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The inhibitory effects of vancomycin on rat bone marrow-derived mesenchymal stem cell differentiation
Kari Hanson1,2, Carly Isder2, Kristen Shogren2
1Departments of1Neurologic Surgery.
Objective:
The use of intrawound vancomycin powder in spine surgery has been shown to decrease the rate of surgical site infections; however, the optimal dose is unknown. High-dose vancomycin inhibits osteoblast proliferation in vitro and may decrease the rate of solid arthrodesis. Bone marrow-derived mesenchymal stem cells (BMSCs) are multipotent cells that are a source of osteogenesis in spine fusions. The purpose of this study was to determine the effects of vancomycin on rat BMSC viability and differentiation in vitro.
Methods:
BMSCs were isolated from the femurs of immature female rats, cultured, and then split into two equal groups; half were treated to stimulate osteoblastic differentiation and half were not. Osteogenesis was stimulated by the addition of 50 µg/mL l-ascorbic acid, 10 mM β-glycerol phosphate, and 0.1 µM dexamethasone. Vancomycin was added to cell culture medium at concentrations of 0, 0.04, 0.4, or 4 mg/mL. Early differentiation was determined by alkaline phosphatase activity (4 days posttreatment) and late differentiation by alizarin red staining for mineralization (9 days posttreatment). Cell viability was determined at both the early and late time points by measurement of formazan colorimetric product.
Results:
Viability within the first 4 days decreased with high-dose vancomycin treatment, with cells receiving 4 mg/mL vancomycin having 40%-60% viability compared to the control. A gradual decrease in alizarin red staining and nodule formation was observed with increasing vancomycin doses. In the presence of the osteogenic factors, vancomycin did not have deleterious effects on alkaline phosphatase activity, whereas a trend toward reduced activity was seen in the absence of osteogenic factors when compared to osteogenically treated cells.
Conclusions:
Vancomycin reduced BMSC viability and impaired late osteogenic differentiation with high-dose treatment. Therefore, the inhibitory effects of high-dose vancomycin on spinal fusion may result from both reduced BMSC viability and some impairment of osteogenic differentiation.
Insights
High-dose vancomycin reduces bone marrow-derived mesenchymal stem cell (BMSC) viability and impairs osteogenic differentiation. This suggests high vancomycin doses may inhibit spinal fusion by affecting BMSCs.
Area of Science:
- Biomedical Engineering
- Orthopedic Surgery
- Stem Cell Biology
Background:
- Intrawound vancomycin powder use in spine surgery can reduce surgical site infections.
- Optimal vancomycin dosage remains unknown.
- High-dose vancomycin may inhibit osteoblast proliferation and solid arthrodesis.
Purpose of the Study:
- To investigate the effects of vancomycin on rat bone marrow-derived mesenchymal stem cell (BMSC) viability and osteogenic differentiation in vitro.
- To determine if vancomycin concentration impacts BMSC function.
Main Methods:
- Rat BMSCs were cultured and treated with osteogenic factors.
- Vancomycin was added at concentrations of 0, 0.04, 0.4, or 4 mg/mL.
- Cell viability, alkaline phosphatase activity, and alizarin red staining were assessed.
Main Results:
- Vancomycin at 4 mg/mL significantly reduced BMSC viability (40%-60% compared to control).
- Increasing vancomycin doses showed a trend toward decreased mineralization (alizarin red staining).
- Vancomycin did not negatively impact alkaline phosphatase activity in the presence of osteogenic factors.
Conclusions:
- High-dose vancomycin impairs BMSC viability and osteogenic differentiation.
- The inhibitory effects of high-dose vancomycin on spinal fusion may be due to reduced BMSC viability and impaired differentiation.
