The Indonesian Biomedical Journal

126-34

Authors: Manchi Vathsalya, Shricharith Shetty, Nikhil Shetty, Jayaprakasha Shetty, Veena Shetty, Sunil Kumar Yeshwanth, Prakash Patil, Mohana Kumar Basavarajappa

BACKGROUND: Melanocyte stem cells (MelSCs) residing in the hair follicle bulge act as melanocyte reservoir for skin and hair, and may serve as an autologous source for treating vitiligo. Therefore, the study aimed to evaluate the in vitro differentiation ability of MelSCs derived from vitiligo patients into melanin-producing melanocytes for potential cellular therapy. METHODS: MelSCs from the vitiliginous (V-MelSCs), non-vitiliginous (NV-MelSCs) regions of vitiligo patients, as well as from control subjects (C-MelSCs) were established for evaluating their differentiation potential into melanin producing cells. The differentiation abilities were compared at the cellular and molecular levels. MelSCs were differentiated in vitro into induced-melanocytes (iMCs) by supplementing the culture medium with melanogenic factors. iMCs were analyzed by quantitative polymerase chain reaction (qPCR) for the expression of key melanogenic markers, including tyrosinase (TYR), tyrosinase-related protein 1 (TYRP1), dopachrome tautomerase (DCT), microphthalmia-associated transcription factor (MITF), c-KIT and S100. iMCs were also stained with L-DOPA to assess TYR activity in cells. Intracellular melanin content in iMCs was evaluated and compared among the vitiligo and control groups. RESULTS: MelSCs induced into iMCs displayed morphological changes with longer dendrites and prominent nuclei. iMCs stained positive for L-DOPA with an average intracellular melanin content of 30 pg/cell. iMCs expressed key melanogenic genes and the relative expression did not differ significantly among the groups. CONCLUSION: NV-MelSCs were unaltered by disease pathogenesis and capable of differentiating into melanocytes compared to V-MelSCs. Hence, these cells might offer a reliable source of melanocytes for vitiligo repigmentation therapy by autologous cellular transplantation.

144-51

Authors: Jusniar Patandung, Ika Yustisia, Rosdiana Natzir, Gita Vita Soraya, Lia Hafiyani, Sulfahri Sulfahri

BACKGROUND: Red macroalgae, such as Eucheuma spinosum, have been found to have potential benefits for bone health due to their unique bioactive compounds, including proteins, polyphenols, polysaccharides, vitamins, and minerals. Therefore, this study was conducted to evaluate the benefits of powdered E. spinosum supplementation in osteoporosis-induced mice. METHODS: Thirty middle-aged mice were divided into 6 groups, namely: healthy control group (HC), negative control group (NC), positive control group (PC), and treatment groups supplemented with 1.25 mg/gBW (T1), 2.5 mg/gBW (T2), and 5 mg/gBW (T3) of E. spinosum powder for twenty days. Mice in NC, PC, T1, T2, and T3 groups were induced with 0.0029 mg/gBW of dexamethasone for 30 days to create osteoporosis mice models. Alkaline phosphatase (ALP) levels were measured by colorimetric methods before and after the intervention. Bone structures were evaluated using X-ray images and histological examination. RESULTS: After the intervention, PC, T1, T2, and T3 groups showed a significant decrease (p≤0.01) in serum ALP levels compared to the NC group, which experienced an increase in ALP levels. The X-ray images revealed that the PC, T1, T2, and T3 showed radiopaque bone density. For bone histology, PC, T2, and T3 showed an improvement with thickened and intact trabeculae, but T1 still had visible osteoporosis cavities. CONCLUSION: Supplementation of 2.5 and 5 mg/gBW of E. spinosum powder were able to improve bone density as well bone histology. Therefore, E. spinosum powder supplementation might potentially improve bone structure in osteoporosis.

162-71

Authors: Deasy Fetarayani, Gatot Soegiarto, Ingrid Suryanti Surono, Anang Endaryanto, Alpha Fardah Athiyyah, Yetti Hernaningsih, Dyah Fauziah, I Ketut Sudiana, Budi Utomo, Cita Rosita Sigit Prakoeswa

BACKGROUND: Airway epithelium constitutes the initial structural defense against inhaled allergens, with its integrity primarily maintained through the formation of tight junctions (TJ) consisting of claudins, occludins, and zonula occludens (ZO). Currently, the effects of probiotics on airway TJ remain unclear. Therefore, this study was conducted to explore the impact of Lactiplantibacillus plantarum IS-10506, a native probiotic from Indonesia, on bronchial epithelial TJ in a mouse model of allergic airway inflammation. METHODS: Twenty-four BALB/c mice were randomly assigned to three groups: negative control, positive control subjected to Dermatophagoides pteryonissinus (Der p) sensitization and challenge, and treatment group exposed to Der p and administered L. plantarum IS-10506 since one week before sensitization up to three weeks after. The expression levels of claudin-18, occludin, and ZO-1 were investigated through immunohistochemistry analysis and quantified based on the immunoreactive score (IRS). RESULTS: The results of this study demonstrated that L. plantarum IS-10506 significantly enhanced the expressions of claudin-18, occludin, and ZO-1 (p<0.05). CONCLUSION: These findings underscore the probiotic's role in fortifying the integrity of airway TJ.

180-8

Authors: Febriyeni Febriyeni, Andon Hestiantoro, Togas Tulandi, Muharam Muharam, Asmarinah Asmarinah, Ferry Sandra

BACKGROUND: The cytokine chemokine ligand 16 (CXCL16) plays an important role in the pathophysiology of endometriosis by regulating the inflammatory response and contributing to the pain-associated endometriosis. Despite this, the impact of epigenetic modifications, such as DNA methylation, on CXCL16 has yet to be fully understood. Therefore, this research was conducted to assess both the mRNA expression and DNA methylation levels of the proinflammatory gene CXCL16 in the endometrium tissue and menstrual blood of patients with and without endometriosis. METHODS: Thirty-five women with and without endometriosis were involved in this research. Subjects' menstrual blood samples were collected using filter paper pads, meanwhile the endometrium tissue were collected by performing biopsy, from which DNA and RNA were extracted. The DNA methylation levels of the CXCL16 were measured using the pyrosequencing method following bisulfite conversion treatment. Meanwhile, the mRNA expression level was measured using the quantitative polymerase chain reaction (qPCR) method and analyzed with the Livak method. RESULTS: The mRNA expression of CXCL16 in menstrual blood of endometriosis subjects was 2.42 times higher compared to control group (p=0.030). Furthermore, the expression of CXCL16 in menstrual blood was identical to that in endometrial tissue (p=0.173). DNA methylation analysis showed that CXCL16 in the menstrual blood of endometriosis subjctes had lower methylation levels compared to controls (p=0.004), indicating hypomethylation. CONCLUSION: Increased mRNA expression and hypomethylation of CXCL16 in the menstrual blood of endometriosis patients could serve as a direct marker for diagnosing endometriosis. However, further study to validate these findings and explore the potential of CXCL16 as a diagnostic tool, and additional research involving larger patient for the cohorts study is necessary.

199-207

Authors: Rini Suswita, Alvarino Alvarino, Eryati Darwin, Jamsari Jamsari

BACKGROUND: Inflammatory bowel disease (IBD) is a prevalent chronic inflammatory disorder of the gastrointestinal tract that may lead to colorectal cancer if untreated. Several medications for IBD have adverse side effects. Thus, there are high interest for searching alternative treatment to treat IBD. Lactococcus lactis D4 isolated from dadih, a traditional fermented buffalo milk product, was investigated for the effect on gut inflammation and microbial composition in the colitis rat model. METHODS: Eighteen male Sprague-Dawley rats were divided into three groups; control rats, colitis-induced rats, and colitis-induced rats treated with L. lactis D4 isolate. The control group received water, while the colitis-induced rats were given azoxymethane (AOM) and dextran sodium sulphate (DSS). Rats’ feces were collected for the analysis of gut microbiota diversity by next-generation sequencing and for the measurement of transforming growth factor (TGF)-β, nuclear factor kappa-B (NF-κB), interleukin (IL)-6, and tumor necrosis factor (TNF)-α colonic expression using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). RESULTS: L. lactis D4 administration was able to reduce inflammation in colitis by decreasing IL-6 expression (0.87 vs. 0.73), while expression of TGF-β, NF-κB, and TNF-α were increased compared to the model group, suggesting a complex immunomodulatory effect. Gut microbiota sequencing revealed a similarity between the control and treatment groups, suggesting L. lactis D4 has great potential to ameliorate gut microbiota dysbiosis in colitis rats. CONCLUSION: L. lactis D4 has a beneficial effect in decreasing pro-inflammatory cytokines and is able to reserve dysbiosis in colitis rat model. Thus, L. lactis D4 might be used as a therapeutic agent for IBD.