,Sng Jun Prk,Je Ho Lee,

aLaboratory of Tissue Engineering,Korea Institute of Radiological and Medical Science,Seoul 139-706,Republic of Korea

bLaboratory of Molecular Medicine,College of Medicine,Dankook University,Seoul 100-380,Republic of Korea

Preparation of chitosan–TPP nanoparticles and their physical and biological properties

Eunsun Leea,Sang Jun Parka,Jae Ho Leeb,Min Sup Kima, Chun-Ho Kima,*

aLaboratory of Tissue Engineering,Korea Institute of Radiological and Medical Science,Seoul 139-706,Republic of Korea

bLaboratory of Molecular Medicine,College of Medicine,Dankook University,Seoul 100-380,Republic of Korea

A R T I C L E I N F O

Article history:

Available online 25 November 2015

Chitosan

Tripolyphosphate

Nanoparticles

DDS

Over the last few decades,several polymeric nanoparticles using biomaterials have been extensively developed for use as drug delivery systems(DDS)and for other medical applications. Therapeutic nanoparticles have the advantage of controlled drug-targeting delivery and release,which improves the stability,enhances the effciency and reduces the side effects of the drug[1].Chitosan is a versatile polymer that has been used in drug delivery system as nanoparticles because of biodegradable,biocompatible,and low toxicity[2].The positive amine group of chitosan can perform the stable nanoparticles with anionic material such as gene,drug,protein and small molecule by electrostatic interaction.

In this study,nanoparticles were prepared by ionic gelation between chitosan and tripolyphosphate(TPP).Their morphologies were observed using scanning electron microscope.Size,polydispersity and zeta potential of nanoparticles were perceived using Zeta-sizer.To evaluate their cell viability,human dermal fbroblasts were treated with various concentrations of chitosan–TPP nanoparticles.After 24 hrs of culture,cell viability was evaluated by CCK-8 assay and normalized against the untreated cells(n=4).To observe cellular uptake of siGFP loaded chitosan–TPP nanoparticles in MDAMB-231/siGFP cells,cells were strained with blue-DAPI for nuclei, green-GFP for siGFP,respectively.The stained cells were observed using a confocal laser-scanning microscope.The biodegradable and biocompatible chitosan–TPP nanoparticles for DDS were formed directly by ionic gelation.Chitosan–TPP nanoparticles formed a sphere shape.Their size and zeta potential were 150～250 nm and 30±5.3 mV,respectively.Size, polydispersity index and zeta potential of chitosan–TPP nanoparticles depended on the molecular weight of chitosan, concentration of TPP,weight ratio of TPP to chitosan,reaction temperature and pH.When nanoparticles concentration was 250 μg/ml,cell viability on human dermal fbroblasts was above80%.Cellular uptake of nanoparticles was signifed using fuorescence microscope on MDA-MB-231.Cellular uptake of chitosan–TPP nanoparticles were increased in proportion to concentration of nanoparticles.In conclusion,chitosan–TPP nanoparticles may be anticipated to contribute to the advancement of gene and drug delivery system.

Fig.1–TEM photograph(A),IR spectra(B),cell viability(C),and cellular uptake(D).

Acknowledgements

This research was supported by the National R&D Program through the Korea Institute of Radiological and Medical Sciences(grant code:1711021779)funded by the Ministry of Science,ICT and Future Planning.

R E F E R E N C E S

[1]Wang JJ,Zeng ZW,Xiao RZ,et al.Recent advances of chitosan nanoparticles as drug carriers.Int J Nanomedicine 2011;6: 765.

[2]Dodane V,Vilivalam VD.Pharmaceutical applications of chitosan.Pharm Sci Technolo Today 1998;1:246.

*E-mail address:chkim@kcch.re.kr.

Peer review under responsibility of Shenyang Pharmaceutical University.

http://dx.doi.org/10.1016/j.ajps.2015.11.065

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/).