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Pharmaceutics 2020, 12, 1044 12 of 19 increasing intensity in the fluorescence of the bacteria treated with AgNPs with increased time while the untreated bacteria had a weak fluorescence (Figure 7). In a similar manner, the fluorescence of the treated bacteria increased with increased concentration of AgNPs in a 1 h exposure period (Figure 8). TPhaurms,acietuwticass20d2e0,d1u2,cxedFOtRhaPtEAERgRNEPVsIEiWnduced an excess generation of ROS in multidrug resistant (M12DofR2)0. Pharmaceutics 2020, 12, x FOR PEER REVIEW 12 of 20 This was based on a time and concentration dependent manner. Figure 7. Changes of reactive oxygen species (ROS) production in AgNPs-treated multidrug-resistant Figure 7. Changes of reactive oxygen species (ROS) production in AgNPs-treated multidrug-resistant FPisgeudreom7.onCahsanagereusgoifnroesactaivt edoixffyegrentspteimcies (iRntOerSv) aplrsoduuncdtieorn filnuAorgeNscPesn-ctreeamteidcrmosuclotipdyruwg-irtehsis×t4a0n0t Pseudomonas aeruginosa at different time intervals under fluorescence microscopy with ×400 Pmsaeugdnoifmicoantaios n.aerNuogtineso:sa(Aa)t tdhieffeurenntrteattimede Pin. taeerrvuaglsinousna dwerithflouuotreosbcesnercveabmleicrfoluscoorepsycenwcieth. (B×4–0F0) magnification. Notes: (A) the untreated P. aeruginosa without observable fluorescence. (B–F) mFluagonreifsiceanticoeno. bNseorvteast:io(nAo)f the buancterreiatetrdeaPte.daweriuthgiAnogsNa Pws iatthdouiftferoebnsteprvoainbtlse oflu0.o5r,e0s.c7e5n, c1e, .1.5(Ba–nFd) Fluorescence observation of the bacteria treated with AgNPs at different points of 0.5, 0.75, 1, 1.5 and 2 F2luho,rreesscpenecteivoeblsye,rvinadtiiocnatoinfgthtehbaatcAtegriNaPtrseaitneduwceithROASgNpPrsodaut dctiifofenreint paotiinmtse-odfe0p.5e,n0d.7e5n,t1m, 1a.5nnaenrd) h, respectively, indicating that AgNPs induce ROS production in a time-dependent manner) adapted 2adha,ptredspwecittihveplye,rminidssicioantinfgrotmhat[1A12g]N, PDsovinedMucediRcOalSPprerossd,u2c0ti1o9n. iAnbbaretivmiaet-idoneps:enAdgeNntPsm—asninlveerr) with permission from [112], Dove Medical Press, 2019. Abbreviations: AgNPs—silver nanoparticle; andanapotpeadrtiwcliet;hROpeSr—mrisesaicotnivefroxmyg[e1n12sp],ecDieosv.e Medical Press, 2019. Abbreviations: AgNPs—silver ROS—reactive oxygen species. nanoparticle; ROS—reactive oxygen species. Figure 8. Changes of ROS production in multidrug-resistant Pseudomonas aeruginosa exposed to different Figure 8. Changes of ROS production in multidrug-resistant Pseudomonas aeruginosa exposed to concentrations of AgNPs under fluorescence microscopy with ×400 magnification. Notes: (A) the F d i i g f f u e r r e e n 8 t . c Co n h c a e n n g t e r a s t i o o f n s R Oo f S A p g r No d P u s c u t i n o d n e r i n f l u m o u r l e t s i c d e r n u c g e - r me s i i c s r t o a s n c t o P p s y e u w d i o t mh o × n 4 a 0 s 0 a me r a u g g n i n i o f i s c a a t e i o x n p . o s N e o d t e t s o : untreated P. aeruginosa without observable fluorescence. (B–E) Fluorescence observation of the bacteria d ( A i f ) f e t r h e e n t u n c o t r n e c a e t n e d t r a P t i . o a n e s r u o g f i n A o s g a N w P i s t h u o n u d t e o r b f s l u e r o v r a e b s c l e e n f c l u e o m r e i s c c r e o n s c c o e . p ( y B w– E i t ) h F × l u 4 o 0 r 0 e s m c e a n g c n e i f i o c b a s t e i o r n v . a t N i o o n t e o s f : exposed to 5.625, 11.25, 22.5 and 45 μg/mL AgNPs, respectively, adapted with permission from [112], ( t Ah e ) b t h a c e t e u r n i a t r e e x a p t e o d s e P d . t a o e r 5 u . 6 g 2 i n 5 o , s 1 a 1 . w2 5 i t , h 2 o 2 u . 5 t a o n b d s e 4 r 5 v μ a b g l / e m f L l u A o g r e N s c P e s n , c r e e s . p ( B e c – t E i v ) e F l y l u , o a r d e a s p c t e e n d c e w o i t b h s p e r e v r a m t i i o s s n i o o n f Dove Medical Press, 2019. Abbreviations: AgNP, silver nanoparticle; ROS, reactive oxygen specie. t f h r o e m b a [ c 1 t 1 e 2 r ] i , a D e o x p v e o s M e d e d t o i c 5 a . l 6 P 2 r 5 e , s 1 s 1 , . 2 2 0 5 1 , 9 2 . 2 A . 5 b a b n r d e v 4 i 5 a t μ i o g n / ms : L A A g N g N P , P s s i , l v r e e s r p n e a c n t i o v p e a l y r t , i a c l d e a ; p R t O e d S , w r e i t a h c t p i v e e r m o x i s y s g i o e n n fsrpoemcie[1. 12], Dove Medical Press, 2019. Abbreviations: AgNP, silver nanoparticle; ROS, reactive oxygen specie. 4.3. DNA Damage 4.3. DNA Damage The DNA of any organism stores genetic information about the cell. Damage to the DNA can eitheTr hresuDlNt iAn mofuatantyioonrgoranceislml destaothre.sCgueineettaicl. i[n1f1o3r]mstautdioinedatbhoeumt tehcehacnelils.mDoafmthageebatocttehreiciDdNalAeffceacnt eoifthAerurNesPusltoinmEu.tactoiloin(oArTcCelCl de1a1t7h7.5C) utihertoaulg. h[11t3r]asntsucdripedtotmheicmaencdhanpirsomteofmthice baapcpteroriaccidheasl.efTfehcet o a n f t i A b a u c N t e P r s i a l o mn e E c h . a c n o i l s i m ( A o f T AC C u N 1 P 1 s 7 w7 5 a ) s t f h o r u o n u d g h t o t b r e a n d s u c e r i t p o t o ( mi ) i a c c h a n a n d g e p r i n o t me o e m m i b c r a a n p e p r p o o a t c e h n e t i s a . l a T n h d e ainhtibaitcitoenrioafl AmTePchaaseniascmtivoiftiAesutNhaPtslewdatsofaoudnedcretoasbeeinduAeTtPol(eiv)eal cthuasngaereindumcteimonbrinanmeeptaobteonlitsimal andPDF Image | Bactericidal Antibacterial Mechanism of Plant Nanoparticles
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