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Agung Putra
Faculty of Medicine Universitas Islam Sultan Agung, Semarang
Agriculture, Biological Sciences & Forestry Biochemistry, Genetics & Molecular Biology Chemistry Computer Science & IT Decision Sciences, Operations Research & Management Dentistry Earth & Planetary Sciences Economics, Econometrics & Finance Education Health Professions Immunology & microbiology Materials Science & Nanotechnology Medicine & Pharmacology Neuroscience Nursing Physics Public Health Social Sciences Other

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Hypoxia-Induced Mesenchymal Stem Cell Exosomes Modulate Protein Kinase A and VEGFR Expression in Ultraviolet B-Induced Hyperpigmentation in Mice Andavania, Sheila Jessica; Syamsunarno, Mas Rizky; Putra, Agung; Setiawan, Eko
Molecular and Cellular Biomedical Sciences Vol 9, No 2 (2025)
Publisher : Cell and BioPharmaceutical Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21705/mcbs.v9i2.594

Abstract

Background: Hyperpigmentation is often exacerbated by ultraviolet-B (UVB) exposure through oxidative stress and activation of pathways like mitogen-activated protein kinase (MAPK) and vascular endothelial growth factor receptor (VEGFR). Current treatments carry risks and necessitate safer alternatives. This study investigated the therapeutic potential of hypoxia-induced mesenchymal stem cell (MSC) exosomes in reducing protein kinase-A (PKA) and VEGFR expression in UVB-induced hyperpigmentation.Materials and methods: A post-test-only control group design was used with 30 male C57BL/6 mice divided into five groups: Healthy group, 0,9% NaCl-treated group, retinol-treated group, and two treatment groups (200 µL Exosomes-treated group and 300 µL Exosomes-treated group. UVB-induced hyperpigmentation was established with 180 mJ/cm² exposures over two weeks. Treatment was administered via subcutaneous injections for seven days. PKA and VEGFR mRNA levels were analyzed using qRT-PCR.Results: PKA expression was significantly lower in the 200 µL Exosomes-treated group (0.34±0.05) and 300 µL Exosomes-treated group (0.21±0.04) groups compared with the 0,9% NaCl-treated group (1.12±0.08) (p<0.001). VEGFR expression similarly decreased in 200 µL Exosomes-treated group (0.32±0.05) and 300 µL Exosomes-treated group (0.18±0.04) versus the 0,9% NaCl-treated group (1.48±0.09) (p<0.001). Both exosome doses achieved reductions comparable to baseline levels observed in the Healthy group.Conclusion: Hypoxia-induced MSC exosomes reduced PKA and VEGFR expression in UVB-induced hyperpigmentation, with the 300 µL dose showing greater efficacy. These findings suggested exosome therapy as a promising alternative for hyperpigmentation treatment. Keywords: hyperpigmentation, MSC, PKA, VEGFR, melanin
Mesenchymal Stem Cell-Derived Exosomes Enhance FGF-1 and SDF-1 Expression in Rats with Second Degree Burns Hariani, Nova Putri; Putra, Agung; Subchan, Prasetyowati; Setiawan, Eko
Molecular and Cellular Biomedical Sciences Vol 9, No 2 (2025)
Publisher : Cell and BioPharmaceutical Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21705/mcbs.v9i2.635

Abstract

Background: Second-degree burns cause extensive damage to the skin and pose significant health challenges, with current treatments facing limitations such as donor skin shortages and complications. Fibroblast growth factor 1 (FGF-1) and stromal-derived growth factor 1 (SDF-1) are critical for tissue repair. Emerging evidence suggests that mesenchymal stem cell-derived exosomes (E-MSCs) are a promising cell-free therapeutic option for enhancing wound healing through the modulation of FGF-1 and SDF-1. This study investigated the effect of E-MSCs on the expression of FGF-1 and SDF-1 genes in rats with second-degree burns.Materials and methods:  This experimental study used a second-degree burn model in Wistar rats, treated with subcutaneous injections of E-MSCs at doses of 100 µL and 200 µL. Gene expression of FGF-1 and SDF-1 was quantified using qRT-PCR. Histological validation confirmed burn severity, and flow cytometry was used to characterize E-MSCs and exosomes.Results: An increase in FGF-1 and SDF-1 expression was observed in exosome-treated groups compared to the NaCL-treated group. The 200 µL E-MSCs-treated group showed the most significant enhancement in both growth factors, with statistically significant differences (p<0.05). These findings underline the efficacy of E-MSCs in modulating critical genes involved in wound healing.Conclusion: E-MSCs significantly upregulate FGF-1 and SDF-1 expression, promoting tissue repair and regeneration in second-degree burn models. This study highlights the potential of E-MSCs as a non-invasive therapeutic approach.  Keywords: exosomes, FGF-1, mesenchymal stem cells, SDF-1
Effector Cytokine Profiles of Ex Vivo Expanded CTLs in Colorectal Cancer HCT-116 Cells Co-Culture Models Ibrahim, Sugeng; Putra, Agung; Hidayah, Nurul; Cahyani, Dini
Indonesian Journal of Medical and Pharmaceutical Science Vol. 4 No. 2 (2025)
Publisher : Sultan Agung Islamic University of Semarang

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.30659/ijmps.v4i2.450

Abstract

Background: Colorectal cancer (CRC) remains a major cause of cancer-related mortality worldwide, with limited benefit from conventional therapies in advanced disease. Cytotoxic T lymphocytes (CTLs, CD8⁺ T cells) are critical mediators of anti-tumor immunity, primarily through direct cytotoxicity and cytokine secretion. However, the dual roles of CTL-derived cytokines, particularly interferon-γ (IFN-γ), tumor necrosis factor-α (TNF-α), and interleukin-6 (IL-6), in CRC progression remain incompletely understood. This study aims to analyze the cytokine profile (IFN-γ, TNF-α, and IL-6) in a CTL–CRC cell direct co-culture model. Methods: CD8⁺ T cells were isolated from peripheral blood mononuclear cells (PBMCs) of CRC patients using magnetic negative selection and activated for 5 days with anti-CD3/CD28 beads and IL-2. Purity and viability were assessed by flow cytometry and morphology. Activated CTLs were co-cultured with the HCT116 CRC cell line at effector-to-target (E:T) ratios of 1:1, 5:1, and 10:1 under direct contact for 48 h. Supernatants were collected and cytokine levels (IFN-γ, TNF-α, and IL-6) were quantified using validated sandwich ELISA kits. Results: Direct co-culture with HCT116 cells significantly increased cytokine secretion in an E:T ratio-dependent manner. IFN-γ secretion rose from 3044.6±120 pg/mL at 1:1 to 4882.1±198 pg/mL at 5:1, plateauing thereafter. TNF-α levels remained relatively constant (628.6±67 pg/mL at 1:1 vs. 674.0±91 pg/mL at 10:1). IL-6, nearly undetectable at 1:1 (0.4±0.1 pg/mL), increased dose-dependently to 5.1±0.9 pg/mL at 10:1. Conclusion: Ex vivo expanded CTLs from CRC patients exhibit a distinct cytokine secretion profile characterized by robust IFN-γ release, stable TNF-α production, and a dose-dependent increase in IL-6 across different effector-to-target ratios.
Protective Effect of Black Rice Extract Cream on Ultraviolet B-Induced Skin Hyperpigmentation in Mice Maryanti, Maryanti; Putra, Agung; Subchan, Prasetyowati
Althea Medical Journal Vol 12, No 3 (2025)
Publisher : Faculty of Medicine Universitas Padjadjaran

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.15850/amj.v12n3.4192

Abstract

Background: Hyperpigmentation is a common sign of skin aging caused by prolonged ultraviolet B (UVB) exposure. Black rice (Oryza sativa L. var glutinosa), known for its high antioxidant content, has moisturizing and regenerative properties that may support skin health. This  study aimed to evaluate the effect of black rice extract cream on transforming growth factor beta (TGF-β) and tumor necrosis factor alpha (TNF-α) expression in a UVB-induced hyperpigmentation mouse model.Method: An in vivo experimental study with post test only control group design was conducted in 2024 at the Stem Cell and Cancer Research Laboratory, Semarang, Indonesia. Twenty-eight male C57BL/6 mice were randomly divided into four groups: healthy control (K1),  UVB-exposed negative control group (K2), UVB-exposed group treated with  7.5% (K3) and 15 % (K4) black rice extract cream  for14 days. On day 15, TGF-β and TNF-α expression levels were analyzed using the RTq-PCR,  normalized to GAPDH. Data were analyzed using One-way ANOVA followed by post-hoc testing.Results: TGF-β gene expression was the highest in K4 (1.87±0.23), followed by K3 (1.52±0.42l) which was statistically significant different between groups (p=000); whereas TNF-α gene expression was the lowest in K4 (1.92±1.02) compared with K3 (5.40±2.28), and the difference between groups was also statistically significant (p=000).  Conclusion: Black rice extract cream increase TGF-β expression and reduces TNF-α expression in UVB-induced hyperpigmentation. These findings suggests its potential as a natural topical agent to mitigate UVB-induced skin damage and premature aging.
Exosome Hypoxic-MSCs, Glutathione, and Vitamin C: Effect on IL-10 Levels and CD-163 Expression Utami, Wulan Dyah; Muhar, Adi Muradi; Sumarawati, Titiek; Putra, Agung; Setiawan, Eko; Ibrahim, Sugeng; Taskworo, Dodik; Haitamy, Mohammad Nurrizki
Indonesian Journal of Pharmaceutical Science and Technology Vol 12, No 3 (2025)
Publisher : Indonesian Journal of Pharmaceutical Science and Technology

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.24198/ijpst.v12i3.60941

Abstract

Hyperpigmentation of the skin is a result of ultraviolet B (UVB) exposure, which causes oxidative stress due to increased reactive oxygen species (ROS), leading to various skin problems, including melanin accumulation. Exosomes can affect melanocyte activity. Exosomes, as small vesicles released by cells, can affect melanocyte activity and play an important role in various hyperpigmentation processes. The study aims to determine the effect of exosome mesenchymal stem cell hypoxia (EH-MSC) and glutathione with vitamin C on IL-10 levels and CD163 expression. IL-10 gene expression was measured using qRT-PCR, while CD163 expression was analyzed via immunohistochemical staining. A total of 30 male C57BL/6 mice were used and randomly assigned to five different treatment groups. The highest expression of IL-10 was observed in the EH-MSCs-treated group (K4), although the difference was not statistically significant compared to the control (p = 0.135). In contrast, the group receiving a combination of EH-MSCs with glutathione and vitamin C (K5) exhibited the highest percentage of CD163 expression, with a statistically significant difference (p = 0.00). These findings demonstrate that the administration of EH-MSC and glutathione with vitamin C significantly increased the expression of CD163, but insignificantly increased IL-10 in C57BL/6 mice with a UVB-induced hyperpigmentation model.
Potential Use of the Gel Extract of Butterfly Pea Flower as Topical Therapy to Prevent Photodamage by Downregulating TNF-α and Caspase-3 Expression Levels in UVB-Exposed Rats Cahyani, Elvana; Putra, Agung; Subchan, Prasetyowati
Makara Journal of Health Research Vol. 27, No. 1
Publisher : UI Scholars Hub

Show Abstract | Download Original | Original Source | Check in Google Scholar

Abstract

Background: Prolonged exposure to UVB radiation causes DNA damage in skin cells by raising the levels of reactive oxygen species, resulting in the production of inflammatory factors and skin issues. Plant extracts are frequently used to counteract photodamage due to their antioxidant properties. One example is the floral extract of the butterfly pea plant, which contains flavonoid antioxidants. However, the effect of the extract on inflammatory factors is unknown. This study investigated how tumor necrosis factor-alpha (TNF-α) and caspase-3 expression changed when a butterfly pea flower extract gel was applied topically to UVB-exposed animals. Methods: Experimental and control groups were tested. The healthy group was not exposed to UVB. The negative controls and treatments 1 and 2 were exposed daily for 5 days at a minimal erythema dose of 160 mJ/cm2 and then treated with a gel-based extract containing 5% and 10% of the extract, respectively. A 96% ethanol solution was used during the maceration step for the extraction. Real-Time Quantitative Reverse Transcription PCR was used to examine gene expression levels in the skin tissue on day 14. Results: The expression levels of TNF-α and caspase-3 decreased in the treatment group, and higher doses of the extract had a greater effect. Conclusions: The gel extract considerably reduced the UVB-induced TNF-α and caspase-3 production in rats.
Secretome hypoxia-mesenchymal stem cells decrease tumor necrosis factor-α and interleukin-18 in kidney of type 2 diabetes mellitus model rats Irawan, Risky Chandra Satria; Putra, Agung; Setyo, Trisnadi; Ghaisani, Shabrina Syifa; Hidayah, Nurul
Universa Medicina Vol. 42 No. 3 (2023)
Publisher : Faculty of Medicine, Universitas Trisakti

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2023.v42.320-328

Abstract

Background Type 2 diabetes mellitus (T2DM) is a chronic disease that affects millions of people worldwide and associated with an increased risk of kidney damage caused by prolonged inflammation. Secretome hypoxia- mesenchymal stem cells (SH-MSCs) have been investigated as a potential therapy for kidney inflammation in T2DM, due to their immunomodulatory properties and ability to promote tissue repair. In this study, we investigated the effects of SH-MSCs on tumor necrosis α (TNF-á) and interleukin-18 (IL-18) in the kidney of the T2DM model rats. MethodsA post-test-only control group involving 24 male Wistar rats. The rats were treated with a high-fat diet (HFD) for 4 weeks and streptozotocin-nicotinamide with sucrose solution for 5 days to induce T2DM animal models. Rats were randomly divided into four groups: healthy, control, and groups treated with SH-MSCs T1 and T2, with doses of 250 µL and 500 µL, respectively. TNF-α and IL-18 gene expression was measured by real time polymerase chain reaction (RT-PCR). One Way ANOVA and post-hoc LSD tests were used to determine the significant difference against all groups based on their quantitative measurement. ResultsAdministration of the SH-MSCs at a dose of 500µL (T2) was able to significantly reduce TNF-α and IL-18 gene expression when compared to control (T2DM rat without treatment) (p<0.05), but not significantly when compared to healthy and SH-MSC at a dose of 250µL (T1) group (p>0.05). ConclusionThis study demonstrated that the SH-MSCs decreased the levels of proinflammatory cytokines TNF-α and IL-18 gene expression in the kidney of T2DM model rats.
The Role of Hypoxic Mesenchymal Stem Cells Conditioned Medium in Increasing Vascular Endothelial Growth Factors (VEGF) Levels and Collagen Synthesis to Accelerate Wound Healing Sunarto, Hadi; Trisnadi, Setyo; Putra, Agung; Sa'dyah, Nur Anna Chalimah; Tjipta, Arya; Chodidjah, Chodidjah
Indonesian Journal of Cancer Chemoprevention Vol 11, No 3 (2020)
Publisher : Indonesian Society for Cancer Chemoprevention

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.14499/indonesianjcanchemoprev11iss3pp134-143

Abstract

Full-thickness wound are areas damage of skin associated with loss of epidermis and dermis. The wound healing mechanism consists proliferation, migration and remodeling. Hypoxic conditional medium of mesenchymal stem cells (HMSCs-CM) contains lots of soluble molecules, such as protein growth factor and cytokine anti-inflammation. The soluble molecule of HMSCs-CM plays a critical role in wound healing by upregulation of VEGF and collagen synthesis. The objective of this study was to evaluate the effect of HMSCs-CM on VEGF and collagen concentrations in rats with incised wounds. The methods of this study were an experimental animal study with post-test only control group design was performed involving 24 Wistar rats. The rats were randomized into four groups consisting of sham, control and two treatment groups (gel of HMSCs-CM at doses of 200 μL and 400 μL). The VEGF levels and collagen density were analyses using ELISA assay and Masson-trichome specific staining, respectively. One-way ANOVA and Post Hoc LSD were used to analyses the data. The results of this study showed that a VEGF levels was significant increased on day 6 with doses-dependent manner. Interestingly, the VEGF levels gradual decrease on day 9. In addition, the decreased of VEGF levels on day 9 in this study in line with our findings in which we found there was a trend in the decreased of collagen density, it indicated the completion of remodeling phase and there has been an acceleration in wound healing. This study demonstrated that HMSCs-CM were able to regulate VEGF levels and collagen synthesis in accelerate wound healing. The role of HMSCs-CM stimulate cutaneous wound healing should be clarified further.Keywords: hypoxic conditional medium of mesenchymal stem cells (HMSCs-CM), vascular endothelial growth factor, collagen synthesis, paracrine factors
Hypoxia-Exosome Mesenchymal Stem Cells Therapy Reduces Interleukin-6 Levels and CD86 Expression Isfandiari, Adelia Bayu; Putra, Agung; Setiawan, Eko
Molecular and Cellular Biomedical Sciences Vol 9, No 3 (2025)
Publisher : Cell and BioPharmaceutical Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21705/mcbs.v9i3.660

Abstract

Background: UVB exposure activates type 1 macrophages (CD86) and increases IL-6, both contributing to collagen loss. exosome hypoxia mesenchymal stem cells (EH-MSC) has anti-inflammatory properties, suggesting its potential role in modulating CD86 activation and IL-6 secretion. This study examines the effects of EH-MSC injections on CD86 and IL-6 levels in UVB-exposed skin with collagen loss.Materials and methods: Experimental research post-test only control group design was conducted with 30 male Wistar rats (Rattus norvegicus, strain: Wistar Han) divided into five groups. G1: healthy rats, G2: UVB-exposed with a subcutaneous injection of 0.9% NaCl, G3: UVB-exposed with Hyaluronic Acid, and G4 & G5: UVB-exposed with EH-MSC injections of 200 µL and 300 µL, respectively. IL-6 and CD86 levels were analysed using ELISA and qRT-PCR at 14 days post-treatment continue with statistical analysis.Results: IL-6 analysis showed that levels in G4 (107.70±47.86 pg/mL) and G5 (58.68±25.37 pg/mL) were notably lower than in G2 and G3 (p<0.05). Compared to G1 (32.28±14.65 pg/mL), G4 exhibited a statistically distinct increase (p<0.05), whereas G5 showed no significant difference (p>0.05). Meanwhile, CD86 data analysis showed that G4 (0.53±0.14 pg/mL) were lower than in G1 (1.03±0.01 pg/mL) and G2 (1.47±0.43 pg/mL), but not significantly different from G3 (0.87±0.1 pg/mL) and G5 (0.36±0.08 pg/mL). Similarly, CD86 expression in G5 decreased relative to G1 and G2 (p<0.05) but remained similar to G3 and G4.Conclusion: EH-MSC injections (200 µL and 300 µL) significantly reduced IL-6 and CD86 levels in collagen loss rats, supporting its potential as a therapeutic approach for UVB-induced skin damage.Keywords: collagen loss, CD86, EH-MSC, IL-6, UVB
Exosome Therapy from Hypoxia-treated Mesenchymal Stem Cells Reduces TNF-α and Increases VEGF Levels in Fluconazole-Induced Alopecia Model Sulistami, Siska Marlina; Mulyani, Sri Priyantini; Putra, Agung; Setiawan, Eko
Molecular and Cellular Biomedical Sciences Vol 9, No 3 (2025)
Publisher : Cell and BioPharmaceutical Institute

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.21705/mcbs.v9i3.642

Abstract

Background: Alopecia is a condition with partial or complete hair loss, leading to psychological distress. Current treatments, such as minoxidil and finasteride, have limited efficacyand side effects. Recent studies suggest that mesenchymal stem cells (MSCs)-derived exosomes offer regenerative potential by modulating inflammation and enhancing hair follicle regeneration, though optimal dosage remains unclear. Tumor necrosis factor-alpha (TNF-α)  inhibits hair follicle growth, while vascular endothelial growth factor (VEGF) promotes hair regrowth. This study evaluates  exosome therapy from hypoxia (Hypo-Exo)-treated MSCs in modulating TNF-α and VEGF in a fluconazole-induced alopecia-like model..Materials and methods: An experimental post-test only control group design was used with 30 male Wistar rats, divided into five groups:  Healthy group, 0.9% NaCl-treated group, 5% Minoxidil-treated group, 100 μg/mL Hypo-Exo MSCs-treated group, and 200 μg/mL Hypo-Exo MSCs-treated group. TNF-α and VEGF levels were analyzed using ELISA on day 14 post-treatment. Results: The highest TNF-α level was found in the 0.9% NaCl-treated group (307.46 ± 20.68 pg/mL) and significantly reduced (p<0.05) in 100 μg/mL Hypo-Exo MSCs-treated group (65.38±15.05 pg/mL) and 200 μg/mL Hypo-Exo MSCs-treated group (37.16±7.14 pg/mL). VEGF levels were the highest in the 200 μg/mL Hypo-Exo MSCs-treated group (189.11±9.75 pg/mL) and 100 μg/mL Hypo-Exo MSCs-treated group (158.50±5.33 pg/mL), compared to the 0.9% NaCl-treated group (69.60±15.39 pg/mL). Conclusion: Hypo-Exo MSCs significantly reduced TNF-α and increased VEGF levels, supporting their potential as a novel regenerative therapy for alopecia. Keywords: alopecia, TNF-α, VEGF, exosome, hypoxia, mesenchymal stem cells 
Co-Authors Agus Widyatmoko, Agus Alif, Iffan Amalina, Nur D. Amalina, Nur Dina Amin, Mustafa M. Andavania, Sheila Jessica Andreas, Yana Antari, Arini Dewi Ati, Sri Umi Azizah Retno Kustiyah Cahyani, Dini Cahyani, Elvana Cahyono, Erwin Budi Candra Satria Irawan, Risky Catharina Suharti Chodidjah Chodidjah Darlan, Dewi M. Daulay, Rini S. Delfitri Munir Dirja, Bayu T. Dirja, Bayu Tirta Djannah, Durrotul Edi Dharmana Eko Setiawan Eko Setiawan Erna Mutiara Evita Mayasari Fahreza, Rakha Fatmawati, Dian Fikriya Novita Sari Firman Alamsyah Fristiani, Yeni Ghaisani, Shabrina Syifa Ginting, Andi R. Hadi Sarosa Haitamy, Mohammad Nurrizki Handoyo, Frigi Eko Hariani, Nova Putri Hasanal, Ihdina Hanifa Hasannuri, Tarrayuana Rhamadia HS, Zakariya Husain, Sofian A. Hutabarat, Nenny Lynda Caroline Hutagalung, Ananta Ibrahim, Sugeng Ignatius Riwanto, Ignatius Ikawati, Muthi' Intan, Yulice Soraya Nur Irawan Mangunatmadja Irawan, Risky Chandra Irawan, Risky Chandra Satria Isfandiari, Adelia Bayu Iskandar Japardi Kuntardjo, Novalia Lusiana Lusiana Mardiana, Ana Maryanti Maryanti Minidian Fasitasari Muchaeroni, Isa Anshori Muhar, Adi Muradi Noka, Isara Abda Nur Anna Chalimah Sadyah Nurul Hidayah NURUL HIDAYAH Pasongka, Zenitalia Pramukarso, Dodik Tugasworo Pramukarso Prasetio, Ardi Prasetyowati Subchan, Prasetyowati Prawitasari, Salindri Purwaningsih, Hesti Rarastoeti Pratiwi Riris Istighfari Jenie Rozi, Muhammad F. Rusda, Muhammad Sa’dyah, Nur Anna C Sari, Shintia Devi Arum Selamat Budijitno Setiawan , Eko Setyo Trisnadi Setyo, Trisnadi Sisca Silvana, Sisca Sitompul, Faya Nuralda Sri Priyantini Sri Priyantini Mulyani Sri Sofyani, Sri Subchan, Prastyowati Sulistami, Siska Marlina Sulistyo, Sona Sunarto, Hadi Sutrisman, Intan Permatasari Suwandi Ng Syamsunarno, Mas Rizky Syamsunarno, Mas Rizky A.A Taskworo, Dodik Taufiq R Nasihun, Taufiq R Titiek Sumarawati Tjipta, Arya Utami, Wulan Dyah Yasmine Azzahara, Salma Yayun Siti Rochmah Yustianingsih, Vivi