The inhibitory effects of vancomycin on rat bone marrow-derived mesenchymal stem cell differentiation

Kari Hanson, Carly Isder, Kristen Shogren, Anthony L. Mikula, Lichun Lu, Michael J. Yaszemski, Benjamin D. Elder

Research output: Contribution to journalArticlepeer-review


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.

Original languageEnglish (US)
Pages (from-to)931-935
Number of pages5
JournalJournal of Neurosurgery: Spine
Issue number6
StatePublished - Jun 2021


  • Arthrodesis
  • Differentiation
  • Mesenchymal stem cells
  • Osteogenesis
  • Vancomycin

ASJC Scopus subject areas

  • Surgery
  • Neurology
  • Clinical Neurology


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