The EGCG and α-Mangosteen Stimulate SHED-IL10 and SHED-LL37 Metabolite Concentration.

OBJECTIVE: Stem cells of human exfoliated deciduous teeth (SHED) metabolites are secreted molecules from SHED, namely cytokines, chemokines, and growth factors. The metabolite can be used in various regenerative therapy based on cell-free immunomodulatory potential effects, like interleukin 10 (IL-10) and LL37. This molecule can stimulate with epigallocatechin gallate (EGCG) and α-mangosteen and has been proven to have anti-inflammatory and antibacterial effects. This study aimed to identify the effect of EGCG and α-mangosteen to SHED metabolite, called SHED-IL10 and SHED-LL37, from six passages to obtain the optimum stimulation and able to use as periodontitis regeneration treatment. MATERIALS AND METHODS: The six different passages of SHED were prepared in Dulbecco's modified Eagle medium and added with EGCG 80% (10 µM), EGCG 95% (10 µM), or α-mangosteen (10 µM). After a 24 hours incubation, each passage was measured with the metabolite concentration, SHED-IL10 and SHED-LL37, with human IL-10 and LL37 using enzyme-linked immunosorbent assay. Each different concentration was then analyzed statistically. RESULTS: The addition of EGCG 95% is able to stimulate the SHED-IL10 optimum concentration in passage 1 (p < 0.01). But, in the different conditions, the addition of EGCG 80%, EGCG 95%, and α-mangosteen was able to stimulate the SHED-LL37 optimum concentration in passage 2 (p < 0.001). CONCLUSION: The addition of EGCG and α-mangosteen can stimulate the SHED-IL10 and SHED-LL37 concentrations. These two metabolites are promising as regenerative therapy through anti-inflammatory and antibacterial properties.

Antibacterial effect of a gingival patch containing nano-emulsion of red dragon fruit peel extract on Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Fusobacterium nucleatum assessed in vitro.

Background: The most common bacteria causing periodontitis were Porphyromonas gingivalis, Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans. At present, plants are considered a valuable source of natural materials for use in the development of antimicrobial, anti-inflammatory and antioxidant agents. Hylocereus plyrhizus or red dragon fruit peel extract (RDFPE) contains terpenoids, and flavonoids can be an alternative. The gingival patch (GP) has been designed to ensure drug delivery and absorption into tissue targets. Objective: To assess the effect of a mucoadhesive gingival patch containing nano-emulsion of red dragon fruit peel extract (GP-nRDFPE) in inhibiting Porphyromonas gingivalis, Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans as compared to control groups. Material and method: Inhibition by diffusion method was carried out in Porphyromonas gingivalis, Fusobacterium nucleatum and Aggregatibacter actinomycetemcomitans medium. The test material was gingival patch mucoadhesive containing nano-emulsion red dragon fruit peel extract (GP-nRDFPR), gingival patch mucoadhesive containing red dragon fruit peel extract (GP-RDFPE), gingival patch mucoadhesive containing doxycycline (GP-dcx) and blank gingival patch (GP), conducted in four replications. The differences in the inhibition were analyzed using ANOVA and post hoc tests (p < 0.05). Result: GP-nRDFPE showed higher inhibition in Porphyromonas gingivalis, Aggregatibacter actinomycetemcomitans, and Fusobacterium nucleatum compared to GP-RDFPE at the concentrations 3.125% and 6.25% (p < 0.05). Conclusion: The GP-nRDFPE showed better anti-periodontic bacteria to Porphyromonas gingivalis, Fusobacterium nucleatum, and Aggregatibacter actinomycetemcomitans in line with its concentration. This is assumed that the GP-nRDFPE can use as periodontitis treatment.

Gingival Mesenchymal Stem Cells Metabolite Decreasing TRAP, NFATc1, and Sclerostin Expression in LPS-Associated Inflammatory Osteolysis In Vivo.

OBJECTIVE: Bone is a dynamic tissue that undergoes remodeling. During bone remodeling, there are transcription factors such as nuclear factor-activated T cells-1 (NFATc1), sclerostin, and tartrate-resistant acid phosphatase (TRAP) that are released for bone resorption. Metabolite from gingival mesenchymal stem cells (GMSCs) has the ability to activate proliferation, migration, immunomodulation, and tissue regeneration of bone cells and tissues. Furthermore, the aim of this study is to investigate the metabolite of GMSCs' effect on expression of NFATc1, TRAP, and sclerostin in calvaria bone resorption of Wistar rats. MATERIALS AND METHODS: Twenty male healthy Wistar rats (Rattus norvegicus), 1 to 2 months old, 250 to 300 g body were divided into four groups, namely group 1 (G1): 100 µg phosphate-buffered saline day 1 to 7; group 2 (G2): 100 µg lipopolysaccharide (LPS) day 1 to 7; group 3 (G3): 100 µg LPS + 100 µg GMSCs metabolite day 1 to 7; and group 4 (G4): 100 µg GMSCs metabolite day 1 to 7. Escherichia coli LPS was used to induce inflammatory osteolysis on the calvaria with subcutaneous injection. GMSCs metabolite was collected after passage 4 to 5, then injected subcutaneously on the calvaria. All samples were sacrificed on the day 8 through cervical dislocation. The expression of TRAP, NFATc1, and sclerostin of osteoclast in the calvaria was observed with 1,000× magnification. STATISTICAL ANALYSIS: One-way analysis of variance and Tukey honest significant different were conducted to analyze differences between groups (p < 0.05). RESULTS: The administration of GMSCs metabolite can significantly decrease TRAP, NFATc1, and sclerostin expression (p < 0.05) in LPS-associated inflammatory osteolysis calvaria in Wistar rats (R. norvegicus). There were significantly different TRAP, NFATc1, and sclerostin expressions between groups (p < 0.05). CONCLUSION: GMSCs metabolite decrease TRAP, NFATc1, and sclerostin expression in LPS-associated osteolysis calvaria in Wistar rats (R. norvegicus) as documented immunohistochemically.

Anti-Periodontopathogenic Ability of Mangrove Leaves (Aegiceras corniculatum) Ethanol Extract: In silico and in vitro study.

OBJECTIVE: Mangrove (Aegiceras corniculatum) is an abundant natural marine resource of Indonesia, which can be explored for treating periodontal disease due to its potential as immunoregulatory, antibacterial, and antioxidant properties. The objective of this study was to investigate the active compound from Indonesian mangrove leaf extract (A. corniculatum) (MLE) for developing a herbal-based mouthwash through in silico and in vitro studies. MATERIALS AND METHODS: Phytochemistry and liquid chromatography-high resolution mass spectrometry (LC-HRMS) were done to explore the active compounds in MLE. Chemistry screening and interaction, absorption, distribution, metabolism, and excretion (ADME), molecular docking simulation, and visualization of MLE active compounds as anti-inflammatory, antioxidant, and antibacterial were investigated in silico The inhibition zone of MLE against Aggregatibacter actinomycetemcomitans (Aa), Porphyromonas gingivalis (Pg), and Fusobacterium nucleatum (Fn) as periodontopathogenic bacterias was performed by diffusion method. Doxycycline 100 mg was used as a positive control, as a treatment group, there were five groups, namely 0%, 25%, 50%, 75%, and 100% MLE. RESULTS: Alkaloid, saponin, flavonoid, triterpenoid, steroid, tannin, and quinone were detected in MLE. A high concentration of (-)epicatechin and coumaric acid (CA) were found in MLE. MLE in 100% concentration has the most effective ability to inhibit Fn, Pg, Aa growth in vitro. (-)-Epicatechin has a higher negative binding affinity than CA that can enhance heat shock protein (HSP)-30, HSP-70, HSP-90, interleukin-10, and FOXP3 and also inhibit interleukin-6, peptidoglycan, flagellin, and dectin in silico. CONCLUSION: MLE of A. corniculatum has antioxidant, anti-inflammatory, and antibacterial activities that can be a potential raw material for developing a herbal-based mouthwash.

The use of mucoadhesive oral patches containing epigallocatechin-3-gallate to treat periodontitis: an in vivo study.

Objectives: The application of topical drugs such as mucoadhesive oral patches (MOPs) do not irritate the mucosa and are able to increase the permeability of drugs to oral tissue. Epigallocatechin-3-gallate (EGCG) is an active ingredient that exhibits significant antibacterial and anti-inflammatory effects. The purpose of this study was to analyze the therapeutic potential of a mucoadhesive oral patch containing EGCG (MOP-EGCG) in a model of periodontitis and investigate its effects on the expression of osteoprotegerin (OPG), receptor activator of nuclear factor-kappa Β ligand (RANKL) and receptor activator of nuclear factor-κB (RANK). Methods: A model of periodontitis was induced in Rattus novergicus used Porphyromonas gingivalis by applying 0.03 ml of bacteria locally with 1 × 1010 colony-forming units (CFU) seven times at 2-day intervals in the central lower incisors. Periodontitis was then treated with MOP (control), a mucoadhesive oral patch containing doxycycline (MOP-doxy) or MOP-EGCG for 1 h/day for 21 days. On days 3, 5, 7, 14 and 21 after treatment, the central lower incisor was biopsied and analyzed by immunohistochemistry for RANK/RANKL and OPG expression in the gingiva tissue. Results: MOP-EGCG extract significantly reduced the expression of RANKL and increased the expression of OPG and RANK (p < 0.05) when compared to the MOP-doxy and MOP groups. Conclusion: MOP-EGCG extract reduced the expression of RANKL and increased the expression of OPG and RANK, thus suggesting that MOP-EGCG can inhibit the loss of alveolar bone in periodontitis.

The ability of mucoadhesive gingival patch loaded with EGCG on IL-6 and IL-10 expression in periodontitis.

Objectives: The mucoadhesive gingival patch is a topical drug delivery process it does not cause any irritation in the mucosa. EGCG (Epigallocatechin-3-gallate) it has potent antioxidant, antiangiogenic and antitumor effects. The present study investigates the ability of mucoadhesive gingival patch loaded with EGCG on periodontitis and its impact on IL-6 and IL-10 expression. Method: Periodontitis model was developed in Wistar rat by induction of Porphyromonas gingivalis. Application of mucoadhesive gingival patch loaded with EGCG (GP-EGCG), mucoadhesive gingival patch loaded with doxycycline (GP-doxy) and blank patch, was done for treated periodontitis 1 h each day during 21 days. Indirect immunohistochemical analysis of IL-6 and IL-10 expressions were analyzed in the mandibular preparation of the anterior incisive region of animal. Results: The GP-EGCG treatment for 3 days until 21 days, consistently increased the IL-10 expression in periodontitis (p < 0.05). In other hand, GP-EGCG treatment lowered the IL-6 expression after 7, 14 and 21 days (p < 0.05). Conclusion: The GP-EGCG is promising for the periodontitis treatment by interfere the IL-6 and IL-10 expression.

The Clinical Pattern and Prevalence of Streptococcus mutans and Streptococcus sobrinus among Adult and Children Patients with Dental Caries

Abstract Objective: To explore the clinical pattern, host factors, and presentation of Streptococcus mutans related to caries incidence among children and adults visiting Universitas Airlangga dental clinic. Material and Methods: This was an observational study with a cross-sectional approach with 50 patients in each group of carious children (6-12 years) and adults (18-35 years). Dental decay samples were taken by sterile excavator, put in a BHI's transport medium, and directly incubated overnight at 37 ºC. The next day, they were sub-cultured microbiologically in Tryptone Yeast Cystine Sucrose Bacitracin (TYCSB) selective medium. Bacterial species and serogroups were examined by PCR. All patient's data were collected from medical records and direct observation. Results: Caries were mostly media type in both children and adults. Oral hygiene (OHIS) in children was higher than in adults but not significantly different according to their DMFT. The highest scores for decay, missed and filled teeth were 16, 8 and 7, with an average of 6.82, 1.22 and 0.63, considered quite high. Conclusion: The prevalence of S. mutans was higher in children's caries than in adults, but among the adult patients the co-incidence of S. mutans and S. sobrinus was associated with higher DMFT. The mutans serotypes e, f, and d were more prevalent among children than adults.

Sucrose, Lactose, and Xylitol Exposures Affect Biofilm Formation of Streptococcus mutans

Abstract Objective: To determine the level of biofilm formation of S. mutans after being exposed to 5% sucrose, 8% lactose, or 1% xylitol. Material and Methods: This research was a laboratory-based experimental study with post-test only control group design. S. mutans was grown in test tubes containing tryptose soy broth (TSB) medium supplemented with 1% glucose. They were incubated at 37° C for 24 hours to grow the biofilms. The culture was then exposed to 5% sucrose, 8% lactose or 1% xylitol, incubated for 24 hours at 37° C, and examined using ELISA at a wavelength of 625 nm. The statistical analysis was performed using a one-way analysis of variance followed by the least significant difference test (a=0.05). Results: There were some differences in the biofilm formation of S. mutans after exposure to 5% sucrose, 8% lactose, or 1% xylitol (p<0.05). An LSD test indicated significant differences among the biofilm formations after exposure to 5% sucrose and 8% lactose and between 5% sucrose and 1% xylitol. In comparison, there were no significant differences (p>0.05) between 8% lactose and 1% xylitol. Conclusion: Sucrose, lactose and xylitol can form biofilms and the formation of lactose biofilms is the same as xylitol.

Cytokines and Chemokines in Periodontitis.

Periodontitis is a common inflammatory periodontal disease affecting a wide range of population all over the world. The causing bacteria releases chemicals which activate the innate immune system to release proinflammatory cytokines contributing to more progression. This activates the acquired immune system leading to more progression of periodontitis. As the immune response goes on, released cytokines and chemokines can damage the periodontal ligaments, gingiva, and alveolar bone. There are many types of cytokines and chemokines in periodontitis. Cytokines are peptide mediators who are responsible for cell signaling and communication. Chemokines are a large subfamily of cytokines having the ability to coordinate leukocyte recruitment and activation. This paper is a narrative review of the literature.This review ensures that inflammatory mediators in the case of periodontitis can cause a noticeable damage in the whole apparatus of the periodontium. It causes soft tissue inflammation and bone damage affected by the mediators of both innate and acquired immune system.The inflammatory process is accompanied by large network of cytokines and chemokines. There is high expression of proinflammatory cytokines such as interleukin (IL)-1α, IL-1ß, IL-6, IL-12, tumor necrosis factor (TNF)-α, and regulatory cytokines such as IL-4, IL-1(RA) receptor antagonist, IL-10, and induced protein (IP)-10. There is also increased production of cytokines IL-10, IL-12, interferon-γ, IP-10, IL-1RA, and IL-4. Cytokines IL-17, IL-6, IL-1ß, TNF-α, macrophage colony-stimulating factor, and prostaglandin E2 trigger the osteoclast activity causing bone resorption.

Effect of lipopolysaccharide derived from surabaya isolates of Actinobacillus actinomycetemcomitans on alveolar bone destruction.

BACKGROUND: Actinobacillus actinomycetemcomitans' lipopolysaccharide (LPS) has a high virulence factor. It interacts with serum protein through receptors on the epithelial cell surface, thereby increasing both interleukin (IL)-1ß, and IL-6 which results in damage to periodontal tissue. AIM: The aim of the study was to identify and evaluate the effect of LPS derived from local isolates (A. actinomycetemcomitans) on the destruction of alveolar bone by means of several biomarkers, including; the number of osteoblasts and osteoclasts, the expression of IL-6, matrix metallopeptidase 1 (MMP-1), and receptor activator of nuclear factor kappa-Β ligand (RANKL). MATERIALS AND METHODS: The isolation of LPS from A. actinomycetemcomitans was calculated using phenol, while purification was performed using Sephadex C-18 column chromatography. 40 Wistar rats were divided into four groups of 10. Each treatment was divided into two groups which were 0.9% NaCl and LPS induced for 7 and 14 days, respectively. Gingival and alveolar bones were further introduced into the induction area, followed by the measuring of osteoblast and osteoclast with hematoxylin-eosin staining, IL-6, MMP-1 and RANKL expression with immunohistochemical. RESULTS: Reduced numbers of osteoblasts at the 7th and 14th day of treatment were detected, while those of osteoclasts increased. There was an increased expression of IL-6, MMP-1, and RANKL in the 7th and 14th-day treatment group. Treatment of LPS from A. actinomycetemcomitans over 7 and 14 days resulted in damage to periodontal tissue and alveolar bone in Wistar rats. CONCLUSION: LPS of A. actinomycetemcomitans administration for 7 and 14 days causes periodontal and alveolar tissue destruction in Wistar rats.