Scientific Dental Journal Vol. 2 No. 1 (2018): January
Publisher : Faculty of Dentistry, Trisakti University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26912/sdj.v2i1.2290

Abstract

Background: Adequate biomechanical preparations, antibacterial irrigants, and intracanal medications to promote the elimination of bacteria and their products are required to succeed root canal treatment. Enterococcus faecalis with its biofilm is known as an important etiological agent in endodontic treatment failures. Chitosan, as a natural product, has an antibacterial activity and is considered less toxic to the periapical tissue than other irrigants. However, the use of this natural product needs to be examined to determine its effectiveness as a root canal irrigant in endodontic treatment; this can be done by comparing it with the most common endodontic irrigant (NaOCl 5.25%) as a positive control. Objective: The objective of the study was to compare the effectiveness between 1–3% chitosan acetate (CA) and 1–3% chitosan citrate (CC) against E. faecalis biofilm formation after treatment for 15, 30, and 60 minutes. Methods: The study was conducted using 12 groups, including 1–3% CA, 1–3% CC, and control groups. E. faecalis biofilms in 96-well plates were exposed to each sample for 15, 30, or 60 minutes. Subsequently, the biofilms were stained with crystal violet solution, and the optical density value was measured using a microtiter plate reader at a wavelength of 600 nm. Results: CA and CC were effective in eradicating E. faecalis biofilm. However, the levels of effectiveness of CC and CA depended on the concentration and application time. Three-way analysis of variance (ANOVA) showed a significant difference between the irrigants (p <0.05) and three application times (p <0.05). The CA was effective in eradicating biofilm after 15 minutes of application, whereas the CC was more effective after 30 and 60 minutes of application. Conclusion: CC and CA are both effective in eradicating E. faecalis biofilm. 
Enzymatic Activity of Bromelain Isolated Pineapple (Ananas comosus) Hump and Its Antibacterial Effect on Enterococcus faecalis Dewi Liliany; Armelia Sari Widyarman; Erni Erfan; Janti Sudiono; Melanie Sadono Djamil
Scientific Dental Journal Vol. 2 No. 2 (2018): May
Publisher : Faculty of Dentistry, Trisakti University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26912/sdj.v2i2.2540

Abstract

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 anti-inflammatory and anticancer properties and exhibit antibacterial activity. Objective: 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) tests using diffusion and dilution techniques tested the antibacterial activity of the bromelain extract on 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.
Pomegranate Juice Inhibits Periodontal Pathogens Biofilm in vitro Armelia Sari Widyarman; Olivia Puspita Suhalim; Dhara Nandary; Citra Fragrantia Theodorea
Scientific Dental Journal Vol. 2 No. 3 (2018): September
Publisher : Faculty of Dentistry, Trisakti University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26912/sdj.v2i3.2572

Abstract

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 biofilm (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.
Isolation and Identification of Indonesian Lactobacillus reuteri strain from Saliva of Young Adults Armelia Sari Widyarman; Stella Pranoto; Citra Fragrantia Theodorea; Endang Winiati Bachtiar; Boy Muchlis Bachtiar
Scientific Dental Journal Vol. 2 No. 2 (2018): May
Publisher : Faculty of Dentistry, Trisakti University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26912/sdj.v2i2.2840

Abstract

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.