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Gene Expression in MC3T3-E1 Cells Treated with Diclofenac and Methylprednisolone
Tomasz P Lehmann1, Ewa Iwańczyk-Skalska1, Jerzy Harasymczuk2
1Department of Biochemistry and Molecular Biology, Poznań University of Medical Sciences, Święcickiego 6, 60-781 Poznan, Poland.
Abstract:
Nonsteroidal anti-inflammatory drugs (NSAIDs) and glucocorticoids (GCs) are often used to treat articular-skeletal disorders. The extended use of NSAIDs and GCs have adverse effects on bone metabolism, reducing bone quality and impairing fracture healing. In the present study, we used mouse pre-osteoblast cells MC3T3-E1 to demonstrate the effects of diclofenac (DF) and methylprednisolone (MP) on cell proliferation and gene expression. Cells were incubated with three doses of DF or MP: 0.5 µM, 5 µM, and 50 µM. MP decreased cell viability even after 24 h, but DF inhibited cell viability after only seven days of treatment. The cells were lysed after one, two, three, and seven days of treatment, and gene expression was analyzed by reverse transcription and quantitative PCR (RT-qPCR) assays. DF did not significantly affect the expression of the osteogenic marker genes. MP modified the expression of Osx, Runx, and Col1a1. We concluded that MP is a more potent inhibitor of mouse pre-osteoblast differentiation and viability than is DF. Our results suggest that prolonged DF treatment could be less harmful to osteoblasts than MP treatment.
Insights
Methylprednisolone (MP) more potently inhibits pre-osteoblast viability and differentiation than diclofenac (DF). Prolonged DF treatment may be less harmful to bone metabolism than MP treatment.
Area of Science:
- Biochemistry
- Cell Biology
- Pharmacology
Background:
- Nonsteroidal anti-inflammatory drugs (NSAIDs) and glucocorticoids (GCs) are commonly prescribed for articular-skeletal disorders.
- Prolonged use of NSAIDs and GCs can negatively impact bone metabolism, leading to reduced bone quality and impaired fracture healing.
Purpose of the Study:
- To investigate the effects of diclofenac (DF) and methylprednisolone (MP) on mouse pre-osteoblast (MC3T3-E1) proliferation and gene expression.
- To compare the inhibitory potential of DF and MP on osteoblast function.
Main Methods:
- MC3T3-E1 cells were treated with varying doses (0.5 µM, 5 µM, 50 µM) of DF or MP.
- Cell viability was assessed at 24 hours and 7 days.
- Gene expression of osteogenic markers was analyzed using RT-qPCR at multiple time points (1, 2, 3, and 7 days).
Main Results:
- MP significantly decreased cell viability within 24 hours, while DF's effect was observed after 7 days.
- DF did not substantially alter the expression of key osteogenic marker genes.
- MP significantly modified the expression of osteoblast-related genes, including Osx, Runx, and Col1a1.
Conclusions:
- Methylprednisolone (MP) exhibits a stronger inhibitory effect on mouse pre-osteoblast differentiation and viability compared to diclofenac (DF).
- These findings suggest that extended diclofenac (DF) treatment may pose a lower risk to osteoblasts than methylprednisolone (MP) treatment.
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