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Journal : Universa Medicina

Ethanol extract of Abrus precatorius L. leaves diminishes inflammatory responses in nicotine-treated human gingival fibroblasts: an in vitro study Kesumaningtias, Raden Roro Widorini; Kusuma, Indra; Suciati, Yulia; Sari, Wening
Universa Medicina Vol. 43 No. 3 (2024)
Publisher : Faculty of Medicine, Universitas Trisakti

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2024.v43.272-279

Abstract

BACKGROUNDNicotine induces oxidative stress in human gingival fibroblasts (HGF) and stimulates the production of cytokines that trigger inflammation. Abrus precatorius L. (AP) leaves contain antioxidants with anti-inflammatory properties that can prevent the formation of free radicals and reduce tissue damage due to inflammation. This study aimed to determine the effect of ethanolic extract of AP leaves (EAP) on interleukin (IL-6) levels and cyclooxygenase-2 (COX-2) gene expression in gingival fibroblasts exposed to nicotine. METHODSCells were randomized into six treatment groups and clustered into the non-treatment control group (NTC), solvent control (SC), nicotine control (NC), and groups treated with nicotine and EAP at doses of 9.375 µg/mL, 18.75 µg/mL, and 37.5 µg/mL, respectively, for 24 hours. IL-6 levels were examined using the Elisa method, while COX-2 gene expression was assessed using PCR. Data were analyzed using Oneway ANOVA and the Kruskal Wallis test. RESULTSIL-6 levels and COX-2 expression were considerably higher in the nicotine control group. Conversely, the cell groups treated with nicotine and EAP had substantially decreased levels of both inflammatory markers IL-6 and COX-2 (p=0.029) across all EAP dose levels compared to the nicotine control group. The highest reduction in response was observed at the dose of 9.375 ìg/mL EAP. CONCLUSIONThese results highlight the potential of Abrus precatorius L. in relieving nicotine-induced inflammation in smokers. By suppressing the production of inflammatory mediators IL-6 and COX-2 in HGF, EAP presents a promising avenue for further in vitro research.
Comparative analysis of KnockOut serum replacement and fetal bovine serum for in vitro culture of human dermal fibroblasts Hidayati, Nur Ilham Risma; Kusuma, Indra; Poerwanto, Eko
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.v42.283-293

Abstract

BACKGROUNDHuman dermal fibroblast (HDF) cultures can be used as a regenerative agent for wound healing. Fetal bovine serum (FBS) as a culture supplement is derived from animals, therefore not being constant in composition, causes variations in research results, thus requiring a substitute such as KnockOut serum replacement (KOSR). This study evaluated the defined KOSR as FBS substitute for HDF culture by measuring the relative expression of basic fibroblast growth factor (bFGF) and keratinocyte growth factor (KGF) messenger RNA (mRNA), HDF cell proliferation, and HDF migration. METHODSHuman dermal fibroblast culture was divided into 2 intervention groups receiving KOSR 5% and KOSR 10%, respectively, and a control group receiving FBS 10%. Reverse transcription polymerase chain reaction (RT-PCR) was used for bFGF and KGF mRNA relative expression at the fifth passage (P5). Cell counting kit-8 (CCK-8) reagent was used for the HDF cell proliferation assay at P5 and the scratch assay was used for HDF cell migration at P6. Data were analyzed using dependent t-test, One-way ANOVA, or Kruskal-Wallis test. RESULTSThere were no significant differences in bFGF and KGF mRNA relative expression and HDF migration velocity between the intervention and control groups (p>0.05 and p>0.05, respectively). The doubling time of the KOSR 5% group showed no significant difference (p>0.05), but KOSR 10% and FBS 10% showed significant differences between treatment days 2-6 and treatment days 6-10 (p<0.05). CONCLUSIONSThe KOSR 10% was comparable to FBS 10% in supporting bFGF and KGF mRNA relative expression, HDF cell proliferation, and HDF cell migration in HDF culture.
Nicotine reduces cell viability and induces oxidative stress in human gingival fibroblasts Azmi, Sabrina; Hadi, Restu Syamsul; Kusuma, Indra; Suciati, Yulia; Sari, Wening
Universa Medicina Vol. 43 No. 1 (2024)
Publisher : Faculty of Medicine, Universitas Trisakti

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18051/UnivMed.2024.v43.20-30

Abstract

BackgroundNicotine, as the main component of cigarettes, is known to interfere with the proliferation of human gingival fibroblasts (HGFs) and can trigger oxidative stress. This study aimed to analyze the impact of nicotine on viability, expression of the antioxidant Nrf2, levels of the product of oxidative stress malondialdehyde (MDA), and the migration capacity of HGFs. MethodsAn experimental laboratory study used fibroblasts isolated from healthy human gingiva. The cells were grouped into the non-treatment control group (NTC), the solvent control (SC), and the treatment groups, exposed to nicotine at various concentrations for twenty-four hours. Cell viability was assesed using the cell counting kit-8 (CCK-8), Nrf2 expression was examined using ELISA, MDA level was measured using an MDA kit, and migration capacity was assessed using a scratch assay. Statistical analysis used one-way Anova or Kruskal-Wallis test. A p-value of <0.05 was expressed statistically significant. ResultsThe Cell viability was substantially reduced in the nicotine group compared to the untreated group, accompanied by changes in cell morphology. In contrast, Nrf2 expression increased significantly (p=0.010) in the 5 mM nicotine group compared with the control group. The MDA levels were not significantly distinct across groups (p=0.056). Cell migration was delayed significantly in the 5 mM nicotine group at 72 hours after scratching compared to the control group. ConclusionNicotine decreased HGFs viability and increased Nrf2 expression significantly in a dose-dependent manner. Nicotine at 5 mM concentration did not alter MDA levels but delayed cell migration.
Co-Authors Agus Sulistiawan Ahmad Sulaeman Ajib Susanto Alida Harahap Alvita, Nini Aridestama, Raihan Aulia Arief Boediono Arief, Ilham Ascobat, Purwantyastuti Asnita, Rika Aufa, Akhdan Aulia, Maila Rizqi Avliya Quratul Marjan Azmi, Sabrina Azzahra, Sabna Bahri, Syukrini Bambang Kiranadi Bambang Purwoko Dewi, Hanik Anggraeni Djoko Sasono, Djoko Edy Supriyadi Eko Poerwanto Elsye Souvriyanti, Elsye Endang Purwaningsih Endy Muhammad Astiwara Erlina Burhan Erlina, Andi Erna Harfiani Evy Latifah Fachry Abda El Rahman Fadilah, Khonsa Fitri Fahrudin Fahrudin Fajrian, Muhammad Adhif Fasli Jalal Fauzi, Achmad Ma’ruf Fikriana, Chika Nur Fitria Athayya Desvianti Galih Saputra, Galih Gusnanto, Arief Hadi, Restu S. Hambani, Susy Hidayati, Anggi Puspa Nur Hidayati, Nur Ilham Risma Idris, Muthoharoh Ilmi, Ibnu Malkan Bakhrul Jemmy Rumengan Juniarti . Kesumaningtias, Raden Roro Widorini Lating, Noval Ali Lina Lasminingrum Muchtiar, Mulyadi Mulyadewi, Amelia Luthfiyah Mulyono, Ibnu Utomo Wahyu Mulyono, Ibnu Utomo Wahyu Noeriwan, Noeriwan Olivia, Prisca Pangestu, Hendri Pelangi Eka Yuwita Pratama, Muhammad Abel Priyono, Harim Putra, Andika Chandra Putri, Pingkan Permata Restu Syamsul Hadi Restuti, Ratna Dwi Rianda Dirkareshza Rianto Setiabudy Rifqatussaadah, Rifqatussaadah Rika Bur Rika Yuliwulandari Riliani, Marisa Ros Sumarny Samsul Mustofa, Muhammad Sapto Mei Budiyanto, Sapto Mei Sophie Yolanda Sulistiawati, Indah Syarifah, Lensy Khosinatus Taupiqqurrahman Tjandra Yoga Aditama Trijaya, Dewi Sekarsari Trinovita Andraini Utami Wahyuningsih Utomo, Ahmad Rusdan H Wening Sari Yati Sumiyati, Yati Yudertha, Andreo Yulia Suciati Yunazar Manjang Yurika Sandra Zannah, Fahma Hayyul