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Yanni Dirgantara
Prodia Stem Cell Laboratory Jl. Kramat 7 No.11, Jakarta
Biochemistry, Genetics & Molecular Biology Public Health

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Proliferation of Peripheral Blood-derived Endothelial Progenitor Cells from Stable Angina Subjects Yudi Her Oktaviono; Djanggan Sargowo; Mohammad Aris Widodo; Yanni Dirgantara; Angliana Chouw; Ferry Sandra
The Indonesian Biomedical Journal Vol 6, No 2 (2014)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v6i2.34

Abstract

BACKGROUND: A population of circulating Endothelial Progenitor Cells (EPCs) has been reported to play important role in maintaining endothelial function and integrity. Since EPCs culture is crucial and an optimized medium is currently available. Therefore we conducted a study to investigate whether stable angina subjects peripheral blood-derived EPCs could be cultured in this medium. Here, we performed study to detect EPCs characteristics and extracellular signalregulated kinase (Erk)1/2 Mitogen-Activated Protein Kinase (MAPK) pathway as possible underlying pathway for EPCs proliferation.METHODS: Peripheral blood EPCs from 8 stable angina subjects were cultured in an optimized medium with/without addition of supplement for 1 or 3 days. Then, the membrane of cultured EPCs were detected with immunofluorescence method for CD34, Vascular Endothelial Growth Factor Receptor 2 (VEGFR-2) and CD133. Colony forming unit (CFU) enumeration was performed. XTT Cell proliferation assay was performed to assess EPCs growth after 1 and 3-days culture. The western blot analysis was performed to detect possible activation of Erk1/2 MAPK.RESULTS: Number of EPCs and CFU cultured for 3 days were significantly higher than the ones cultured for 1 day (p=0.012). EPCs membrane markers from stable angina subjects were detected as well as CFUs were formed. There were significant increase of EPCs number, CFUs number and phosphorylated-Erk2 amount when the groups with and without supplement were compared (p<0.05). Meanwhile U0126, a MAPK Erk1/2 (MEK1/2) inhibitor, significantly inhibited the supplement-induced EPCs number, CFUs number and phosphorylated-Erk2 amount (p<0.05).CONCLUSION: Our results showed that ERK2 MAPK signaling pathway might play an important role in supplement-induced peripheral blood EPCs proliferation in subjects with stable angina.KEYWORDS: endothelial progenitor cell, EPC, p42, Erk2, proliferation
Macerated-Pineapple Core Crude Extract-derived Bromelain Has Low Cytotoxic Effect in NIH-3T3 Fibroblast Dewi Liliany Margaretta; Angliana Chouw; Yanni Dirgantara; Melanie Sadono Djamil; Ferry Sandra
The Indonesian Biomedical Journal Vol 7, No 2 (2015)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v7i2.75

Abstract

BACKGROUND: Bromelain is a sulfhydryl proteolytic enzyme that can hydrolyze protein, protease or peptide. Bromelain can be found in pineapple stem, fruit and core. Bromelain is composed of 212 amino acid residues with cysteine-25 forming a polypeptide chain that can hydrolyze peptide bonds by H2O. In medicine, bromelain has been developed as antibiotic, cancer drug, anti-inflammatory agent and immunomodulator. In dentistry, bromelain has potential to reduce plaque formation on the teeth and to irrigate root canal.METHODS: Pineapple core was dried for 3 days to get simplicia. Then simplicia was extracted with water solvent for 24 hours. After that, the macerated-pineapple core crude extract-derived bromelain (PCB) was separated by Sodium Dodecyl Sulphate-Polyacrylamide Gel Electrophoresis (SDS-PAGE) followed by Coomassie Brilliant Blue (CBB) staining to ensure the presence of bromelain. In cytotoxic test, NIH-3T3 fibroblast cultures were treated with extracts in various concentrations to for 24 or 48 hours. Number of fibroblasts was calculated using 3-(4,5-dimethylthiazol-2- yl)-2,5-Diphenyltetrazolium bromide (MTT) assay.RESULTS: Pineapple core extraction using maceration method produced relative high yield (concentration: 1.5424 g/mL) of bromelain, which was confirmed by CBB staining results with the molecular weight of 33 kDa. Based on cytotoxic test results of PCB on NIH-3T3 fibroblasts, 24-hours-incubation LD50 was 95.7 g/L, while 48-hours-incubation LD50 was 51.1 g/L.CONCLUSION: PCB has low cytotoxic effect in NIH-3T3 fibroblasts.KEYWORDS: bromelain, pineapple, extract, cytotoxic, MTT
Co-Authors Angliana Chouw Dewi Liliany Margaretta, Dewi Liliany Djanggan Sargowo Ferry Sandra Melanie Sadono Djamil Mohammad Aris Widodo Yudi Her Oktaviono, Yudi Her