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All Journal Jurnal Kedokteran Brawijaya Majalah Kedokteran Bandung Universa Medicina
Agung Putra
Faculty of Medicine Universitas Islam Sultan Agung, Semarang
Health Professions Immunology & microbiology Medicine & Pharmacology Public Health

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Hypoxia enhances self-renewal properties and markers of mesenchymal stem cells Yustianingsih, Vivi; Sumarawati, Titiek; Putra, Agung
Universa Medicina Vol 38, No 3 (2019)
Publisher : Faculty of Medicine, Trisakti University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (668.593 KB) | DOI: 10.18051/UnivMed.2019.v38.164-171

Abstract

BackgroundMesenchymal stem cells (MSCs) are multipotent stromal cells that express CD73, CD90, and CD105 surface markers, but not CD14, CD45, CD34, CD11b, and HLA-DR. MSCs under hypoxic conditions have the essential role of maintaining the stemness capacity by releasing several growth factors into their medium, known as hypoxia conditioned medium (HCM). This study was performed to compare the effect of percentage of HCM to normoxic medium (NM) in increasing MSC proliferation marked by proliferation rate and surface marker expression.MethodsThis study was of post-test only control group design using human umbilical cord-MSCs (hUC-MSCs) as subjects. The HCM treatment group was obtained by culturing MSCs under 5% O2, whereas the NM control group was grown under 20% O2. The hUC-MSCs were divided into 4 groups with different dose ratios of HCM to NM (25%:75%; 50%:50%; 75%:25% for P1, P2 and P3, respectively and 100% of NM for the controls). All of these groups were maintained at 37oC and the data was collected after 72 hours incubation. MSC marker expression of CD73, CD90 and CD105 was analyzed using flow cytometry and MSC proliferation by trypan blue assay. ResultThere were significant differences in MSC marker expression of CD73, CD90 and CD105 and proliferation at all dose ratios of HCM to NM (p<0.05).ConclusionLow oxygen concentration promotes MSC proliferation and stemness thus it might be beneficial for maintaining the MSC physiologic niche in-vitro.
Peran Mesenchymal Stem Cells dalam Regulasi PDGF dan Sel Islet pada Diabetes HS, Zakariya; Putra, Agung
Jurnal Kedokteran Brawijaya Vol 30, No 2 (2018)
Publisher : Fakultas Kedokteran Universitas Brawijaya

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21776/ub.jkb.2018.030.02.4

Abstract

Diabetes merupakan kelompok penyakit metabolik yang ditandai dengan peningkatan kadar glukosa darah sebagai akibat kerusakan sel β pankreas dan atau resistensi insulin. Penelitian klinik menunjukan bahwa transplantasi islet dapat memperbaiki gejala terkait diabetes, meskipun demikian keterbatasan pendonor menjadi masalah serius. Hasil penelitian terkini melaporkan bahwa Mesenchymal Stem Cells (MSCs) berpotensi tinggi dalam meregenerasi kerusakan jaringan pankreas termasuk sel islet dengan melibatkan berbagai growth factor terutama PDGF. Tujuan penelitian ini adalah untuk mengetahui pengaruh MSCs tali pusat dalam meningkatkan jumlah sel islet dan gambaran kadar PDGF pada hari ke 44 fase remodelling jaringan pankreas yang rusak. Penelitian ini menggunakan 20 mencit (Mus musculus) Balb-C yang diinduksi streptozotocin hingga diabetes. Terdapat 4 kelompok penelitian (n=5/kelompok) terdiri atas kontrol (K) dan perlakuan (P). Kelompok kontrol diberi PBS sementara P1, P2, dan P3 diberi MSCs dengan dosis berbeda, yaitu: P1=1,5x105, P2= 3x105, dan P3=6x105 sel secara intraperitoneal. Pada hari ke 44 dilakukan pemeriksaan ELISA pada sampel darah untuk mengetahui kadar PDGF dan histopatologi jaringan untuk penghitungan sel islet pankreas. Hasil penelitian didapatkan peningkatan jumlah sel islet pankreas secara signifikan (p<0,05) dan kadar PDGF sesuai dengan kelompok kontrol. Penelitian eksperimental pada mencit (Mus musculus) inimengesankan bahwa MSCs mampu meningkatkan jumlah sel islet pankreas dan meregulasi kadar PDGF.
TNF-α-ACTIVATED MSC-CM TOPICAL GEL EFFECTIVE IN INCREASING PDGF LEVEL, FIBROBLAST DENSITY, AND WOUND HEALING PROCESS COMPARED TO SUBCUTANEOUS INJECTION COMBINATION Kuntardjo, Novalia; Dharmana, Edi; Chodidjah, Chodidjah; Nasihun, Taufiq R; Putra, Agung
Majalah Kedokteran Bandung Vol 51, No 1 (2019)
Publisher : Faculty of Medicine, Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15395/mkb.v51n1.1479

Abstract

Mesenchymal stem cells (MSCs) are multipotent stromal cells that have the capacity to regenerate tissue damage. However, they have several limitations. MSC-CM as a new approach treatment is widely used to solve the limitation of MSC in wound healing. The aim of this study was to evaluate the effectiveness of TNF-?-activated MSC-CM topical gel compare to topical-subcutaneous injection combination on wound healing acceleration. This study was conducted between April and August 2018 at the Stem Cell and Cancer Research Laboratory (SCCR), Faculty of Medicine, Sultan Agung Islamic University, Semarang. Experimental post-test only control group design was performed by involving 36 animal models randomly divided into six groups; T1, T2 (MSC-CM in topical gel 100 ?L; 200 ?L); ST1, ST2 (MSC-CM in subcutaneous injection : topical gel = 80 ?L:20 ?L; 160 ?L:40 ?L); CT (200 ?L medium free TNF-?); CST (PBS in subcutaneous injection : topical gel = 160 ?L :40 ?L). The measurement of PDGF level on day 3 and 6 was conducted using ELISA assay while the fibroblast density was analyzed by light microcopy. It was found that there was was a significant increase in PDGF and fibroblast density on day 6 in the topical group when compared to the combination group (p<0,05). It is concluded that the MSC-CM topical gel is more effective than combination of topical-subcutaneous injection.Key words: Combination, fibroblast, MSC-CM, PDGF, subcutaneous MSC-CM, topical MSC-CM MSC-CM Topikal yang diaktivasi TNF-? Efektif Dalam Peningkatan Level PDGF, Densitas Fibroblast, dan Mempercepat Penyembuhan Luka dibanding dengan Kombinasi Injeksi SubkutanMesenchymal stem cells (MSCs) merupakan sel stroma multipoten yang memiliki kemampuan untuk meregenerasi kerusakan jaringan. Namun, MSC memiliki beberapa keterbatasan. MSC-CM sebagai terapi pendekatan baru digunakan untuk mengatasi keterbatasan MSC dalam penyembuhan luka. Tujuan penelitian ini adalah menilai efektivitas MSC-CM topikal yang diaktvasi TNF-? dibandingdengan kombinasi topikal-injeksi subkutan pada percepatan penyembuhan luka. Penelitian ini dilaksanakan pada bulan April?Agustus 2018 di Laboratorium Stem Cell and Cancer Research (SCCR), Fakultas Kedokteran, Universitas Islam Sultan Agung, Semarang. Penelitian menggunakan eksperimen laboratorium dengan rancangan post-test only control group, menggunakan 36 tikus galur wistar yang dibagi secara acak menjadi 6 kelompok; T1, T2 (MSC-CM gel topikal 100?L; 200?L); ST1, ST2 (MSC-CM injeksi subkutan : gel topikal = 80 ?L:20 ?L; 160 ?L:40 ?L); CT (200 ?L medium tanpa TNF-?); CST (PBS injeksi subkutan: gel topikal = 160 ?L :40 ?L). Pengukuran kadar PDGF pada hari ke-3 dan ke-6 mengunakan ELISA, sedangkan jumlah fibroblas dilihat mengunakan mikroskop cahaya. Hasil penelitian ini menunjukkan peningkatan kadar PDGF dan jumlah fibroblas yang signifikan dihari ke-6 pada MSC-CM gel topikal dibanding dengan kombinasi topical-injeksi subukutan (p <0.05). Simpulan penelitian ini adalah pemberian MSC-CM secara topical lebih efektif dibanding dengan kombinasi topikal-injeksi subkutan.Kata kunci: Fibroblas, konditional medium, MSC-CM kombinasi, MSC-CM topikal, PDGF
PERAN INDUKSI TNF-α SERIAL DOSES DALAM PENINGKATAN VEGF DAN PDGF MESENCHYMAL STEM CELLS Putra, Agung; Hutagalung, Ananta; Hasanal, Ihdina Hanifa; Trisnadi, Setyo; Djannah, Durrotul; Cahyono, Erwin Budi; Intan, Yulice Soraya Nur
Majalah Kedokteran Bandung Vol 50, No 2 (2018)
Publisher : Faculty of Medicine, Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1316.352 KB) | DOI: 10.15395/mkb.v50n2.1278

Abstract

Mesenchymal stem cell (MSC) mempunyai kemampuan immunoregulasi dan regenerasi melalui supresi pelepasan mediator proinflamasidan peningkatan molekul proliferasi terutama vascular endothelial growth factor (VEGF) dan platelet-derived growth factor (PDGF). Mesenchymal stem cell yang diaktivasi TNF-? dengan dosis tertentu mampu meningkatkan sekresi VEGF dan PDGF, namun dosis optimal TNF-? yang mampu memaksimalkan ekspresi molekul tersebut belum diketahui secara pasti. Variasi dosis TNF-? digunakan pada penelitian ini dengan tujuan mengetahui dosis optimal, rendah, dan tinggi TNF-? dalam memaksimalkan ekspresi VEGF dan PDGF. Penelitian ini mengunakan post-test only control group design dengan 5 kelompok penelitian, terdiri atas satu kelompok kontrol (K) dan 4 kelompok perlakuan (P) (TNF-?= 5, 10, 40, 80 ng/mL) yang diinduksikan pada MSC dengan inkubasi 24 jam, kemudian kadar PDGF dan VEGF diukur dengan metode ELISA. Penelitian ini dilakukan antara bulan September?November 2017 di Laboratorium Stem Cell and Cancer Research (SCCR), Fakultas Kedokteran, Universitas Islam Sultan Agung, Semarang. Hasil penelitian menunjukkan peningkatan kadar PDGF dan VEGF secara signifikan (p<0,05) dimulai dari dosis TNF-? 5 ng/mL, optimal padadosis 10 ng/mL dan mulai terjadi penurunan pada dosis 40 ng/mL. Induksi TNF-? pada MSC mampu memaksimalkan kadar VEGF dan PDGF pada dosis 10 ng/mL.Kata kunci: MSC, PDGF, TNF-?, VEGF Effect of TNF-? Serial Doses Inducition on Increasing VEGF dan PDGF in Mesenchymal Stem Cells Mesenchymal Stem Cells (MSCs) have immunoregulation and regeneration capabilities through suppression of proinflammatory mediator release and increase of proliferative molecules, particularly the vascular endothelial growth factor (VEGF) and platelet-derived growth factor (PDGF) TNF-? activated MSC in a certain dose has the ability to increase VEGF and PDGF levels; however, the exact optimum dose of TNF-? to optimize the levels of VEGF and PDGF is unclear. In this study, TNF-? dose variations were used to determine the optimum, low, and high doses of TNF-? in optimizing VEGF and PDGF expression. This was a post-test only control group study with five study groups consisting of one control (K) and four treatment groups(P). The treatment groups were treated with 5, 10, 40 and 80 ng/mL of TNF-? for 24 hours. PDGF and VEGF levels were measured using ELISA. This study was conducted between September?November 2017 at the Stem Cell and Cancer Research Laboratory (SCCR), Faculty of Medicine, Sultan Agung Islamic University, Semarang. The results show significant increased in PDGF and VEGF levels (p<0.05) starting from TNF-? 5 ng/mL as the initiation dose to 10 ng/mL as the optimum dose and reduction was seen starting from 40 ng/mL dose. TNF-induced MSCs have the ability to increase the VEGF and PDGF levels with an optimum dose of 10 ng/mL.Key words: MSC, PDGF, TNF-?, VEGF
Hypoxia-preconditioned mesenchymal stem cells attenuate peritoneal adhesion through TGF-β inhibition Trisnadi, Setyo; Muhar, Adi Muradi; Putra, Agung; Kustiyah, Azizah Retno
Universa Medicina Vol 39, No 2 (2020)
Publisher : Faculty of Medicine, Trisakti University

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (1708.691 KB) | DOI: 10.18051/UnivMed.2020.v39.97-104

Abstract

BackgroundPeritoneal adhesions (PAs) are generally described as fibrous bands between intra-abdominal organs following an abdominal surgical operation. The definitive treatments of PAs are currently ineffective yet. Hypoxia-mesenchymal stem cells (H-MSCs) have a higher capability to survive at the site of injury than normoxia-MSCs (N-MSCs) to repair injured tissue without fibrosis. This study aimed to analyze the effect of H-MSCs in controlling formation of PAs by reducing TGF-β level in a rat model. Methods A study of post-test only control group design was conducted, involving eighteen PA rat models weighing 250 ± 25 g that were randomly assigned into 3 groups, comprising control group (C), and groups T1 and T2 receiving H-MSC treatment at doses of 3 x 106 and 1.5 x 106, respectively. To induce H-MSCs, MSCs were incubated in hypoxic conditions at 5% O2 and 37oC for 24 hours. Expression level of TGF-β was analyzed by enzyme-linked immunosorbent assay (ELISA) at 450 nm and adhesion formation was described macroscopically. The Kruskal-Wallis variance analysis was used to analyze significant differences among the groups.ResultsThe results of this study showed that H-MSCs in group T1 inhibited TGF-β expression significantly on day 8 (p&lt;0.001) and day 14 (p&lt;0.05). Moreover, there was almost no adhesion apparent following H-MSC administration in group T1. ConclusionsBased on this study, we conclude that H-MSCs may attenuate PA formation following inhibition of TGF-β expression in the PA rat model.
Suppression of transforming growth factor-β by mesenchymal stem-cells accelerates liver regeneration in liver fibrosis animal model Sa’dyah, Nur Anna C; Putra, Agung; Dirja, Bayu Tirta; Hidayah, Nurul; Yasmine Azzahara, Salma; Candra Satria Irawan, Risky
Universa Medicina Vol. 40 No. 1 (2021)
Publisher : Faculty of Medicine, Universitas Trisakti

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2021.v40.29-35

Abstract

IntroductionLiver fibrosis (LF) results from the unregulated chronic wound healing process in liver tissue. Transforming growth factor-beta (TGF-β) is the major contributing cytokine of LF promotion through activation of quiescent hepatic stellate cells (HSCs) into myofibroblasts (MFs) and increased extracellular matrix (ECM) deposition such as collagen leading to scar tissue development. Mesenchymal stem cells (MSCs) have an immunomodulatory capability that could be used as a new treatment for repairing and regenerating LF through suppression of TGF-β. This study aimed to examine the role of MSCs in liver fibrosis animal models through suppression of TGF-β levels without scar formation particularly in the proliferation phase.MethodsIn this study, a completely randomized design was used with sample size of 24. Male Sprague Dawley rats were injected intraperitoneally (IP) with carbon tetrachloride (CCl4), twice weekly, for eight weeks to induce LF. Rats were randomly assigned to four groups: negative control, CCl4 group, and CCL4 + MSC-treated groups T1 and T2, at doses of 1 x 106 and 2x106 cells, respectively. TGF-β levels were analyzed by enzyme-linked immunosorbent assay (ELISA). One-way ANOVA and a least significant difference (LSD) was used to analyse the data. ResultsThe TGF levels of LF rat models decreased on day 7 after MSC administration. The levels of TGF-β in both MSC groups T1 and T2 decreased significantly compared with the control group (p<0.05). The TGF-β suppression capability of T2 was optimal and more significant than that of T1.ConclusionMSCs can suppress TGF levels in liver fibrosis induced rats.
Co-Authors Azizah Retno Kustiyah Cahyono, Erwin Budi Candra Satria Irawan, Risky Chodidjah Chodidjah Dirja, Bayu Tirta Djannah, Durrotul Edi Dharmana Hasanal, Ihdina Hanifa HS, Zakariya Hutagalung, Ananta Intan, Yulice Soraya Nur Kuntardjo, Novalia Muhar, Adi Muradi NURUL HIDAYAH Sa’dyah, Nur Anna C Setyo Trisnadi Taufiq R Nasihun, Taufiq R Titiek Sumarawati Yasmine Azzahara, Salma Yustianingsih, Vivi