Banun Kusumawardani PhD , Muhammad Nurul Amin PhD , Yani C. Rahayu drg , Desi S. Sari PhD , Morin I. Altariq , Arini H. Putri , Amara Kanya , Chiquita Prahasanti PhD , Mohammed A. Aljunaid MDS
{"title":"人牙龈间充质干细胞-溶糖体可减轻过氧化氢诱导的氧化应激对成骨细胞的不利影响","authors":"Banun Kusumawardani PhD , Muhammad Nurul Amin PhD , Yani C. Rahayu drg , Desi S. Sari PhD , Morin I. Altariq , Arini H. Putri , Amara Kanya , Chiquita Prahasanti PhD , Mohammed A. Aljunaid MDS","doi":"10.1016/j.jtumed.2024.05.002","DOIUrl":null,"url":null,"abstract":"<div><h3>Objective</h3><p>To determine total protein content, antioxidant activity, and protective ability of lyophilized human gingival mesenchymal stem cells (hGMSCs)-secretome in hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) induced oxidative stress model.</p></div><div><h3>Methods</h3><p>Human GMSCs were cultured to obtain a conditioned medium (secretome), then lyophilized to produce lyosecretome. Total protein was determined by bicinchoninic acid assay (BCA) and SDS-PAGE to improve protein measurements. Antioxidant concentration was measured by ABTS assay, while the protective ability of lyosecretome against oxidative stress was determined by the metabolic activity of osteoblast cells. The study group was divided into a control group (culture medium) and a lyosecretome treatment group (0.0; 0.157, 0.313, 0.625, 1.25, 2.5, 5, and 10 mg/mL + H<sub>2</sub>O<sub>2</sub>).</p></div><div><h3>Results</h3><p>Lyosecretome had a protein concentration of 2086.00 ± 0.20 μg/ml, with a molecular weight of 174, 74, 61, 55, and 26 kDa, which are thought to facilitate cell migration, as well as bind cytokines and growth factors. Lyosecretome also provided the highest antioxidant activity of 93.51% at a concentration of 4.8 mg/ml, with an IC<sub>50</sub> value of 2.08 mg/ml. The highest cell metabolic activity (79.53 ± 2.41%) was shown in the 1.25 mg/ml lyosecretome treatment group. All concentrations of hGMSC-lyosecretome attenuate the adverse effect of H<sub>2</sub>O<sub>2</sub>-induced oxidative stress.</p></div><div><h3>Conclusion</h3><p>Lyosecretome obtained from hGMSCs can maintain metabolic activity in osteoblast cells as protection against H<sub>2</sub>O<sub>2</sub> oxidative stress.</p></div>","PeriodicalId":46806,"journal":{"name":"Journal of Taibah University Medical Sciences","volume":null,"pages":null},"PeriodicalIF":1.5000,"publicationDate":"2024-05-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.sciencedirect.com/science/article/pii/S1658361224000404/pdfft?md5=cf5d01de8430e90bf05d363674e52fa4&pid=1-s2.0-S1658361224000404-main.pdf","citationCount":"0","resultStr":"{\"title\":\"Human gingival mesenchymal stem cells-lyosecretome attenuates adverse effect of hydrogen peroxide-induced oxidative stress on osteoblast cells\",\"authors\":\"Banun Kusumawardani PhD , Muhammad Nurul Amin PhD , Yani C. Rahayu drg , Desi S. Sari PhD , Morin I. Altariq , Arini H. Putri , Amara Kanya , Chiquita Prahasanti PhD , Mohammed A. Aljunaid MDS\",\"doi\":\"10.1016/j.jtumed.2024.05.002\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><h3>Objective</h3><p>To determine total protein content, antioxidant activity, and protective ability of lyophilized human gingival mesenchymal stem cells (hGMSCs)-secretome in hydrogen peroxide (H<sub>2</sub>O<sub>2</sub>) induced oxidative stress model.</p></div><div><h3>Methods</h3><p>Human GMSCs were cultured to obtain a conditioned medium (secretome), then lyophilized to produce lyosecretome. Total protein was determined by bicinchoninic acid assay (BCA) and SDS-PAGE to improve protein measurements. Antioxidant concentration was measured by ABTS assay, while the protective ability of lyosecretome against oxidative stress was determined by the metabolic activity of osteoblast cells. The study group was divided into a control group (culture medium) and a lyosecretome treatment group (0.0; 0.157, 0.313, 0.625, 1.25, 2.5, 5, and 10 mg/mL + H<sub>2</sub>O<sub>2</sub>).</p></div><div><h3>Results</h3><p>Lyosecretome had a protein concentration of 2086.00 ± 0.20 μg/ml, with a molecular weight of 174, 74, 61, 55, and 26 kDa, which are thought to facilitate cell migration, as well as bind cytokines and growth factors. Lyosecretome also provided the highest antioxidant activity of 93.51% at a concentration of 4.8 mg/ml, with an IC<sub>50</sub> value of 2.08 mg/ml. The highest cell metabolic activity (79.53 ± 2.41%) was shown in the 1.25 mg/ml lyosecretome treatment group. All concentrations of hGMSC-lyosecretome attenuate the adverse effect of H<sub>2</sub>O<sub>2</sub>-induced oxidative stress.</p></div><div><h3>Conclusion</h3><p>Lyosecretome obtained from hGMSCs can maintain metabolic activity in osteoblast cells as protection against H<sub>2</sub>O<sub>2</sub> oxidative stress.</p></div>\",\"PeriodicalId\":46806,\"journal\":{\"name\":\"Journal of Taibah University Medical Sciences\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":1.5000,\"publicationDate\":\"2024-05-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.sciencedirect.com/science/article/pii/S1658361224000404/pdfft?md5=cf5d01de8430e90bf05d363674e52fa4&pid=1-s2.0-S1658361224000404-main.pdf\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Journal of Taibah University Medical Sciences\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S1658361224000404\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"MEDICINE, GENERAL & INTERNAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Journal of Taibah University Medical Sciences","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S1658361224000404","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MEDICINE, GENERAL & INTERNAL","Score":null,"Total":0}
Human gingival mesenchymal stem cells-lyosecretome attenuates adverse effect of hydrogen peroxide-induced oxidative stress on osteoblast cells
Objective
To determine total protein content, antioxidant activity, and protective ability of lyophilized human gingival mesenchymal stem cells (hGMSCs)-secretome in hydrogen peroxide (H2O2) induced oxidative stress model.
Methods
Human GMSCs were cultured to obtain a conditioned medium (secretome), then lyophilized to produce lyosecretome. Total protein was determined by bicinchoninic acid assay (BCA) and SDS-PAGE to improve protein measurements. Antioxidant concentration was measured by ABTS assay, while the protective ability of lyosecretome against oxidative stress was determined by the metabolic activity of osteoblast cells. The study group was divided into a control group (culture medium) and a lyosecretome treatment group (0.0; 0.157, 0.313, 0.625, 1.25, 2.5, 5, and 10 mg/mL + H2O2).
Results
Lyosecretome had a protein concentration of 2086.00 ± 0.20 μg/ml, with a molecular weight of 174, 74, 61, 55, and 26 kDa, which are thought to facilitate cell migration, as well as bind cytokines and growth factors. Lyosecretome also provided the highest antioxidant activity of 93.51% at a concentration of 4.8 mg/ml, with an IC50 value of 2.08 mg/ml. The highest cell metabolic activity (79.53 ± 2.41%) was shown in the 1.25 mg/ml lyosecretome treatment group. All concentrations of hGMSC-lyosecretome attenuate the adverse effect of H2O2-induced oxidative stress.
Conclusion
Lyosecretome obtained from hGMSCs can maintain metabolic activity in osteoblast cells as protection against H2O2 oxidative stress.
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