Nanoscale Stolephorus sp. powder fabrication using high-energy milling for bioactive materials in dentistry.

BACKGROUND: The application of natural products in dentistry has been widely explored. Anchovy (Stolephorus in Latin) has been examined for its bioactive content (calcium, phosphorus and fluoride) as an agent for bone stimulation and tooth development, topical fluoridation and pulp capping. Ball milling has been used to prepare calcium oxide nanoparticles from snakehead fish bone. OBJECTIVES: The aim of the study was to reduce the particle size of Stolephorus sp. powder to the nanoscale using high-energy ball milling for 8, 12 and 24 h, and to analyze the optimal milling time by comparing the powder characteristics. MATERIAL AND METHODS: The Stolephorus sp. were oven-dried at 50°C for 6 h, after which the entire fish were crushed into powder. The fish powder was produced by blending the material for 5 min and passing it through a 200-mesh sieve. The remaining dried fish was blended again for 5 min until it passed through the sieve. The top-down approach to the particle size reduction was performed using high-energy milling at 3 distinct time points (8, 12 and 24 h). The characteristics of the powder were evaluated using a particle size analyzer, a Fourier-transform infrared spectrometer (FTIR) and scanning electron microscopy-energy dispersive spectroscopy (SEM-EDS). RESULTS: The Stolephorus sp. powder contained 64.50% protein, 7,420 mg/kg sodium, 28,912 mg/kg calcium, and 1,924 mg/kg magnesium. The high-energy milling process resulted in a reduction of the particle size from the microscale to the nanoscale. The analysis of the average particle size and polydispersity index indicated that 24 h of milling showed the most optimal results. Furthermore, the functional groups exhibited no significant alteration at 3 milling times (p ≥ 0.05, FTIR analysis). CONCLUSIONS: The high-energy milling method has the potential to reduce the particle size of Stolephorus sp. powder to the nanoscale at the 8- and 24-h milling periods. The powder resulting from the 24-h milling process had a size of 789.3 ±170.7 nm, smooth size distribution, good size uniformity, a polydispersity index of 0.763, no significant change in organic and inorganic compound content, and a calcium/phosphorus ratio that was the closest to that of hydroxyapatite (HAp).

Mixing ratio of nano hydroxyapatite and Epigallocatechin-3-gallate (EGCG) towards viscosity and antibacterial effect as a potential pulp capping Material: An experimental study.

Background: Finding a new natural scaffold is challenging due to crucial impact on long-term treatment outcomes in pulp capping. In this context, nano hydroxyapatite (nano-HA) is a potential candidate having similar properties to bone tissue in the body. The compound is often synthesized with Epigallocatechin-3-gallate (EGCG) which offers anti-inflammatory and antibacterial properties. Therefore, this study aims to contribute novel insights into the development of effective pulp capping materials by determining the viscosity ratio of the combination of nano-HA and EGCG applied to the cavity according to standard pulp capping material, as well as proving the antibacterial effect against Lactobacillus acidophilus. Methods: The combination of nano-HA - EGCG is divided into three treatment groups, (G1) 1:1 ratio, (G2) 1:1.5 ratio, (G3) 1:2 ratio, as well as control group G4 (Ca(OH)2 and aquadest) with a ratio of 1:1. Meanwhile, each group is tested for viscosity using a Brookfield viscometer. The well diffusion method is used to determine the antibacterial activity by measuring the diameter of the inhibition zone for each treatment, with C1 (Ca(OH)2 and aquadest) as control group at a ratio of 1:1, and three treatment groups (nano-HA - EGCG), (C2) 0.5:1 ratio, (C3) 1:1 ratio, and (C4) 2:1 ratio. Results: The results show that there is a difference in the viscosity of each group with G3 having a viscosity of 12.0183 cP, which is closest to control. Furthermore, significant differences are also reported in antibacterial activity between control and treatment groups. Conclusion: The ratio of 1:2 (G3) has a viscosity that closely matches the standard of pulp capping materials. The combinations of nano-HA and EGCG are proven to have antibacterial power against Lactobacillus acidophilus.

The Inhibitory Ability of Cocoa Pod Husk Extract on Enterococcus faecalis Glucosyltransferase Enzyme Activity.

BACKGROUND: The presence of Enterococcus faecalis in root canal is considered as one of the factors causing root canal treatment failure since the bacteria are capable of producing glucosyltransferase enzymes that play a role in forming endodontic biofilms. Hence, the bacteria are resistant to antibiotics. On the other hand, cocoa pod husk extract which is rich in chemical components especially flavonoids, tannins, and saponins, is thought to have an ability to inhibit Enterococcus faecalis glucosyltransferase enzyme activity. AIM: The aim of this research is to analyze the inhibitory ability of cocoa pod husk extract against E. faecalis glucosyltransferase enzyme activity. MATERIALS AND METHODS: A total of 27 research samples were divided into three groups, namely, positive control (chlorhexidine gluconate 2%), negative control (aquades), and cocoa pod husk extract 3.12%. Next the enzymatic activity of each sample group was calculated based on the size of the fructose area read by high-performance liquid chromatography (HPLC) expressed in percent (%) and then converted to µmol/mL fructose which was considered as 1 unit of glucosyltransferase enzyme activity. Subsequently, the data were analyzed statistically using Kruskal-Wallis test. RESULTS: The results of data analysis using the Kruskal-Wallis test showed significant differences between groups of samples (p <0.05). CONCLUSION: Cocoa pod husk extract of 3.12% has inhibitory effect on E. faecalis glucosyltransferase enzyme activity. CLINICAL SIGNIFICANCE: The use of cocoa pod husk extract meets the requirements and is proven useful as an irrigation agent in the treatment of root canals, because it contains antibacterial properties against E. faecalis.

Effect of Calcium Hydroxide Combinations with Green Tea Extract and Cocoa Pod Husk Extract on p38 MAPK and Reparative Dentine.

AIM AND OBJECTIVE: The aim of this research is to analyze the effect of calcium hydroxide combinations with green tea extract and the combination of calcium hydroxide with cocoa pod husk extract on the activation of p38 MAPK and wide area of reparative dentin in mice dental. MATERIALS AND METHODS: This study used 36 rats that were randomly divided into three treatment groups: positive control group was applied calcium hydroxide and aquades (group I), the test group was applied calcium hydroxide combined with cocoa pod husk extract (group II), and the next test group was applied using calcium hydroxide combined with green tea extract (group III); all the cavities were restored with RMGIC. On day 7 and 28, experimental animals from each treatment group were killed by peritoneal injection to see the activation of p38 MAPK, while reparative dentin was only seen on day 28. RESULTS: The result of data analysis using Multiple Comparison Tukey HSD test showed significant difference between the positive control group and the test groups for the average p38 MAPK activation value on day 7 and 28. But there was no significant difference between two test groups. The same thing was obtained in the calculation of the average area of reparative dentin, where group I had the lowest value compared to groups II and III on day 28 with a significant difference. There was no significant difference between groups II and III. CONCLUSION: The use of combination calcium hydroxide with green tea extract and combination calcium hydroxide with cocoa pod husk extract have significant effect on p38 MAPK activation and wide area of reparative dentin in mice dental. CLINICAL SIGNIFICANCE: The use of combination calcium hydroxide with green tea extract and combination calcium hydroxide with cocoa pod husk extract have been proven to activate more p38 and form a wider reparative dentin.

East java extract propolis as potential intracanal medicament in experimentally induced chronic apical periodontitis.

INTRODUCTION: A persistent infection after cleaning and shaping root canal is the main etiology of root canal treatment failure. Enterococcus faecalis has been considered as one of the most resistant species in root canal treatment. E. faecalis can stimulate receptor activator of nuclear factor-kappa B ligand (RANKL) which can increase nuclear factor of activated T-cell (NFATc1) in chronic apical periodontitis. East Java propolis has antibacterial effects and is biocompatible with in vitro effects. AIM: This study is aimed to analyze the East Java propolis extract as potential intracanal medicament in chronic apical periodontitis caused by E. faecalis bacterial infection. MATERIALS AND METHODS: This study used 30 Wistar rats divided into three groups. In Group I, the first upper right molar tooth as healthy tooth was used for negative control group. In Group II, the first upper right molar tooth was used for a prepared root canal, and 10 ml brain heart infusion broth containing E. faecalis ATCC29212 106 CFU was injected into the canal and restored with glass-ionomer cement (GIC) for the experimentally induced chronic apical periodontitis group. In Group III, after root canal preparation, E. faecalis ATCC 29212 106 CFU was injected, and then, 10 µl propolis applied and tooth restored with GIC. It took 21 days for the periapical lesions to develop after pulp infection. The rats were then sacrificed to conduct immunohistochemical examinations in order to measure the expressions of RANKL and NFATc1. RESULTS: The average of RANKL and NFATc1 expression in Group III was significantly lower than those in the experimentally induced chronic apical periodontitis group (P < 0.05). CONCLUSION: It can be concluded that East Java propolis extract is a potential intracanal medicament through the study of experimentally induced chronic apical periodontitis caused by E. faecalis infection in Wistar rats.

Cleaning Efficacy of Root Canal Irrigation with Positive and Negative Pressure System.

INTRODUCTION: The purpose of this study was to analyze the differences between irrigant replacement in the positive and negative pressure irrigation systems regarding root canal cleaning efficacy. METHODS AND MATERIALS: A total of 27 extracted single-root mandibular premolars with 18-20 mm root canal length were decoronated and equally divided into three groups (n=9) based on the irrigation system used: positive irrigation with side-vented needle as the control group (C), positive irrigation with an open-ended needle as the first group (T1) and negative irrigation as the second group (T2). The root canals were irrigated with 2.5% NaOCl between each instrumentation, followed by a final irrigation with 5 mL of sterile distilled water. The irrigation replacements were monitored by means of computational fluid dynamic (CFD), while a scanning electrone microscope (SEM) was used to observe the smear layers and plug evaluations after the teeth had been sectioned longitudinally and buccolingually halves subsequently cut in apical third area. The result was analyzed using the Kruskal Wallis, Mann Whitney and Spearman correlation tests. The level of significance was set at 0.05. RESULT: Irrigant replacement in the negative pressure irrigation system tends to produce a greater effect in reaching the apical end compared to in the positive pressure irrigation system. This resulted in significantly superior smear layer removal in the apical third area (P<0.05). CONCLUSION: The irrigation solution exchange of the negative pressure irrigation system is more capable of reaching the apical end compared to the positive pressure irrigation system, resulting in a higher sanitation level in the apical third of the root canal.

Expression of Osteoprotegrin and Osteoclast Level in Chronic Apical Periodontitis Induced with East Java Propolis Extract.

INTRODUCTION: The objective of this animal study was to promote East Java propolis as a potential natural intracanal medicament for periapical chronic apical periodontitis bone resorption through evaluating the expression of osteoprotegrin (OPG) and osteoclast level. METHODS AND MATERIALS: Propolis extract was produced using a maceration procedure. Thirty Wistar rats were divided into three groups. In group I, the control group, the first upper right molar constituted a healthy tooth. In group II, containing rodents with experimentally chronicapical periodontitis, infection with Enterococcus faecalis ATCC29212 106 CFU was performed. In group III, the treatment group, after being injected with E faecalis, 10 µL propolis was applied. It required 21 days to induce post-pulp chronic apical periodontitis infection. The rats were euthanised for immunohistochemical examination in order to measure the expression of OPG and to count histologically the number of osteoclast. RESULT: The expression of OPG and osteoclast constituted 17.5±1.58 and 6.4±0.96 in group I, 10±2 and 16.2±1.31 in group II and 17±1.69 and 7.5±1.08 in group III. Group I presented the highest level of OPG expression but the lowest level of osteoclast expression. There were significant differences between groups II and III and group I regarding OPG and osteoclast expression (P<0.05). CONCLUSION: East Java Propolis was a potential intracanal medicament promoting an increase in osteoprotegerin expression and a decrease in the number of osteoclasts thereby inhibiting osteoclastogenesis.