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Presoaking reticular polypropylene meshes in antiseptic solution inhibits the growth of Staphylococcus aureus in vitro Pérez-Köhler B1, García-Moreno F1, Bayon Y2, Pascual G3, Bellón JM1. 1Department of Surgery, Medical and Social Sciences, University of Alcalá, Spain. 2Covidien-Sofradim Production, Trévoux, France. 3Department of Medicine and Medical Specialties, University of Alcalá, Spain. 1,3Networking Research Center on Bioengineering, Biomaterials and Nanomedicine (CIBER-BBN), Spain. Poster nº 31 Background. As a preventive measure to avoid the post-operative infection of a prosthetic mesh used for hernia repair, the mesh may be soaked in antibacterial solution immediately before its implantation in the host. This study assesses the use of the antiseptics, chlorhexidine (CHX) and allicin (natural antiseptic not previously used for this purpose) vs the antibiotic vancomycin for the pretreatment of a knitted polypropylene (PP) mesh in an in vitro model of contamination by Staphylococcus aureus (Sa). Methods. Fragments (1x1cm) of the PP mesh Surgipro™ were dipped in solutions of saline (0.9%), vancomycin (40μg/mL), allicin (1,000μg/mL), CHX (0.05%) or allicin-CHX (900μg/mL-0.05%) and incubated for 24/48/72 h on contaminated agar plates (106 CFU Sa ATCC25923) (n=7 per time point and treatment) in the agar diffusion test. The antimicrobial mesh DualMesh® Plus (DM+) was used as a positive control. At each time point, inhibition halos were measured and bacterial adhesion to the meshes determined by sonication (n=5) and scanning electron microscopy (n=2). As a measure of treatment cytotoxicity, cell viability was examined on rabbit skin fibroblasts. The Mann-Whitney U test was used for statistical analysis (mean SEM ). u Fig 1. Zones of inhibition created by the pretreated meshes after 72 h of the contamination. t Fig 2. Diameter of the inhibition zones (mm) created by the meshes at the different time points. #: PP + allicin-CHX vs PP control, PP + allicin at 24/48/72 h (p<0.01); vs PP + CHX at 24/48/72 h (p<0.05). ƒ: PP + CHX vs PP control, PP + allicin at 24/48/72 h (p<0.01). φ: PP + vancomycin vs PP control at 24/48/72 h (p<0.01); vs PP + allicin at 24/48/72 h (p<0.05). δ: DM+ vs PP control, PP + allicin at 24/48/72 h (p<0.01). ζ: PP + vancomycin vs PP + allicin-CHX 24 h (p<0.05); vs CHX at 72 h (p<0.05). Results. At each time point, the largest inhibition halos were observed for the mixture allicin-CHX (p<0.05). CHX was more effective than vancomycin (p<0.05), and q Fig 3. Bacterial colonization of the meshes allicin allowed bacterial colonization of the inhibition after 72 h of the contamination (x2,000). zone 24 h after Sa contamination [Figs. 1, 2]. 3.15 x 10 CFU/mesh 2.19 x 10 CFU/mesh As observed with DM+, none of the PP meshes presoaked in vancomycin, CHX or allicin-CHX were colonized by bacteria on their surface. In contrast, the materials treated with saline and allicin showed the highest bacterial counts (p<0.01) [Fig. 3]. Vancomycin was the most innocuous treatment towards fibroblasts (p<0.001). Although allicin and CHX were shown to be cytotoxic, their toxicity was significantly reduced when combined (p<0.001) [Figs. 4, 5]. 9 7 u Fig 4. Percentage of the alamarBlue® reduction from cells treated with the antibacterial solutions vs non-treated cells, after 24h. #: Fb + vancomycin vs Fb + allicin, Fb + CHX, Fb + allicin-CHX (p<0.001). ƒ: Fb + allicin-CHX vs Fb + allicin, Fb + CHX (p<0.001). u Fig 5. Cultured skin fibroblasts after exposition to the antibacterial treatments for 24h (x100). Conclusion. The pretreatment of prosthetic meshes with antiseptics such as CHX, alone or combined with allicin, inhibited bacterial adhesion to the mesh surface in vitro over at least 72 h, as did vancomycin and DM+. The antiseptic combination allicin-CHX could be useful as prophylactic drugs to prevent bacterial adhesion to a prosthetic material without compromising the initial stages of hernia repair.