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Molecules 2020, 25, 2023 9 of 14 therefore surface functionalization before application is important to prevent aggregation and enhance their killing efficacy in biofilms, and reduce their cellular uptake and cytotoxicity. So natural and environmentally benign compounds, such as extracellular fungal compounds used in this work for the nanoparticle synthesis, could constitute effective stabilizing agents enhancing bacterial killing in biofilms and reducing cytotoxicity of silver nanoparticles [14]. According to the results, the biogenic nanoparticles synthesized in this work were able to eradicate the E. coli and C. albicans biofilms (Figure 10). Future works will include optimization of these biofilm eradication experiments, eMvoalelcualetsi2n0g20t,h25e, xeffFOecRtPoEfERdRiffEeVrIeEnWt treatment condition for biofilms, including incubation tim1e0 aofn1d5 nanoparticle dose. Figure 9. Confocal Raman spectroscopy images obtained by combination of stretching C-H bands from Molecules 2020, 25, x FOR PEER REVIEW 11 of 15 mFigcuroroerg9.anCiosmnfo(rceadl)RaanmdatnhespAegc-tNrobscaonpdysaimssaignesedobtotatihnedPcbhyNcPosm(bliunea)t:io(an)oEf.csotlrie,t(cbh)inCg.aClb-iHcanbsa.nds from microorganism (red) and the Ag-N bands assigned to the PchNPs (blue): (a) E. coli, (b) C. albicans. 2.7. Antibiofilm Activiy of PchNPs The biofilm biomass of E. coli and C. albicans was measured with crystal violet stain. Comparison of the basal measures (biofilm without PchNP) with treated biofilm allowed analyzing the nanoparticles ability to eradicate biofilms. Reports on the antibiofilm activity of silver nanoparticles have been promising; showing that exposure to silver nanoparticles produced changes to the structural biofilm conformation of Candida albicans [27] and effectively inhibited E. coli and Pseudomonas aeruginosa biofilm formation [24]. Unfortunately, silver nanoparticles are prone to aggregation, reducing their antimicrobial efficacy and therefore surface functionalization before application is important to prevent aggregation and enhance their killing efficacy in biofilms, and reduce their cellular uptake and cytotoxicity. So natural and environmentally benign compounds, such as extracellular fungal compounds used in this work for the nanoparticle synthesis, could constitute effective stabilizing agents enhancing bacterial killing in biofilms and reducing cytotoxicity of silver nanoparticles [14]. According to the results, the biogenic nanoparticles synthesized in this work were able to eradicate the E. coli and C. albicans biofiFlmigsu(rFei1g0u. rAec1ti0v)i.tyFuoftuPrcehNwPosrakgsawinisltl(ian)c: lEu.dcoelioapntdim(bi)z:aCti.oanlbiocafnths beisoefiblmiosf.ilDmifferaendticleattieorsnrexppresreinmt ents, Figure 10. Activity of PchNPs against (a): E. coli and (b): C. albicans biofilms. Different letters represent evalusiagtninifigcatnhtediefffefreecntceosfadt piffPDF Image | Biofilm Eradication Using Biogenic Silver Nanoparticles
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