Molecular and Cellular Biomedical Sciences (E-Journal)
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The μDrop Method Enhances Melanin Content Measurement in the in vitro Melanogenesis Model Using B16F10 Cell Line
No full textBackground: The B16F10 cell line is a cell frequently used in melanin content assays. However, reports on cell models using B16F10 are limited, particularly as the robust model cell in the Indonesian cosmetics industry. We found measuring melanin content using microplate spectrophotometry to be challenging, so this research was conducted to develop a method using μDrop spectrophotometry.Materials and methods: In this in vitro study, the B16F10 melanoma cell line was cultured in Roswell Park Memorial Institute (RPMI) medium containing 5% fetal bovine serum (FBS). The cells were categorized into control, stimulated, and treated groups. Melanogenesis stimulation was achieved using 1μM α-melanocyte-stimulating hormone (α-MSH), while inhibition using 800 μg/ml kojic acid. After treatment, the cells were incubated for 48 hours. Their melanin content was then measured using an ELISA reader with a μDrop method and compared with the microplate method. Statistical analysis used a one-way ANOVA test with Turkey’s Post Hoc analysis.Results: The μDrop method increased the melanin signal into the linear range of machine readings, while the signals from the microplate method fell far below this range. The B16F10 melanoma cell lines stimulated by α-MSH exhibited increased melanin production compared with the control group, while kojic acid treatment significantly reduced (p<0.05) melanin content in the stimulated group.Conclusion: The μDrop method significantly outperformed the microplate method in measuring melanin content within melanogenesis cell models, offering enhanced accuracy and particularly excelling at quantifying low content of melanin. Keywords: μDrop, microplate, melanin, melanogenesis, B16F10 cell line, RPM
Mesenchymal Stem Cell-Derived Exosomes Enhance FGF-1 and SDF-1 Expression in Rats with Second Degree Burns
No full textBackground: 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-
Endophytic Bacteria in Acalypha indica L. Leaves and Their Antimicrobial Activity Against Staphylococcus aureus and Candida albicans
No full textBackground: The anting-anting plant (Acalypha indica L.) is used in herbal medicine in the treatment of various diseases. The leaf extract of this plant is known for its antimicrobial activity, but the antimicrobial properties of the endophytic bacteria within its leaves have never been reported. This research aims to determine the antimicrobial activity of endophytic bacteria from the leaves of the anting-anting plant. Materials and methods: The isolation of endophytic bacteria was performed using the spread plate method on nutrient agar (NA) media. Following isolation, the bacterial isolates were characterized through macroscopic and microscopic examination, as well as biochemical tests, which included indole production, hydrogen sulfide (H2S) production, motility, Simmons citrate utilization, methyl red-Voges-Proskauer (MR-VP) test, catalase test, and triple sugar iron agar (TSIA) test. Identification of the bacterial isolates was conducted according to Bergey's Manual of Systematic Bacteriology. Additionally, the antimicrobial activity of the isolates was assessed using the diffusion methodResults: Fourteen isolates of anting-anting leaf endophytic bacteria were obtained (coded as BEDA 1 to BEDA 14). The BEDA 5 isolate exhibited the largest inhibitory zone diameter against Staphylococcus aureus (31.48 mm), while BEDA 9 showed a significant inhibitory zone diameter against Candida albicans (17.84 mm). Conclusion: The two isolates (BEDA 5 and BEDA 9) exhibited significant antimicrobial activity, indicating their potential as promising candidates for alternative antimicrobial agents. These results suggest that endophytic bacteria from Acalypha indica may play an essential role in combating antibiotic resistance and in the development of new therapeutic strategies.Keywords: endophytic bacteria, characterization, antimicrobial activity, Acalypha indic
Vitamin D Deficiency is Associated with Hypocalcemia in Preterm Infants
No full textBackground: Vitamin D deficiency results in various problems, like rickets, osteomalacia, heart problems, cancer, diabetes, and autoimmune diseases. Hypocalcemia is a common disorder among preterm infants, indicating vitamin D deficiency. This study was conducted to analyze the association of vitamin D deficiency with hypocalcemia in preterm infants.Materials and methods: A cross-sectional study was performed in preterm infants born in our hospital from December 2022 to May 2023. Venous blood was collected within the first 24 hours to assess vitamin D and calcium levels. Chi-square test and logistic regression analysis were used to assess the association of gestational age, sex, birth weight, and vitamin D with the incidence of hypocalcemia. The significance was determined with p<0.05.Results: There were 40 preterm newborns, comprising 37.5% moderately preterm, 20% very preterm, and 42.5% extremely preterm. Most subjects were female (52.5%). Low birth weight, very low birth weight, and extremely low birth weight occurred in 55%, 27.5%, and 17.5%, respectively. Vitamin D insufficiency and deficiency were observed in 20% and 80% subjects, respectively. Most subjects had hypocalcemia (62.5%). Chi-square test obtained a significant association of vitamin D deficiency with hypocalcemia (p=0.029).Conclusion: Vitamin D deficiency is significantly associated with the incidence of hypocalcemia in preterm infants.Keywords: Vitamin D, hypocalcemia, preterm neonate
Angiotensin-Converting Enzyme Genetic Polymorphism rs4343 as Risk of Diabetic Nephropathy in Jambi-Malay Population
No full textBackground: Diabetic nephropathy (DN) is one of the frequent complications of type II diabetes mellitus (T2DM) in Jambi province. Controlling blood glucose and blood pressure does not guarantee DN prevention, since genetic factors may also contribute to this disease. Multi-ethnic studies showed that one of the strongest genetic factors associated with DN was single nucleotide polymorphism rs4343 of angiotensin-converting enzyme (ACE) gene. Study regarding phenotype-genotype association of ACE rs4343 and DN has not yet been performed in Jambi Province, which is dominated by Malay ethnicity. This study was conducted to reveal the association between ACE rs4343 and the risk of DN in the Jambi-Malay population.Materials and methods: This was a cross-sectional study involving 75 subjects (44 with DN and 31 without DN) who suffered from T2DM and hypertension. DN was defined as albumin to creatinine ratio (ACR) ≥30 mg/g. Genotyping was performed with one-step tetra amplification refractory mutation system-polymerase chain reaction (PCR) using specific primer for ACE rs4343. Bivariate and multivariate analyses were performed to analyze the genetic risk for DN.Results: The bivariate analysis showed the proportion of DN subjects was higher than non-DN within the AG genotype (11:1) than within the AA (33:30) genotype. This difference was statistically significant (p=0.012; OR (95% CI): 10.00 (1.22-82.15)). Multivariate analysis showed that AG genotype (p=0.047; OR (95% CI): 10.04 (1.03-97.31)) and uncontrolled blood pressure (p=0.001; OR (95% CI): 6.72 (2.08-21.71)) were the risk factors of DN in the Jambi-Malay population.Conclusion: Polymorphism of ACE rs4343 is a risk factor of DN in the Jambi-Malay Population.Keywords: rs4343, angiotensin-converting enzyme gene, diabetic nephropathy, Malay, Jamb
Mitochondrial Dynamics: An Attractive Therapeutic Target for Ischemia-Reperfusion Injury in the Heart
No full textMyocardial infarction is one of the leading causes of death worldwide. Current treatments do not compensate for the loss of cardiomyocytes, thus progression to heart failure is often inevitable. In myocardial infarction, the occlusion of coronary arteries and sudden restoration of blood flow give rise to ischemia-reperfusion injury, which leads to cardiomyocyte death. Mitochondria are not only involved in the bioenergetic aspect of the heart but also play a pivotal role in cell death during ischemia-reperfusion injury. Their morphology dynamically changes via fusion and fission in a balanced manner to maintain cellular health. However, ischemia-reperfusion injury triggers excessive mitochondrial fission, which is pathological to the myocardium. This review article discusses the association between myocardial ischemia-reperfusion injury and mitochondrial dynamics, serving as a rationale for a novel therapeutic strategy for myocardial infarction. Strategic modulation of mitochondrial dynamics under this pathological setting has been shown to be effective for cardioprotection. Increasing mitochondrial fusion or reducing excessive mitochondrial fission in the myocardial tissue could prevent cardiomyocyte death, thereby reducing infarct size. Proof-of-concept studies have utilized small molecules and peptides to implement this strategy into in vivo myocardial ischemia-reperfusion injury models. However, there remains a need to address the issues of specificity, bioavailability, and potency of these pharmacological agents before future application in cardiovascular therapeutics. Nevertheless, there has been growing interest in this therapeutic strategy in recent years, rendering it an attractive approach for ischemia-reperfusion injury in the heart.Keywords: mitochondria, heart, ischemia-reperfusion, cardioprotectio
The Prospect of Probiotics to Treat Metabolic Syndrome
No full textMetabolic syndrome remains as a major health problem in the world today, with a prevalence of 23.4% in people aged 26-82 years. A high-fat, high-carbohydrate diet and lack of physical activity are considered as one of the triggers for metabolic syndrome. Dysbiosis is a condition where there is an imbalance between pathogenic and non-pathogenic bacteria in the human gut. Currently, an association has been found between dysbiosis and metabolic syndrome. Dysbiosis causes the generation of fermentation products in the form of active metabolites that can modulate hormones and other physiological functions. In metabolic syndrome, low-grade inflammation, energy metabolism, and disruption of the gut brain axis are thought to be the main mechanisms of the development of metabolic syndrome due to dysbiosis. Probiotics may be a promising therapeutic agent in the treatment of metabolic syndrome, by improving dysbiosis to eubiosis. Based on previously conducted clinical trials, it is currently known that probiotics can improve lipid profiles, fasting blood glucose, homeostatic model assessment for insulin resistance (HOMA-IR), vascular cell adhesion molecule 1 (VCAM-1), glucagon-like peptide-1 (GLP-1), peptide YY (PYY), and body mass index (BMI). However, the results found are still varied, so a dose ranging study is needed to determine the duration, bacterial composition and dose of probiotics as a therapeutic agent for metabolic syndrome. Keywords: insulin resistance, dysbiosis, gut-brain axi
T Allele of FOXO3 rs2802292 Increases CCL2 Concentration and Slightly Decreases TGF-β Concentration in Indonesian Elderly
No full textBackground: Cellular senescence and the senescence-associated secretory phenotype (SASP) are pivotal factors influencing aging and age-related diseases. SASP secretes cytokines, chemokines, metalloproteinases, and growth factors that cause chronic inflammation. C-C ligand 2 (CCL2) and transforming growth factor-beta (TGF-β) are SASP markers secreted by senescent cells. This study investigated the relationship between the FOXO3 variant rs2802292 and SASP markers, focusing on CCL2 and TGF-β.Materials and methods: A cross-sectional study involving 72 elderly individuals from Jakarta was conducted. A sandwich enzyme-linked immunosorbent assay (ELISA)was used to quantify CCL2 and TGF-β concentrations. Random blood glucose, blood pressure, and FOXO3 rs2802292 genotyping data were obtained from a previous study. Differences in CCL2 and TGF-β concentrations between genotype groups were analyzed using one-way ANOVA and the Kruskal-Wallis test. Meanwhile, differences in CCL2 and TGF-β concentrations between allele groups were analyzed using the Mann-Whitney test.Results: The CCL2 and TGF-β concentrations of the subjects were 66.5 (10.58-190.9) pg/mL and 6,319 (2,379-13,846) pg/mL, respectively. There were significant differences in CCL2 concentrations among the FOXO3 rs2802292 genotypes (p=0.041). However, there were no significant differences in TGF-β concentrations among FOXO3 rs2802292 genotypes (p=0.955). Subjects with the G allele had significantly lower CCL2 concentrations compared with those with the T allele (p=0.033). TGF-β concentrations did not significantly differ between G and T alleles (p=0.771).Conclusion: CCL2 concentrations are associated with the FOXO3 variant rs2802292 in the elderly population. The T allele of FOXO3 rs2802292 increased CCL2 concentration and slightly decreased TGF-β concentration in elderly individuals.Keywords: aging, SASP, CCL2, TGF-β, SNP, FOXO3, rs280229
Potency of Peripheral Blood- and Umbilical Cord Blood-derived Dendritic Cells and Their Secretomes as Vaccines for Cancer
No full textDendritic cell (DC) vaccines, as immunotherapy agents, can gather up and transport cancer-related antigens to T lymphocytes, activating anti-tumor effector responses. After being activated by DC, cytotoxic T lymphocyte cells (CTL) will secrete the cytolytic granzyme B that can effectively induce rapid apoptosis of target cells. On the other hand, DC also secrete several cytokines and a large number of exosomes, which together operate as a whole antigen-presenting entity. The efficacy of the vaccine’s treatment may be affected by the sources used for DC vaccines. Umbilical cord blood (UCB) from healthy donors can be employed when autologous cancer patient’s peripheral blood (PB) cannot be used as a source for isolating DC due to genetic abnormalities. Comparing UCB to other sources, there is a painless method of collecting sources as opposed to PB, which necessitates a venipuncture or leukapheresis procedure to isolate the blood. Many studies related to the use of PB-DC have been carried out, but research on potential comparisons between PB-DC and UCB-DC is still very limited. In this review, the potential of PB- and UCB-derived DC and their secretomes for cancer will be discussed.Keywords: dendritic cells, vaccines, umbilical cord blood, peripheral bloo
Evasion of the Immune System by Glioblastoma Multiforme: An Obstacle to Achieving Effective Therapies
No full textGlioblastoma multiforme (GBM), a highly aggressive and malignant form of brain cancer, continues to pose a significant challenge in the field of oncology. Despite ongoing advancements in treatment strategies, the prognosis for GBM patients remains grim, with a 5-year survival rate hovering around 5%. The management of GBM involves multiple therapeutic approaches, including immunotherapy, but optimal treatment outcomes in terms of overcoming tumor recurrence and resistance have not been achieved. A key factor contributing to therapy resistance and the progression of GBM is the tumor's ability to evade the immune system, referred to as immune escape from cancer. This phenomenon reflects the tumor cells' efforts to adapt and survive the body's immune response. The release and expression of molecules like TGF-ß, IL-10, PD-L1, and NKG2DL by GBM cells impact the activation, recognition, and elimination of tumor cells by the immune system. Additionally, the involvement of cells such as MDSCs, Tregs, and TAMs plays a role in inhibiting the immune system's function, thereby promoting the development of GBM cells. A better comprehension of GBM's immune escape, supported by technological advances, will significantly aid in the future management of GBM patients' treatment.Keywords: glioblastoma multiforme, GBM, cancer immunity, immune evasion, immune escape, immunotherap