Journal of Shenyang Pharmaceutical University

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1 Journal of Shenyang Pharmaceutical University Vol. 29 No. 12 Dec p PEG HeLa 1 1* monomethoxy polyethylene glycol-polyactic coglycolic acid-nanoparticles mpeg-plga-nps HeLa mpeg-plga-nps mpeg-plga-nps zeta mpeg-plga-nps -6 coumarin 6 HPLC HeLa mpeg-plga-nps ± 4. 3 nm Zeta ± mv PLGA- NPs 35. 5% mpeg-plga-nps 92. 5% PLGA-NPs 3 HeLa mpeg-plga-nps PLGA-NPs 2 HeLa PLGA-NPs mpeg-plga-nps mpeg- PLGA-NPs HeLa mpeg-plga-nps PLGA-NPs P < HeLa R 94 A human papilloma virus HPV mucoadhesive polyethylene glycol PEG mucus penetrating particles MPP polyactic-coglycolic acid PLGA PEG PLGA 1 nanoparticles NPs PLGA-NPs mpeg-plga-nps 7-6 coumarin 6 PLGA-NPs mpeg-plga-nps HeLa ZR2009CM qlf _hi@ 126. com * Tel ldh@ ytu. edu. cn

2 Agilent 1100 Agilent Nicomp 380ZLS PSS A 5 g L - 1 NICOMP JY92-Ⅱ 10 g L - 1 B 20 g L - 1 S g L Olympus FV C PM Olympus B mg L - 1 C Nikon Mircro 21R - 20 min Thermo Forma Series II 750 nm PM Thermo Synergy HT 1 d BioTek Nalge Nunc 2. 3 SEM PLGA-NPs mpeg-plga-nps 5% CO S-4800 SEM PM 3 3 d - PLGA Mr = m LA m GA = PLGA-NPs mpeg-plga-nps 1 ml - - mpeg-plga 1 g L - 1 PM 1 mg PBS 1 mpeg ml 37 1 h 4 - pig gastric mucin PM -6 Sigma RPMI Medium 1640 GiBco MTT r min min - PM 1 PM % = PM PM HeLa 100% ml 1 g L ml 2. 1 mpeg-plga 25 mg 2 ml ph r min h 30 ml 1 ml r min h 0. 2% 60 HPLC mpeg-plga-nps PLGA-NPs mg -6-6 % = % 2 mpeg-plga-nps PLGA-NPs 2. 6 NICOMP 380 ZLS HeLa Zeta MTT assay -6 HeLa HeLa 10% RPMI h PLGA-NPs mpeg-

3 12 PEG HeLa 919 PLGA-NPs mlpbs μl HPLC g L μl 4 h 96 BSA dimethyl sulfoxide DMSO 150 μl 10 min BSA nm μg 10 μl CBB 200 μl % = 5 min % 3 nm BSA HeLa HeLa μl Lab - Tek r min min h 5 μl -6 5 μl CBB 200 μl PLGA-NPs mpeg-plga-nps 100 μl 5 min 595 nm 100 mg L - 1 C mg L h HeLa PBS 3 4% 30 min % = /520 nm / HeLa 100% HeLa HeLa 24 h3. 1 zeta -6-6 PLGA-NPs mpeg-plga-nps 200 μl PLGA-NPs mpeg-pl- 200 mg L - 1 C6 GA-NPs zeta mg L h mg L Bradford 3 24 h MTT μl 10 μl 3 1 PBS ml Table 1 Physico-chemical properties and PM anti-adhesive rate of nanoparticles n = 3 珔 x ± s 1 PM n = 3 珔 x ± s Formulation d /nm Polydispersity index V zeta /mv EE /% PM anti-adhesive rate /% PLGA-NPs ± ± ± ± ± 1. 9 mpeg-plga-nps ± ± ± ± ± 1. 4 mpeg-plga-nps mpeg-plga-nps zeta PLGA-NPs PEG 10 mv mpeg-plga PEG -6 > 70%

4 PLGA-NPs mpeg-plga-nps 3. 2 scanning electron microscope SEM SEM mpeg-plga-nps 1 mpeg-plga-nps 100 nm Nicomp 380ZLS 3. 5 HeLa 24 h -6 < 12% 6 h < 8% -6 MTT PLGA-NPs mpeg-plga-nps -6 HeLa 24 h 3 Fig. 1 1 SEM images of mpeg-plga nanoparticles mpeg-plga SEM 3. 3 PM 50 ~ 500 mg L - 1 Fig. 3 A = ρ r 2 = PLGA-NPS mpeg-plga-nps Fig. 2 In vitro release of coumarin-6 loaded PLGA and PEG-PLGA nanoparticles n = 3 珔 x ± s 2-6 PLGA-NPs mpeg-plga-nps n = 3 珔 x ± s In vitro inhibition of PLGA and mpeg-plga nanoparticles in HeLa cells n = 3 珔 x ± s RSD 0. 5% 3 PLGA-NPs mpeg-plga-nps HeLa RSD 1. 1% n = 3 珔 x ± s 1 PLGA- HeLa NPs PM 35. 5% mpeg-pl- 80% GA-NPs PM 92. 5% PLGA-NPs 3 mpeg-plga-nps 200 mg L - 1 HeLa 24 h > 90% 3. 4 HPLC HeLa 4 HeLa -6 2 PLGA-NPs 2 h HeLa mpeg-plga-nps HeLa mpeg-plga-nps PL- GA-NPs HeLa 4 h 6 h He- La 3. 7 HeLa BSA 0 ~ 5 μg A = m r 2 =

5 12 PEG HeLa 921 HeLa 5 4 Fig. 4 PLGA-NPs mpeg-plga-nps Fig. 5 Uptake of PLGA and mpeg-plga nanoparticles in HeLa cell line n = 3 珔 x ± s A Coumarin-6 solution B PLGA-NPs C mpeg-plga-nps Confocal images of HeLa cells coincubated withnanoparticles HeLa PLGA-NPs mpeg-plga-nps 5 HeLa PLGA-NPs mpeg-plga-nps n = 3 珔 x ± s 5 HeLa mpeg-plga-nps PLGA-NPs 6 h HeLa PLGA-NPs 1 CU Y BOOTH C J SALTZMAN W M. In vivo distribution of surface-modified PLGA nanoparticles following 2. 49% mpeg-plga-nps 4. 71% PLGA-NPs 2 intravaginal delivery J. J Control Release J. 4 b. mpeg-plga-nps HeLa mpeg- PLGA-NPs PLGA- NPs PEG PEG a. mpeg-plga-nps 3 LAI S K O'HANLON D E HARROLD S et a1. Rapid PLGA-NPs transport of large polymeric nanoparticles in fresh undiluted human mucus J. Proceedings of National Acade- PEG my of Sciences CU Y SALTZMAN W M. Controlled surface modification with poly ethylene glycol enhances diffusion of PL- 10 mpeg-plga-nps GA nanoparticles in human cervical mucus J. Molecu-

6 lar Pharmaceutics YONCHEVA K LIZARRAGA E IRACHE J M. Pegylated nanoparticles based on poly methyl vinyl ether-comaleic anhydride preparation and evaluation of their bioadhesive properties J. Eur J Pharm sci YANG M LAI S K HANES J et al. Biodegardable nanoparticles composed entirely of safe materials that rapidly penetrate human mucus J. Angew Chem Int Ed J PLGA J CONE R A. Barrier properties of mucus J. Adv Drug Deliv Rev PAMUJULA S HAZARI S BOLDEN G. Cellular delivery of PEGylated PLGA nanoparticles J. Journal of Pharmacy and Pharmacology A549 J SADZUKA Y KISHI K HIROTA S et al. Effect of polyethyleneglycol PEG chain on cell uptake of J. PEG-Modified liposomes J. Journal of Liposome Research ZHANG Y KOHLER N ZHANG M Q. Surface modification of superparamagnetic magnetite nanoparticles and their intracellular uptake J. Biomaterials 2002 Anti-adhesive activity and uptake by HeLa cells of mpeg- PLGA nanoparticles QIN Li-fang 1 LIN Dong-hai 1* LI Gang 2 XIE Xin-xin 1 WANG Jun-teng 1 WEN Zhen 1 1. School of Pharmacy Yantai University Yantai China 2. School of Life Sciences Yantai University Yantai China Abstract Objective To prepare mpeg-plga nanoparticles discuss its properties and in vitro anti-adhesive activity and investigate the cellular uptake of them in human cervical cancer cells HeLa. Methods mpeg- PLGA-NPs w ere prepared by a solvent diffusion method. The particle size distribution and Zeta potential of nanoparticles w ere measured by light scattering. The anti-adhesive activity of nanoparticles w as evaluated by pig gastric mucin PM binding experiments. Coumarin 6 was incorporated into nanoparticles as fluorescent marker. The cellular uptake of PLGA-NPs and PEG-PLGA-NPs by HeLa cell lines w ere determined by laser scanning confocal microscope and HPLC. Results The mean size of mpeg-plga-nps was ± 4. 3 nm Zeta potential was ± mv. The PM anti-adhesive rate of PLGA-NPs and PEG-PLGA- NPs was 35. 5% and 92. 5% respectively. The anti-adhesive activity of mpeg-plga-nps w as three times higher than PLGA-NPs. All nanoparticles showed low cytotoxicity. At the same incubation time the uptake of mpeg-plga-nps was two times higher than that of unmodified PLGA-NPs. The cellular uptake of PL- GA-NPs and mpeg-plga-nps w as dependent on the incubation time. Conclusions mpeg-plga-nps has much higher anti-adhesive activity. The uptake of mpeg-plga-nps by HeLa cells is much higher compared to the PLGA-NPs P < The mpeg-plga-nps shows better cell affinity than PLGA-NPs. Key words mpeg-plga nanoparticle anti-adhesive activity HeLa cell line cellular uptake