Antimicrobial sol-gel data collection

Published: 10 November 2023| Version 1 | DOI: 10.17632/53yh35mrfd.1
sarah forbes


Raw and processed data. Sol-gel UPEC 2023


Steps to reproduce

Preparation of biocide impregnated sol-gel polymers The sol was prepared as according to Nichol et al 2021. Batch biofilm viability assay 10 µl of 10 mg/ml antimicrobial containing sol-gel solution was added to the base of each well within a 96-well microtiter plate prior to polymerisation at room temperature for 18 hrs. 150 µl of diluted bacterial inoculum (OD600 0.008) was deposited into the well and plates were incubated at 37oC for 18 hrs. After incubation, culture was aspirated, wells were washed with 100 µl of sterile PBS prior to addition of 100 µl of BacTiter-Glo reagent (Promega, USA) and incubation for 5 minutes at room temperature. Drip flow reactor biofilm catheter model Silicone Rush Foley catheter sections (Teleflex Urology Care, UK) were cut into segments (1 cm) halved longitudinally, autoclaved and placed inside a drip-flow reactor continuous culture system either uncoated or after dip-coating in 10 mg/ml antimicrobial impregnated sol-gel. Catheter sections were added to the reactor and initially inoculated with 25 ml of bacterial culture (OD600 0.008) prior to static incubation for 6 hrs at 37 ºC. Subsequently the reactor was angled at a 10º decline and artificial urine was pumped into the chambers at 0.83 ml/min for 72 hrs at 37ºC. Catheter sections were aseptically removed from the system, rinsed in 2 x 10 ml of PBS and vortexed for 3 x 60 seconds to remove adhered bacteria. Biofilms were serially diluted in sterile PBS and plated onto MHA plates in triplicate, plates were incubated for 24 hrs at 37ºC and CFU/ml were determined. Agar overlay assay In brief, 2.4 x 106 L929 cells in 10 ml of Eagle minimum essential medium supplemented with 10% foetal bovine serum were seeded into 60 mm diameter cell culture plates. Cells were incubated for 48 hrs at 37 ºC and 5 % CO2 to form a monolayer on the base of the dish. After incubation the medium was removed by aspiration and cells were washed twice in 10 ml of PBS. 10 ml of EMEM containing 1 % agar was added to each dish and was allowed to solidify at room temperature. After the agar solidified, 10 ml of a 0.1 % neutral red solution was added to the centre of each plate, which was then rotated to evenly distribute the dye, left for 15 min, and excess solution was removed by aspiration. One 100 µg antimicrobial containing sol-gel disc was placed in the centre of each cell culture dish. Plates were incubated for 24 hrs at 37 °C and 5% CO2 before being checked for cell lysis. Cytotoxicity was characterised by a white colourless zone of dead cells around the implanted region. The diameter of the lysis zone was measured in mm. Disc diffusion assay Overnight cultures of bacteria were diluted to OD600 0.008 and 100 µl of diluted inoculum was spread across the surface of an MHA plate. Sol-gel discs containing 100 µg of the biocides/QSIs of interest were placed onto the centre of inoculated plates and incubated at 37 ºC for 24 hrs. Zones of inhibition were measured in mm.


Sheffield Hallam University


Antimicrobial Evaluation