Chitosan was modified by conjugating coupling with linolenic acid through the l-ethyl-3-( 3-dimethylami-nopropyyl) carbodiimmide (EDC)-mediated reaction. The degree of substitution 1.8% (i.e. 1.8 linolenic acid group per 100anhydroglucose units) was measured by 1H NMR. The critical aggregation concentration (CAC) of the self-aggregate of hy-drophobically modified chitosan was determined by measuring the fluorescence intensity of the pyrene as a fluorescent probe.The CAC value in phosphate-buffered saline (PBS) solution (pH7.4) was 5 × 10-2 mg mL-1. The average particle size of self-aggregates of hydrophobically modified chitosan in PBS solution (pH7.4) was 210.8 nm with a unimodal size distributionranging from 100 to 500 nm. Transmission electron microscopy (TEM) study showed that the formation of near sphericalshape nanoparticles has enough structural integrity. The loading ability of hydrophibically modified chitosan (LA-chitosan)was investigated by using bovine serum albumin (BSA) as the model. The loading capacity of self-aggregated nanoparticlesincreases (19.85% ±0.04% to 37.57% ±0.25%) with the concentration of BSA (0.1-0.5mgmL-1).
LIU ChenguangDesai Kashappa Goud H.CHEN XiguangPark Hyun-Jin
Oleoylchitosans (O-chitosans) with different degrees of substitution (DS) were synthesized by reacting chitosan with oleoyl chloride. The chemical structures of the products were characterized by 1H NMR and FT-IR. These results suggested the for- mation of an amide linkage between the amino groups of chitosan and the carboxyl groups of oleic acid. The viscosity of O-chitosan sharply increased with the increase of concentration, whereas that of unmodified chitosan rose only slightly. This increase became larger as the DS increased. All of the O-chitosans could reduce surface tension slightly. The critical aggregation concentration (CAC) of O-chitosans with DS 5%, 11%, and 27% were 79.43 mgL-1, 31.6 mgL-1, and 10 mgL-1, respectively. Nanoparticles were prepared using an O/W emulsification method. The mean diameters of the polymeric amphiphilic nanoparticles of O-chitosans with DS 5% and 11% were around 327.4 nm and 275.3 nm, respectively.
The captopril/Chitosan-gelatin net-polymer microspheres (Cap/CGNPMs) were prepared using Chitosan(CS) and gelatin(Gel) by the methods of emulsification. A cross linked reagent alone or in combination with microcrystalline cellulose(MCC) was added in the process of preparation of microspheres to eliminate dose dumping and burst phenomenon of microspheres for the improvement of the therapeutic efficiency and the decrease of the side effects of captopril(Cap). The results indicate that Cap/CGNPMs have a spherical shape , smooth surface morphology and integral inside structure and no adhesive phenomena and good mobility,and the size distribution is mainly from 220 to 280 μm. Researches on the Cap release test in vitro demonstrate that Cap/CGNPMs are of the role of retarding release of Cap compared with Cap ordinary tablets (COT), embedding ratio (ER) ,drug loading (DL), and swelling ratio (SR), and release behaviors of CGNPMS are influenced by process conditions of preparation such as experimental material ratio (EMR) , composition of cross linking reagents. Among these factors , the EMR(1/4),CLR (FOR+TPP) and 0.75% microcrystalline cellulose (MCC) added to the microspheres are the optimal scheme to the preparation of Cap/CGNPMs. The Cap/CGNPMs have a good characteristic of sustained release of drug, and the process of emulsification and cross-linking process is simple and stable. The CGNPMs is probable to be one of an ideal sustained release system for water-soluble drugs.
The cytotoxicity profile of three chitosan derivatives with different affinity to water was evaluated in vitro.The derivatives selected were carboxymethylated-chitosan(CMCH),linoleic acid modified- chitosan(LACH)and deoxycholic acid modified-chitosan(DACH),respectively,and the results of FTIR and NMR confirmed the successful modification.Cytotoxicity of these polymers was investigated via the red blood cell lysis assay and the MTT assay.The red blood cell lysis test showed that CH elicited a certain level of red blood cell toxicity,while CMCH,LACH and DACH all displayed low membrane damaging effects, with the hemolysis rates of 2.385%,1.560% and 4.404%,respectively,which comes well within permissible limit(5%).The MTT assay revealed that CH exhibited significant inhibitory effect on fibroblast proliferation at higher concentration,while its three derivatives showed no cytotoxicity.CMCH had stimulatory effects on fetal mouse fibroblast proliferation.Differences in cytotoxicity of CH and its derivatives may result from the specific chemical modifications leading to the alteration of molecular charge density and type of the cationic functionalities,structure and sequence,and conformational flexibility.
