Chitosan Oligosaccharide/Salicylic Acid Nanoparticles for Doxorubicin Delivery

Transcription

1 / 1,2 2 2* 1* ( ) / / / MTT / / 16.92% µg ml 1 Zeta nm 48.6 mv Hep-G µg ml 1 10% / 8.52% 93.15% nm 33.6 mv ph / ph / / R943 B (2014) Chitosan Oligosaccharide/Salicylic Acid Nanoparticles for Doxorubicin Delivery HUANG Peiwen 1,2, CHEN Suping 2, WEI Xiaohong 2*, SHI Senlin 1* (1.College of Pharmaceutical Science, Zhejiang Chinese Medical University, Hangzhou , China; 2.Department of Pharmaceutical, School of Medicine, Hangzhou Normal University, Hangzhou , China) ABSTRACT: OBJECTIVE To investigate the probability of chitosan oligosaccharide (CSO)/salicylic acid (SA) nanoparticles to encapsulate alkalized doxorubicin and to evaluate the physicochemical properties and in vitro profile of drug delivery system. METHODS CSO/SA conjugates were synthesized by a reaction between amino-groups of CSO and carboxyl groups of SA using 1-ethyl-3-(3-dimethyla-minopropyl) carbodiimide as a coupling agent. The amino substitution degree(sd%) of CSO/SA was determined by a 2, 4, 6-trinitrobenzene sulfonic acid (TNBS) test. The CSO/SA blank nanoparticles were prepared by ultrasonic dispersion method. The critical aggregation concentration of nanoparticles was obtained by the fluorescence pyrene study. Dynamic light scattering method was used to determine the size and Zeta potential of nanoparticles. MTT assay was used to evaluate the cytotoxicity of nanoparticles. Alkalizated doxorubicin was used as the model drug, drug-loaded CSO/SA nanoparticles were prepared by dialysis method. RESULTS The amino substitution degree was 16.92%, and the critical aggregation concentration of blank nanoparticles was µg ml 1. The particle size and Zeta potential of blank nanoparticle were nm and 48.6 mv, respectively. The IC 50 of the nanoparticles on cell Hep-G2 was µg ml 1. The drug content of nanoparticles was 8.52% and the loading efficiency was 93.15% when the amount of drug fed was 10%, the particle size and Zeta potential of drug-loaded nanoparticles were nm and 33.6 mv, respectively. In vitro release results illustrated that drug-loading nanoparticles had ph sensitivity, and the drugs were released mostly in the way of erosion. CONCLUSION CSO/SA can encapsulate alkalized doxorubicin to form uniform, sustain drug delivery nanoparticles. The drug-loaded nanoparticles present ph-dependent property. CSO/SA nanoparticles may be a potential candidate to be a drug carrier material. KEY WORDS: chitosan oligosaccharide/salicylic acid; alkalized doxorubicin; nanoparticles; in vitro release [1] ( ) (2001R10059) Tel: huangpeiwen@sina.com * Tel: (0571) xiaohongwei1@foxmail.com Tel: (0571) pjstone@163.com Chin J Mod Appl Pharm, 2014 January, Vol.31 No.1 61

2 [2-4] [5-7] [8] 60 (Hydrotrope) [9-10] π π [11] [12-16] / [15] π / / / ( ) (EDC ) ( Sigma-Aldrich) 2, 4, 6- (TNBS Sigma ) ( U110803) Hep-G2( ) DMEM( Gibco) ( ) (MTT Sigma- Aldrich) ( Hyclone) 1.2 DF-101S ( ) SM-3 ( ) Modulyod-230 ( Thermo Electron Corporation) JY ( ) Sorvall fresco ( Biofuge Primo R) JF 2004 ( ) Fluoromax-4 ( Horiba Scientific) HZ-8812S ( ) ( Thermo) UV-2550 ( Shimadzu) HS-3120 ( ) zs90 ( Malvern) SPA 3800N (TEM)( SEIKO) Forma 3111 CO 2 ( Thermo) imark ( 62 Chin J Mod Appl Pharm, 2014 January, Vol.31 No

3 Bio-Rad) / [15] EDC / 1 2 EDC g g 30 ml A 0.5 g(mw=9 000) 25 ml B B 25 A 53 h ( 7 000) 10 h / 2.2 / TNBS [16] / / 10 mg 10 ml 1.0 mg ml µl 3.0 ml 4% 2.0 ml 0.1% TNBS 2.0 ml 37 2 h 2.0 mol L ml nm 1.0 mg ml 1 / 300 µl 3.0 ml / 9 000/ 50% ( )% 2.3 / 1 mg ml 1 (400 W 2 s 4 s 40 ) 2.4 / (critical aggregation concentration CAC) [17] CAC 15 mg 250 ml 60 µg ml 1 1 ml 50 ml 1.2 µg ml µg ml ml 8 10~4 000 µg ml 1 5 ml h 339 nm 350~450 nm ( 3 nm 5 nm) 1 I 1 (374 nm) 3 I 3 (385 nm) CAC 9000/ 50% CAC µg ml / 50% CAC Fig 1 CAC of CSO 9000 /SA 50% 2.5 / / Hep-G2 Hep-G2 10% DMEM ( 100 U ml 1 ) ( 37 5%CO 2 ) Hep-G µl 96 CO 2 24 h / 200 µl 100~2 400 µg ml h 5 mg ml 1 MTT 20 µl 4 h 150 µl DMSO 15 min 570 nm Chin J Mod Appl Pharm, 2014 January, Vol.31 No.1 63

4 A (%) = (1 A 570( treated ) 570( control) ) 100% A 570(treated) A 570(control) / Hep-G2 IC µg ml [18] DMSO 10 g L h ( 1 000) 12 h DMSO / [18-19] / / 10 mg 3 ml (400 W 2 s 4 s 20 ) 5 ml 2 mg ml 1 / DMSO 1.0 mg ml 1 DMSO 1 ml 10% (400 W 2 s 4 s 40 ) 12 h ( 7 000) 12 h DMSO r min min mg ml 1 DMSO (DMSO =99 1) µg ml 1 (EX=468 nm EM=566 nm 5 nm 700 mv) y= x r= DMSO = min (entrapment rate EE) (drug-loading rate DL) C V EE = ' C' V (%) DL(%) = Wd Wc + C' V C V W d W c 10% / 8.52% 93.15% nm(pdi 0.802) ( )mV nm(pdi 0.601) ( )mV mg ml 1 / 1 2% (TEM) [20] 2( ) 2 A B Fig 2 Image of nanoparticles A blank nanoparticles; B DOX-loaded nanoparticles µg ( 7 000) 5 ml ph 64 Chin J Mod Appl Pharm, 2014 January, Vol.31 No

5 (ph ph ) 47 µg DMSO 5 ml r min h ph PBS 3 3 / A / B Fig 3 Loads of adriamycin shell oligosaccharide/salicylic acid of drug-loading nanoparticles and free adriamycin in vitro release in different buffer A DOX-loaded CSO 9000 /SA 50% nanoparticle; B DOX from DOX/DMSO 3 ph ph=5.0 ph=7.4 [21-22] Mt Q = M n = kt M Mt t n k n= <n<1 1 1 Tab 1 Equation fitting of the in vitro release behavior of the formed drug delivery systems in different buffer r PBS (ph=5.0) Q=0.034t PBS (ph=6.8) Q=0.021t PBS (ph=7.4) Q=0.015t n=0.5 3 EDC / CAC ( CAC 1.0 mg ml 1 [23] ) 1 CAC 48.6 mv / / π π π π / Hep-G2 IC µg ml Chin J Mod Appl Pharm, 2014 January, Vol.31 No.1 65

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