Cinnamaldehyde ca

We purchased a panel of essential oils (Plant Guru, Plainfield, NJ, USA) and cinnamaldehyde (CA) (Sigma-Aldrich, St. Louis, MO, USA). The essential oils from Plant Guru company are tested by third party laboratory using GC/MS, and the GC/MS report can be found on their website [19 ]. Dimethyl sulfoxide (DMSO)-soluble essential oils were prepared at 10% (v/v) in DMSO as stock solution, which was then added with seven-day old stationary phase cultures at ration of 1:50 to achieve 0.2% of essential oils in the mixture. The 0.2% essential oils were further diluted to the stationary phase culture to get the desired concentration for evaluating anti-borrelia activity. DMSO-insoluble essential oils were directly added to B. burgdorferi cultures, then vortexed to form aqueous suspension, followed by immediate transfer of essential oil aqueous suspension in serial dilutions to desired concentrations and then added to B. burgdorferi cultures. Doxycycline (Dox), cefuroxime (CefU), (Sigma-Aldrich, St. Louis, MO, USA), and daptomycin (Dap) (AK Scientific, Union City, CA, USA) were prepared at a concentration of 5 mg/mL in suitable solvents [20 ,21 ], then filter-sterilized by 0.2 μm filter and stored at −20 °C as stock solutions.

Gentamicin (GEN, potency:
50–60 mg/mL) and cinnamaldehyde (CA) were purchased from Sigma-Aldrich,
Darmstadt, Germany. Poly(vinyl alcohol) (PVA), M_w: 89,000–98,000 (99+% hydrolyzed), gelatin from bovine
skin (GEL, gel strength ∼225 g bloom, Type B), glutaraldehyde
solution (GA, 50% wt, M_w: 100.12 g/mol),
Tween 80, phosphate buffer saline (PBS, pH = 7.4), Mueller–Hinton
Agar, and Luria Bertani (LB) broth were bought from Sigma-Aldrich
(St. Louis, MO). Crystal violet was obtained from Merck. Mouse embryonic
fibroblast (MEF) cells (SCRC-1040, ATCC), DMEM high glucose with 4.5
g/L d-glucose, l-glutamine, and sodium pyruvate
(NutriCulture, Eco Biotech), phosphate buffer saline (PBS) (Eco Biotech),
fetal bovine serum (Gibco), 1% penicillin–streptomycin (PAN
Biotech), DiOC6 (3,3′-dihexyloxacarbocyanine iodide) (Thermo
Fisher), and propidium iodine (Sigma-Aldrich) were used in the study.

U87, U251 and H4 cell lines were obtained from ATCC (Manassas, VA, USA). The U87 and H4 cells were grown in Dulbecco’s Minimal Essential Medium (DMEM) (Corning, Manassas, VA, USA) supplemented with 10% fetal bovine serum, 1% Penicillin-Streptomycin and 2 mM L-glutamine. U251 cells were cultured in Eagle’s Minimal Essential Medium (EMEM) (Corning, Manassas, VA, USA) supplemented with 10% fetal bovine serum, 1% Penicillin-Streptomycin and 2 mM L-glutamine. Cultures were incubated at 37 °C in an atmosphere of 5% CO₂. U87 cells expressing enhanced green fluorescent protein (U87eGFP) was derived as reported previously [4 (link)] and were used in the present study. Cinnamaldehyde (CA) (cat# W228613), trans-Cinnamaldehyde (TCA) (cat# 239968) and 2-Methoxycinnamaldehyde (MCA) (cat# W318101) were purchased from Sigma Aldrich (St. Louis, MO, USA). A stock solution of concentration of 100 mM of each compound was made in dimethyl sulfoxide. For flow cytometry, a Muse Cell analyzer and Muse assay kits were purchased from Luminex Corporation (Austin, TX, USA).