Gibco pbs

Tobramycin (Tob, molecular weight (Mw) 467.5 Da), ciprofloxacin (Cipro, Mw 331.3 Da), itaconic acid (IA, Mw 130.1 Da), peptone, Tween20 and casein were obtained from Merck. Salts and organic compounds of analytical grade (NH₄Cl, KCl, tris-HCl, chloroform, glycerol, glucose, formic acid and MgSO₄·7H₂O) were obtained from Merck and VWR. Yeast extract was obtained from Fluka. Bacto™ Tryptone were obtained from BD Biosciences. Luria Bertani (LB) agar was obtained from Carl Roth. Gibco^® PBS was obtained from Life Technologies. Purified water was produced by Milli-Q water purification system (Merk Millipore, Billerica, MA, USA)
The Pseudomonas aeruginosa strain PA14 wild type (wt) was obtained from ATCC or DSMZ, the German Collection of Microorganism and Cell Cultures GmbH (PA14 = DSMZ19882), stored in glycerol stocks at −80 °C. Bacteria were cultured in a minimal proteose peptone glucose ammonium salt (PPGAS) medium. PPGAS was composed of 1 g/L NH₄Cl, 1.5 g/L KCl, 19 g/L Tris-HCl, 10 g/L peptone, 5 g/L glucose and 0.1 g/L MgSO4·7H₂O. The medium was adjusted to pH 7.2 ± 0.2 and sterilized before use. Agar solution was composed of 15.5 g/L LB agar, 10 g/L peptone, 5 g/L NaCl and 5 g/L yeast extract, the solution was sterilized before being plated.

The genomic DNA of each of the 100 A. baumannii isolates was extracted from a single colony grown in TSB (Oxoid, UK) culture media by using the ZR Fungal/Bacterial kit [29 ]. The manufacturer’s instructions were followed with some modifications; instead of 50 to 100 mg of cells, 1 000 μl of overnight broth was used, centrifuged (Eppendorf 5417C, Germany) and the pellet was resuspended in phosphate buffered saline (PBS) solution (pH 7.2, Gibco® PBS, Life Technologies Corporation, USA) and 600 μl of the lysis solution were used instead of the prescribed 750 μl. The extracted genomic DNA was stored at -20 °C (Defy Ltd, Multimode, SA) until further analysis.

All chemicals were purchased from Sigma-Aldrich unless otherwise specified. Tetrahydrofuran (HPLC grade) (THF), ethanol, absolute (HPLC grade) (EtOH), ethyl acetate (analytical grade reagent), and Formic Acid Optima LCMS were purchased from Fisher Scientific. Acetonitrile and methanol (MeOH) were obtained from VWR Chemicals. Yeast extract was obtained from Fluka. Bacto™ Tryptone was obtained from BD Biosciences. Luria Bertani (LB) agar was obtained from Carl Roth. Gibco® HBSS (1x) Hanks' Balanced Salt Solution and Gibco® PBS was obtained from Life Technologies. Purified water was prepared by a Milli-Q water purification system (Merck Millipore, Billerica, MA, USA) (called water in the manuscript).

All devices were primed by submerging the device under distilled water and applying a vacuum of approximately 75 kPa below atmosphere for 10 min. Air was then slowly released into the vacuum chamber while the devices were submerged, priming the channels (including dead-ends) with distilled water. After priming, data collection was carried out on a benchtop in room air. Unless otherwise specified, all fluidic connections were made with 0.03-inch-inner-diameter fluorinated ethylene propylene (FEP) tubing (1520XL, IDEX-HS) and PEEK fittings purchased from IDEX Health & Sciences. The various tubular fluidic resistors were made using 0.01-inch-inner-diameter FEP tubing (1527L, IDEX-HS). The specific resistor lengths and other component details for each circuit are provided in Extended Data Table 1. Computer-controlled pressure sources (LineUp FlowEZ, Fluigent) were used to supply pressures for characterization of the microfluidic devices. Unless otherwise specified, all reservoirs for the pressure sources (P-CAP, Fluigent) were filled with 1× phosphate-buffered saline (PBS; Gibco PBS, Fisher Scientific). All pressure measurements were made using Honeywell pressure sensors (ABPDRRV015PDAA5) and logged on a computer using MATLAB. All flow measurements were made using Sensirion flow meters (SLI-1000).

Blood was collected with informed consent from three healthy voluntary donors (age 28–51 years) through needle prick and it was subsequently suspended in phosphate-buffered saline solution (Gibco PBS, Fisher Scientific, Schwerte, Germany). Following collection, samples were centrifuged for 5 min at 3000  $\times$  g. The sedimented RBCs were washed three times with PBS and final RBC samples were adjusted at a hematocrit concentration of $1\mathrm{\%}\mathrm{Ht}$ in a PBS solution that contained $1\mathrm{g}/\mathrm{l}$ bovine serum albumin (BSA, Sigma-Aldrich, Taufkirchen, Germany). The PBS/BSA mixture is a Newtonian fluid with a shear viscosity of roughly $\eta =1.2\mathrm{mPa}\mathrm{s}$ .^13,42 (link)
Blood withdrawal, sample preparation, and microfluidic experiments were performed according to the guidelines of the Declaration of Helsinki and approved by the ethics committee of the “Ärztekammer des Saarlandes” (permission number 51/18).

Blood was collected into EDTA-containing tubes ( $1.6$ mg/mL EDTA, SARSTEDT, Nümbrecht, Germany) with informed consent from three healthy male voluntary donors (age 28–31 years). It was centrifuged for 5 min at 3000× g to separate RBCs and plasma. Sedimented RBCs were washed three times with a phosphate-buffered saline solution (Gibco PBS, Fisher Scientific, Schwerte, Germany). Finally, a hematocrit concentration of $1\%\mathrm{Ht}$ was adjusted in a PBS solution that contained $1$ g/L bovine serum albumin (BSA, Sigma-Aldrich, Taufkirchen, Germany). The viscosity of the PBS/BSA solutions at $20°$ C was approximately $1.2$ mPa s [56 (link)], similar to the viscosity of human blood plasma. Since we do not observe significant inter-individual variations in the results, data were averaged between the three donors.
Blood withdrawal, sample preparation, and microfluidic experiments were performed according to the guidelines of the Declaration of Helsinki and approved by the ethics committee of the “Ärztekammer des Saarlandes” (permission number 51/18).

Prototypes were investigated by KYKY EM3200 scanning electron microscope (KYKY Technology Development Ltd., China) to study the morphology of scaffolds and cells. Scaffolds were washed with Gibco PBS (Fisher Scientific GmbH, Germany) at day 14 of cell culture and fixed with %1.5 glutaraldehyde (Sigma-Aldrich Solutions, Germany) solution followed by dehydration using gradient ethanol (Sigma-Aldrich Solutions, Germany). To visualize the internal regions, scaffolds were fractured in liquid nitrogen (Arian Gas Co., Iran), sputter coated with gold, and examined by the SEM eventually. To analyze the pore structure of scaffold prototypes, ImageJ software (NIH Image, Bethesda, MD, USA) was utilized.