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

Abstract

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.

Comparison of the differentiation abilities of bone marrow‑derived mesenchymal stem cells and adipose‑derived mesenchymal stem cells toward nucleus pulposus‑like cells in three‑dimensional culture

“…The present results indicated that BMSCs and ADSCs differentiated toward an NPC-like phenotype, and that ADSCs had a greater ability to differentiate into NPC-like cells than BMSCs. Previous studies demonstrated that both BMSCs ( 53 , 58 ) and ADSCs ( 27 ) could differentiate toward NPC-like cells under the same conditions with a differentiating medium.…”

“…Isolation of BMSCs and ADSCs. BMSCs were isolated and purified using the whole bone marrow cell repeated adherent culture method (27)(28)(29)(30). All animal experiments were approved by the Animal Experiment and Ethics Committee of Kunming Medical University (Kunming, China; approval no.…”

Effect of Vancomycin Applied to the Surgical Site on Fracture Healing in a Diabetic Rat Model

“…For instance, Hanson et al 15 demonstrated that exposure of rat bone marrow–derived mesenchymal stem cells to 4000 µg/mL vancomycin for 4 days reduced cell viability by >60% and impaired calcium deposition. Braun et al 5 determined that only vancomycin concentrations >1000 µg/mL will decrease cell viability and ALP activity in human osteoblasts.…”

Antimicrobial peptide-based materials: opportunities and challenges

“…Although, it is reported that antibiotic treatment controls infection, it inhibits the osteoblast proliferation and viability, resulting in poor bone regeneration. 71,72 Several AMPs are identified with the capability to differentiate MSCs to osteoblast and to facilitate bone repair. Li et al proposed that AMPs activate transcription of bone morphogenetic protein-2 (BMP-2), which further activates SMAD1/5/8 and mitogen-activated protein (MAP) kinase pathways, leading to phosphorylation of SMAD1/5/8 proteins.…”