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A rapid screening method to evaluate the impact of nanoparticles on macrophages

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Academic year: 2021

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(11) . "!&"   Plate J774.1. Add NP in. Add NP in 24 h. Day -1. time. 23 h. 0h. staining of cells for flow cytometry. untreated. NP1 (AuNP). NP2 (NH2PVA). NP3 (NH2PVA-noCy5). NP4 (NH2PVA-AuNP). NP5 (NH2PVA-AuNP-noCy5). collect supernatant for ELISA.

(12). . . . . Toxicity/uptake staining    . . & )%*. )*%!*%. ( )--. +**",(%

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(27). .  LPS-treated cells.      . "

(28) ",. NP added at 0 h. LPS-treated cells. staurosporine-treated cells.  . . . . Plate 2 incubated at 4°C. NP added after 23 h. . "!&" 0)!'! !% !! ! H. I0ED %*'(! +(*/$$"&EDD4$"&MJ</$$$,<',,'%($,/*+"&$$,!/$$+8,*'.*&" !, "&-,"'&5 %"-% /+ *($ /",! EDD 4$ *+! %"-% >+)-*+? '* ID 4$ *+! %"-% & ID 4$ < '&,"&"& %"-%>',+?8'"&!"",,".-(,#5($,F/+"&-,,HA8,*FG!"&-,"'&5ID4$.

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

(30) . 40. *. 20. 60 40 20. 80 60 40 20. tre un. tre. NH. NH. un. tre. 0. at e A N u d 2P H N VA 2 P P -A V A u st NP au ro. 0. at Au e d N 2P H N VA 2 P P -A V A u st NP au ro. 0. un. *. at Au e d N 2P H N VA 2 P P -A V A u st NP au ro. 60. 80. NH. 80. 100 % of PI+/AnnV-. 100 % of PI+/AnnV+. % of PI-/annV+. 100. "!&" 1)!

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(32) 100. 1h on untreated cells. 50. 1h on stauro-treated cells 24h. P -A. uN. VA 2P. NH. 2P. VA. NH. N Au. at un. tre. P. 0. ed. % of Cy5+ cells. . "!&" 2)!&4+

(33) !0"&"!! !"   KKH8E%*'(! +/*"&-,'*FH!/",!'-,>/!",?'*/",!> *1?+,-*'+('*"&'*"& '*  F('$1%*'*,!$+,!'-*'"&-,"'&8+('+",".'&,*'$5%*'(! +/*"&-,'*FH!/",! '* F('$1%*>$#?8(,#/+&$1+1$'/1,'%,*1,* ,"& '&$".>&&0"&; '-$< & ,".?$$+8*&, '1IQ$$+"+(",8!**(*+&,+%&S',$+,H"&(&&, 0(*"%&,+8 . .

(34) J774.1. 40 20 0. un. tre. 4 2 0. tre. at e NH A NH uN d ho 2 P m ho V 2 P P o- m A V N o- - A A H N u 2 P H NP VA 2 P -A V A u st NP au ro. 0. un. MFI (fold change). 20. 60. at e NH A NH uN d ho 2 P m ho V 2 P P o- m A V N o- - A A H N u 2 P H NP VA 2 P -A V A uN P LP S. 40. 80. 6. tre. 60. C. 100. un. 80. B. at e NH A NH uN d ho 2 P m ho V 2 P P o- m A V N o- - A A H N u 2 P H NP VA 2 P -A V A uN P. % of live cells. 100. % of Cy5+ cells. A. MSC-2. 80 60 40 20. NH N 2 -P H u n EG2 PV tre NH /P A Aate C 2 VA u d C C OO - P E A N P O O H G uN O O -P A P H H V u -P - A N EG PE A P /P G uN VA Au P AUNP N P. 0. E. 3 MFI (fold change). 100. 2 1 0. NH N 2 -P H u n EG2 PV tre NH /P A Aate C 2 VA u d C C OO - P E A N P O O H G uN O O -P A P H H V u -P - A N EG PE A P /P G uN VA Au P AUNP N R P 84 8. % of live cells. D. "!&" 3)&!!%!&("!"!#!!"! "

(35) $! !!& * KKH8E><?%*'(! '*<F%1$'"+-((*++'*$$$"&+>5?/*"&-,'*FH!,GKA/",! "*&,'*%-$,"'&+'-'*('$1%*+'*$'/1,'%,*1&$1+"+8,(-*&, '$".>&&0"& Q. ; '-$<& ,".?$$+8,-(*&, '1I $$+&,(- <. %"&$-'*+&"&,&+",1> 6 '$!& '%(*,'-&,*,+%($?8<7!**(*+&,+%&S',$+,G"&(&&, 0(*"%&,+857!**(*+&,+%&',*"($",+''&0(*"%&,8.

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