Membrane Structure Alteration of DOPC Liposome Induced by Interaction with Gene Carrier Polyethyleneimine†

doi:10.7503/cjcu20170273

Abstract

Abstract:

To get a better understanding on the molecular basis of the cytotoxicity of PEI which has been considered as “golden standard” for polymeric gene delivery carriers. Dynamic light scattering, fluorescence spectra, zeta-potential measurement and isothermal titration calorimetry were conducted to reveal the mechanism of interaction between PEIs(average molecular weight of 25000, 10000 and 1800) and 1,2-dioleoyl-sn-glycero-3-phosphatidylcholine(DOPC) liposome. The influence on the polarity of microenvironment and the permeability of liposome bilayer were also investigated. The result showed that PEI bound to DOPC vesicles via hydrogen bond or van Waals interactions between the amide groups and the phosphorylcholine heads. The complex formation with PEI induced aggregation and increase in zeta potential of liposomes at low PEI concentration up to 0.075 mg/mL. A further increase in PEI concentration made little change on the surface potential, however reduced the aggregation of the vesicle due to the repulsion between the adsorbed PEI chains. PEI binding decreased the packing density of hydrocarbon chain of lipid molecules and the hydrophobicity in the bilayer membrane, and thus resulted in an enhanced permeability of calcein and quercetin through the membrane. The polymer size played an important role in PEI-DOPC liposome interaction. PEI with higher molecular weight was more favorable to interact with DOPC and more efficient to perturb the structural properties of the membrane.

Key words: Polyethyleneimine, Phosphatidylcholine, Liposome, Interaction, Membrane structure

CLC Number:

O648

TrendMD:

WEI Yanshan, WEI Bangzhi, HUANG Aimin, MA Lin. Membrane Structure Alteration of DOPC Liposome Induced by Interaction with Gene Carrier Polyethyleneimine^†[J]. Chem. J. Chinese Universities, 2017, 38(12): 2271.

Figures/Tables 9

Fig.1 PEI concentration dependence of hydrodynamic diameter(Dhy) of DOPC LUVs in PBS cDOPC=50 μmol/L.

Fig.2 Fluorescence spectra of DOPC LUVs labeled with NBD-PE and Rh-PE in PBS(pH=7.4) containing 0(a), 0.05(b) and 0.1(c) mg/mL PEI25kInsert: relative intensity of fluorescence emission of NBD-PE and Rh-PE(F520/F590) in DOPC LUVs in PBS containing different concentrations of PEI25k. λex=463 nm, c(DOPC)=50 μmol/L.

Fig.3 Variation of zeta potential of DOPC LUVs in PBS(pH=7.4) containing PEIs c(DOPC)=50 μmol/L.

Fig.4 ITC profiles for titration of PEI25k into DOPC LUVs in PBS solution at 25 ℃ and pH=7.4(A) Exothermic heat release upon injection of PEI25k into DOPC LUVs dispersion; (B) integrated heat data associated with the interaction of PEI25k and DOPC LUVs. c(PEI25k)=5.0 mg/mL, c(DOPC)=750 μmol/L.

Fig.5 Schematic representation of concentration-dependent interaction of PEI with DOPC LUVs

Fig.6 PEI concentration dependence of ANS fluorescence emission maximum in PBS(pH=7.4)in the absence(dash line) and presence(solid line) of DOPC LUVsλex=388 nm; c(ANS)=2.5 μmol/L; c(DOPC)=50 μmol/L.

Fig.7 Time course leakage of calcein from DOPC LUVs in PBS containing PEI25k c(DOPC)=2.5 μmol/L.

Fig.8 PEI concentration dependence of maximum leakage of calcein from DOPC LUVs c(DOPC)=2.5 μmol/L.

Fig.9 PEI concentration dependence of fluorescence intensity of quercetin at 564 nm(F564)(A) In the absence(solid line) and presence(dash line) of DOPC LUVs in PBS(pH=7.4); (B) difference in the fluorescence emission in the absence and presence of DOPC LUVs. λex=465 nm, c(DOPC)=50 μmol/L, c(Que)=5 μmol/L.

References 36

[1]	Yin H., Kanasty R. L., Eltoukhy A. A., Vegas A. J., Dorkin J. R., Anderson D. G., Nat. Rev. Genet., 2014, 15, 541—555
[2]	Giacca M., Zacchigna S., J. Control. Release, 2012, 161, 377—388
[3]	Tiera M. J., Shi Q., Winnik F. M., Fernandes J. C., Curr. Gene Ther., 2011, 11, 288—306
[4]	Neu M., Fischer D., Kissel T., J. Gene Med., 2005, 7, 992—1009
[5]	Sawant R. R., Sriraman S. K., Navarro G., Biswas S., Dalvi R. A., Torchilin V. P., Biomaterials,2012, 33, 3942—3951
[6]	Yamano S., Dai J., Hanatani S., Haku K., Yamanaka T., Ishioka M., Takayama T., Yuvienco C., Khapli S., Moursi A. M., Montclare J. K., Biomaterials,2014, 35, 1705—1715
[7]	Chen L. N., Wang Y., Zhu Y., Sun Y. X., Wang Y. X., Chem. J. Chinese Universities, 2013, 34(3), 720—725
	(陈丽娜, 王颖, 朱莹, 孙一新, 王幽香. 高等学校化学学报, 2013, 34(3), 720—725)
[8]	Liu Y. J., Zhang P., Du J. W., Wang Y. X., Chem. J. Chinese Universities, 2016, 37(5), 1003—1009
	(刘亚杰, 张鹏, 杜建委, 王幽香. 高等学校化学学报, 2016, 37(5), 1003—1009)
[9]	Yang S., Guo Z., Yang X., Xie J., Robert J. L., Jiang D., Teng L., Chem. Res. Chinese Universities, 2015, 31(3), 401—405
[10]	Zhong D., Jiao Y., Zhang Y., Zhang W., Li N., Zuo Q., Wang Q., Xue W., Liu Z., Biomaterials,2013, 34, 294—305
[11]	Larsen A. K., Malinska D., Koszela-Piotrowska I., Parhamifar L., Hunter A. C., Moghimi S. M., Mitochondrion,2012, 12, 162—168
[12]	Moghimi S. M., Symonds P., Murray J. C., Hunter A. C., Debska G., Szewczyk A., Mol. Ther., 2005, 11, 990—995
[13]	Won Y. Y., Sharma R., Konieczny S. F., J. Control. Release, 2009, 139, 88—93
[14]	Gabrielson N. P., Pack D. W., J. Control. Release, 2009, 136, 54—61
[15]	Parhamifar L., Larsen A. K., Hunter A. C., Andresen T. L., Moghimi S. M., Soft Matter., 2010, 6, 4001—4009
[16]	Yasuhara K., Tsukamoto M., Tsuji Y., Kikuchi J., Colloids Surf. A: Physicochem. Eng. Aspects,2012, 415, 461—467
[17]	Sabín J., Vázquez-Vázquez C., Prieto G., Bordi F., Sarmiento F., Langmuir,2012, 28, 10534—10542
[18]	Haouz A., Mohsni S. E., Zentz C., Merola F., Alpert B., Eur. J. Biochem., 1999, 264, 250—257
[19]	Stewart J. C. M., Anal. Biochem., 1980, 104, 10—14
[20]	Torchilin V.P., Weissig V.; translated by Deng Y. H., Xu H., Liposomes: A Practical Approach, Chemical Industry Press, Beijing, 2007, 89—92
	(Torchilin V.P., Weissig V.著; 邓意辉, 徐晖[译]. 脂质体, 北京: 化学工业出版社, 2007, 89—92)
[21]	Struck D., Hoekstra D., Pagano R. E., Biochem., 1981, 20, 4093—4099
[22]	Morgan C. G., Thomas E. W., Yianni Y. P., Biochim. Biophys. Acta, 1983, 728, 356—362
[23]	Ross P. D., Subramanian S., Biochem., 1981, 20, 3096—3102
[24]	Pfau A., Schrepp W., Horn D., Langmuir,1999, 15, 3219—3225
[25]	Rao Z., Taguchi T., Colloids Surf. B: Biointerf., 2012, 97, 248—253
[26]	Lakowicz J.R., Principles of Fluorescence Spectroscopy, 2nd Ed., Plenum Press, New York, 1999, 185—486
[27]	Montesano G., Bartucci R., Belsito S., Marsh D., Sportelli L., J. Biophys., 2001, 80, 1372—1383
[28]	Marsh D., J. Biophys., 2001, 81, 2154—2162
[29]	Tribet C., Vial F., Soft Matter., 2008, 4, 68—81
[30]	Tirosh O., Barenholz Y., Katzhendler J., Priev A., J. Biophys., 1998, 74, 1371—1379
[31]	Kozlova N. O., Bruskovskaya I. B., Okuneva I. B., Melik-Nubarov N. S., Yaroslavov A. A., Kabanov V. A., Menger F. M., Biochim. Biophys. Acta, 2001, 1514, 139—151
[32]	Evans K. O., Laszlo J. A., Compton D. L., Biochim. Biophys. Acta, 2014, 1838, 445—455
[33]	Tammela P., Laitinen L., Galkin A., Wennberg T., Heczko R., Vuorela H., Slotte J. P., Vuorela P., Arch. Biochem. Biophys., 2004, 425(2), 193—199
[34]	Wolfbeis O. S., Begum M., Geiger H., Z. Naturforsch., 1984, 39 b , 231—237
[35]	Falkovskaia E., Sengupta P. K., Kasha M., Chem. Phys. Lett., 1998, 297, 109—114
[36]	Sengupta P. K., Kasha M., Chem. Phys. Lett., 1979, 68(2/3), 382—385