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All Journal The Indonesian Biomedical Journal Scientific Dental Journal
Muhammad Ihsan Rizal, Muhammad Ihsan
Deparment Of Biochemistry And Molecular Biology, Division Of Oral Biology, Faculty Of Dentistry, Trisakti University
Biochemistry, Genetics & Molecular Biology Dentistry Education Health Professions Immunology & microbiology Public Health

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Brucea javanica Leaf Extract Activates Caspase-9 and Caspase-3 of Mitochondrial Apoptotic Pathway in Human Oral Squamous Cell Carcinoma Muhammad Ihsan Rizal; Ferry Sandra
The Indonesian Biomedical Journal Vol 8, No 1 (2016)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v8i1.198

Abstract

BACKGROUND: We previously reported Brucea javanica leaf extract (BJLE) induced apoptosis in human oral squamous cell carcinoma (HSC2) cells by attenuation of mitochondrial membrane permeability. However, further underlying mechanism is not known yet. Therefore, we conducted a study to investigate activation of Caspases related to attenuation of mitochondrial membrane permeability in BJLE-treated human oral squamous cell carcinoma.METHODS: B. javanica leaves were collected, identified, minced, dried, extracted with distilled ethanol at room temperature for 24 hours, filtered and evaporated. Resulted BJLE was stored at 4°C. HSC-2 and HSC-3 cells were fasted for 12 hours and treated with BJLE in various concentrations for 24 hours. Treated HSC-2 and HSC-3 cells were lysed and subjected to western blot, to detect cleaved-Caspase-9, cleaved-Caspase-3 and β-actin. All visualized bands were captured and quantified.RESULTS: Low numbers and morphological alterations of adherent HSC-2 and HSC-3 cells were observed in the group of cells treated with 500, 100 and 10 μg/mL BJLE. Numbers of adherent HSC-2 and HSC-3 cells treated with BJLE were shown decreased along with the increase of BJLE concentrations. Meanwhile, numbers of floating HSC-2 and HSC-3 cells were increased. Bands of cleaved-Caspase-9 and cleaved-Caspase-3 were observed in HSC-2 and HSC-3 cells treated with 500 and 100 μg/mL BJLE. Higher-density bands of cleaved-Caspase-9 and cleaved-Caspase-3 were observed in HSC-2 and HSC-3 cells treated with 500 μg/mL BJLE than 100 μg/mL BJLE. CONCLUSION: BJLE could induce apoptosis by activation of Caspase 9 and Caspase 3 of mitochondrial apoptotic pathway in human oral squamous cell carcinoma. KEYWORDS: Brucea javanica, leaf, apoptosis, HSC-2, HSC-3, Caspase 9, Caspase 3
Level of Salivary Uric Acid in Gingivitis and Periodontitis Patients Muhammad Ihsan Rizal; Stiefani Vega
Scientific Dental Journal Vol. 1 No. 1 (2017): September
Publisher : Faculty of Dentistry, Trisakti University

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.26912/sdj.v1i1.1912

Abstract

Background: Periodontal disease is common chronic adult condition. Antioxidants are present in the body fluid as protection against free radical. Uric acid is one of antioxidants that can be found in saliva. Moreover, the relationship among the antioxidant enzymes activities and clinical periodontal status were investigated. Objectives: The aim of the study was to observe uric acid level activities in the saliva of gingivitis and periodontitis patients. Methods: Six patients with gingivitis and six patients with periodontitis in Dental Hospital Trisakti University were included in the study. Clinical condition of each subject, the plaque index, and probing depth were determined. The salivary uric acid level was measured using the Folin-Wu method. Result: Salivary uric acid levels in the periodontitis patients with a mean ± SD  7.40 ± 0.31  (p = 0.004) were found to be higher compared to the gingivitis patients (mean ± SD = 6.84 ± 0.19). In addition, there were no significant differences in salivary uric acid levels between gender (p = 0.641). Conclusion: Uric acid levels in periodontitis patients were found to be higher than in gingivitis patients. Moreover, uric acid has more role on periodontitis than in gingivitis as an antioxidant agent.
Caffeic Acid Induces Apoptosis in MG-63 Osteosarcoma Cells via Protein Kinase C Delta (PKCδ) Translocation and Mitochondrial Membrane Potential Reduction Ferry Sandra; Muhammad Ihsan Rizal; Caecilia Caroline Aliwarga; Jenifer Christy Hadimartana; Maria Celinna
The Indonesian Biomedical Journal Vol 14, No 4 (2022)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v14i4.2089

Abstract

BACKGROUND: Caffeic acid has been reported to activate caspases in MG-63 osteosarcoma cells, which can lead to apoptosis via both extrinsic and intrinsic apoptotic pathways. Translocation of protein kinase C delta (PKCδ), which reduces mitochondrial membrane potential (ΔΨm), is involved in apoptosis. The role of PKCδ translocation and ΔΨm alteration in caffeic acid-induced MG-63 cell apoptosis are largely unknown. Present study investigated the effect of caffeic acid on PKCδ translocation and ΔΨm in MG-63 cells.METHODS: MG-63 cells were cultured and starved, followed by pretreatment with or without Z-VAD-FMK and treatment with or without 10 μg/mL caffeic acid. MG-63 cells were collected, lysed, and processed to obtain cytosolic and mitochondrial fractions. Each fraction was subjected to immunoblotting analysis by using anti-PKCδ antibody. Mitochondrial membrane potential (ΔΨm) was measured using flow cytometry.RESULTS: Cytosolic PKCδ levels were higher than mitochondrial PKCδ levels in untreated and 1 h caffeic acid treatment groups. Inversely, cytosolic PKCδ levels were lower than the mitochondrial PKCδ levels after 6 and 12 h caffeic acid treatment. By Z-VAD-FMK pretreatment, cytosolic PKCδ levels were higher than mitochondrial PKCδ after 6 and 12 h caffeic acid treatment. After 6 h treatment with caffeic acid, ΔΨm was slightly shifted. More shifting occurred in MG-63 cells treated with caffeic acid for 12 h. The ΔΨm shifting was inhibited by Z-VAD-FMK pretreatment.CONCLUSION: Caffeic acid could trigger apoptosis of MG-63 osteosarcoma cells by inducing PKCδ translocation to mitochondria and reducing ΔΨm, which might cause MMP.KEYWORDS: caffeic acid, MG-63, osteosarcoma, PKCδ, mitochondrial membrane potential, mitochondrial membrane permeabilization, Z-VAD-FMK
Caffeic Acid Induces Intrinsic Apoptotic Pathway in MG-63 Osteosarcoma Cells Through Bid Truncation and Cytochrome c Release Ferry Sandra; Muhammad Ihsan Rizal; Ayasha Hajjar Audreyandra Wahid; Monica Andajana; Maria Celinna
The Indonesian Biomedical Journal Vol 14, No 3 (2022)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v14i3.2032

Abstract

BACKGROUND: Caffeic acid has been reported to induce apoptosis in MG-63 osteosarcoma cells via caspases activation. However, apoptotic pathway that is involved in the caffeic acid-induced apoptosis is still unclear. Present study aimed to investigate the role of cytochrome c (Cyt c) release and BH3-interacting death (Bid) activation in caffeic acid-induced apoptosis in MG-63 osteosarcoma cells.METHODS: MG-63 cells were cultured, pretreated with/without Z-VAD FMK and treated with/without 10 μg/mL caffeic acid. Treated MG-63 cells were then lysed, homogenized, and processed further to prepare cell lysate and mitochondrial fraction. Immunoblotting method was used to measure the amount of Bid and truncated Bid (t-Bid) as well as mitochondrial and cytosolic Cyt c.RESULTS: The amount of Bid and mitochondrial Cyt c in MG-63 cells decreased in a time-dependent manner, while the amount of t-Bid and cytosolic Cyt c increased in a time-dependent manner. By pretreatment of 100 μM Z-VAD-FMK for 2 h, the amount of Bid and mitochondrial Cyt c was significantly higher, while the amount of t-Bid and cytosolic Cyt c was significantly lower after caffeic acid treatment for 6 and 12 h compared to MG-63 cells that were not pretreated.CONCLUSION: Caffeic acid could induce Cyt c release through the activation of Bid in MG-63 osteosarcoma cells.KEYWORDS: caffeic acid, osteosarcoma, MG-63 cells, Bid, t-Bid, cytochrome c, Z-VAD-FMK
Caffeic Acid Inhibits Swelling, Bone Loss, and Osteoclastogenesis in Adjuvant-induced Arthritis Rats Ferry Sandra; Muhammad Ihsan Rizal; Nurrani Mustika Dewi; Toshio Kukita
The Indonesian Biomedical Journal Vol 14, No 3 (2022)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v14i3.2033

Abstract

BACKGROUND: Increase in inflammatory cytokine levels promotes pathological osteoclast differentiation. Caffeic acid has anti-inflammatory properties and can inhibit osteoclast bone resorption. In vitro studies have reported the ability of caffeic acid in inhibiting osteoclastogenesis pathways, however the in vivo study is rarely conducted. The aim of this study is to examine the role of caffeic acid in reducing inflammation and inhibiting osteoclastogenesis in Adjuvant-Induced Arthritis (AIA) rats.METHODS: Rats were injected with Freund’s Complete Adjuvant (CFA) and mineral oil. One day after injection, various concentration (0, 5, 25, 125 mg) of caffeic acid were given gastro-intestinally. Swelling degree in rats’ ankle joints was determined by measuring height and width of each ankle joint. Bone loss level was examined with soft X-ray, and then bone density was calculated. To examine osteoclastogenesis, ankle joints were stained with Tartrate-Resistant Acid Phosphatase (TRAP) and evaluated microscopically. RESULTS: Ankle joints of AIA rats had severe swelling before treated, yet the swelling was reduced based on concentration-dependent after receiving caffeic acid. Severe bone loss in AIA rats’ ankle joints were also observed, however the treatment of 125 mg caffeic acid showed remarkable inhibition effect toward rats’ bone loss. Osteoclastogenesis in AIA rats’ ankle joints were higher than the normal ones, as indicated with high TRAP-positive Multinucleated Cells (MNCs). But low number of TRAP-positive MNCs was observed in ankle joint of AIA rats that received 125 mg caffeic acid.CONCLUSION: Administration of caffeic acid can reduce the degree of swallowing, inhibit bone loss, and inhibit osteoclastogenesis in ankle joint of arthritis-induced rats.KEYWORDS: caffeic acid, osteoclastogenesis, bone loss, swelling, inflammation, RANKL, TNF-α
Co-Authors Ayasha Hajjar Audreyandra Wahid Caecilia Caroline Aliwarga Ferry Sandra Jenifer Christy Hadimartana Maria Celinna Monica Andajana Nurrani Mustika Dewi Stiefani Vega Toshio Kukita