Molecular and Cellular Biomedical Sciences (E-Journal)
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n-Hexane Fraction of Cucumis melo L. Cultivar Gama Melon Parfum: An in vitro Study in MCF7 and T47D Cells Line
No full textBackground: Cucumis melo a melon species, typically has a sweet taste. Some cultivars are known for their distinctive bitter flesh due to its higher levels of cucurbitacin. Cucurbitacin is semipolar compound that has anticancer properties. However, the anticancer effects of cucurbitacin from gama melon parfum (GMP) have not been widely studied. The use of n-Hexane as a non-polar solvent in GMP melon fractionation is to dissolve the non-polar parts of the plant. However, Cucurbitacin was found in the n-hexane fraction of Cucurbita pepo L. Therefore, this study will investigate the presence of Cucurbitacin in the n-Hexane fraction and its effects on breast cancer cells T47D and MCF7.Materials and methods: Dry simplicia of GMP melon fruit were macerated using methanol and fractionated using n-hexane. The presence of cucurbitacin was detected using the high-performance liquid chromatography (HPLC) method. Cell cytotoxicity tests were assessed using the MTT assay, with concentrations of 7.8125, 15.625, 31.25, 62.5, and 125 µg/mLResults: Cucurbitacin compounds were detected in the n-hexane fraction at a concentration of 7.6 µg/mL per 10 mg of n-hexane fraction. MCF7 cell viability was lower than that of T47D cells across all concentrations tested. MCF7 cell viability was below 50% at a concentration of 62.5 µg/mL. In contrast, T47D cell viability remained at 100% even at the highest concentration of 125 µg/mL. The IC50 value of MCF7 cells was 43.5 µg/mL.Conclusion: The cucurbitacin content in the n-Hexane fraction was 7.6 µg/mL per 10mg fraction. At this concentration, it moderately inhibits the proliferation of MCF7 cells.Keywords: gama melon parfum, cucurbitacin, HPLC, T47D, MCF
Apoptotic Effects Sulfated Polysaccharides of Caulerpa racemosa Extract on Colorectal Cancer Cells through Caspase-3
No full textAbstractBackground: Colorectal cancer originates from progressive genetic alterations in colorectal epithelial cells. While current therapies (surgery, radiotherapy, and chemotherapy) remain cornerstone treatments, chemotherapy often induces systemic toxicity, adverse effects, and acquired resistance. Sulfated polysaccharides (SPs) from the green alga Caulerpa racemosa demonstrate higher sulfate content than red algal derivatives, correlating with enhanced bioactivity. Despite their potential, SPs from green algae remain understudied compared to brown and red algal counterparts. This study evaluated the anticancer potential of C. racemosa SPs against colorectal cancer through viability inhibition and apoptosis induction.Materials and Methods: SPs were extracted via Microwave-Assisted Extraction (MAE) and characterized using iodine testing and FTIR spectroscopy. Cytotoxicity was evaluated in WiDr colorectal cancer cells using MTT assay after 24-hour exposure. Apoptotic mechanisms were investigated through in silico molecular docking targeting Caspase-3 activation.Results: SPs were confirmed by a blue color change and FTIR absorption at 1232 cm-1. At 100 µg/mL, low toxicity was observed based on abundant formazan crystals. Concentrations of 200–400 µg/mL showed predominant viable cells, whereas 500 µg/mL caused significant growth inhibition and cell death. In silico analysis demonstrated that SPs may induce apoptosis by Caspase-3 activation.Conclusion: SPs of C. racemosa inhibit colorectal cancer cell viability at a concentration of 500 µg/mL and may induce apoptosis via Caspase-3 activation.Keywords: apoptosis, Caulerpa racemosa, colorectal cancer, sulfated polysaccharide
Elevated Procalcitonin Levels in Pediatric Severe Bacterial Pneumonia Caused by Klebsiella pneumoniae
No full textBackground: Klebsiella pneumoniae is a major pathogen in pediatric pneumonia. Procalcitonin (PCT) distinguishes between Gram positive and Gram negative infections but lacks data on K. pneumoniae infection's relationship with PCT levels in children. Therefore, this study was conducted to investigate the serum PCT levels in children with K. pneumoniae infection.Materials and methods: A cross-sectional study was conducted on 61 pediatric subjects with the age of 2-59 months with severe bacterial pneumonia. Blood and sputum samples were collected and analyzed for PCT and cultured for 24 hours. PCT levels and K. pneumoniae infection were statistical analyzed with chi-square and logistic regression. Receiver operating characteristic (ROC) analysis was used to determine the PCT cut-off specific to K. pneumoniae. Results: K. pneumoniae was identified in 33%. Median PCT levels were significantly higher in the K. pneumoniae group (p<0.05). A PCT cut-off of 0.725 ng/mL yielded 70% sensitivity, 70.7% specificity, negative predictive value (NPV) of 82.9%, and area under the receiver operating characteristic curve (AUROC) of 0.74. Elevated PCT was significantly associated with K. pneumoniae infection with Odds ratio (OR) of 12.08, 95% Confidence Interval (CI): 2.54-57.36; p=0.002).Conclusion: Along with K. pneumoniae infection, serum PCT levels was elevated, supporting its potential as a biomarker for early diagnosis. Keywords: K. pneumoniae, procalcitonin, bacterial pneumonia, pediatri
Exosomes from Hypoxic Mesenchymal Stem Cells Enhance TGF-β Expression and Promote Collagen Regeneration in Wistar Rats with Collagen Loss
No full textBackground: Ultraviolet B (UVB) exposure triggers reactive oxygen species (ROS) formation, inhibits procollagen synthesis via the transforming growth factor-beta (TGF-β)/Smad pathway, and leads to collagen loss. Exosomes derived from hypoxia-conditioned mesenchymal stem cells (EH-MSCs) are more effective than those from normoxic MSCs due to their higher content of miRNAs and cytokines with anti-inflammatory and regenerative properties. This study aims to evaluate the effect of EH-MSC injection on TGF- β levels and collagen density in Wistar male rats with UVB-induced collagen loss. Materials and methods: This in vivo experimental study used a post-test only control group design with 30 rats, divided into five groups: Healthy group, 0.9% NaCl-treated group, hyaluronic acid (HA)-treated group, 200μL EH-MSC-treated group, and 300μL EH-MSC-treated group. TGF-β levels were analyzed using ELISA, while collagen density was assessed with Masson trichrome staining.Results: Highest mean TGF-β levels were observed in the 300μL EH-MSC-treated group (155.56±99.84 pg/mL), while the highest collagen density was found in the Healthy group (23.07±1.81 pg/mL). Mann-Whitney test indicated a significant increase (p=0.008) in TGF-β levels in treatment groups compared to the 0.9% NaCl-treated group. Post Hoc LSD Tamhane test for collagen density also showed a significant increase (p=0.000) in treatment groups compared to the 0.9% NaCl-treated group. Conclusion: EH-MSC injection significantly increased TGF-β levels and collagen density, indicating its potential for promoting skin repair in UVB-induced collagen loss.Keywords: EH-MSC, collagen loss, TGF-β, collagen density
UC-MSCs Secretome Induces Proliferation of CD4+ T Cells, CD8+ T Cells, NK Cells, and Increases sPD-1 Levels in Severe COVID-19’s Whole Blood
No full textBackground: Clinical features of severe coronavirus disease 2019 (COVID-19) predominantly include respiratory symptoms and exacerbated multi-organ complications, especially in patients with comorbidities. Cellular immunity, including lymphocytes, is a critical factor in combating SARS-CoV-2 infection. However, immune dysregulation occurs in severe COVID-19 patients, characterized by cytokine storm and lymphopenia. The effectiveness of mesenchymal stem cell (MSC) therapies for COVID-19 is being assessed. The secretome released by MSC functions similarly to the cells themselves as an immunomodulator, offering potential advantages in terms of safety and cost-effectiveness. This study was conducted to assess the effect of umbilical cord MSC-derived (UC-MSC) secretome treatment on lymphocyte count and soluble programmed cell death-1 (sPD-1) levels in severe COVID-19 patient's whole blood.Materials and methods: Twelve whole blood samples from healthy individuals and severe COVID-19 patients were analyzed for lymphocyte count and functional activation using flow cytometry, along with sPD-1 level measurement in pre-treatment and post-secretome conditions.Results: The lymphocyte count in severe COVID-19 patients was significantly decreased, particularly for T cells and NK cells, indicating lymphopenia. Following secretome treatment, CD4+ T cell counts significantly increased compared to pre-treatment, although this change was not significant in the negative control group. Additionally, there was a minimal reduction in B cell count and an increase in sPD-1 levels. Elevated sPD-1 may alleviate T cell exhaustion by interfering with PD-1 binding to programmed death-ligand 1 (PD-L1).Conclusion: Administration of UC-MSC secretome to the whole blood of severe COVID-19 patients suggested immune improvement, with significant increases in CD4+ T cell counts, enhanced B cell survival, and elevated sPD-1 levels. Keywords: COVID-19, cellular immunity, lymphocytes, secretome, MS
Exosome Therapy from Hypoxia-treated Mesenchymal Stem Cells Reduces TNF-α and Increases VEGF Levels in Fluconazole-Induced Alopecia Model
No full textBackground: 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
Hypoxia-Induced Mesenchymal Stem Cell Exosomes Modulate Protein Kinase A and VEGFR Expression in Ultraviolet B-Induced Hyperpigmentation in Mice
No full textBackground: 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, melani
SARM Rad140 Increases Osteoblasts, Muscle Fiber Size, Myonuclei, and Reduces Osteoclasts in Orchidectomized Wistar Rats
No full textBackground: Orchidectomy is a surgical androgen deprivation therapy (ADT) for prostate cancer patients to achieve castrate testosterone levels. Selective androgen receptor modulators (SARMs) are used to mitigate the adverse effects of ADT, including elevated risk of osteoporosis, and reduced skeletal muscle mass. Rad140 is a novel SARMs that has high affinity for the androgen receptors. This study was conducted to determine the effects of SARM Rad140 on the number of osteoblasts, osteoclasts, muscle fiber cross-sectional area (CSA), muscle size, and number of myonuclei in rats underwent orchidectomy.Materials and Methods: An experimental study was conducted using a randomized post-test only control group design. Following orchidectomy, SARM Rad140 was administered orally for six weeks at various doses. Osteoblasts, osteoclasts, muscle fiber CSA, muscle size, and number of myonuclei were measured. Quantitative analysis was performed using one-way ANOVA.Results: There were significant differences in the effects of SARM Rad140 at different doses on osteoblast and osteoclast cells. At higher doses, the osteoblast cell counts in rats tended to increase, while the osteoclast counts tended to decline. The treatment group also showed significant results in the CSA of the gastrocnemius muscle fibers, as well as in the number of myonuclei of the gastrocnemius muscle.Conclusion: SARM Rad140 significantly increased the number of osteoblasts, muscle fiber CSA, and gastrocnemius muscle myonuclei, while decreasing osteoclasts. SARM Rad140 is a promising therapy for osteoporosis and muscle weakening due to ADT.Keywords: SARM Rad140, orchidectomy, osteoblasts, osteoclasts, muscle fiber cross sectional area, myonucle
Increased expression of pap and sfa Genes in Biofilm-Forming Uropathogenic Escherichia coli Associated with Urinary Tract Infections
No full textBackground: Urinary tract infections cover a broad spectrum of infectious syndromes and affect the urinary tract from the urethra to the kidneys. Generally caused by uropathogenic Escherichia coli (UPEC), and their pathogenesis is greatly influenced by biofilm formation, which results in persistent and recurrent infections. UPEC uses filamentous adhesive structures such as pili or fimbriae, pyelonephritis-associated pili, and S fimbriae, which are regulated by the pap and sfa operons, respectively. The purpose of the study was to detect the effects of two virulence genes, (pap 7 and sfa 9) on biofilm-forming UPEC associated with urinary tract infections.Materials and methods: A total of 123 UPEC isolates were collected from clinical microbiology laboratory section of a general hospital in Surabaya, Indonesia. Urine samples yielded UPEC with significant counts (≥105 CFU/ml), and the biofilm development was analyzed using the Congo red agar method. The presence of pap 7 and sfa 9 genes in the isolates was determined using PCR assay. Results: Among the 123 UPEC isolates, 66 isolates were able to form biofilms, as determined using the Congo red agar (CRA) method. Biofilm-forming UPEC isolates exhibited a high positivity frequency for the pap 7 gene (65.85%), while the positivity frequency for the sfa 9 gene was significantly lower (14.63%). Conclusion: An increase in th expression of pap 7 and sfa 9 are is associated with the ability to form biofilms, which could serve as a diagnostic marker for biofilm formation potential vaccine target.Keywords: biofilm, pap, sfa, uropathogenic, Escherichia coli, UT
Hypoxia-Exosome Mesenchymal Stem Cells Therapy Reduces Interleukin-6 Levels and CD86 Expression
No full textBackground: 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 (p0.05). 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, UV