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Armelia Sari Widyarman
Department of Microbiology, Division of Oral Biology, Faculty of Dentistry, Trisakti University
Biochemistry, Genetics & Molecular Biology Dentistry Education Health Professions Immunology & microbiology

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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.
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.
Co-Authors Anastasia E Prahasti Boy Muchlis Bachtiar Citra Fragrantia Theodorea Dewi Liliany Endang Winiati Bachtiar Erik Idrus Erni Erfan, Erni Janti Sudiono Melanie Sadono Djamil Shirley Trisna Yunita Stella Pranoto Stephanie Brigitta Widjaja Tien Suwartini Tjokro Prasetyadi Uppalavanna Witedja