Nanoparticle-Based Rice Husk Liquid Smoke as Periodontitis Therapy through OPG, RANK, and RANKL Expression.

Introduction: Periodontitis therapy employing nanomaterials with submicron sizes holds promise for enhancing osteogenesis and facilitating periodontal cell proliferation. This study aims to assess the potential of nanoparticle-based rice husk liquid smoke (n-RHLS) in an animal model of periodontitis by evaluating the expression of osteoprotegerin (OPG), receptor activator of nuclear factor-kß (RANK), and receptor activator of nuclear factor-kß ligand (RANKL). Methods: Twenty-eight male Wistar rats were inoculated with 109 CFU/ml of Porphyromonas gingivalis in the sulcus mandibular incisor region to create periodontitis and subsequently treated with n-RHLS while the control with saline. Immunohistochemical staining was performed on the mandibular incisor to assess OPG, RANK, and RANKL expression 2 and 7 days after treatment. Results: OPG expression exhibited a significant increase at both 2 and 7 days, while RANKL expression decreased notably after 7 days of treatment using n-RHLS (p < 0.05). In contrast, RANK expression did not show significant differences compared to the control groups (p > 0.05). Conclusion: Nanostructured liquid smoke derived from rice husk nanoparticles (n-RHLS) demonstrates potential as a therapeutic agent for periodontitis, especially on OPG/RANK/RANKL expression, by modulating OPG and RANKL expression to support periodontal tissue health.

6-Octadecenoic and Oleic Acid in Liquid Smoke Rice Husk Showed COVID-19 Inhibitor Properties.

In recent years, liquid smoke rice husk (LSRH) has shown its therapeutic potency to diabetes, wound healing, stomatitis, and periodontitis. The phenol, 6-octadecenoic acid, oleic acid, and 9-octadecanoic acid were responsible for their therapeutic effect. The LSRH also demonstrated their potential for infectious diseases such as coronavirus disease (COVID-19). Therefore, the molecular dynamics (MDs) simulation and pharmacophore analysis was performed to analyse the binding stability of 6-octadecenoic and oleic acid. Based on MD simulation, 6-octadecenoic and oleic acids seemed to retain their interactions with Ser144 and Thr24, respectively, with hydrogen bond distance less than 2.9 Å. This interaction was stable during the simulation and has hydrophobic and hydrogen bonds/acceptors. The 6-octadecenoic acid and oleic acid were confirmed to have great potency as inhibitors for COVID-19. These compounds also showed that the existence of hydrophobic and hydrogen bonds/acceptors could increase biological activity.

Adipose Mesenchymal Stem Cell Metabolites Oral Gel Enhance Pro-Angiogenic Factors Expression, Angiogenesis, and Clinical Outcome of Oral Ulcer Rat Model.

OBJECTIVE: Enhancing wound healing capacity is one of the main principles in oral ulcer management. Efficient oral ulcer management will accelerate clinical symptom amelioration and prevent complications. Adipose mesenchymal stem cell metabolites (AdMSCM), a novel biological product, contains a plethora of bioactive mediators that can induce a series of processes in wound healing. This study will analyze the clinical outcome, angiogenesis, and expression of FGF-2 and VEGFA in the oral ulcer rat model after AdMSCM oral gel application. MATERIALS AND METHODS: Twenty healthy male Wistar rats (Rattus novergicus) were used to create oral ulcer animal models. AdMSCM oral gel treatment was performed three times daily for 3 and 7 days. Clinical outcome was assessed by measuring the major diameter of the ulcer; the angiogenesis was evaluated through histological assessment; the expression of VEGFA and FGF-2 was assessed using the immunohistochemistry method. STATISTICAL ANALYSIS: This study uses parametric comparative analysis using one-way analysis of variance (ANOVA) and post-hoc Tukey's HSD test RESULTS: The application of AdMSCM oral gel in an oral ulcer rat model significantly enhanced the clinical outcome (p < 0.05). In addition, similar results were shown in the histologic assessment of angiogenesis and supported by the significant increase of VEGFA and FGF-2 expression. CONCLUSIONS: AdMSCM oral gel accelerates oral ulcer healing processes, proven by the enhancement of angiogenesis, pro-angiogenic factors expression, and clinical outcomes.

Nanoparticles of Liquid Smoke Rice Husk Inhibit Porphyromonas gingivalis.

OBJECTIVE: Utilization of liquid smoke rice husk can be used as an alternative treatment because of the antimicrobial properties. Advances in drug delivery systems are increasingly developing to increase the bioavailability of drugs and reduce the side effects of these drugs, namely nanoparticles. In this study, nanoparticles of liquid smoke rice husk (nLSRH) were tested the antimicrobial against Porphyromonas gingivalis. MATERIALS AND METHOD: This type of research is an experimental in vitro laboratory using Porphyromonas gingivalis culture. nLSRH contained liquid smoke rice husk concentration of 1, 2.5, 5, 7.5, 10, 12.5, 15, and 17.5%. The antibacterial was performed using the dilution methods. RESULTS: The nLRSH concentration of 1% showed clearest medium. The highest number of colonies Porphyromonas gingivalis was observed at nLSRH concentration of 1% (40.3 colony-forming unit [CFU]) and decreased at a concentration of 2.5% (11.3 CFU); other concentration or no bacterial colony growth was found. The nLSRH concentration of 2.5% can be determined as the minimum inhibitory concentration and nLSRH concentration of 5% can be determined as the minimum bactericidal concentration. CONCLUSION: nLSRH have antimicrobial activity against Porphyromonas gingivalis. This finding able to drive the next research to develop nLSRH as gingival and periodontitis disease is caused by Porphyromonas gingivalis.

Osteoblast Viability of Liquid Smoke Rice Hull and Nanoparticles Form as Periodontitis Treatment.

OBJECTIVE: Rice husk liquid smoke nanoparticles have the potential to be developed as a drug because they have anti-inflammatory effects that can modulate the process of osteoblast stimulation through osteoblast stimulation by thorough small size and enter cells easily. The osteoblast is the key of alveolar regeneration in periodontitis treatment. This present study analyzed the differences of liquid smoke rice husk and nanoparticles of liquid smoke rice husk on osteoblast viability as periodontitis treatment MATERIALS AND METHODS: The liquid smoke rice husk was obtained from the pyrolysis process. The nanoparticles were made with chitosan, maltodextrin, and difference of concentration of liquid smoke rice husk (such as 1, 2.5, 5, 7.5, 10, 12.5, 15, and 17.5%). The viability of osteoblast was analyzed by 2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. STATISTICAL ANALYSIS: The data were analyzed using independent t-test to analyze the differences between liquid smoke rice husk and nanoparticles of liquid smoke rice husk, the significant was set a p<0.05. RESULT: The nanoparticles of liquid smoke rice husk showed higher osteoblast viability compared liquid smoke rice husk. The nanoparticles' concentration of 5, 7.5, and 10% showed higher osteoblast viability compared liquid smoke rice husk (p = 0.002, 0.000, and 0.001, respectively). CONCLUSION: The nanoparticles of liquid smoke rice husk showed higher viability of osteoblast. This confirmed that the nanoparticles were able to reduce the toxicity in the higher concentration of liquid smoke of rice husk.

In Silico Study of Liquid Smoke Rice Husk against COVID-19.

OBJECTIVES: Liquid smoke rice husk has been researched and proved to consist of antibacterial, analgesic, anti-inflammatory, and growth factor stimulatory properties. By these complex properties, the liquid smoke rice hull is able to purpose as a novel coronavirus disease 2019 (COVID-19) inhibitor. The research was conducted to analyze the role of the dominant compound in rice husk liquid smoke against one of the main proteases in complex with inhibitor N3 of COVID-19 and 6LU7 protein data bank (PDB) ID. MATERIAL: AND METHODS: The Molecular Operating Environment (MOE) 2020.0901 (Chemical computing group) was used to analyzed the interaction. The molecular structure test, including phenol, mequinol, 2-methoxy-phenol, 6-octadecenoic acid, oleic acid, 9-cctadecenoic acid, was chosen. The lopinavir as positive control and 6LU7 as COVID-19 protein were chosen. All the protein analyses were conducted using docking molecular. RESULT: The phenol, 2-methoxy-phenol, mequinol and 9-octadecenoic acid have higher binding free energy that causes difficult to bind to the active site of protein 6LU7 (-3.4758, -3.5509, -3.6845, and -5.0173 kcal/mol, respectively). The minor component of liquid smoke, such as 6-octadecenoic acid and oleic acid, has the binding free energy (-5,5327 and -5,4208 kcal/mol) and more factor of binding presumably as active COVID-19 inhibitor. CONCLUSION: The liquid smoke rice husk has active component like 6-octadecenoic acid and oleic acid are presumably as active COVID-19 inhibitor.

The expression of interleukin-1ß and nuclear factor erythroid-2 in the periodontitis after treatment of liquid smoke rice hull.

Nonsurgical treatment such as scaling and root planing has been the main therapy of periodontal management. However, some individuals are nonresponsive with only mechanical therapy. Rice hull liquid smoke (RH-LS) with higher of phenolic compound supposed to interfere with inflammation process. Thus, the purpose of this study is to investigate the anti-inflammation properties of RH-LS for chronic periodontitis adjunct therapy by observing the expression of nuclear factor erythroid-2 (Nrf-2) and interleukin-1ß (IL-1ß). Two groups of Wistar rats were used. Control group was induced by 1 × 109 colony-forming unit (CFU)/ml Porphyromonas gingivalis (PG) and treated with aquadest for 2 and 7 days. Meanwhile, the other group was induced by 1 × 109 CFU/ml PG and treated with RH-LS for 2 and 7 days. Two central incisive/mandibulae were dissected and stained with immunohistochemistry for analyzed the expression of Nrf-2 and IL-1ß. Data were then analyzed statistically using independent t-test (P = 0.05). Treatment with RH-LS for 7 days showed lower expression of IL-1ß (4.00 ± 1.58) than control group (13.60 ± 2.70) and showed higher expression of Nrf-2 (11.60 ± 1.95) than control group (4.20 ± 1.64) (P = 0.000) (P < 0.05). Treatment of RH-LS for 2 days showed higher expression of Nrf-2 (2.20 ± 1.31) than control group (6.80 ± 1.92) (P = 0.003) (P < 0.05). RH-LS showed anti-inflammation properties by elevating the expression of Nrf-2 and depressing expression of IL-1ß in periodontitis induced by PG.

Growth factor stimulation for the healing of traumatic ulcers with liquid rice hull smoke.

OBJECTIVE: The healing process of a traumatic ulcer requires growth factors to rebuild the lost tissue after the inflammatory process has been completed. Liquid rice hull smoke (LR-HS) has shown unique anti-inflammatory properties. This study analyses the role of LR-HS in growth factor stimulation for the healing of traumatic ulcers, such as fibroblast growth factor (FGF), vascular endothelial growth factor (VEGF), platelet-derived growth factor (PDGF), and collagen type 1 (COL-1) expression. METHODS: We obtained LR-HS from the pyrolysis of rice hulls. Traumatic ulcers were created in the labial fornix incisive inferior of Wistar rats and treated with LR-HS once a day for 3, 5, and 7 days. The control group was treated with sterile water. Each animal was sacrificed after treatment, and its labial fornix incisive inferior tissues were biopsied and immunohistochemically stained to examine FGF, VEGF, PDGF, and COL-1 expression. RESULT: The treatment of traumatic ulcers with LR-HS showed an increase in FGF, VEGF, PDGF, and COL-1 expression. VEGF expression increased under LR-HS treatment compared with the control 7-day treatment groups (p < 0.000). FGF and COL-1 expression increased under LR-HS treatment compared with the control 5- and 7-day treatment groups (p < 0.000). PDGF expression increased after treatment with LR-HS for 3, 5, and 7 days (p < 0.000). CONCLUSION: This study has demonstrated that LR-HS can induce the expression of growth factors during the healing of a traumatic ulcer using immunohistochemical staining. We suggest that LR-HS can be used as a herbal medicine for oral ulcer therapy.

The Effects of Rice Husk Liquid Smoke in Porphyromonas gingivalis-Induced Periodontitis.

OBJECTIVES: The purpose of this study was to analyze the effects of rice husk liquid smoke in Porphyromonas gingivalis-induced periodontitis in the inflammatory and proliferation marker such as nuclear factor kappa ß (NF-kB), tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), transforming growth factor-ß (TGF-ß), fibroblast growth factor 2 (FGF2), collagen type 1 (COL-1) expression, and the number of macrophages, lymphocytes, and fibroblasts. MATERIALS AND METHODS: Rice husk liquid smoke is obtained by the pyrolysis process. Porphyromonas gingivalis-induced periodontitis in 20 µL phosphate-buffered saline containing 1 × 109 CFU was injected into the lower anterior gingival sulcus of Wistar rats. The periodontitis was then treated with 20 µL/20 g body weight of rice husk liquid smoke once a day for 2 and 7 days, respectively. After treatment, the bone and lower anterior gingival sulcus were analyzed with immunohistochemistry and hematoxylin-eosin staining. RESULTS: The treatment of periodontitis with rice husk liquid smoke showed a lower NF-kB, TNF-α, and IL-6 expression and a higher TGF-ß, FGF2, and COL-1 expression than the control after treatment for 2 and 7 days (p < 0.05), respectively. The number of macrophages and fibroblasts was also higher when compared with the control group (p < 0.05), but the number of lymphocytes was lower than the control (p < 0.05). CONCLUSION: Rice husk liquid smoke showed its effects on Porphyromonas gingivalis-induced periodontitis with a decrease in inflammatory markers and an increase in proliferation markers. The development of a rice husk liquid smoke periodontitis treatment is promising.

The Role of Rice Hull Liquid Smoke in the Traumatic Ulcer Healing.

OBJECTIVE: The aim of this study was to prove the role of rice hull liquid smoke (RH-LS) on lymphocytes, macrophages, fibroblasts, interleukin 6 (IL-6), and transforming growth factor ß (TGF-ß) expression during traumatic ulcer healing. MATERIALS AND METHODS: The RH-LS was obtained from the pyrolysis process. Traumatic ulcers were made 10 mm along the labial fornix incisive inferior of Wistar rat using a round stainless-steel blade. In control group, traumatic ulcers were treated using sterile water, and meanwhile in experimental group were treated using RH-LS once a day for 3, 5, and 7 days. After treatment, animal was terminated and their labial fornix incisive inferior tissues were biopsy and stained using hematoxylin and eosin staining to determine lymphocytes, macrophages, and fibroblasts. The IL-6 and TGF-ß expressions were analyzed used immunohistochemistry staining. RESULT: The lymphocytes, macrophages, and fibroblasts were higher in the RH-LS group for 3-, 5-, and 7-day treatment (p < 0.05). The IL-6 expression was higher only in the 5-day treatment, and the TGF-ß expression was higher in the 3- and 7-day treatment. CONCLUSION: The RH-LS able to accelerated the traumatic ulcer healing by increasing the number of lymphocytes, macrophages, fibroblasts, IL-6, and TGF-ß expression.

Concanavalin A Enhanced Proliferation and Osteogenic Differentiation of Dental Pulp Stem Cells.

OBJECTIVE: Dental pulp stem cells (DPSCs) can be used as a component in the formation of regenerative dentine during direct pulp capping therapy. Concanavalin A (ConA) is a type of lectin with a molecular weight of 26 kDa derived from the Canavalia ensiformis plant. Lectins possess strong proliferation and differentiation abilities in various animal cells including lymphocytes, osteoblasts, and chondrocytes. The aim of study was to determine the effect of ConA on the proliferation and osteogenic differentiation of DPSCs in vitro. MATERIALS AND METHODS: In this in vitro study, DPSCs were isolated from third molars before ConA induction was performed at concentrations of 5 and 10 µg/mL. The proliferation assay was determined by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay. Osteogenic differentiation was determined by means of mineralization. STATISTICAL ANALYSIS: Data were analyzed using analysis of variance and a Student's t-test. The p-value was set at 0.05. RESULTS: The addition of 5 and 10 µg/mL of ConA to DPSCs can significantly increase the proliferation and osteogenic differentiation of DPSCs (p ≤0.05). CONCLUSION: ConA can increase the proliferation and osteogenic differentiation of DPSCs.

Potential of Distilled Liquid Smoke Derived from Coconut (Cocos nucifera L) Shell for Traumatic Ulcer Healing in Diabetic Rats.

OBJECTIVE: Distilled liquid smoke (DLS) is a result of coconut processing by-product that not only serves as a natural food preservative but also has a promising therapeutic effect. The healing potential of DLS derived from coconut (Cocos nucifera L) shell was investigated on a traumatic ulcer with the diabetic rat. MATERIALS AND METHODS: DLS was analyzed the component by gas chromatograph mass spectrometry. Diabetic condition was induced by alloxan in 55 male Wistar rats. Ten mm of traumatic ulcer was made along the labial fornix incisive inferior after the diabetic condition was confirmed. Then DLS coconut shell, benzydamine hydrochloride, and sterile distilled water were applied topically for 3, 5, and 7 days. The potential healing was evaluated based on the expression of nuclear factor kappa beta (NFκB) and tumor necrosis factor alpha (TNF-α) on macrophages using immunohistochemical staining and the amount of collagen using Masson Trichome staining. The difference between each group was analyzed using one-way analysis of variance. The least significant difference test is used to determine the significant difference (p < 0.05). RESULTS: The major compounds found were phenol (36.6%), 2-methoxyphenol (guaiacol) (25.2%), furfural (17.8%), and 4-ethyl-2-methoxyphenol (3.5%) with 28 other minor constituents. The lowest NFκB and TNF-α expression on macrophage was observed by topical application of DLS derived from coconut shell for 3, 5, and 7 days of treatment. The amount of collagen was increased and indicated by the highest result of DLS compared to others. CONCLUSION: The DLS derived from coconut (Cocos nucifera L) shell was able to improve traumatic ulcer healing in a person with diabetes.