Tobramycin

Human BMSCs were purchased from Cyagen (USA) and cultured in human BMSCs basal media (Cyagen, USA) with 10% fetal bovine serum (Cyagen, USA), 1% penicillin-streptomycin (Cyagen, USA), and glutamine (Cyagen, CA, USA). And osteogenic induction was conducted by BMSCs osteogenic differentiation media (Cyagen, USA) with 10% fetal bovine serum (Cyagen, USA), glutamine (Cyagen, CA, USA), 1% penicillin-streptomycin (Cyagen, USA), ascorbic acid (Cyagen, CA, USA), β-Glycerolphosphate disodium (Cyagen, USA), and dexamethasone (Cyagen, USA). All the cells were cultured at 37°C in saturated humidity containing 5% CO₂. The media were changed every two days. The BMSCs were digested using 0.25% trypsin (Solarbio, China), when they grew to a confluence of 70–80%. To investigate the effect of topical antibiotics, BMSCs were cultured without antibiotics (Con group) or with antibiotics, which further subdivided into three subgroups: Van group: cultured with vancomycin (Solarbio, China), (1g per person11 (link),12 (link) = 14.28μg/mL), Tob group: cultured with tobramycin (Solarbio, China), (2g per person3 (link),13 (link)=28.57μg/mL), Van+Tob group (vancomycin 14.28μg/mL combined with tobramycin 28.57μg/mL).

Linolenic acid (Sigma-Aldrich), Azocasein (Sigma-Aldrich), AlarmaBlue cell viability assay kit (KeyGen BioTECH, China), Tobramycin (Solarbio, China), Crystal violet (Solarbio, China), LIVE/DEAD BacLight Bacterial viability kit (Molecular Probes, USA), Biozol Total RNA extraction reagent (BioFlux, China), TransScript Green Two-Step qRT-PCR SuperMix kit (Transgen Biotech, China), Trans2 K DNA ladder (Transgen Biotech, China), 2X Taq PCR MasterMix (TianGen, China), Ethidium bromide (Sigma, China), Cetrimide agar base (Xiya, China), Dimethyl sulfoxide (AMRESCO, Ohio-USA), and any other reagents were of analytical grade.