人牙龈间充质干细胞裂解物分泌组减轻过氧化氢诱导的氧化应激对成骨细胞的不良影响。

Human gingival mesenchymal stem cells-lyosecretome attenuates adverse effect of hydrogen peroxide-induced oxidative stress on osteoblast cells.

作者信息

Kusumawardani Banun, Nurul Amin Muhammad, Rahayu Yani C, Sari Desi S, Altariq Morin I, Putri Arini H, Kanya Amara, Prahasanti Chiquita, Aljunaid Mohammed A

机构信息

Department of Biomedical Sciences, Faculty of Dentistry, University of Jember, Indonesia.

Department of Oral Biology, Faculty of Dentistry, University of Jember, Indonesia.

出版信息

J Taibah Univ Med Sci. 2024 May 13;19(3):687-695. doi: 10.1016/j.jtumed.2024.05.002. eCollection 2024 Jun.

DOI:10.1016/j.jtumed.2024.05.002

PMID:38831997

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11145533/

Abstract

OBJECTIVE

To determine total protein content, antioxidant activity, and protective ability of lyophilized human gingival mesenchymal stem cells (hGMSCs)-secretome in hydrogen peroxide (HO) 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 + HO).

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 IC 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 HO-induced oxidative stress.

CONCLUSION

Lyosecretome obtained from hGMSCs can maintain metabolic activity in osteoblast cells as protection against HO oxidative stress.

摘要

目的

在过氧化氢（H₂O₂）诱导的氧化应激模型中，测定冻干人牙龈间充质干细胞（hGMSCs）分泌组的总蛋白含量、抗氧化活性和保护能力。

方法

培养人GMSCs以获得条件培养基（分泌组），然后冻干制成冻干分泌组。通过二辛可宁酸测定法（BCA）和十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳（SDS - PAGE）测定总蛋白，以改进蛋白质测量。通过ABTS测定法测量抗氧化剂浓度，而冻干分泌组对氧化应激的保护能力则通过成骨细胞的代谢活性来确定。研究组分为对照组（培养基）和冻干分泌组治疗组（0.0；0.157、0.313、0.625、1.25、2.5、5和10 mg/mL + H₂O₂）。

结果

冻干分泌组的蛋白质浓度为2086.00±0.20 μg/ml，分子量分别为174、74、61、55和26 kDa，这些分子量的蛋白质被认为有助于细胞迁移，以及结合细胞因子和生长因子。冻干分泌组在浓度为4.8 mg/ml时还具有最高的抗氧化活性，为93.51%，IC₅₀值为2.08 mg/ml。在1.25 mg/ml冻干分泌组治疗组中显示出最高的细胞代谢活性（79.53±2.41%）。所有浓度的hGMSC - 冻干分泌组均减轻了H₂O₂诱导的氧化应激的不利影响。