Low Molecular Weight Water-soluble Chitosan Research Articles

Induction of permeability and apoptosis in colon cancer cell line with chitosan

The effect of chitosan on tight junctions (TJs) permeability on Caco-2 cell monolayer intestinal model was investigated. We have also investigated the effects of low molecular weight water-soluble chitosan (LMWC) on apoptosis in HT-29 cell. The changes in barrier properties of Caco-2 cell monolayers, including transepithelial electrical resistance (TEER) and permeability to Lucifer yellow, were assessed in response to chitosan treatment. High molecular weight chitosan (HMWC) was found to cause up to 50% dose-dependent reduction in TEER of Caco-2 cell monolayer without damage to the cell membrane under lower concentration. The effect of HMWC on TJs was confirmed by increased permeability of lucifer yellow when cells were treated with 0.001-0.0001% HMWC for 120 min compared to control cells. Results showed that HMWC did not affect the F-actin of cytoskeleton. LMWC was proven to be an antitumor compound as shown by inducing apoptosis as a function of DNA fragmentation. These results suggest that HMWC is an useful drug delivery agent in paracellular pathway and LMWC has potential value in colon cancer therapy.

Anticancer Effect of Intracellular-Delivered Doxorubicin Using a Redox-Responsive LMWSC-g-Lipoic Acid Micelles

To induce a quick-drug release, lipoic acid (LA) was introduced to amine group of low molecular weight water-soluble chitosan (LMWSC) by coupling agent. The disulfide bond (-S-S-) of lipoyl group from LMWSC-grafted lipoic acid (LL) can response to glutathione (GSH) at cytoplasm with reducing environment, where LL can rapidly be disassembled via dissociation of disulfide bond (-S-S-) by GSH. The doxorubicin (DOX)-encapsulated LL (LLDOX) was prepared by dialysis method, which allowed quick release of DOX by destabilization of inner-hydrophobic core of LL when disulfide bond (-S-S-) was dissociated by GSH. To demonstrate a redox-responsive release behavior of LLDOX, its drug release was accomplished under PBS buffer (pH 7.4) and GSH (10 mM) condition. In addition, the particle size and morphology of LL and LLDOX were respectively confirmed by DLS and TEM. The particle sizes of LL30% and LL60% were indicated as 268.8±30.9 and 308.1±11.9 nm, respectively. In addition, particle size of LLDOX was decreased more than that of LL. Also, surface charge of LLDOX was displayed to strong positive charge (LLDOX30%: 12.6±0.5 mV, LLDOX60%: 9.6±1.1 mV). Moreover, their morphological structure has a spherical shape. The cytotoxicity LL and LLDOX was confirmed by using MTT assay. Besides, to investigate the intracellular uptake of DOX from LLDOX against HeLa and AGS cell lines, fluorescence image was observed by using fluorescence microscopy. These results suggest that LL is an excellent as a drug carrier due to high anticancer effect by rapid drug release with redox-responsive effect.

Evaluation of hyaluronic acid-combined ternary complexes for serum-resistant and targeted gene delivery system

Branched polyethylenimine (bPEI) was well known as high transfection agent, which has many amine group. However, utilization of bPEI was limited due to high toxicity. To solve these problems, bPEI was introduced to low molecular weight water-soluble chitosan (LMWSC) with coupling agent. In addition, hyaluronic acid (HA), one of natural anion polymer, was introduced to binary complex of pDNA/bPEI-grafted LMWSC (LMPEI) to target the specific cancer cell and impart the serum resistant. Ternary complexes of pDNA/LMPEI/HA were prepared by electrostatic charge interaction and their binding affinity and DNase protection assay were conducted by gel retardation assay. Particle size of ternary complexes showed that had each 482 ± 245.4 (pDNA/LMPEI2%/HA, 1:16:1, w/w/w) and 410 ± 78.5 nm (pDNA/LMPEI4%/HA, 1:16:2, w/w/w). Moreover, to demonstrate serum-resistant effect of ternary complexes, particle size of them was measured according to incubated time (0–10 h) under serum condition. Transfection assay of ternary complexes showed that their transfection efficiency in CD44-receptor overexpressed HCT116 cell was higher than CD44-receptor negative CT26 cell. Additionally, intracellular uptake of ternary complexes with propidium iodide (PI)-labeled pDNA was observed to confirm targeting effect and cellular internalization by fluorescence microscopy. These results suggest that ternary complexes are superb gene carrier with excellent serum-resistant and high gene transfection.

Evaluation of disulfide bond-conjugated LMWSC-g-bPEI as non-viral vector for low cytotoxicity and efficient gene delivery

For efficient gene delivery, non-viral vectors should have high cellular uptake, excellent endosomal escape, and the ability to rapidly release the gene into the cytoplasm. Here, we developed a disulfide bond-conjugated bioreducible LMWSC-g-bPEI (LCP) composed of low molecular-weight water soluble chitosan (LMWSC), bPEI, and cystamine (Cys). The developed LCP had advantages such as low toxicity, great endosomal escape, and rapid release of pDNA into the cytoplasm. The polyplexes with LCP showed higher uptake into the nucleus and greater transfection efficiency than that without disulfide bond. Moreover, LCP polymer and polyplexes with LCP indicated lower cytotoxicity than bPEI 25kDa. In addition, a gel retardation assay and particle size were analyzed to demonstrate the reduction-sensitive gene delivery system. Besides, intracellular uptake pathway of polyplexes was investigated by various endocytosis inhibitor and confirmed to internalization into cell via macropinocytosis. These results suggest that bioreducible LCP is a superb non-viral vector for efficient gene delivery.

"저분자량 수용성 키토산과 para-aminosalicylic acid를 이용한 고분자 약물의 제조 및 특성 분석

"저분자량 수용성 키토산과 para-aminosalicylic acid를 이용한 고분자 약물의 제조 및 특성 분석

Antitumor effect of ternary complex with gene, peptide, and transferrin-grafted LMWSC for cancer therapy

Event Abstract Back to Event Antitumor effect of ternary complex with gene, peptide, and transferrin-grafted LMWSC for cancer therapy Jae-Woon Nah1, Gyeong-Won Jeong1, Won-Seok Kim1 and Changyong Choi1 1 Sunchon National University, Department of Polymer Science and Engineering, Korea Introduction: It is well known that gene therapy can cure serious diseases, such as cancer, infection, innate immunodeficiency, cystic fibrosis, and cardiovascular diseases. However, gene delivery within one's own body decreases transfection efficiency because the gene is degraded by enzymes in body[1]. Among the polymer-based non-viral gene carriers, chitosan is a good candidate to interact with DNA or RNA. Therefore the psiRNA-bcl2 gene and (KLA)4 peptide coated by LMWSC with targeting ligand was delivered by receptor-mediated endocytosis. The delivered compounds were denoted ternary complexes. These complexes effectively inhibited cancer cell growth via the co-activity of psiRNA-bcl2 and (KLA)4 peptide. (KLA)4 peptide of cationic, induced apoptosis by disrupting mitochondrial membrane, is fused to deliver with gene by electrostatic interaction. In addition, polyplexed genes and peptides can be degraded by enzymes in-vivo[2]. Therefore, low molecular weight-water soluble chitosan (LMWSC) as a non-viral vector which is a linear cationic polysaccharide, has good biocompatibility, antibacterial activity, solubility, and stability. In addition, LMWSC can act as a gene carrier because it has many free amine-groups and LMWSC used in this study was modified with PEGylated transferrin. In this study, a combination therapy with peptides and genes effectively inhibited cancer growth via mitochondrial apoptosis and gene silencing. To deliver the peptide and gene, ternary complexes with targeting ligand-grafted LMWSC were prepared. The ternary complexes were then investigated to confirm the ability to condense with genes and undergo transfection, as well as for cytotoxicity and anticancer activity. Experimental Methods: DNA/(KLA)4/LMWSC-TfP ternary complexes were prepared by electrostatic charge interaction in aqueous reactions. To prepare DNA/(KLA)4 polyplexes, (KLA)4 was dissolved in distilled water. The solution of (KLA)4 peptide was then added to the pDNA solution and incubated for 30 min at room temperature. Finally, the LMWSC-TfP solution was added to the polyplex solution and incubated for 30 min at room temperature. Binding affinity, stability from Dnase, and releasing assay of ternary complexes was conducted by gelretardation assay. Also, transfection efficacy of ternary complexes was accomplished in PC3 cell lines. In addition, cytotoxicity and antitumor activity of that was confirmed by MTT assay in L929 and PC3 cell lines. Results and Discussion: To deliver effectively gene and peptide in cancer cell, targeting-ligand grafted ternary complex was prepared by electrostatic interaction. The delivered gene and (KLA)4 peptide showed that psiRNA exerted RNA-silencing action in the nucleus, while the (KLA)4 peptide inhibited cancer growth through mitochondrial apoptosis. Gene transfection of ternary complex showed that targeting-ligand grafted ternary complex was transfected more effectively than that without targeting-ligand at all ratio (1:1:64, 1:2:64, 1:4:64) because cellular uptake of it was increased by targeting-ligand. Also, antitumor activity of ternary complex was indicated that the more amount of psiRNA was increased, the more cell viability was decreased. It means that ternary complex was entered into PC3 cells and tumor growth was inhibited by gene scilencing of psiRNA and mitochondrial apoptosis of KLA peptide, respectively. Conclusion: In this study, we prepared ternary complex with targeting-ligand grafted LMWSC by electrostatic interaction. The transfection of ternary complex was increased by targeting-ligand. Moreover, antitumor activity showed that was increased by gene and peptide co-action. These results suggest that ternary complex can be effectively inhibiting tumor growth by delivered gene and peptide. This work was supported by National Research Foundation of Korea (NRF) grant funded by the Ministry of Science, ICT & Future Planning. (No. NRF-2014R1A2A1A10053027).

Algicidal effects of free-amine water-soluble chitosan to marine harmful algal species

Harmful algal blooms (HABs) commonly cause significant damage to aquatic systems due to toxin production and depletion of oxygen. In this study, the low molecular weight water-soluble chitosan (LMWSC) was investigated as a new algicidal compound by first investigating the potential of LMWSC in selective combating HABs. The LMWSCs disrupted algal cell and chloroplast membranes within 1h against HABs, leading to efflux of the intracellular components. For co-cultivation of harmful and harmless algal, LMWSC selectively kill the harmful algal. Based on these results, LMWSC is a promising novel material for harmful algal bloom control.

소수성 항균 펩타이드 전달체로 응용하기 위한 데옥시콜릭산이 결합된 저분자량 수용성 키토산 나노입자의 제조와 특성

소수성 항균 펩타이드 전달체로 응용하기 위한 데옥시콜릭산이 결합된 저분자량 수용성 키토산 나노입자의 제조와 특성

Antibacterial activity of water-soluble chitosan produced from non-traditional marine resources

Two different chitosan polymers were obtained from chitin extracted from the crab Polybius henslowii through different methods: (1) water-soluble chitosan (WSC) through N-acetylation with addition of acetic anhydride and (2) low molecular-weight water-soluble chitosan (LWSC) by the use of H2O2. WSC and LWSC polymers were characterized by FTIR and GPC analysis and investigated for antibacterial activity against a range of Gram-negative bacteria (Escherichia coli 1103, Escherichia coli 10503, Salmonella sp.) and Gram-positive bacteria (Staphylococcus aureus, Bacillus subtilis and Lactobacillus planctarum). This study is a contribution to economic valorization of Polybius by-catch and sustainable development of west coast Portuguese fisheries communities. Financial support was obtained from European Fisheries Fund – Axis 4 managed by Fishery Local Action Group Oeste-Portugal.

Water-soluble chitosan and herbal honey compound alleviates atopic dermatitis-like lesions in NC/Nga mice

Atopic dermatitis (AD) is a chronic inflammatory skin disease that was influenced by complex interactions via genetic, environmental, immunologic, and biochemical factors, though the cause of AD is still unknown. It characterized by elevated serum immunoglobulin E (IgE) levels, immunological abnormalities, and eosinophilia in the tissues and peripheral blood. In the present study, we applied an antimicrobial moisturizing cream containing low-molecular weight water-soluble chitosan and herbal honey (AMCH) to remedy AD-like lesions. The inhibiting effect of AMCH on NC/Nga mice, that AD-like lesion was induced by 1-chloro-2,4-dinitrobenzene (DNCB), was evaluated by examining sensory evaluation scores, scratching behavior, immune cells in blood, serum IgE level, infiltration of mast cells, and skin histology. The total sensory evaluation scores, scratching behavior, the level of serum IgE, interlukin-4 (IL-4), and IL-12 in AD mouse model were significantly reduced by AMCH. Moreover, its suppressing effect resulted in decreased mast cell infiltration. Our results suggest that AMCH might be beneficial as a potent agent for treatment of AD-like lesion.

A novel protocol for the oxidative degradation of chitosan with hydrogen peroxide catalyzed by peroxomolybdate in aqueous solution

A novel pathway is reported to obtain low molecular weight, water-soluble chitosan (LMWSC) via oxidative degradation of chitosan with a catalyst containing active Mo(O2) sites. H2O2 was found to play a fatal role in the regeneration of Mo(O2). Moreover, (TBA)3{PO4[MoO(O2)2]4} can be recovered and reused four times without loss of reactivity.

PH-dependent mode of antibacterial actions of low molecular weight water-soluble chitosan (LMWSC) against various pathogens

This study examined the antibacterial mode of action of low molecular weight water-soluble chitosan (LMWSC; MW1, MW3, MW5 and MW10) using a combination of approaches, including antibacterial assays, culture in media with different pH values, bactericidal kinetics, cellular leakage measurements, depolarization, and electron microscopy, as well as an in vivo study utilizing a wound infection and wound healing model. The antibacterial activity of LMWSC was related inversely to pH, with higher activities being observed at lower pH values. In addition, evaluation of the effect of bactericidal concentrations of LMWSC on the morphology of Bacillus megaterium and Escherichia coli O–157 revealed that it induces filamentation. Furthermore, the degree of depolarization (in Escherichia coli, pH 7.4) and calcein leakage (lipid composition; L-α-phosphatidylethanolamine (PE)/L-α-phosphatidyl-dl-glycerol (PG)=7/3 (w/w), pH 5.4 and 7.4) were evaluated. Moreover, cells cultured at various pHs were evaluated by confocal microscopy (pH 5.4 and 7.4). These results showed that treatment with LMWSC with different molecular weights had different effects on bacteria. In particular, LMWSC (MW10) cause more visible damage to the bacterial cell membrane than lower molecular weight LMWSC, which is due most likely to the penetration of cells by the lower weight molecules. Interestingly, MW5 was found to attack the membrane and penetrate the cells. In addition, scanning electron microscopy showed that MW10 caused significant morphological changes to the surface of the bacteria. Finally, an in vivo study utilizing a wound infection or healing model showed that LMWSC had the potential for use as a lead compound for the development of a novel anti-infective or healing compound.

Characterization and preparation of core–shell type nanoparticle for encapsulation of anticancer drug

The aim of this study is to prepare delivery vehicles of paclitaxel using low molecular weight water-soluble chitosan (LMWSC) and evaluate them as an anticancer drug delivery system. LMWSC was modified with methoxy polyethylene glycol (LMWSC-MPEG, ChitoPEG), and then it was conjugated with cholesterol (LMWSC-MPEG-Chol). Core–shell type LMWSC-MPEG-Chol nanoparticles (LMWSC-NPs) were prepared by the dialysis method, and the core–shell structure was confirmed by 1H NMR analysis. To this polymer, paclitaxel was encapsulated and core–shell type nanoparticles were prepared. The release tests indicated that release of paclitaxel from the core–shell type nanoparticles and its transport across the dialysis membrane was slower than dialysis of free paclitaxel. In a cytotoxicity study using CT26 cell, the paclitaxel-encapsulated core–shell type nanoparticles (LMWSC-NPs) showed a toxicity against tumor cells similar to paclitaxel itself. The results of a tumor inhibition test with CT26 implanted upon mouse tumor models in vivo indicated that the application of a dose of 10 mg/kg of LMWSC-NPT showed a superior survival rate, and a slower tumor growth than when paclitaxel alone was administered, although the tumor growth and survival rate were not significantly changed at a dose of 2 mg/kg. The LMWSC-NPT dose above 10 mg/kg showed a superior antitumor activity.

Insulin-incorporated chitosan nanoparticles based on polyelectrolyte complex formation

Insulin-incorporated nanoparticles were prepared by polyelectrolyte complex formation using low-molecular weight water soluble chitosan (LMWSC). Insulin-incorporated nanoparticles showed spherical shapes with a particle size of approximately 200 nm. The average particle size, drug content and loading efficiency increased with increasing insulin feed. The zeta potential decreased with increasing insulin feed, indicating that the electrostatic interaction increased with increasing insulin feed. At 1H nuclear magnetic resonance (NMR) spectra, insulin and LMWSC had intrinsic peaks while the peaks of the nanoparticles were similar to LMWSC itself at D2O. However, the specific peaks for insulin appeared as a result of the breakdown of nanoparticles using DCl. When polyelectrolyte complex had dissociated, fluorescein isothiocyanate (FITC)-labeled insulin in the nanoparticles showed similar UV absorption peaks to FITC-labeled insulin only. At the release study, insulin was released continuously from the nanoparticles over 120 h at all formulations. Higher insulin contents induced a decrease in the rate of insulin released from nanoparticles. Consequently, LMWSC and insulin successively formed polyelectrolyte complexes as a nanocarriers and LMWSC can be considered a good candidate for insulin delivery. Open image in new window

Chitosan: a novel platform in proton-driven DNA strand rearrangement actuation

Nanometre-scaled DNA machine based on molecular recognition properties of DNA has now become a powerful tool in nanodevices, miniaturized structure, and nanofabrication. The common principle behind designing a DNA nanomachine is DNA strand exchange or rearrangement, which is solely controlled by the stabilization through associative and dissociative forces arising from base pair interaction of DNA molecules. Thus, highly effective DNA reaction actuator will make DNA nanomachine more flexible, controllable, and powerful device. Here, we report the novel polymer-mediated platform in proton-driven DNA strand rearrangement actuation. This cationic low molecular weight water-soluble chitosan (LMWSC) exhibited pH-dependent complexation with oligodeoxynucleotides (ODN). It formed complex with ODN only at low pH and accelerated the DNA strand exchange (or rearrangement) reaction between dsDNA and its complementary ssDNA at pH 5.0. However, no complexation was observed between LMWSC and ODN at neutral pH. We assume that at physiological pH, LMWSC is not protonated enough for formation of complex with ODN. Therefore, it can not diminish the electrostatic repulsion among the negatively charged DNA strands of the three-stranded intermediate formed during the strand exchange reaction. In contrast, LMWSC becomes positively charged at acidic pH, and it stabilizes the three-stranded intermediate by spreading out the accumulated counter-ions and increasing the entropy of the system. This fascinating observation prompted us to believe that this intelligent proton-driven DNA reaction actuator has a potential for the precise control of DNA nanomachine and would be applied for operating and controlling nanoscaled machine.

Enhance of Tumor Targeting by Receptor-Mediated Endocytosis Using Low Molecular Water-Soluble Chitosan Nanoparticles Loaded with Anticancer Agent

In this study, we prepared using low molecular weight water-soluble chitosan nanoparticle loaded paclitaxel (LMWSC-NPT) and investigated the potential as a drug carrier which is able to accumulate in the tumor site. In the experiment of receptor-mediated endocytosis, LMWSC-NPT was treated with sodium azid (NaN3) as an inhibitor of endocytosis process. As results, the antitumor activity of LMWSC-NPT treated with sodium azid didn’t show but LMWSC-NPT was shown the high antitumor activity. Therefore, LMWSC-NPs modified with hydrophobic group will be useful anticancer agent carrier via receptor-mediated endocytosis.

Chito-oligosaccharides as an Alternative to Antimicrobials in Improving Performance, Digestibility and Microbial Ecology of the Gut in Weanling Pigs

A total of 126 crossbred weanling pigs (average body weight of 6.3±0.3 kg) were used to investigate the effect of chito-oligosaccharide (COS) on growth performance, nutrient digestibility, pH of gastro-intestinal tract (GI), intestinal and fecal microflora of young piglets. Pigs were allocated to three dietary treatments based on body weight and gender in a single factorial arrangement. Treatments were control (No COS), T1 (0.2% COS during starter (6-13 kg) and 0.1% COS during grower (13-30 kg) phases, and T2 (0.4% COS during starter (6-13 kg) and 0.3% COS during grower (13-30 kg) phases, respectively. Each treatment had 3 replicates and 14 pigs were raised in each pen. COS is a low molecular weight water-soluble chitosan that can be obtained from chitin of the crab shell after deacetylation with concentrated sodium hydroxide at high temperature and then further decomposition by chitosanase enzyme in the presence of ascorbic acid. For the starter and grower periods, there were no significant differences (p>0.05) in average daily gain (ADG) and feed to gain ratio among treatments. However, during the overall period (6-30 kg), T2 showed better (p<0.05) feed to gain ratio than other treatments. A digestibility study was conducted at the end of grower phase which showed improvement (p<0.05) in DM and crude fat digestibility in T2 over the control. At 25 kg body weight, 6 pigs per treatment (2 per replicate) were sacrificed to determine the effect of diets on pH and microbial count at different sections of the GI tract. The pH of the cecal contents in pigs fed 0.1% COS was higher (p<0.05) than in the other treatments. Total anaerobic bacterial number increased from cecum to rectum in all treatments. The weekly total bacterial counts showed higher (p<0.05) in feces of pigs fed COS than that of untreated pigs at the 8 th week. The number of fecal E. coli in untreated pigs at 4 th wk was 7.35 log CFU/g compared to 6.71 and 6.54 log CFU/g in 0.1 and 0.3% COS-treated pigs, respectively. Similarly, at 8 th wk, fecal clostridium spp. were lower in pigs fed 0.3% COS (5.43 log CFU/g) than in untreated pigs (6.26 log CFU/g). In conclusion, these results indicated that chito-oligosaccharide could improve feed efficiency in young pigs and inhibited the growth of harmful bacteria.

Low molecular weight water‐soluble chitosans: Preparation with the aid of cellulase, characterization, and solubility

AbstractPreparation of water‐soluble chitosan (WSC) was made by treating partially N‐deacetylated chitosan with acetic anhydride in aqueous acetic acid. The optimal conditions of preparing WSC were determined on the basis of orthogonal tests. Low molecular weight WSC with broad molecular weight (600–1.5 kDa) were obtained by the depolymerization of WSC using cellulase at optimum condition of pH 4.5 and 60°C. The solubility of WSC in water and aqueous organic solvents was investigated in detail. Weight–average molecular weight (Mw) and molecular weight distribution (Mw/Mn) of samples were measured by gel permeation chromatography. The structure of WSC and its degraded products were characterized by XRD, FTIR, and MALDI‐TOF MS. The decrease of molecular weight led to transformation of crystal structure and the increase of solubility, but the chemical structures of residues were not modified compared to WSC, which was not hydrolyzed. The solubility of the WSC in water and aqueous organic solvents increased with the decrease of molecular weight. The solubility of the WSC with low molecular weight was rather high even in aqueous dimethylacetamide and dimethylsulfoxide. © 2006 Wiley Periodicals, Inc. J Appl Polym Sci 102: 1098–1105, 2006

Antioxidant activity of half N-acetylated water-soluble chitosan in vitro

Different molecular weight water-soluble chitosan (half N-acetylated chitosan) was prepared and the structure of water-soluble chitosan was characterized by FT-IR. The pH dependence of water solubility of water-soluble chitosan was evaluated from turbidity. Total antioxidant activity, reducing power, superoxide anion radical and hydroxyl radical quenching assay, metal chelating activity, and H2O2 scavenging activity were used for the evaluation of different molecular weight half N-acetylated chitosan in vitro. Low-molecular weight water-soluble chitosan (WSC4) exhibited high reductive capacity and expressed good inhibition of linoleic acid peroxidation in the linoleic acid model system. WSC4 (0.25 mg/mL) scavenged 78.8% of superoxide radical. At 5 mg/mL, scavenging percentage of WSC1, WSC2, WSC3, and WSC4 against hydroxyl radical was 49.3%, 66.8%, 77.1%, and 83.7%, respectively. These results indicate that water-soluble chitosan is an ideally natural antioxidant, and its antioxidant activity depends on its molecular weight.

All-trans retinoic acid-associated low molecular weight water-soluble chitosan nanoparticles based on ion complex

The purpose of this study is to develop novel nanoparticles based on polyion complex formation between low molecular weight water-soluble chitosan (LMWSC) and all-trans retinoic acid (atRA). LMWSC nanoparticles encapsulating atRA based on polyion complex were prepared by mixing of atRA into LMWSC aqueous solution using ultrasonication. In FTIR spectra, the carbonyl group of atRA at 1690 cm−1 disappeared or decreased when ion complexes were formed between LMWSC and atRA. In1H NMR spectra, specific peaks of atRA disappeared when atRA-encapsulated LMWSC (RAC) nanoparticles were reconstituted into D2O while specific peaks both of atRA and LMWSC appeared in D2O/DMSO (1/3, v/v) mixture. XRD patterns also showed that the crystal peaks of atRA were disappeared by encapsulation into LMWSC nanoparticles. LMWSC nanoparticles encapsulating atRA have spherical shapes with particle size below 200 nm. The mechanism of encapsulation of atRA into LMWSC nanoparticles was thought to be an ion complex formation between LMWSC and atRA. LMWSC nanoparticles showed high atRA loading efficiency over 90% (w/w). AtRA was continuously released from nanoparticles over 10 days. Inin vitro cell cytotoxicity test, free atRA showed higher cytotoxic effect against CT 26 colon carcinoma cell line on 1 day. However, RAC nanoparticles showed similar cytotoxicity against CT 26 cells on 2 day. These results suggest the potential for the introduction of LMWSC nanoparticles into various biomedical fields such as drug delivery.