Xanthine-Xanthine Oxidase Modeling Service for Osteoblast
Oxidative stress is the sudden production of a large amount of reactive oxygen species (ROS) due to the abnormal metabolism of the body caused by endogenous or exogenous stimulation, or the damage of the antioxidant defense system, the destruction of the dynamic balance of the production and clearance of oxygen free radicals, and the accumulation of reactive oxygen species, resulting in oxidative damage. Current studies have found that the pathogenesis of chronic inflammatory diseases and bone resorption related diseases is related to oxidative stress caused by reactive oxygen species.
As a risk factor of disease, oxidative stress has been paid more and more attention. Applying certain stimulating factors to cells can produce a large amount of ROS in cells. Establishing cell oxidative stress model is a common method to study the mechanism of oxidative stress injury and screen antioxidant drugs in vitro. Creative Bioarray established the oxidative stress model of human osteoblast like cells by stimulating MG63 cells with superoxide anion produced by xanthine / xanthine oxidase (X / XO).
Technology Principles
Figure 1. Pathways of xanthine oxidase. (Dash et al., 2021)
Osteoblasts are important cells involved in bone remodeling. The functional state of osteoblasts is an important factor affecting the health of periodontal tissue.
We recommend using X / XO model to establish osteoblast like cell oxidative stress model and stimulate MG63 cells to produce oxidative stress in vitro. Compared with X / XO, although hydrogen peroxide is cheap, its concentration changes significantly due to unstable potency, poor sealing and long storage time. Therefore, there is a large difference in the concentration when H2O2 is used to establish the oxidation damage model. Xanthine oxidase can catalyze the oxidation of hypoxanthine to xanthine and then to uric acid. Molecular oxygen is used as electron acceptor to reduce molecular oxygen to O2-， This is also one of the production pathways of endogenous ROS in cells, which can better simulate the state of oxidative stress.
What We Do
We have successfully used MG63 cell line for modeling, and have found the best X / XO concentration and induction time.
Osteoblast culture and modeling of the given strains
Primary cultured human osteoblasts were established by standard technology and identified by bone nodules and alkaline phosphatase. The cells were cultured in the incubator. According to the changes of cell growth density and medium pH, the medium is changed every 2 ~ 3 day. The experiment was carried out when the cells grew to the logarithmic growth stage.
We have found the best concentration and time of X / XO induction. However, if you choose other cell lines, we will conduct a small-scale grouping experiment before modeling to determine the best modeling conditions.
Modeling quality evaluation system
The production of intracellular ROS was detected by flow cytometry.
The production of ROS in cells was detected by oxidation sensitive fluorescent probe combined with flow cytometry.
The morphological structure of cells was observed by inverted phase contrast microscope.
MTT assay was used to observe and study the cell proliferation activity.
Drug administration and evaluation services
We can also adjust the modeling process according to the specific needs of customers, provide administration experiment, pharmacology, efficacy evaluation and pharmacokinetic analysis, and assist customers to study osteoblast stress and / or pathological significance, which has important theoretical significance for clarifying orthopaedic related diseases (such as osteogenesis imperfecta and osteofluorosis).
Model Application
The model can be used to study the effects of oxidative stress on the morphology and proliferation of human osteoblast like cells, and provide a theoretical basis for further studying the mechanism of oxidative stress in chronic bone resorption diseases such as periodontitis and periimplant inflammation.
Creative Bioarray is dedicated to providing high-quality products, comprehensive services, and tailored solutions to support and facilitate life sciences and pharmaceutical research and development. If you have any questions or needs, please contact us, and our customer service staff will help you at the first time.
Reference
- Dash, S., Pattanayak, S., Jena, B., Panda, M. K., & Singh, Y. D. (2021). Xanthine oxidase perspective in human health. Current Biotechnology, 9(4), 255-262..
All services and products are for scientific use only, not for medical use！
