Thoms Kissel

aSchool of Pharmacy,Shenyang Pharmaceutical University,Wenhua Road 103,110016 Shenyang,China

bDepartment of Pharmaceutics and Biopharmacy,Phillips-Universit?t of Marburg,Ketzerbach 63,D-35032 Marburg,Germany

Layer-by-layer nanostructured protein loaded nanoparticles:A feasibility study using lysozyme as model protein and chitosan as coating material

Cuifang Caia,*,Shirui Maoa,Thomas Kisselb

aSchool of Pharmacy,Shenyang Pharmaceutical University,Wenhua Road 103,110016 Shenyang,China

bDepartment of Pharmaceutics and Biopharmacy,Phillips-Universit?t of Marburg,Ketzerbach 63,D-35032 Marburg,Germany

A R T I C L E I N F O

Article history:

Available online 25 November 2015

Chitosan

Nanoparticles

Lysozyme

Electrostatic interaction

Protein carrier

Layer-by-layer

Stabilization of proteins in delivery devices and design of appropriate protein carriers are major research issues due to the extreme sensitivity of proteins.Previously,negatively charged nanoparticles,consisting of poly(lactic-co-glycolic acid)(PLGA) and poly(styrene-co-4–styrene-sulfonate)(PSS),showed considerably high loading capacity for positively charged model protein lysozyme depending on the surface charge density of nanoparticles.Chitosan coated lysozyme loaded layer-bylayer protein delivery nanocarrier were prepared and the infuence of chitosan composition,concentration and initial protein loading on the release and stability of nanoparticles were investigated.

Nanoparticles were prepared by a solvent displacement method.Loading with oppositely charged model protein lysozyme and followed by chitosan coating were completed through electrostatic interaction between oppositely charged nanoparticle-protein,and nanoparticle-chitosan.Morphology of nanoparticles was investigated using scanning electron microscopy(SEM)and transmission electron microscopy(TEM). At a ratio of PLGA to PSS 10%and initial theoretical lysozymeloading 10%,the effect of chitosan(CS)(150 kDa)concentration on the properties of the lysozyme loaded nanoparticles is shown in Fig.1.The results indicate that the concentration of CS had a signifcant infuence on the particle size and zetapotential of all the resulting formulations investigated.More specifcally,the size of the CS-coated nanoparticles linearly increased upon addition of increasing amounts of CS from 244 nm to 606 nm(R2=0.9671)within the range of chitosan concentrations from 0 to 1.92 mg/ml.The SEM and AFM images of nanoparticles were shown in Fig.2.In addition,dissociation of lysozyme was dependent on polymer composite,irrespective of initial protein loading.Moreover,with this polymer coating more stable particles were obtained in PBS,without burst release within 24 hours.In light of these results,these novel polymer coated protein loaded nanoparticles with layerby-layer nanostructure could be suggested as promising new protein delivery systems with benefcial properties,i.e. protection of protein,controlled release,stability,and combined with potential properties of coated polymer.

Fig.1–Size(Z-Ave)(A)and zeta-potential(ZP)(B)of lysozyme-loaded nanoparticles after incubation with chitosan(150 kDa) solution at different concentrations(mean±S.D.,n=3)(A)and SEM images of nanoparticles at the chitosan concentration of 60(A)and 120 μg/ml(B),respectively.TEM images of uncoated nanoparticles(C)and coated nanoparticles(D)at the chitosan concentration of 120 μg/ml,respectively(B).

R E F E R E N C E S

[1]Osuna B,Vauthier C,Farabollini A,et al.Mucoadhesion mechanism of chitosan and thiolated chitosan-poly(isobutyl cyanoacrylate)core-shell nanoparticles.Biomaterials 2007;28:2233–2243.

[2]Zhang L,Sun M,Guo R,et al.Chitosan surface-modifed hydroxycamptothecin loaded nanoparticles with enhanced transport across Caco-2 cell monolayer.J Nanosci Nanotechnol 2006;6:2912–2920.

[3]Prego C,Fabre M,Torres D,et al.Effcacy and mechanism of action of chitosan nanocapsules for oral peptide delivery. Pharm Res 2006;23:549–556.

*E-mail address:bbpharmcai@sina.com.

Peer review under responsibility of Shenyang Pharmaceutical University.

http://dx.doi.org/10.1016/j.ajps.2015.10.046

1818-0876/?2016 Production and hosting by Elsevier B.V.on behalf of Shenyang Pharmaceutical University.This is an open access article under the CC BY-NC-ND license(http://creativecommons.org/licenses/by-nc-nd/4.0/).