EFFECT OF VEILLONELLA INFANTIUM ON BIOFILM FORMATION OF ORAL STREPTOCOCCUS

Objective: Therefore, we aimed in this study to evaluate the effect of Veillonella infantium on the biofilm formation of oral Streptococcus species.Methods: Dual-species biofilm was formed between V. infantium and oral Streptococcus using the wire method, and it was then incubated at 37°C underanaerobic condition for 5 days. Biofilm formation was determined by measuring the DNA concentration. Single species biofilm of oral Streptococcuswas generated under the same conditions and was used as a control group.Result: The presence of V. infantium decreased the biofilm formation of Streptococcus mutans, where, in contrast, the formation of biofilm inStreptococcus sanguinis was increased by the presence of V. infantium (p<0.05).Conclusion: The presence of V. infantium decreased the biofilm formation of S. mutans

Association between Volatile Sulfur Compounds Prevotella intermedia and Matrix Metalloproteinase-8 Expression

Objective: To determine the correlation between levels of methyl mercaptan (CH3SH) hydrogen sulfide (H2S), the proportion of Prevotella intermedia (Pi), and matrix metalloproteinase-8 (MMP-8) gene expression levels in periodontitis patients accompanied by halitosis. Material and Methods: Samples were obtained from gingival crevicular fluid (GCF) in the deepest pocket and by swabbing in the tongue coating area in patients with periodontitis presenting with halitosis (n = 23) and healthy subjects as controls (n = 7). The values of CH3SH and H2S were obtained using Oral Chroma. The proportion of Pi and MMP-8 expression levels were evaluated using PCR-RT. All the result was statistically analyzed using SPSS software. Results: The levels of CH3SH and H2S in participants with PD ≥ 6 mm showed a robust negative correlation with the proportion of P. intermedia in GCF and tongue coating. No statistically significant association was detected between CH3SH and H2S levels and MMP-8 expression levels (p>0.05). Conclusion:  There is no association between CH3SH and H2S levels, the proportion of P. intermedia, and MMP-8 expression in patients with periodontitis accompanied by halitosis

Isolation and Identification of Indonesian Lactobacillus reuteri strain from Saliva of Young Adults

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

BIOFILM FORMATION OF CANDIDA ALBICANS EXPOSED TO ETHANOL EXTRACT OF PROPOLIS

Objective: Propolis extract showed an excellent in vitro performance against yeast and was additionally found to be fungistatic and fungicidal. Propolisextract is also used for treatment and prevention of fungal infections. However, its effectiveness against Candida albicans biofilm formation requiresinvestigation. The study evaluated the ability of propolis to inhibit C. albicans while the fungus is growing as a biofilm in vitro.Methods: Two reference strains, C. albicans ATCC 25923 and a clinical strain (laboratory stock), were used in this study. For the biofilm experiment,the fungi were cultured in Tryptic Soy Broth medium with 1% sucrose and incubated at 37°C for 24 h, and different concentrations of ethanol extractof propolis were used as the inhibitor agents. Biofilm assays were performed in 96-well microtiter plates, quantification of the total biofilm biomasswas performed using a crystal violet staining method, and the Student’s t-test was chosen for statistical analyses.Results: Our data showed that 3 h incubation with propolis did not affect the biomass in the experimental group compared to the control. When theincubation time was extended to 18 h, the biomass increased significantly compared to the control.Conclusion: This study showed that several concentrations of propolis did not inhibit biofilm. However, in each incubation time, we observed nohyphal morphology in the biofilm mass. Propolis might attenuate the opportunistic virulence of fungus growing as a biofilm in vitro. Further studiesare necessary to confirm this phenomenon

ANALISIS MORFOLOGI KOLONI DAN KERAGAMAN GENOTIP STREPTOCOCCUS SANGUINIS YANG BERASAL DARI PLAK GIGI DAN SALIVA PENDERITA PENYAKIT JANTUNG KORONER: ANALYSIS OF COLONY MORPHOLOGY AND GENOTYPE DIVERSITY OF STREPTOCOCCUS SANGUINIS FROM THE DENTAL PLAQUE AND SALIVA OF CORONARY HEART DISEASE PATIENTS

Penelitian sebelumnya menunjukkan akumulasi kalkulus supragingiva pada pasien dengan Penyakit Jantung Koroner(PJK) terbentuk lebih cepat dan lebih banyak dibandingkan pasien non PJK. Streptococcus sanguinis dikenal bukan sajadalam hubungannya dengan pembentukan karang gigi, tetapi juga dengan endokarditis bakterial yang dapat menimbulkankematian. Penelitian awal ini bertujuan untuk mengidentifikasi S.sanguinis yang berasal dari plak gigi dan saliva pasienPJK. Sampel berasal dari plak gigi dan saliva 17 pasien PJK yang disebar pada plat Agar Mitis Salivarius dan diinkubasidalam suasana mikroaerofilik. Total sampel adalah 50,35 berasal plak gigi, 15 dari saliva, yang diasumsikan sebagai S.sanguinis atas`dasar morfologi koloninya yang jelas, lekat, berbentuk bintang, diambil dan dikonfirmasi dengan caramenumbuhkan setiap koloni kedalam agar darah bernutrisi. Bacterial chromosomal DNA diekstraksi dan dikonformasisebagai S. sanguinis dengan menggunakan primer PCR amplifikasi spesifik regio intergenik (475-bp) dan strain standardS. sanguinis ATCC 10556. Diversiti genotip diobservasi menggunakan primer OPA2. Didapatkan 4 dari 50 isolat (8%), 1koloni dari saliva, dan 3 koloni dari plak gigi dikonformasi sebagai S. sanguinis. Sebagai kesimpulan, morfologi koloni S.sanguinis jelas, lekat, berbentuk bintang, didapatkan 4 keragaman genotip S. sanguinis di dalam plak gigi dan salivapasien PJK

Expression of Toll-Like Receptor 4 and Matrix Metalloproteinase 8 in Gingival Crevicular Fluid in Patients with Periodontitis

Objective:To determine the expression of TLR4 and MMP8 in gingival crevicular fluid [GCF] in patients with periodontitis. Material and Methods:Clinical samples were collected from 23 gingival crevicular fluid of periodontal disease subjects (n = 14) and healthy periodontal subjects (n=9). Measurement of Clinical parameters of probing pocket depth (PPD), bleeding on probing (BOP), and clinical attachment loss (CAL) were included as diagnostic criteria. Pocket Depth (PD) and CAL were defined as present if the PPD was ≥ 4 mm and the CAL ≥ 1 mm. Expression of TLR4 and MMP8 in the gingival crevicular fluid of deep pockets (PD≥ 6mm), shallow pockets (PD 4-5 mm) and healthy periodontal sulcus (0-3 mm) were evaluated by quantitative real-time polymerase chain reaction (qRT-PCR). Statistical analysis to compare the pocket was using Independent t-test and Mann-Whitney test. Correlation between mRNA expression and clinical parameters was analyzed using Spearman’s correlation test. Results:Expression of TLR4 was higher in shallow pockets compared to the control group, but the difference was not statistically significant (p>0.05). The expression of MMP8 was higher in shallow pockets compared to the control group, but the difference was not statistically significant (p>0.05) either. There is no significant correlation between TLR4 and MMP8 with clinical periodontal parameters. Conclusion:TLR4 and MMP8 mRNA expression levels should not be used as a clinical biomarker in periodontitis diagnostic tools