Synthesis and Structural Characterization of Four Different Concentrations of Ant Nest (Myrmecodia pendens) Collagen Membranes with Potential for Medical Applications.

Purpose: The purpose of this study was to synthesize and structurally characterize four ant nest membranes in four different concentrations and determine the best concentration that could potentially be used as an alternative material for the production of new collagen barrier membranes. Materials and Methods: Membranes were created by mixing ant nest extracts at various concentrations of 0.5%, 1%, 1.5%, and 2%, as well as collagen, chitosan, and Polyvinyl Alcohol (PVA) using a film casting. A Universal Testing Machine (UTM) was used to evaluate mechanical properties including elastic modulus, tensile strength, maximum elongation, elongation at break, and maximum force. Water absorption was performed, FTIR was used for functional group identification, and morphology was examined using SEM. Additionally, EDS was used to identify the composition and distribution of elements in membranes. Statistical analysis was conducted using ANOVA (analysis of variance) and post hoc testing with a significance level of p <0.01 for quantitative data. Results: The results showed that the mechanical properties produced the following mean (standard deviation): elastic modulus 0.87 Mpa (0.11), tensile strength 16.32 N/mm2 (2.46), maximum elongation 4.96% (1.72), elongation at break 5.23% (1.87), and maximum force 22.50 N (5.06). The average water absorption capacity of all four membranes had a p-value <0.01. FTIR spectrum showed various peaks corresponding to functional groups, while SEM results indicated a homogeneous mixture. EDS analysis confirmed that the addition of ant plant extract at 0.5%, 1%, and 1.5% resulted in the presence of elements C, O, and Ca. Meanwhile, membranes prepared with 2% concentration had a different composition, namely C, O, Ca, and Na. Conclusion: Increasing the concentration of ant nest affects the values of the membrane's mechanical properties parameters, including the elastic modulus (0.87 Mpa), tensile strength (16.32 N/mm2), maximum elongation (4.96%), elongation at break (5.23%), and maximum force (22.50 N). The average membrane absorption of water (p value <0.01) was also affected. SEM images showed homogeneous mixing, and membrane EDS results consisted of C, O, and Ca composition. However, there was no effect on FTIR functional groups. The anthill membrane with a 1% concentration has the potential to serve as an alternative membrane in guided tissue regeneration.

Pharmacological Activity of Chemical Compounds of Potato Peel Waste (Solanum tuberosum L.) in vitro: A Scoping Review.

Introduction: The potato (Solanum tuberosum L.) is a short-lived tuber plant with a round to oval shape and varying colors, depending on the variety. It is known that only the inside of the potato is used, while the peel is generally discarded. However, recent studies have shown that potato peels contain many health-beneficial compounds. Purpose: This study aimed to investigate the compounds present in potato peels and their in vitro activities. Methods: A scoping review following the PRISMA guidelines was conducted. The selection process involved identifying articles of in vitro research published within the last 10 years (2012-2022). Electronic searches were conducted using the portals Scopus, ScienceDirect, EBSCOhost, and Portal Garuda by using the keywords "potato" or "Solanum tuberosum" and "peel" or "skin". The search was limited to articles in English with full text availability. Results: The screening process resulted in a total of 4773 articles from the four search engines; 14 articles were obtained that met the requirements for the review, most of which use extract preparations in their research. Extracts of flavonoids, phenols, and glycoalkaloids are the most frequently studied compounds, and their antioxidant and anti-inflammatory activity have undergone extensive research. Conclusion: The potential compounds contained in potato peels, including flavonoids, phenols, and glycoalkaloids, are highly abundant and offer numerous benefits. Provides opportunities for further research to prove the potential pathway activity of the compound. These compounds have been the subject of extensive research, suggesting their significance in the context of health and nutrition.

The Potential of 24-Propylcholestrol as Antibacterial Oral Bacteria of Enterococcus faecalis ATCC 29212 and Inhibitor Biofilms Formation: in vitro and in silico Study.

Introduction: Uncontrolled biofilm can cause several diseases such as dental caries, gingivitis, and periodontitis. Dental caries is a disease caused by the accumulation of plaque-containing pathogenic bacteria, including Enterococcus faecalis. These bacteria infect the root canals of teeth and colonize to form biofilms. Biofilm inhibition is carried out by interfering with cell wall formation metabolism. MurA enzyme has a role in peptidoglycan biosynthesis of cell walls. Enterococcal surface protein (Esp) is the main contributor of E. faecalis to form biofilms. In addition, inhibition of biofilms by interfering with the quorum sensing (QS) system, suppressing gelatinase virulence factors by blocking autoinducers gelatinase biosynthesis-activating pheromone (GBAP). Purpose: Knowing the potential of Piper betel Linn. compounds as antibacterial in vitro and antibiofilm agents against E. faecalis in silico. Patients and Methods: The compounds were purified by a bioactivity-guided chromatographic method. Antibacterial activity was tested by disc diffusion method, in vitro studies. In silico study, compound P. betel L. was used as the test ligand and compared with positive control fosfomycin, ambuic acid, quercetin, and taxifolin. The proteins used MurA, Esp, GBAP, and gelatinase were docking with the Autodock Vina PyRx 0.8 followed by the PYMOL program and visualized with the Discovery Studio 2020 program. Results: An antibacterial compound was identified 24-propylcholesterol which can inhibit the activity of E. faecalis ATCC 29212 with MIC value of 78.1 µg/mL and MBC value of 156.3 µg/mL. Molecular docking results showed the binding affinity of 24-propylcholesterol with MurA, ESP, GBAP, and gelatinase enzymes was -7.6, -8.7, -5.3, and -7.9 kcal/mol. Conclusion: 24-propylcholesterol has potential as an antibacterial against E. faecalis and as an antibiofilm through in silico inhibition of QS. However, further research is needed in vitro and in vivo to determine the effectiveness of these compounds as antibacterial and antibiofilm.

Ocimum Species: A Review on Chemical Constituents and Antibacterial Activity.

Infection by bacteria is one of the main problems in health. The use of commercial antibiotics is still one of the treatments to overcome these problems. However, high levels of consumption lead to antibiotic resistance. Several types of antibiotics have been reported to experience resistance. One solution that can be given is the use of natural antibacterial products. There have been many studies reporting the potential antibacterial activity of the Ocimum plant. Ocimum is known to be one of the medicinal plants that have been used traditionally by local people. This plant contains components of secondary metabolites such as phenolics, flavonoids, steroids, terpenoids, and alkaloids. Therefore, in this paper, we will discuss five types of Ocimum species, namely O. americanum, O. basilicum, O. gratissimum, O. campechianum, and O. sanctum. The five species are known to contain many chemical constituents and have good antibacterial activity against several pathogenic bacteria.

Mode action prediction of catechin from Uncaria gambir Roxb. against UDP-N-acetylenolpyruvyl-glucosamine reductase (MurB enzyme) of Streptococcus mutans: In silico study.

The prevalence of oral health problems in the global population is still high, especially dental caries, which is considered a multifactorial disease involving the role of bacteria, namely Streptococcus mutans. Gram-positive bacteria metabolize carbohydrates and sugars and convert them into lactic acid, causing dental caries. The peptidoglycan (PG) layer at the outer surface of the bacteria acts as protection. MurB enzyme is known for its contribution to PG biosynthesis. Gambir (Uncaria gambir Roxb.) is famous for many efficacies. Previous studies show that catechin from herb plants such as U. gambir has antibacterial activity. This study aimed to evaluate and predict the antibacterial activity of catechin from U. gambir against the MurB enzyme, which contributes to forming the bacteria PG, with an in silico approach. The structure of the MurB enzyme was collected from UniProt, and the ligands (catechin and chlorhexidine) structures were obtained from PubChem. The AutoDock software was used to dock both ligand and MurB enzyme visualized using PyMOL and analyzed using BIOVIA. The results showed that catechin has a binding affinity of more than - 7 kcal/mol against the MurB enzyme, and chlorhexidine has a higher binding affinity than catechin. Both catechin and chlorhexidine have similar amino acids attachment by hydrogen bonds. The results showed that catechin has competitive antibacterial activity against chlorhexidine in inhibiting the MurB enzyme.

Prediction Mechanism of Nevadensin as Antibacterial Agent against S. sanguinis: In vitro and In silico Studies.

BACKGROUND: Streptococcus sanguinis can contribute to tooth demineralization, which can lead to dental caries. Antibiotics used indefinitely to treat dental caries can lead to bacterial resistance. Discovering new antibacterial agents from natural products, like Ocimum basilicum, will help combat antibiotic resistance. In silico analysis (molecular docking) can help determine the lead compound by studying the molecular interaction between the drug and the target receptor (MurA enzyme and DNA gyrase). It is a potential candidate for antibacterial drug development. OBJECTIVES: The research objective is to isolate the secondary metabolite of O. basilicum extract that exhibits activity against S. sanguinis through in vitro and in silico analysis. METHODS: n-Hexane extract of O. basilicum was purified by combining column chromatography with bioactivity-guided fractionation. The in vitro antibacterial activity against S. sanguinis was determined using the disc diffusion and microdilution method, while molecular docking simulation of nevadensin (1) with MurA enzyme and DNA gyrase was performed by using PyRx 0.8 program. RESULTS: Nevadensin from O. basilicum was successfully isolated and characterized by spectroscopic methods. This compound showed antibacterial activity against S. sanguinis with MIC and MBC values of 3750 and 15000 µg/mL, respectively. In silico analysis showed that the binding affinity to MurA was -8.5 Kcal/mol, and the binding affinity to DNA gyrase was -6.7 Kcal/mol. The binding of nevadensin-MurA is greater than fosfomycin-MurA. Otherwise, Nevadensin-DNA gyrase has a weaker binding affinity than fluoroquinolone-DNA gyrase and chlorhexidine-DNA gyrase. CONCLUSION: Nevadensin showed potential as a new natural antibacterial agent by inhibiting the MurA enzyme rather than DNA gyrase.

Action mode of ursolic acid as a natural antioxidant and inhibitor of superoxide dismutase: In vitro and in silico study.

Recently, the antioxidant is applied for the teeth bleaching treatment as an alternative of toxic material of hydrogen peroxide that is used in teeth bleaching. One of natural sources antioxidant is Uncaria gambir those containing active antioxidant agents. To be applied as a new bioactive constituent in teeth bleaching treatment, a preexperimental study is performed. The aim of the study is to identify the antioxidant constituent of U. gambir and predict their activity including action mode as an inhibitor of enzyme superoxide dismutase (SOD) through in vitro and in silico method. Combination of chromatography methods and spectroscopic analysis is used for isolated bioactive antioxidant constituent. The antioxidant activity was evaluated by in vitro assay against diphenylpicrylhydrazyl (DPPH)and SOD, respectively, while prediction of action mode of the active compounds as SOD-mutant enzyme inhibitor was conducted by in silico study using AutoDock 4.2 program. Antioxidant of ursolic acid was isolated from U. gambir with inhibitory concentration50 values 1721 ± 30.6 and 392 ± 53.57 µg/mL, respectively, against DPPH and SOD. By in silico study presented that ursolic acid inhibited SOD enzyme with a binding affinity of - 5.4 kcal/mol those higher than a quercetin as a positive control. The ursolic acid was identified as a potential natural antioxidant with potentially activity to inhibit SOD mutant.

Bio-Mechanism of Catechin as Pheromone Signal Inhibitor: Prediction of Antibacterial Agent Action Mode by In Vitro and In Silico Study.

The utilization of medicinal plants has long been explored for the discovery of antibacterial agents and the most effective mechanisms or new targets that can prevent and control the spread of antibiotic resistance. One kind of bacterial cell wall inhibition is the inactivation of the MurA enzyme that contributes to the formation of peptidoglycan. Another approach is to interfere with the cell-cell communication of bacteria called the Quorum sensing (QS) system. The blocking of auto-inducer such as gelatinase biosynthesis-activating pheromone (GBAP) can also suppress the virulence factors of gelatinase and serine protease. This research, in particular, aims to analyze lead compounds as antibacterial and anti-QS agents from Gambir (Uncaria gambir Roxburgh) through protein inhibition by in silico study. Antibacterial agents were isolated by bioactivity-guided isolation using a combination of chromatographic methods, and their chemical structures were determined by spectroscopic analysis methods. The in vitro antibacterial activity was evaluated by disc diffusion methods to determine inhibitory values. Meanwhile, in the in silico analysis, the compound of Uncaria gambir was used as ligand and compared with fosfomycin, ambuic acid, quercetin, and taxifolin as the standard ligand. These ligands were attached to MurA, GBAP, gelatinase, and serine proteases using Autodock Vina in PyRx 0.8 followed by PYMOL for combining the ligand conformation and proteins. plus programs to explore the complex, and visualized by Discovery Studio 2020 Client program. The antibacterial agent was identified as catechin that showed inhibitory activity against Enterococcus faecalis ATCC 29212 with inhibition zones of 11.70 mm at 10%, together with MIC and MBC values of 0.63 and 1.25 µg/mL, respectively. In the in silico study, the molecular interaction of catechin with MurA, GBAP, and gelatinase proteins showed good binding energy compared with two positive controls, namely fosfomycin and ambuic acid. It is better to use catechin-MurA (-8.5 Kcal/mol) and catechin-gelatinase (-7.8 Kcal/mol), as they have binding energies which are not marginally different from quercetin and taxifolin. On the other hand, the binding energy of serine protease is lower than quercetin, taxifolin, and ambuic acid. Based on the data, catechin has potency as an antibacterial through the inhibition of GBAP proteins, gelatinase, and serine protease that play a role in the QS system. This is the first discovery of the potential of catechin as an alternative antibacterial agent with an effective mechanism to prevent and control oral disease affected by antibiotic resistance.

Effectiveness of Bioactive Compound as Antibacterial and Anti-Quorum Sensing Agent from Myrmecodia pendans: An In Silico Study.

BACKGROUND: antibiotic resistance encourages the development of new therapies, or the discovery of novel antibacterial agents. Previous research revealed that Myrmecodia pendans (Sarang Semut) contain potential antibacterial agents. However, specific proteins inhibited by them have not yet been identified as either proteins targeted by antibiotics or proteins that have a role in the quorum-sensing system. This study aims to investigate and predict the action mode of antibacterial compounds with specific proteins by following the molecular docking approach. METHODS: butein (1), biflavonoid (2), 3″-methoxyepicatechin-3-O-epicatechin (3), 2-dodecyl-4-hydroxylbenzaldehyde (4), 2-dodecyl-4-hydroxylbenzaldehyde (5), pomolic acid (6), betulin (7), and sitosterol-(6'-O-tridecanoil)-3-O-ß-D-glucopyranoside (8) from M. pendans act as the ligand. Antibiotics or substrates in each protein were used as a positive control. To screen the bioactivity of compounds, ligands were analyzed by Prediction of Activity Spectra for Substances (PASS) program. They were docked with 12 proteins by AutoDock Vina in the PyRx 0.8 software application. Those proteins are penicillin-binding protein (PBP), MurB, Sortase A (SrtA), deoxyribonucleic acid (DNA) gyrase, ribonucleic acid (RNA) polymerase, ribosomal protein, Cytolysin M (ClyM), FsrB, gelatinase binding-activating pheromone (GBAP), and PgrX retrieved from UniProt. The docking results were analyzed by the ProteinsPlus and Discovery Studio software applications. RESULTS: most compounds have Pa value over 0.5 against proteins in the cell wall. In nearly all proteins, biflavonoid (2) has the strongest binding affinity. However, compound 2 binds only three residues, so that 2 is the non-competitive inhibitor. CONCLUSION: compound 2 can be a lead compound for an antibacterial agent in each pathway.

Potential Fatty Acid as Antibacterial Agent Against Oral Bacteria of Streptococcus mutans and Streptococcus sanguinis from Basil (Ocimum americanum): In vitro and In silico Studies.

BACKGROUND: Streptococcus mutans and Streptococcus sanguinis are Gram-positive bacteria that cause dental caries. MurA enzyme acts as a catalyst in the formation of peptidoglycan in bacterial cell walls, making it ideal as an antibacterial target. Basil (Ocimum americanum) is an edible plant that is diverse and has been used as a herbal medicine for a long time. It has been reported that basil has a pharmacological effect as well as antibacterial activity. The purpose of this study was to identify antibacterial compounds in O. americanum and analyze their inhibition activity on MurA enzyme. METHODS: Fresh leaves from O. americanum were extracted with n-hexane and purified by a combination of column chromatography on normal and reverse phases together with in vitro bioactivity assay against S. mutans ATCC 25175 and S. sanguinis ATCC 10556, respectively, while in silico molecular docking simulation of lauric acid (1) was conducted using PyRx 0.8. RESULTS: The structure determination of antibacterial compound by spectroscopic methods resulted in an active compound lauric acid (1). The in vitro evaluation of antibacterial activity in compound 1 showed Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) values of 78.13 and 156.3 ppm and 1250 and 2500 ppm against S. sanguinis and S. mutans, respectively. Further analysis and in silico evaluation determined lauric acid (1) as MurA Enzyme inhibitor. Lauric acid (1) showed a binding affinity of -5.2 Kcal/mol, which was higher than fosfomycin. CONCLUSION: Lauric acid showed the potential as a new natural antibacterial agent through MurA inhibition in bacterial cell wall biosynthesis.

Potential Allylpyrocatechol Derivatives as Antibacterial Agent Against Oral Pathogen of S. sanguinis ATCC 10,556 and as Inhibitor of MurA Enzymes: in vitro and in silico Study.

BACKGROUND: Streptococcus sanguinis is Gram-positive bacteria that contribute to caries. Many antibacterial agents are resistant against bacteria so that the discovery of new antibacterial agents is a crucial issue. Mechanism of antibacterial agents by disrupting cell wall bacteria is a promising target to be developed. One of the enzymes contributing to the cell wall is MurA enzyme. MurA is an enzyme catalyzing the first step of peptidoglycan biosynthesis in the cell wall formation. Inhibiting MurA is an effective and efficient way to kill the bacteria. Source of bioactive compounds including the antibacterial agent can be found in natural product such as herbal plant. Piper betle L. was reported to contain active antibacterial compounds. However, there is no more information on the antibacterial activity and molecular mechanism of P. betle's compound against S. sanguinis. PURPOSE: The study aims to identify antibacterial constituents of P. betle L. and evaluate their activities through two different methods including in vitro and in silico analysis. MATERIALS AND METHODS: The antibacterial agent was purified by bioactivity-guided isolation with combination chromatography methods and the chemical structure was determined by spectroscopic methods. The in vitro antibacterial activity was evaluated by disc diffusion and dilution methods while the in silico study of a compound binds on the MurA was determined using PyRx program. RESULTS: The antibacterial compound identified as allylpyrocatechol showed inhibitory activity against S. sanguinis with an inhibition zone of 11.85 mm at 1%, together with MIC and MBC values of 39.1 and 78.1 µg/mL, respectively. Prediction for molecular inhibition mechanism of allylpyrocatechols against the MurA presented two allylpyrocatechol derivatives showing binding activity of -5.4, stronger than fosfomycin as a reference with the binding activity of -4.6. CONCLUSION: Two allylpyrocatechol derivatives were predicted to have a good potency as a novel natural antibacterial agent against S. sanguinis through blocking MurA activity that causes disruption of bacterial cell wall.

Fabrication and Characterization of Chitosan-Collagen Membrane from Barramundi (Lates Calcarifer) Scales for Guided Tissue Regeneration.

OBJECTIVES: The aim of this study was to evaluate the synthesis, mechanical strength, and morphology of chitosan-collagen membranes from barramundi scales for guided tissue regeneration technique. MATERIALS AND METHODS: Collagen was extracted from barramundi scales by immersion in acetic acid. The resulting wet collagen was later dried. The membrane was fabricated by mixing chitosan with collagen from barramundi scales. Membrane characterization parameters were measured using Fourier transform infrared (FTIR), scanning electron microscopy (SEM) and mechanical property. RESULTS: The FTIR spectrum showed the typical peak of the mixture of chitosan and collagen. The tensile strength and elongation at break of the membrane in dry condition were 0.28 MPa and 8.53%, respectively, while in the wet condition these were 0.12 MPa and 25.6%. The membrane porosity test result was 38.85%; SEM result showed a porous membrane surface with size varying around 16 to 100 µm. CONCLUSION: The chitosan-collagen membrane from the barramundi scale showed the fibrous membrane surface that has ideal porous size as guided tissue regeneration membrane and the lower mechanical strength.

The effect of the chitosan-collagen membrane on wound healing process in rat mandibular defect.

BACKGROUND: Collagen and chitosan are potential biomaterials for medical applications; chitosan-collagen membranes are used as a barrier membrane in guided tissue regeneration and guided bone regeneration. AIMS: The purpose of this study was to analyze the effect of the chitosan-collagen membrane on wound healing in rat mandibular defect by counting the number of fibroblasts and new blood vessels. MATERIALS AND METHODS: As much as 24 male Wistar rats were divided into two groups, the treatment and control group. Bone defects were made In the rat mandible with diamond bur with a diameter of 2 mm, then the defect was covered with a chitosan-collagen membrane, and the control group was covered without application of chitosan-collagen membrane. After the 3rd, 7th, 14th, and the 21st day, the defect site was analyzed histologically. The number of fibroblasts and blood vessels was counted under a light microscope, at five fields with ×1000 and ×400 microscope magnification. STATISTICAL ANALYSIS USED: This study was done by using analysis of variance and unpaired t-test. RESULTS: The average number of fibroblasts and blood vessels in the treatment group was higher than the control group. There was a significant difference in the number of fibroblasts on the 3rd and 7th day (P = 0.001; P = 0.001) and the number of blood vessels on the 3rd day (P = 0.04). CONCLUSION: The chitosan-collagen membrane was able to increase the number of fibroblasts and new blood vessels in the wound healing process.

MIC, MBIC, MBEC analyses of garlic extract (Allium sativum) from Indonesian variety against Streptococcus mutans

Garlic (Allium sativum) has been used for traditional medicine since ages. It has many uses such as an antioxidant, antiseptic, anti-viral, anti-fungal, anti-cancer, anti-aging, reduce the risk of heart disease, and antibacterial1. Aim: This study was designed to evaluate the inhibition and eradication ability for garlic against Streptococcus mutans, the bacteria that cause dental caries. Methods: Garlic extract was obtained from Ciwidey garlic maseration process in etanol 96%. MIC was a test to measure the inhibition ability of garlic extract against Streptococcus mutans in planktonic form, and MBIC in biofilms form. MBEC was a test to measure the eradication ability of garlic extract against Streptococcus mutans in biofilms form. Statistic analysis using ANOVA followed by post hoc with ρ value <0.05. Results: The garlic extract showed MIC value at 9.39 % and MBIC value at 37.5 % but was not observed to have eradication activity against Streptococcus mutans in biofilms form up to concentration 37.5%. Conclusion: Garlic extract inhibit Streptococcus mutans both in planktonic and biofilm form. Garlic extract has no eradication activity against Streptococcus mutans in biofilms form up to concentration 37.5% (AU)