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Abstract Aptamers that bind live bacterial cells have been widely investigated, but their potential to inhibit Candida albicans biofilm formation needs to be further explored. The aims of this study were to evaluate the binding of C. albicans to RNA aptamers and to examine the potential of aptamers to inhibit C. albicans biofilm formation in vitro. In this study, RNA aptamers selected against yeast cells of C. albicans ATCC 10231 were developed using the systematic evolution of ligands by exponential enrichment (SELEX) technique. The binding affinity of the resulting aptamers was then determined by an aptamer‐linked immobilized sorbent assay (ALISA), and a colorimetric (MTT) assay was used to measure the metabolic activity of Candida biofilms. After 11 rounds of SELEX, two candidate aptamers, Ca‐apt‐1 and Ca‐apt‐12, were identified. The Ca‐apt‐1 aptamer also recognized C. albicans isolated from clinical specimens but did not recognize other oral microorganisms (i.e., Streptococcus mutans and Saccharomyces cerevisiae). The ALISA results showed that the binding affinity of these aptamers was comparable to that of an anti‐C. albicans monoclonal antibody. In addition, Ca‐apt‐1 could inhibit biofilm and hyphal formation of C. albicans in vitro, as demonstrated using biofilm assays. This study shows that RNA aptamers could potentially be used in diagnostic and therapeutic applications for C. albicans‐related disease in the future.

Introduction: Early Childhood Caries (ECC) affects a significant proportion of the child population worldwide. Salivary proteins contain a host defense system that plays a role in the caries pathogenesis. Aim: To analyse the protein profile and salivary viscosity from ECC and caries-free subjects. Materials and Methods: The number of subjects included in the study was 16 ECC and 16 caries-free. Salivary viscosity was assessed visually and salivary protein profile was analysed using SDS PAGE. Results: The five most prominent protein bands with molecular mass 15 kDa, 25 kDa, 60 kDa, 65 kDa and 95 kDa were found in ECC patients with the frequency of occurrences in order 8, 3, 16, 6, 2 and caries-free subjects 14, 5, 16, 16, 9. The saliva viscosity of ECC patient was higher than the caries-free subjects. Conclusion: In ECC patients, the frequency of occurrences of salivary protein profile 15 kDa, 25 kDa, 65 kDa and 95 kDa were found less than in caries-free subjects. Meanwhile, protein profile 60 kDa has the same frequency of occurrence in ECC and caries-free subjects. Viscosity of ECC patient was higher than the caries-free subject.

Background: The aim of this study was to analyze the synergistic relationship between Candida albicans and Streptococcus mutans in children with early childhood caries (ECC) experience. Methods: Dental plaque and unstimulated saliva samples were taken from 30 subjects aged 3-5 years old, half with (n=15, dmft > 4) and half without (n=15) ECC. The abundance of C. albicans and S. mutans and relative to total bacteria load were quantify by real-time PCR (qPCR). This method was also employed to investigate the mRNA expression of glycosyltransferase ( gtfB) gene in dental plaque. Student’s t-test and Pearson’s correlation were used to perform statistical analysis. Results: Within the ECC group, the quantity of both microorganisms were higher in the saliva than in dental plaque. The ratio of C. albicans to total bacteria was higher in saliva than in plaque samples (p S. mutans (p < 0.05). The different value of C. albicans and S. mutans in saliva was positively correlated, and negatively correlated in dental plaque. Transcription level of S. mutans gtfB showed a positive correlation with C. albicans concentration in dental plaque. Conclusion: C. albicans has a positive correlation with cariogenic traits of S. mutans in ECC-related biofilm of young children.

Background/purpose: In the oral environment, Candida albicans interacts with many bacteria, including Enterococcus faecalis. We investigated the susceptibility of C. albicans biofilm development to the presence of unencapsulated E. faecalis cps2 in comparison with reference strains (E. faecalis ATCC 29212) or their respective spent medium (collected at 6 hours). Material and methods: Crystal violet stain was used to measure the total biofilm mass, whereas quantitative real-time polymerase chain reaction was used to analyze the change in expression of the mRNA of hypha morphology (ALS1 and ALS3) and biofilm maturation (EFB1). Results: At the intermediate stage, C. albicans resisted the presence of each E. faecalis strain tested and their spent medium. However, at the maturation stage, the unencapsulated strain was stronger in reducing C. albicans biofilms than the reference strain (P

This study was carried out to investigate the phenotype and genotype of E. faecalis isolated from the root canal and saliva of primary endodontic patients with periapical lesions. Eighteen adult male and female individuals suffering from primary endodontic infection, either had or had not periapical lesions, were involved in this study. Root canal scraping and saliva were collected from each subject and used for bacterial quantitation using a real-time polymerase chain reaction (RT-PCR). Enterococci were isolated using ChromAgar medium and then identified using both biochemical (Gram staining and catalase tests) and molecular biology (conventional PCR) methods. Gelatinase activity, polysaccharide capsul profile and mRNA ace expression level were determined using microbiological, biochemical and molecular biology approach, respectively. Genotype of E. faecalis was determined based on nucleotide sequence of ace and gelE genes analyzed using web-based 3730xl DNA Analyze software. The results showed that high proportion of E. faecalis found in both root canal and saliva of is related to the incidence of periapical lessions in the primary endodontic patients. This is contrast to the insignificant relationship found between Cps polymorphism, gelatinase activity, and mRNA ace expression with periapical lesions in the patients, respectively.DOI: 10.14693/jdi.v23i1.960

Heat Shock Proteins (HSP) are highly conserved immunoreactive group of proteins found in microorganisms and animal/human tissue. In addition to heat, other stressful conditiions also induce stressed proteins, especially anorexia, heavy metal ion, exposure to H2O2 and infection by DNA or RNA viruses. Recent studies suggest the involvement of HSPs as autoantigens in autoimmune diseases, including rheumatoid arthritis, systemic lupus erythematosus, Bechet's syndrome, recurrent oral uclers, oral lichen planus and other. The HSPs 60 - 65 KDa might be involved in the pathogenesis of autoimmune diseases such as Bechet's syndrome, recurrent oral ulcers, and oral lichen planus. This paper will discuss the immunopathogenesis mechanism of those diseases induced by HSPs.

The utilization of cajuput essential oil as a flavor in candy may produce a physiological active added value. Some compounds of cajuput plant (Melaleuca cajuputi L) have been reported for their anti-microbial activities. Candida albicans is a normal commensal organism in human mouth. However, it may become virulent and responsible for oral diseases known as oral candidiasis. This study aimed to determine the effect of cajuput and peppermint oil in cajuputs candy in inhibiting the C. albicans biofilms formation by using in vitro biofilm assay and viability assay. Furthermore, the influence of concentration of cajuput oil on the anti-microbial activities had been analyzed. All the tested concentration of cajuput oil in cajuputs candy was effective to inhibit the viability of C. albicans. The provision of flavor components of cajuput and peppermint oil could produce synergistic effects compared to a single flavor component. The addition of cajuput oil at 0.6% was able to inhibit the viability of C. albicans. The activities of the cajuput oil showed positive correlation to the concentration. The variable of plus and minus 0.1% addition of the cajuput oil concentration, however, produced no significant difference to inhibit the growth of C. albicans in biofilm. Sensory test, hedonic test, was conducted to evaluate the flavor, aroma, and overall attributes, resulting in no significant difference between 0.6 to 0.8% additions of cajuput oil upon the sensory acceptance.

Prevotella intermedia and Porphyromonas gingivalis are two of the specific microorganisms that frequently occurs in chronic periodontitis. As protein plays an important role in bacteria's life and its virulence factor, we tried to explore variation in P. intermedia and P. gingivalis cell protein profile which isolated from different periodontl pocket depth of chronic periodontitis patient. P. intermedia and P. gingivalis were identified using Polymerase Chain Reaction (PCR) technique, and its protein profile were evaluated using Sodium Dodecyl Sulfate - Polyacrylamide Gel Electrophoresis (SDS-PAGE). SDS-PAGE result showed that there were significant difference between protein profile of P. intermedia cell which isolated from slhallow and deep pocket with dominant proteins of molecular weight 200 kDa, 110 kDa, 40 kDa, and 25 kDa, however, relationship between pocket depth and P. intermedia cell protein pofile could not be concluded because unrepresentative number of P. intermedia colonies. In the other hand the variation in P. gingivalis cell protein profile was not influenced by pocket depth with dominant protein called hemin binding protein (HbBp).DOI: 10.14693/jdi.v17i1.110

Xylitol is reported to inhibit the growth of C. albicans. Objectives: Investigating serum factor role in inhibiting the growth of C. albicans and the effect of 1%, 5%, 10% xylitol on C. albicans resistance in serum in vitro. Methods: Identification of C. albicans (oral swab of candidiasis patient) was conducted using CHROMAgar, confirmed by germ tube test. The cultures were serially diluted, inoculated in Saburoud Dextrose Broth (SDB) contained 0% (control), 1%, 5%, or 10% xylitol, and kept for 3 or 7 days. These inoculations were then exposed to either active or inactive serum (Fetal Bovine Serum heated in 65ºC for 30 minutes) for 2 hours in 37ºC. The colony forming unit (CFU) of C. albicans in Saburoud Dextrose Agar (SDA) were counted after 2 days. C. albicans ATCC 10231 strain was used as a comparison. One-way ANOVA with 0.05 was used. Results: After 3 days cultured in media with or without xylitol, the CFU of C. albicans exposed to active serum were significantly lower than those exposed to inactive serum (p=0.032). Although not statistically significant (p=0.689), increased concentration of xylitol lead to increased resistance of C. albicans in active serum. Only 7 day exposure of 10% xylitol resulted in significantly higher growth of C. albicans (p=0.034). No significant difference of C. albicans CFU in active or inactive serum (p=0.404). Conclusion: Serum factor has role in inhibiting C. albicans growth in vitro. Exposure of 1%, 5%, or 10% xylitol for 3 or 7 days has no significant effect on C. albicans resistance in serum.DOI: 10.14693/jdi.v16i2.98

The growth of C. albicans is influenced by glucose intake. Xylitol is commonly used as sugar substitute. Reported effective concentrations of xylitol in reducing C. albicans growth in vitro were varied, 1%, 5%, and 10%. Objectives: Investigate the effect of different concentration and duration of xylitol exposure in inhibiting C. albicans growth in vitro. Method: Identification of C. albicans from oral swab of a male candidiasis patient was conducted using CHROMagar, confirmed by germ tube test. C. albicans suspension (108 cells/μl) were inoculated in SDB contained 1%, 5%, 10% xylitol, and without xylitol (as control), for 3 and 7 days, then incubated in 37oC on SDA and counted for their CFU after 48 hours. The C. albicans ATCC 10231 strain was used as a comparison. Results: After 3 days, increased concentration of xylitol (1%, 5%, 10%) lead to decrease growth of C. albicans, both the ATCC 10231 (125%; 51%; 14% respectively) and the clinical isolate (103%; 81%; 42%), p = 0.044. Significant lower growth of C. albicans compared to control were only seen in those exposed to 10% xylitol (p = 0.024). After 7 days, exposure of 1%, 5%, 10% xylitol did not significantly affect the growth of C. albicans (p = 0.396). Conclusion: The growth of C. albicans could be inhibited by 10% xylitol for 3 days.DOI: 10.14693/jdi.v16i1.12

High carbohydrate intake is one of predisposing factors of oral candidiasis. Objective: Investigating the effect of 1%,5%,10% glucose addition on the growth of C.albicans in vitro. Method: C.albicans sample was taken from oral swab of a male oral candidiasis patient. Identification of C.albicans was conducted using CHROMagar and confirmed by germ tube formation in serum. As a comparison, C.albicans ATCC10231 was used. After 2 days the cultures were serially diluted and inoculated in SDB without glucose, and with 1%,5%,10% addditional glucose, kept for 3 and 7 days in room temperature, then inoculated in SDA. The CFU/ml were counted after 2 days. ANOVA with α0.05 was used. Result: Statisticaly, additional 1% glucose for 3 days lead to significant decreased of growth of both clinical strain and ATCC 10231 C. albicans. However, only additional 5% and 10% glucose in clinical isolate for 7 days increased the growth of C.albicans significantly. Conclusion: The effect of additional glucose on the increased growth of C.albicans in vitro is influenced by the concentration, exposure duration of glucose, and by the strain of C.albicans.DOI: 10.14693/jdi.v16i1.14

Background: Elderly often suffer malnutrition and oral candidiasis. Candida albicans (C. albicans) which is the most prominent cause of oral candidiasis, is one of commensal oral micro-flora. Nutritional status affect the characteristic of saliva. Saliva is the regulator in the development of C. albicans from comensal into pathogen. Purpose: The purpose of this study was to determining the correlation between elderly nutritional status with salivary total protein and its activity in inhibiting C. albicans growth and biofilm formation. Methods: Using mini nutritional assessment 30 elderly were classified into normal and malnutrition groups. Total protein of unstimulated saliva was measured using Bradford protein assay. The colony forming unit (CFU) of C. albicans was counted on 72 hours on SDA cultures without (control) or with 2 hour saliva exposure. Biofilm formation was analyzed from the optical density of 10–5 C. albicans suspension without saliva exposure (control) or with exposure of 10.000 μg/ml saliva and incubated in 37° C for 2 days. The suspension was put into 96 well plates, stained with crystal-violet dye, and analyzed using microplate reader. Differences between groups were analyzed using independent t-test or Kruskall-Wallis. Correlation between variables was analyzed using Spearman test. Results: Salivary total protein of normal elderly (1.113.5 ± 1.1143.3) was higher than those of malnutrition (613.6 ± 253.6) but not statistically significant (p > 0.05). The CFU of C. albicans exposed to saliva of normal samples (2.060 cfu/ml) was significantly lower than control (24.100 cfu/ml) and those exposed to malnutrition saliva (5.513.3 cfu/ml). C. albicans biofilm formation is highest in controls (0.177), lower in those exposed to malnourished saliva (0.151) and lowest in those exposed to saliva of good nourished elderly (0.133). Conclusion: Although does not cause significant decrease of salivary total protein, malnutrition in elderly results in lower capacity of saliva in inhibiting the growth and declining the virulence of C. albicans.Latar belakang: Lansia sering menderita malnutrisi dan kandidiasis oral. Candida albicans yang merupakan penyebab utama terjadinya candidiasis, adalah salah satu mikroflora rongga mulut yang bersifat konvensional. Malnutrisi memengaruhi karakteristik saliva. Saliva merupakan regulator utama perkembangan Candida albicans (C. albicans) dari sifat konvensional menjadi bersifat patogen.Tujuan: Tujuan dari penelitian ini adalah untuk menentukan korelasi antara status gizi lansia dengan total protein dan aktivitas saliva dalam menghambat pertumbuhan dan pembentukan biofilm C. albicans. Metode: Menggunakan mini nutritional assessment, 30 lansia diklasifikasikan menjadi kelompok gizi baik dan gizi buruk. Total protein unstimulated saliva diukur dengan metode Bradford protein assay. Colony forming unit (CFU) dihitung pada kultur C. albicans pada saburaud dextrose agar (SDA) berusia 72 jam yang sebelumnya telah dipaparkan saliva selama 2 jam. Kontrol adalah kultur C. albicans tanpa paparan saliva. Pembentukan biofilm adalah pengukuran optical density suspensi 10–5 C. albicans tanpa paparan saliva (kontrol) atau dengan paparan saliva 10.000 μg/ml dan diinkubasi pada suhu 37° C selama 2 hari. Suspensi tersebut kemudian dimasukkan ke dalam 96 well plates, diberi pewarna crystal violet, dan diukur menggunakan microplate reader. Analisis data menggunakan uji beda t Independen atau Kruskall-Wallis, dan uji korelasi Spearman. Hasil: Total protein saliva lansia gizi baik (1.113,5 ± 1.1143,3) lebih tinggi dari lansia gizi buruk (613,6 ± 253,6) tetapi tidak bermakna secara statistik (p > 0,05). Pembentukan koloni C. albicans yang terpapar saliva lansia gizi baik (2.060 cfu/ml) secara signifikan lebih rendah dari kontrol (24.100 cfu/ml) dan daripada yang terpapar saliva lansia gizi buruk (5.513,3 cfu/ml). Pembentukan biofilm C. albicans tetinggi pada kontrol (0,177), lebih rendah pada yang terpapar saliva gizi buruk (0,151) dan terrendah pada yang terpapar saliva gizi baik (0,133). Kesimpulan: Meskipun malnutrisi tidak menyebabkan penurunan total protein saliva lansia, tetapi menurunkan kapasitasnya dalam menghambat pertumbuhan dan mengurangi virulensi C. albicans.

Experimental studies have shown that dental pulp tissue has potential to regenerate dentine in response to adverse stimuli, such as caries and associated operative procedures. However, the potential of dental pulp regeneration seems to be limited by regenerative capacity of the cell involved. In this study, we report the effect of transfection of a recombinant plasmid containing human BMP-2 gene in proliferation and differentiation of dental pulp tissue in vitro. The regenerative capacity was analyzed by ALP production and calcium content. Results showed that the transfected dental pulp cell was able to differentiate into the odontoblast phenotype, indicating the presence of odontoblast progentitor cells in dental pulp tissue.

Background: Dental caries remains as main problem in Indonesia and its prevalence is high (90.05%). However, there is no appropriate data that can be used to analyze dental caries in toddlers, especially baby bottle tooth decay syndrome (BBTD), though the number of BBTD cases is high in some pediatric dental clinics (90% of patients visiting the clinics). Even though some factors have already been considered to be the risk factor of BBTD, the main risk factor of BBTD is still unknown, especially BBTD in Indonesia. Purpose: This research was aimed to obtain data relating with bottle-feeding habit in 3-5 year old children in Indonesia and its caries risk. Method: The study was an observational research conducted with clinical examination through caries status (deft) of each child deserved by pediatric dentists and through questionnaire distributed to parents to examine the risk factor of BBTD. Observation was conducted on 62 children in the range of age 3 to 5 years old with bottle-feeding habit. Result: The results revealed that status of caries was various. The data showed that the frequency of bottle feeding more than twice could trigger BBTD 2.27 times higher than other factors such as the use of bottle feeding as a pacifier prior sleeping, the period of bottle-feeding, and the breast-feeding experience. Conclusion: though milk as subtract can possibly become a factor triggering caries, the frequency of bottle-feeding is highly considered as main factor. Since it could modulated the bacterial colonization on dental surface, which affects its virulence.Latar belakang: Karies masih menjadi masalah utama di Indonesia. Dalam praktek sehari-hari prevalensi karies masih sangat tinggi (90.05%). Belum ada data yang memadai dalam penelaahan karies yang spesifik pada anak balita selama ini khususnya kasus sindroma karies botol (SKB) sementara itu kasus SKB ditemukan sangat tinggi di beberapa klinik gigi anak (90% dari jumlah pasien yang datang ke klinik). Beberapa faktor menjadi resiko kejadian SKB dan belum diketahui faktor resiko utama kejadian karies khususnya di Indonesia. Tujuan: Penelitian ini dilakukan guna mendapatkan data yang berhubungan dengan kebiasaan minum susu botol pada anak usia 3-5 tahun di Indonesia serta resiko kejadian karies yang ditimbulkannya. Metode: Penelitian ini merupakan penelitian observasional yang dilakukan dengan metode pemeriksaan klinis melalui pencatatan status karies (deft) setiap anak oleh dokter gigi anak serta pengisian kuesioner yang dilakukan oleh orang tua untuk menentukan faktor resiko kejadian SKB. Pengamatan dilakukan pada 62 orang anak usia 3-5 tahun yang mempunyai kebiasaan minum susu botol sesuai dengan kriteria inklusi. Hasil: Hasil pemeriksaan klinis dan kuesioner memberikan gambaran status karies yang bervariasi. Data yang didapat dari penelitian ini menjelaskan, bahwa frekuensi minum susu botol lebih dari dua kali menyebabkan SKB 2.27 kali lebih besar dibandingkan dengan beberapa faktor lain, seperti menjadikannya pengantar tidur, lamanya mengonsumsi, dan riwayat minum ASI. Kesimpulan: Susu sebagai subtrat mungkin dapat dijadikan alasan kejadian karies akan tetapi yang menjadi resiko utama kejadian adalah frekuensi konsumsi susu botol itu sendiri. Kondisi ini dapat dihubungkan dengan modulasi substrat terhadap perkembangan kolonisasi bakteri di permukaan gigi, sehingga secara tidak langsung juga mempengaruhi virulensinya.

Background: Streptococcus mutans are gram positive bacteria classified into viridians group, and have a role in pathogenesis of dental caries. It’s adhesion to the tooth surface is mediated by cell surface proteins, which interact with specific receptor located in tooth pellicle. Glucan binding protein, Glukosyltransferase, and antigen I/II are basic proteins of S. mutans, which have a role in initiating the interaction. A previous study showed that chicken’s IgY can interfere the interaction. Purpose: The objective of this study was to assess the ability of IgY in recognizing the surface molecule of Streptococcus mutans expressed by various serotypes (c, d, e, f) and a strain derived from IPB, Bogor. Method: Western blot was used as a method to determine such capability. Result: The result showed that IgY has a potency to recognize antigen I/II, but not the other proteins on the cell surface of all bacteria tested. Conclusion: The ability of IgY to bind the surface protein, antigen I/II, indicates that this avian antibody could be used as a candidate for anti-adhesion in preventing dental caries.

Quorum sensing systems has been identified as one of mechanism carried out by numerous Gram-positive and Gram-negative bacteria to coordinate virulence and biofilm development. Using quorum sensing bacterial colonies synchronize gene expression and phenotype change allowing them to protect their niche. The purpose of this review is to present a synopsis of the literature on bacterial quorum sensing and we highlight the role of specific signaling molecules that might be used as a target of inhibitor agent in dental preventive perspective.