Development of a thermo-responsive hydrogel as a carrier for biofilm targeting enzymes and antimicrobials for the treatment of orthopaedic device-related infections
(2022) ECCMID 2022 — Location: Lisbonne (23.April.2022)
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- Authors
Buzisa Mbuku, Randy 
- Abstract
- Background Orthopedic device-related infection (ODRI) is a leading cause of failure following arthroplasty or osteosynthesis. In this setting, antibiotic activity against micro-organisms is reduced by biofilm formation. The use of matrix-targeting enzymes is a promising option to restore this activity. However, the ideal drug form for this application is undetermined. We propose the use of a thermoresponsive hydrogel containing a previously described enzyme combination (TEC; Poilvache et al. 2021) associated with vancomycin and describe the release kinetics of TEC and the activity of the TEC and vancomycin combination against S.aureus biofilms. Methods Release kinetics: A solution of 20%(w/v) Pluronic F-127 in a TEC solution (Poilvache et al. 2021) containing 10mg/ml of vancomycin, was prepared at 4°C. 1mL was placed in 15mL tubes and gelled at room temperature. 9ml of Tris-HCl buffer was added and tubes were incubated at 37°C. 0.5mL samples were regularly collected over 7 days and replaced with fresh buffer to maintain total volume. Samples proteins contents were determined using the BCA assay. Antibiofilm activity: ATCC33591 S.aureus biofilms were grown for 24h at 37°C with a continuous 50rpm agitation on Ti-6Al-4V coupons by adding 3ml of a 6log10 suspension in TGN (TSB+1%glucose+2%NaCl) to 12wells plates containing the coupons. Samples were placed in 6wells plates containing 1ml of the hydrogel containing buffer (controls), TEC, vancomycin, or both. 9ml of TGN was added and samples were incubated in identical conditions for 24h. CFU counts were obtained after samples sonication, dilutions, and TSA plating. Results (a)Figure 1. A reproducible release of proteins from the hydrogels was observed following an exponential plateau model, with a rate constant of 0.08h-1, a plateau reached after 72h, and 80% proteins release after 17h30. (b)Figure 2. Exposure of mature S. aureus biofilms to hydrogels containing either TEC or vancomycin reduced CFU counts by respectively 1.2 and 1.3log10 when compared to controls (p>0.05), while the CFU counts in samples exposed to hydrogel containing both TEC and vancomycin were reduced by 3log10 (p<0.05). Conclusions We show that a thermoresponsive hydrogel could be an advantageous carrier for biofilm targeting enzymes and antimicrobials, allowing for a prolonged release while maintaining a good antibiofilm activity.
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Citations
Poilvache, H., Buzisa Mbuku, R., Freyberg, A., Cornu, O., & Van Bambeke, F. (2022). Development of a thermo-responsive hydrogel as a carrier for biofilm targeting enzymes and antimicrobials for the treatment of orthopaedic device-related infections. ECCMID 2022, Lisbonne. https://hdl.handle.net/2078.5/271910